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EPA STAFF EVALUATION AND REVIEW REPORT
www.epa.govt.nz
Application for approval to import Mainman for release
APP202145
9 February 2015
2
Application for approval to import Mainman for release (APP202145)
February 2015
Overview
Application Code APP202145
Application Type To import or manufacture for release any hazardous substance under
Section 28 of the Hazardous Substances and New Organisms Act
1996 (“the Act”)
Application Sub-Type Notified - Category C
Applicant ISK New Zealand Limited
Purpose of the application
To import Mainman insecticide, which contains 500 g/kg of
flonicamid, for the control of various insect pests in potatoes and
other horticultural crops
Date Application Received 27 May 2014
Submission Period 12 June 2014 –24 July 2014
Submissions received Three submissions were received, from:
Gerry Coates on behalf of Te Rūnanga o Ngāi Tahu (“Ngāi Tahu”)
Don MacLeod on behalf of The National Beekeepers’ Association of
New Zealand (“the NBA”)
Philippa Rawlinson on behalf of Federated Farmers of New Zealand
Bee Industry Group (“Federated Farmers”)
Information requests and
time waivers
Further information was requested under section 58 of the Act.
Consequently, a time waiver was applied under section 59 of the Act
to the start of the consideration period
Hearing date 23 Feburary 2015
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Application for approval to import Mainman for release (APP202145)
February 2015
Table of Contents
1. Executive Summary .................................................................................................................... 6
2. Background .................................................................................................................................. 9
3. Process, consultation and notification ..................................................................................... 9
4. Hazardous properties ............................................................................................................... 11
5. Submissions .............................................................................................................................. 11
6. Risk and benefit assessment ................................................................................................... 16
7. Controls ...................................................................................................................................... 22
8. Overall evaluation and recommendation ................................................................................ 25
Appendix A: Submissions received .................................................................................................. 27
Appendix B: EPA standard risk descriptors ..................................................................................... 28
Appendix C: Classification of Mainman ............................................................................................ 31
Appendix D: Active ingredient and metabolites ............................................................................... 73
Appendix E: Staff’s risk identification ............................................................................................. 193
Appendix F: Qualitative risk assessment ....................................................................................... 194
Appendix G: Human health risk assessment ................................................................................. 196
Appendix H: Environmental risk assessment ................................................................................ 204
Appendix I: Controls applying to Mainman .................................................................................... 208
Appendix J: Confidential information ............................................................................................. 212
Appendix K: Standard terms and abbreviations ............................................................................ 213
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Application for approval to import Mainman for release (APP202145)
February 2015
List of tables and figures (listed in order of appearance in the document)
Table 1 Hazard classifications of Mainman as proposed by the applicant and the staff ............. 11
Table 2 Risk descriptors ..................................................................................................................... 28
Table 3 Likelihood Descriptors .......................................................................................................... 28
Table 4 Magnitude of adverse effect .................................................................................................. 29
Table 5 Magnitude of beneficial effect ............................................................................................... 30
Table 6 Applicant and staff classifications of the mixture .............................................................. 31
Table 7 Physical and chemical properties of the mixture ............................................................... 33
Table 8 Identification of flonicamid ................................................................................................... 73
Table 9 Applicant and staff classifications of the active ingredient .............................................. 74
Table 10 Physico-chemical properties of flonicamid ....................................................................... 75
Table 11 Summary of studies for the active ingredient with NOAEL and LOAEL values
and key effects. ................................................................................................................... 122
Figure 1: Proposed metabolic pathway of IKI-220 in aerobic water/sediment systems ............ 134
Figure 2: Degradation pathway of flonicamid in soils ................................................................... 139
Table 12 Potential sources of risks associated with hazardous substances ............................. 193
Table 13 Qualitative assessment of human health risks ............................................................... 194
Table 14 Qualitative assessment of risks to the environment ...................................................... 195
Table 15 Deriving an AOEL for flonicamid ...................................................................................... 196
Table 16 Derivation of dermal absorption value in humans ......................................................... 197
Table 17 Risk Quotients for Mainman: Potato – 80 g ai/ha, 2 applications at 7 days,
ground based ..................................................................................................................... 198
Table 18 Risk Quotients for Mainman: Confidential use 1 – 70 g ai/ha, 2 applications at
14 days, ground based ....................................................................................................... 198
Table 19 Risk Quotients for Mainman: Confidential use 3– 70 g ai/ha, 2 applications at
7 days, ground based ......................................................................................................... 199
Table 20 Risk Quotients for Mainman: Confidential use 5 – 70 g ai/ha, 3 applications at
21 days ................................................................................................................................. 199
Table 21 Risk Quotients for Mainman: Re-entry exposure modelling ......................................... 200
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Application for approval to import Mainman for release (APP202145)
February 2015
Table 22 Risk Quotients for Mainman: Use on Potatoes ............................................................... 201
Table 23 Risk Quotients for Mainman: Confidential uses 1 and 2 – 70 g ai/ha,
2 applications at 14 days, ground based and aerial ....................................................... 202
Table 24 Risk Quotients for Mainman: Confidential uses 3 and 4 – 70 g ai/ha,
2 applications at 7 days, ground based and aerial ........................................................ 202
Table 25 Risk Quotients for Mainman: Confidential use 5 – 70 g ai/ha,
3 applications at 21 days .................................................................................................. 203
Table 26 Summary of environmental fate data on flonicamid and its metabolites . ... 204
Table 27 Summary of ecotoxicological data on flonicamid and its metabolites – ..................... 205
Table 28: Controls for Mainman ....................................................................................................... 208
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Application for approval to import Mainman for release (APP202145)
February 2015
1. Executive Summary
ISK New Zealand has applied to import Mainman, containing 500 g/kg flonicamid, for the control of 1.1.
various insect pests in potatoes and other horticultural crops. It is intended to be applied as a dilute
spray by ground-based and aerial application methods at rates of up to 80 g flonicamid /ha, at a
maximum of three times a year.
The staff of the Environmental Protection Authority (“the staff”) have identified the hazard classifications 1.2.
of Mainman based on product data, the composition of the substance, and the properties of its
components:
Hazard Endpoint EPA classification
Eye irritant 6.4A
Vertebrate ecotoxicity 9.3C
In response to public notification, three submissions were received (Section 5 and Appendix A): One 1.3.
opposing approval of Mainman, one which neither opposed nor supported the application, and one
opposing the application in its current form.
Ngāi Tahu had concerns about the lack of information available to submitters and thought there had 1.4.
been no consideration of Māori issues. They were concerned about the effects on aquatic and
terrestrial invertebrates and wanted tests to be carried out on native species.
The National Beekeepers’ Association was concerned with the effects of Mainman on bees. They 1.5.
wanted to ensure that the EPA had access to all the information overseas regulators had. They wanted
a draft label to be publically available for submitters to comment on. They recommended that the EPA
apply a control stating that Mainman must not be applied to flowering plants within 10 days of flowering
and that the approval be limited to potatoes.
Federated Farmers Bee Interest Group had concerns about bees foraging on weeds in the crop and 1.6.
crop margins. They wanted an enforceable label, clearly listing each warning and how to avoid effects
on bees and other beneficial insects in nearby habitats. If these concerns were addressed, Federated
Farmers would consider withdrawing its opposition.
The staff conducted quantitative human health and environmental risk assessments (Section 6 and 1.7.
Appendixes E-G) based on the evaluation of toxicological and ecotoxicological data for the formulated
substance (Mainman) and the active ingredient. These assessments considered the exposure and
subsequent effects on people and the environment throughout the entire life cycle of the substance.
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Application for approval to import Mainman for release (APP202145)
February 2015
The exposure modelling indicates that risks to operators are acceptable even without the use of 1.8.
personal protective equipment (PPE) and PPE is not triggered by the hazard classifications of
Mainman. However, the staff consider it is appropriate to apply requirements for PPE as its use when
handling agrichemicals is good practice. The exposure assessment also indicates acceptable risks for
re-entry workers even without the use of PPE. However, the staff consider that in order to promote
good practice it is appropriate to include a label statement requiring the use of PPE when entering
treated areas before the spray has dried.
The risk assessment shows that low risks are expected for the environment. However, effects on bees 1.9.
could occur if the bees are directly exposed to the spray. For this reason, the staff recommend that bee
protection controls are applied to Mainman.
Based on the risk assessment, the staff have proposed controls to reduce worker exposure, prevent 1.10.
Mainman entering waterways and prohibit it from being applied to flowering plants or when bees are
present. These controls include:
a maximum application rate of 80 g ai/ha
bee protection controls are applied to Mainman, and must be mentioned on the label
Mainman must not be applied onto or into water
label statement should be added requiring the use of PPE if entering treated areas before the
spray has dried
a limit of 3 g/kg is set for the toxicologically relevant impurity toluene in the active ingredient.
With the suite of proposed controls in place the staff consider that the risks to human health and the
environment are reduced to negligible.
The staff assessed the risks posed by Mainman to the relationship of Māori to the environment and their 1.11.
culture and traditions with their ancestral lands, water, taonga and the principles of the Treaty of
Waitangi (Te Tiriti o Waitangi). The assessment identified potential cultural risks from the impacts on
taonga species, such as native pollinators, and the potential disruption to the mauri of native species
and valued ecosystems. However, given the proposed use in agricultural ecosystems and with the
proposed controls in place, the staff consider the cultural impacts of Mainman can be appropriately
managed.
The staff consider that the approval and subsequent availability of Mainman would give rise to 1.12.
significant (i.e. non-negligible) benefits. These benefits include:
improved crop health and yields
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Application for approval to import Mainman for release (APP202145)
February 2015
the availability of a new active ingredient with a new mode of action for the control of insect
pests in potatoes and other horticultural crops, which will provide a new solution for controlling
insect pests
reduced risk of pests developing resistance to insecticidal active because this will allow new
combinations of actives with different modes of action to be used; effectively managing pest
resistance also prolongs the efficacy of active ingredients
the opportunity to replace more hazardous insecticides that are currently used (e.g.
organophosphates).
With the proposed controls in place, the potential benefits of Mainman will outweigh the risks posed by 1.13.
the substance. Therefore, the staff consider that the application by ISK New Zealand to import
Mainman can be approved in accordance with clause 26 of the Hazardous Substances and New
Organisms (Methodology) Order 1998 (“the Methodology”).
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Application for approval to import Mainman for release (APP202145)
February 2015
2. Background
Mainman is a foliar applied insecticide that is intended to be used to control sucking pests in 2.1.
horticultural crops.
Mainman contains 500 g/kg of flonicamid as the active ingredient. Flonicamid is a new active ingredient 2.2.
to New Zealand, i.e. Mainman is the first product containing flonicamid to be considered for approval in
New Zealand. However, Mainman has been approved for use in other jurisdictions including: Canada,
the European Union, Japan, the United States of America, and South Korea.
The full formulation of Mainman is presented in the confidential Appendix I. 2.3.
A summary of the lifecycle of the substance is as follows: 2.4.
Mainman will be imported into New Zealand fully formulated, packaged and labelled in 500 g to
5 kg high density polyethylene plastic containers. Mainman is not expected to be imported in
bulk containers or repackaged in New Zealand; however, if re-packaging were required (e.g.
due to a damaged shipment) it would take place in facilities approved for that purpose under
the Agricultural Chemicals and Veterinary Medicines Act 1997 (ACVM Act).
Containers of Mainman will be transported by sea, road or rail throughout New Zealand. The
substance will be stored in secure chemical storage facilities.
Mainman will be used by commercial growers and contractors who are familiar with the safe
practices for storing and handling pesticides. Domestic or home-use by untrained users is not
anticipated.
Mainman will be diluted with water and applied as a foliar spray to potatoes and other crops to
treat sucking pests such as aphids and psyllids.
The applicant expects that Mainman will be disposed of by use, eliminating the need to dispose
of unused chemical. However, excess chemical can be disposed of to landfill or waste water
treatment plants according to local hazardous waste requirements. Users will be advised to
rinse the containers which can be disposed of at an approved landfill or recycled via an
appropriate recycling programme.
3. Process, consultation and notification
The application was lodged pursuant to section 28 of the Act on 5 November 2013 and formally 3.1.
received on 27 May 2014.
The Minister for the Environment was advised of the application in writing on 12 June 2014, in 3.2.
accordance with section 53(4) of the Act.
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Application for approval to import Mainman for release (APP202145)
February 2015
The Ministry for the Environment, WorkSafe New Zealand, the Ministry of Health, the Department of 3.3.
Conservation and the Agricultural Compounds and Veterinary Medicines (ACVM) group of the Ministry
for Primary Industries were notified of the application on 12 June 2014 and invited to comment. No
comments were received.
The staff advised the applicant that consultation with Māori would be beneficial for this application. A 3.4.
summary of the application was prepared and distributed on behalf of the applicant to the EPA’s Māori
advisory network, Te Herenga, in April 2014. .Although some feedback was received, it was not
applicable to the application but rather focused on the decision making process so did not alter the
application. The feedback was provided to the applicant to allow them to review the application before
it was formally received.
The application was publicly notified and opened for submissions on 12 June 2014; the submission 3.5.
period closed on 24 July 2014.
The application was publicly notified because the staff considered it to be of significant public interest in 3.6.
that Mainman contains an active ingredient that is new to New Zealand.
Three submissions were received. Te Rūnanga o Ngāi Tahu (“Ngāi Tahu”) opposed the approval of 3.7.
Mainman, the National Beekeepers’ Association (“the NBA”) did not indicate whether they opposed or
supported the application. Federated Farmers Bee Industry Group (“Federated Farmers”) opposed the
application in its current form but was not opposed to the use of Mainman by farmers to control sucking
pests on horticultural crops. The issues raised in these submissions are discussed in section 5. Ngāi
Tahu and the NBA indicated that they wished to be heard in person by the committee.
During the evaluation of the application, the staff required further information to complete the evaluation 3.8.
and risk assessment, which was requested under section 58 of the Act.
In preparing this report, the following documents were used: 3.9.
the application form and confidential material submitted by the applicant (including toxicological
and ecotoxicological studies on the product, the active ingredient and its metabolites, the full
composition of the product, chemical and physical properties of the product, and the purity of
the active ingredient)
the submissions
a report from the EPA’s Māori Advisory Committee, Ngā Kaihautū Tikanga Taiao, and
other available information.
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Application for approval to import Mainman for release (APP202145)
February 2015
4. Hazardous properties
The staff determined the hazard profile of Mainman (Table 1) as described in Appendix C. 4.1.
The classifications determined by the staff are different to those submitted by the applicant (Table 1). 4.2.
The difference in the 6.1 (oral) classification is because the applicant used the toxicity of the active
ingredient, flonicamid, to determine the classification, rather than the results of the acute oral toxicity
test performed with Mainman. The staff determined that the 6.1 classification did not apply to Mainman.
Likewise the applicant anticipated that flonicamid, and therefore Mainman would be classified 6.9B but
the staff determined that this classification should not apply to either flonicamid or Mainman. The
applicant and the study author did not consider that the formulation should be classified as an eye
irritant, based on European Union classification criteria. However the HSNO criteria for 6.4A
classification are met (mean Draize score ≥ 1, but < 3, for corneal opacity). The 9.1D classification was
applied by the applicant because Mainman is intended to be biocidal. However, given that the staff
have assigned the 9.3C classification there is no need for the 9.1D biocidal classification. The staff has
assigned the 9.3C classification based on data submitted by the applicant for flonicamid and data on
other components of Mainman.
Table 1 Hazard classifications of Mainman as proposed by the applicant and the staff
Hazard Endpoint Applicant classification EPA classification
Acute toxicity (oral) 6.1E -
Eye irritation - 6.4A
Target organ systemic toxicity 6.9B -
Aquatic ecotoxicity 9.1D -
Ecotoxic to terrestrial vertebrates - 9.3C
5. Submissions
The submissions received by the EPA are attached as Appendix A. The key points from the 5.1.
submissions have been grouped into related issues and are discussed, alongside the EPA staff
response, below. All submissions were sent to the applicant.
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Application for approval to import Mainman for release (APP202145)
February 2015
Matters related to issues broader than this application
Submissions
Ngāi Tahu expressed dissatisfaction with the level of toxicity and ecotoxicity information that is made 5.2.
publically available with applications; the lack of testing on New Zealand native species; and that the full
composition of the substance, impurities in the active ingredient and other commercially sensitive
information are not disclosed to the public.
EPA staff response to the submissions
The staff note that these matters have been raised by Ngāi Tahu in submissions to previous 5.3.
applications. The Committee has previously stated that these matters are not specific to the
consideration of individual applications but relate to the requirements of the Act itself, and the EPA’s
approach to implementing the Act1. Therefore these matters are not discussed further in this evaluation
report but can be found in the submissions attached as Appendix A. The staff also note that the EPA
has, and continues to, work with Ngāi Tahu and applicants on these issues outside the application
process.
Risks
Submissions
Ngāi Tahu commented that the applicant did not provide any evidence to support their statement that 5.4.
they were “unaware of any significant or adverse impact the importation, release and use of the
substance would have on Māori cultural spiritual, ethical and social-economic values”, and questioned if
the applicant had considered the potential cultural risks of this substance.
The NBA and Federated Farmers have both raised concerns about the potential impacts of Mainman on 5.5.
honey bees. The NBA notes that the active ingredient of Mainman, flonicamid, is systemic i.e. it is
transmitted throughout the plant, and this may result in honey bees being exposed to flonicamid via
nectar or pollen of flowering plants. They request that a control is placed on Mainman to prevent it
being applied to flowering plants or immediately before flowering. Federated Farmers had similar
concerns and note that bees could also be exposed if Mainman was applied to flowering weeds in the
crop or crop margins, even if the crop itself was not in flower. They request that an enforceable label
warning is placed on Mainman to warn users about the risks to bees.
1 Recommendations in the Decision for application APP201999 – Solvigo.
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Application for approval to import Mainman for release (APP202145)
February 2015
Ngāi Tahu have raised concerns about the risks to aquatic and terrestrial species, due to the toxicity 5.6.
values cited in the application form, specifically the ‘low concentrations’ tested (Trout LD502 >100 mg/L;
Quail LD50 >2000 mg/kg/day; bees LD50 (oral) >60 µg/bee, LD50 (contact) >100 µg/bee, soil organisms;
earthworm NOEC3 >1000 mg/kg)
4.
The NBA notes this application only includes use on potatoes (GAP table, section 5.1 of the application 5.7.
form) but that similar substances from the same company are approved for use on brassicas, pome fruit
and stone fruit overseas. The NBA notes use on potatoes is low risk to bees because potatoes are not
a significant source of pollen for bees and do not rely on bees for pollination. However, they are
concerned about market creep and off-label uses – i.e. use on other crops that may be higher risk.
Ngāi Tahu have noted that the applicant did not include information on the metabolites of flonicamid in 5.8.
the application form, and that the DT505 of flonicamid in water was presented (DT50 =34 days), but that
no values for degradation in the soil were presented. They also note that it is impossible for them to
assess the risks of the metabolites because those metabolites are not named in the application form.
EPA staff response to the submissions
The staff note that the applicant sought guidance from members of the EPA’s Te Herenga network, 5.9.
including Ngāi Tahu, to provide assistance in identifying the impact on Māori cultural spiritual, ethical
and social-economic values. The responses received best related to the decision making process,
rather than the application itself. Ngāi Tahu indicated that they preferred to contribute at the public
submissions stage rather than the consultation stage of the application process.
The staff note that the toxicity test values (LD50 and NOEL) cited in the application form are relatively 5.10.
high and that the values for aquatic and soil environments and terrestrial invertebrates (bees) are below
thresholds for the respective HSNO classifications i.e. these classifications are not triggered. The LD50
test determines the dose at which 50% of the test population would be killed. High LD50 values indicate
lower toxicity to the test organism.
The staff note that Mainman has low toxicity to adult bees, hence it does not trigger a 9.4 classification. 5.11.
However, this test for 9.4 classification is only based on the acute effect to adult bees. The EPA’s risk
2 LD50 is the statistically or experimentally determined dose at which 50% of the test population dies.
3 NOEC is the no observed effect concentration.
4 Values quoted in the submission
5DT50 is the time required for 50% of the substances to degrade.
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Application for approval to import Mainman for release (APP202145)
February 2015
assessment considers a broader range of human health and ecological outcomes than those
considered to determine the classification of a substance.
The chronic effects of a substance and effects to other life stages are considered as part of the EPA’s 5.12.
full risk assessment which considers a broader range of human health and ecological outcomes than
those required to determine the classification of a substance. The studies submitted by the applicant
(Appendix C of this report) indicate that flonicamid could affect bee larvae; therefore the staff have
proposed controls to restrict the use of Mainman on flowering plants.
Regarding the LD50 values presented in the application form, the staff note that the concentrations 5.13.
tested are high and correspond to low toxicity. The toxicity testing guidelines allow limit tests to be
conducted when a product is expected to be of low toxicity. In a limit test, a high concentration is tested
first and if the result is not observed then there is no need to repeat the test at lower concentrations.
The staff note that the LD50 values presented in the application form for trout, bees and soil organisms
are below the threshold for triggering the respective hazard classifications.
The staff have noted the broader use pattern of Mainman, and similar products overseas, and have 5.14.
included additional scenarios in their risk assessments to account for possible future uses in New
Zealand.
The staff agree that limited information on metabolites and degradation is provided in the application 5.15.
form, but acknowledge that this is probably because it is not requested in the application form. The
staff also note that sufficient information on the metabolism and environmental fate of Mainman and its
active ingredient were provided with the application to allow the risk assessment to be completed.
Benefits
Submissions
Ngāi Tahu have noted that the benefits listed by the applicant are limited to the substance containing a 5.16.
‘new active ingredient which has a different mode of chemistry’, which will ‘minimise the risk of pest
resistance developing’ and a product with a ‘lesser hazard classification’ than existing insecticides, and
that the application does not provide a persuasive case for the benefits of this substance.
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Application for approval to import Mainman for release (APP202145)
February 2015
EPA staff response to the submissions
The staff agree that limited information was provided about the benefits, but that this is consistent with 5.17.
the level of benefits information typically provided with applications. The staff consider that the benefits
mentioned are valid.
Controls
Submissions
The NBA have indicated that they would like the EPA to apply the E3 control, regulation 49 from the 5.18.
Hazardous Substances (Classes 6, 8 and 9) Regulations 2001. This control prohibits the application of
a substance to flowering plants and plants that are expected to flower within a set period (up to 10
days).
Federated Farmers have similar concerns to the NBA, and are particularly concerned that the product 5.19.
could be applied to flowering weeds in or around a crop even if the crop is not attractive to bees.
Federated farmers have indicated that they would like to see clear enforceable warning about the risks
to bees on product labels. They have suggested the following wording for warning statements that
should appear on the label.
The product should not be used on horticultural crops during daylight hours when bees are
foraging on bee attractive weeds in the crop or flowers in headlands and /or paddock margins.
The product should not be used on horticultural crops during daylight hours when bees are
foraging.
Federated Farmers were also concerned about the warnings on the product label, practically that these 5.20.
were not clear and did not adequately address the risks posed by the substance. They have suggested
that the following warning appear on the label and that each warning is presented on a separate line to
improve clarity.
Harmful – may be harmful [if] swallowed, inhaled, or absorbed through the skin
May cause damage to the hematopoietic system from prolonged or repeated exposure
Designed for biocidal action against specific insects
Avoid spray drift to reduce harmful effects on beneficial insects in nearby habitats
Mainman shall not be applied directly onto, over or into water
Avoid contamination of any water supply with product or empty container
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Application for approval to import Mainman for release (APP202145)
February 2015
EPA staff response to the submissions
The staff agreed with the submitters that the E3 control should be applied. The staff took submitters 5.21.
comments about label directions into consideration when drafting the proposed controls. The staff
consider that the controls recommended in this document will mitigate the risks to people and the
environment.
Other matters
Submissions
Ngāi Tahu also note that the application is incomplete because it is not signed and witnessed as 5.22.
required.
EPA staff response to the submissions
The statutory declaration section of the EPA application form is only required to be completed by 5.23.
applicants seeking a rapid application under section 28A of the Act. This application is a full
assessment under section 28 of the Act and the statutory declaration is not required, and the
application form is appropriately signed.
6. Risk and benefit assessment
The staff conducted a qualitative risk assessment covering all stages of Mainman’s lifecycle, and a full 6.1.
quantitative human health and ecotoxicological risk assessment for the use phase. Full details of those
assessments are shown in Appendices E to G. This section provides a summary of the staff
assessment of the risks and benefits of Mainman.
Risks during manufacture, packaging and importation
The applicant intends to import Mainman packaged for sale, and manufacturing or repackaging in New 6.2.
Zealand is not anticipated. However if manufacturing or packing were to occur in New Zealand, the
HSNO requirement’s for equipment, emergency management and personal protective equipment (PPE)
would apply as well as WorkSafe New Zealand’s health and safety requirements. Assuming
compliance with the default controls and other relevant legislation, the staff consider that the risks to
people during import, manufacture and packaging would be negligible.
Risks during transport and storage
Workers and bystanders will only be exposed to the substance during transport and storage in isolated 6.3.
incidents where spills occur. Once in New Zealand, HSNO controls (e.g. labels, SDS, packaging) and
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Application for approval to import Mainman for release (APP202145)
February 2015
adherence to the Land Transport Rule 45001, Civil Aviation Act 1990 and Maritime Transport Act 1994
(as applicable) will apply. Therefore, the likelihood of people or the environment being exposed to
Mainman while it is being transported is so low that the resulting level of risk is considered negligible.
Risks during use
In addition to the use on potatoes described in the application form, the applicant intends to develop the 6.4.
use of Mainman on other crops in future but has requested that these uses are kept confidential. These
uses have been included in the risk assessment and identified as confidential use 1 to 5.
Human health effects
The quantitative human health risk assessment, detailed in Appendix F, determined that the risks to 6.5.
operators, bystanders and re-entry workers are acceptable (i.e. below the acceptable operator
exposure level (AOEL)) for the proposed uses of Mainman.
The exposure modelling indicated that risks for operators and re-entry workers are acceptable even 6.6.
without the use of PPE, and controls requiring PPE are not triggered by the hazard classifications of
Mainman. However, it is considered good practice to use of PPE when handling agrichemicals
therefore the staff recommend applying a labelling control that states that:
“Personal protective equipment (PPE) should be worn if entering areas treated with the
substance before the spray has dried”
or words to this effect. The minimum of PPE recommended is chemical resistant gloves, long sleeved
trousers and long sleeved shirt.
Environmental effects
Due to the proposed use pattern of Mainman, target and non-target organisms in the environment are 6.7.
expected to be exposed to this substance. Therefore a quantitative risk assessment was completed for
the active ingredient in Mainman, flonicamid. The risk assessment included consideration of the
metabolites of flonicamid.
The environmental risk assessment showed that Mainman poses a low risk to the environment when 6.8.
used according to the proposed application rates and label instructions. It was considered harmful to
terrestrial vertebrates (9.3C) but did not trigger aquatic (9.1), soil (9.2) or terrestrial invertebrates (9.4)
classifications.
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Application for approval to import Mainman for release (APP202145)
February 2015
The potential effect on bees and other beneficial insects has been noted by the submitters. The 6.9.
ecological risk assessment showed Mainman posed a low risk to adult bees and non-target arthropods
(e.g. predatory mites (Typhlodromus pyri), parasitic wasp (Aphidius rhopalosiphi), lacewing
(Chrysoperla carnea), hoverfly (Episyrphus balteatus) and ladybird (Coccinella septempunctata)).
However, several studies submitted with the application reported increased mortality and behavioural 6.10.
impairments (e.g. nervousness, dis-coordinated movement and apathy). In one study the effects were
observed on the day after the substance was applied, when the application occurred during foraging but
not when the substance was applied at the end of the day after foraging had ceased.
Consequently the staff consider that the use of Mainman should be restricted to plants (including weeds 6.11.
within and around the crop) that are not in flower, to minimise the likelihood that foraging bees will be
exposed to the substance. Therefore, the E3 Control6 has been in the draft controls under section 77
(3) of the Act.
It is considered that, with the default and additional controls in place, the risks to the environment from 6.12.
the use of Mainman will mitigate the level of risk to negligible.
Risks during disposal
The applicant indicates that the substance should be disposed of via an appropriate waste facility 6.13.
according to local by-laws, and that containers can be triple rinsed and disposed of at a landfill that is
consented to receive that type of waste or recycled.
Disposal of the substance and used containers in accordance with the requirements of the Hazardous 6.14.
Substances (Disposal) Regulations 2001 will ensure individuals and the environment are not exposed
to the substance. The level of risk during the disposal stage of the lifecycle is considered negligible.
Relationship of Māori to the Environment
The potential effects on the relationship of Māori to the environment have been assessed in accordance 6.15.
with sections 6(d) and 8 of the HSNO Act. Under these sections everyone exercising functions,
powers, and duties under the Act must take into account the relationship of Māori and their ancestral
lands, their water, their taonga and the principles of the Treaty of Waitangi (te Tiriti o Waitangi).
6 Regulation 49 of the Hazardous Substances (Classes 6,8 and 9) Regulations 2001
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In consideration of these requirements, this section provides an evaluation of impacts (positive and 6.16.
negative) to kaitiakitanga and the Treaty of Waitangi.
Kaitiakitanga
Kaitiakitanga7 is the exercise of guardianship of natural and physical resources by the tangata whenua 6.17.
of an area in accordance with tikanga Māori. A kaitiakitanga role has many functions including enabling
the protection of the life-supporting capacity of resources; maintaining the character and spiritual
integrity of those resource; as well as preserving them for future generations.
Members of the EPA’s Te Herenga network were given the opportunity to comment on this application 6.18.
prior to the public notification process. Although no specific information was provided during
consultation, the staff identified some key cultural elements that are relevant to this application.
In particular, the staff note that native pollinators are a taonga species with a critical role for kaitiaki in 6.19.
maintaining native food sources, rongoa and forest regeneration. In addition, the use of chemicals in
the environment always poses the potential for disruption to the mauri of native and valued species and
ecosystems, and consequently the ability of Māori to manage such impacts in their kaitiakitanga role.
Information relating to bees and other invertebrates is outlined elsewhere in this report. Though there is 6.20.
no specific information available on the impacts to native pollinators – including native bee species –
the staff note that Mainman is not intended for use in native ecosystems where such species are likely
to be more abundant. The use of the substance in highly modified horticultural regions also lends to a
conclusion that negative impacts on native species and ecosystems are likely to be minimal, particularly
in light of the ecotoxicology risk assessment outlined elsewhere in this report. .
In addition, with the controls proposed in this report (particularly those relating to labelling) , the likely 6.21.
use pattern, and environmental risk assessment, the staff consider that any potential impact to the
relationship of Māori to the environment will be negligible.
Treaty of Waitangi
In determining the impacts and relevance of the principles of the Treaty of Waitangi, we refer to the 6.22.
principles of partnership, participation and protection. These principles are regularly referenced by the
courts and the Waitangi Tribunal.
7 As defined in the Resource Management Act (1991); kaitiakitanga is not specifically defined in the HSNO Act.
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The principles of partnership and participation refer to the “shared obligation on both the Crown and 6.23.
Māori to act reasonably, honourably and in good faith towards each other to ensure the making of
informed decisions on matters affecting the interests of Māori”.
Additionally, the Waitangi tribunal has previously recommended that “environmental matters, especially 6.24.
as they may affect Māori access to traditional food resource also require consultation with Māori people
concerned.”
The EPA’s Engagement with Māori Policy requires that consultation be undertaken both prior to and 6.25.
throughout the application process where significant impacts to Māori interests are posed. Also, that
such consultation should lead to the effective exchange of information between the applicant and Māori,
as appropriate. Another purpose of consultation in this context is to provide decision-makers with
information to enable them to effectively evaluate the risks, costs and benefits of the substance, and to
make better informed decisions. This is in accordance with the decision-makers legal duty under the
HSNO Act.
In April 2014 and on behalf of the applicant, the EPA sent out summary information on the Mainman 6.26.
draft application to Te Herenga (the EPA’s national network of Māori resource managers, practitioners
and experts in the environment space) requesting enquires or feedback regarding the application. No
feedback was received that would be of benefit to this application.
In June 2014 the application was open for public submissions. Ngāi Tahu provided a submission on the 6.27.
application during this period.
Ngāi Tahu’s submission questioned whether the applicant had provided sufficient evidence to show that 6.28.
Mainman and its breakdown products pose no risks to terrestrial and aquatic environments. Ngāi Tahu
also note that the benefits of releasing further agrichemicals should be explicitly spelt out.
The Crown's settlement with Ngāi Tahu includes recognition on the traditional relationship Ngāi Tahu 6.29.
have with their taonga species. The settlement recognised that Ngāi Tahu and the government have
similar objectives in environmental and conservation management – that being to protect and enhance
what is special about New Zealand for future generations.
Taonga species, under the Ngāi Tahu Claims Settlement Act 1998 (“NTCS Act”) include native birds, 6.30.
plants and animals. The area of cultural significance and importance to Ngāi Tahu is defined by the Te
Rūnanga o Ngāi Tahu Act 1996 as being most of the South Island (excluding a northern segment) and
the islands to the south including Stewart Island (Rakiura) and others.
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Ngā Kaihautū and the submitters also touched on the absence of data from the Mainman application 6.31.
and commented that the data that was made available was extremely limited, lacking reference to any
relevant research. Ngāi Tahu continually comment on the lack of data made available within the
submission period, noting that due to this very fact it was impossible to evaluate the potential for risk.
The approval of an application for a hazardous substance, where significant uncertainty exists regarding 6.32.
the potential for adverse effects on taonga species and traditional Māori values and practices, may be
viewed as being inconsistent with the principle of active protection.
Whilst these frustrations are acknowledged, the staff note that given the risk assessment provided in 6.33.
this report, the level of uncertainty relating to the potential for impact to taonga species, Māori values
and practices is unlikely to be significant.
The staff have also considered that with proposed controls and measures in place, no significant risks 6.34.
are posed. With this in mind, the staff consider that the application is unlikely to be inconsistent with the
principles of the Treaty of Waitangi.
New Zealand’s international obligations
The staff have not identified any international obligations that might be affected by the use of Mainman. 6.35.
Assessment of benefits
The active ingredient of Mainman, flonicamid, has a different mode of action to currently available 6.36.
products. This is important for managing pesticide resistance, because it disrupts the natural
development of resistance to any particular class of active ingredient. The staff agree that the
availability of alternative chemistries is beneficial for growers.
The applicant also notes that Mainman has less hazards and lower hazard classifications than other 6.37.
insecticides used for the same purpose, and can therefore be considered a safer alternative to those
products. The low toxicity of Mainman means that it poses less risk to users than alternative products.
The staff consider that less hazardous products are beneficial to users both from a human health and
ease of use perspective.
The staff consider that Mainman will provide a benefit to growers and regional economic benefits to 6.38.
small organisations. Therefore the staff consider that there will be significant non-negligible benefits
associated with the approval of Mainman.
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The effects of the substance being unavailable
The effects of the substance being unavailable relate to fewer tools for disease management and 6.39.
pesticide resistance management. This could result in an adverse impact on crop quality and flow-on
economic impacts for individual growers.
7. Controls
A set of default controls are specified by regulations under the Act, based on the hazard classification 7.1.
determined for Mainman and form the basis of the controls set out in Appendix H. Based on the risk
assessment, the following additions, variations and deletions are considered applicable to Mainman.
Setting exposure limits
Tolerable Exposure Limits (TELs) and Acceptable Daily Exposure (ADE), Potential Daily Exposure 7.2.
(PDE) values can be set to control hazardous substances entering the environment in quantities
sufficient to present a risk to people. No TELs have been set for any component of Mainman. ADE and
PDE values have been set for flonicamid and the staff consider that these values are applicable to
Mainman; therefore the following PDE values have been applied:
ADE = 0.073 mg/kg bw/day8
PDEfood = 0.0511 mg/kg bw/day
PDEdrinking water = 0.0146 mg/kg bw/day
PDE other = 0.0073 mg/kg bw/day
The EPA typically adopts Workplace Exposure Standard (WES) values listed in WorkSafe New Zealand 7.3.
WES document9 to control exposure in places of work. WES values exist for component G but the staff
recommend that these should not apply to Mainman due to the low concentration of this component in
the substance.
Environmental Exposure Limits (EELs) can be set to limit hazardous substances from entering the 7.4.
environment in quantities sufficient to present a risk to it. The quantitative ecotoxicological risk
assessment (Appendix G) indicated that the risk posed to the soil environment was negligible and the
8 Units are mg of active ingredicent per kilogram bodyweitht per day
9 http://www.business.govt.nz/healthandsafetygroup/information-guidance/all-guidance-items/workplace-exposure-standards-
and-biological-exposure-indices/workplace-exposure-standards-and-biological-indices-2013.pdf
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risks to the aquatic environment were non-negligible. The staff consider that preventing exposure of the
aquatic environment to Mainman is the most effective way to manage the risks to the aquatic
environment. Consequently the proposed controls, set out in Appendix H, restricting the substance
from being applied to water, are recommended. No EELs have been proposed for any component of
Mainman at this time because the risk of adverse effects to the environment has been qualitatively
assessed as negligible provided that users comply with the controls as set out in Appendix H. The
default EEL values have been deleted.
Variation and deletion of controls
No controls have been varied or deleted. 7.5.
Additional controls
As no EEL has been proposed for Mainman, a maximum application rate is not required to be set under 7.6.
regulation 48 of the Hazardous Substances (Classes 6, 8, and 9 Controls) Regulations 2001. However,
the assessment of risk only takes into account the use patterns proposed by the applicant. The risk of
using higher application rates or different use patterns is unknown. Therefore it is considered
appropriate to take a precautionary approach and apply controls to limit the application of Mainman to
the established use-pattern parameters. Consequently a maximum application rate has been set under
section 77A of the Act. The following additional control is applied to Mainman to restrict the level of risk
to the environment:
The maximum application rate of this substance is 80 g of flonicamid/ ha.
The staff note that the active ingredient in Mainman, flonicamid, is associated with the toxicologically 7.7.
significant impurity toluene. When present in high enough concentrations such impurities may cause
adverse effects to people and/or the environment. Imposing a restriction on the maximum amount of
toluene that can be present in the active ingredient used to manufacture Mainman will prevent the
impurity from occurring in concentrations sufficient to cause adverse effects to people or the
environment. Consequently, the following additional control is applied to Mainman:
The flonicamid component of this substance must not contain more than 3 g/kg of toluene.
Due to the proposed use pattern as an insecticide for commercial horticultural crops, Mainman is 7.8.
expected to be applied in a wide dispersive manner, and significant environmental exposure could
occur. Staff note that it is not unusual for horticultural fields to be boarded by ditches, water-races, or
streams which may be inhabited by local and introduced aquatic species, or be used for drinking, stock
or irrigation water. Therefore people or the environment could be affected if Mainman was applied
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Application for approval to import Mainman for release (APP202145)
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directly to water. Staff also note that this substance is not intended to be applied directly onto water
and has therefore not been assessed for such uses in this application. Staff propose applying the
precautionary approach (section 7 of the Act) and limiting the use to this substance to terrestrial
applications. Consequently, the following additional control is applied to Mainman to restrict the level of
risk to the environment:
This substance must not be applied onto, into or over surface water10
The staff note that although Mainman does not trigger a 9.4 (invertebrate ecotoxicity) classification, the 7.9.
ecological risk assessment identified risks to bees. Specifically, several tunnel and field studies
reported bee mortalities and sub-lethal effects on the day of treatment when bees are directly exposed
to the spray of Mainman. Therefore it is considered appropriate to apply controls to limit the application
of Mainman to crops or areas that are not attractive to bees; this includes non-flowering crops if
flowering weeds are present in the crop or field margins.
Control E3 (Section 49 of the Hazardous Substances (Classes 6, 8 and 9) Amendment Regulations 7.10.
2004) includes requirements for the protection of terrestrial invertebrates (including bees) where a
substance is sprayed into the environment. As Mainman is not classified as a 9.4 substance, this
control does not form part of the default controls and therefore the staff recommend applying the control
under section 77(3)(a) of the Act.
The proposed additional controls set certain requirements or restrictions on the users of the substance, 7.11.
such as a maximum application rate. The staff consider that the most effective way to communicate
these requirements to the user is through the substance label. Other communication mechanisms (e.g.
safety data sheets) can be used to convey this type of information, but label statements are more likely
to be read every time a substance is used, so are considered the most effective way of communicating
key requirements for a particular substance. Therefore, the staff recommend that the following
statements, or words to this effect, are provided on the label for Mainman.
“Do not apply this product to any plant likely to be visited by bees at the time of application;
or immediately after application until spray has dried; or in areas where bees are foraging"
The maximum application rate of this substance is 80 g of flonicamid/ ha
10 where ‘water‘ means water in all its physical forms, whether flowing or not, but does not include water in any form while in a pipe,
tank or cistern or water used in the dilution of the substance prior to application or water used to rinse the container after use
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The staff note that the exposure assessment for operators and re-entry workers showed that the risks of 7.12.
the proposed uses was acceptable even without PPE and the default controls requiring PPE were not
triggered. However it is considered good practice to wear PPE when handling agrichemicals and this
should be communicated to users. Therefore the staff recommend adding the following statement, or
words to this effect, on the substance label:
Do not allow entry into treated areas until the spray has dried, unless wearing cotton overalls
buttoned to the neck and wrist (or equivalent clothing) and chemical resistant gloves.
Clothing must be laundered after each day’s use
The staff note that the application of the proposed additional controls will be more effective than the 7.13.
specified (default) controls in terms of its effect on the management, use and risks of Mainman.
Environmental user charges
The staff consider that applying controls to Mainman is an effective means of managing risks associated 7.14.
with this substance. Therefore, it is not considered necessary to apply environmental user charges to
this substance as an alternative or additional means of achieving effective risk management.
Accordingly, no report has been made to the Minister for the Environment.
Review of controls for cost-effectiveness
The staff consider that the proposed controls are the most cost-effective means of managing the 7.15.
identified potential risks and costs associated with this substance.
8. Overall evaluation and recommendation
The benefits assessment indicated that there were non-negligible benefits of Mainman to human health, 8.1.
the environment and the local economy.
The risk assessment indicates that there is a negligible level of risk to society, community and the local 8.2.
economy when using Mainman with the proposed controls in place.
The cultural risk assessment indicated that there were potential impacts on kaitiakitanga, but that the 8.3.
default and additional controls could appropriately manage the cultural impacts of this substance, and
that approval of this application is unlikely to be inconsistent with the principles of the Treaty of
Waitangi.
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With the proposed controls in place, the overall level of benefit provided by the availability of Mainman 8.4.
will outweigh the overall risk. Therefore, it is recommended that the application be approved with the
controls documented in Appendix H, in accordance with clause 26 of the Methodology.
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Appendix A: Submissions received
The submissions received for Mainman are available on the EPA website at http://www.epa.govt.nz/search-
databases/Pages/applications-details.aspx?appID=APP202145#
(Click on the tag “Documents” of the webpage)
For future reference, the ‘keyword’ reference number for this application is APP202145.
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Appendix B: EPA standard risk descriptors
Table 2 shows the risk descriptors used by the staff. Risk descriptors are determined by a combination of
the magnitude of risk and the likelihood of it occurring. Table 3 shows the likelihood descriptors used, and
Tables 4 and 5 the magnitude descriptors used to assess risks and benefits respectively.
These tables are taken from Decision Making. A Technical Guide to Identifying, Assessing and
Evaluating Risks, Costs and Benefits (March 2009), ER-TG-05-02 03/09, further information about these
risk descriptors can be found in this document.
Table 2 Risk descriptors
Likelihood
Magnitude
Minimal Minor Moderate Major Massive
Highly improbable Negligible Negligible Negligible Low Low
Very unlikely Negligible Negligible Low Low Medium
Unlikely Negligible Low Low Medium Medium
Likely Low Low Medium Medium High
Highly likely Low Medium Medium High High
Table 3 Likelihood Descriptors
Descriptor Description
Highly improbable Almost certainly not occurring but cannot be totally ruled out
Very unlikely Considered only to occur in very unusual circumstances
Unlikely (occasional) Could occur, but is not expected to occur under normal operating conditions
Likely A good chance that it may occur under normal operating conditions.
Highly likely Almost certain, or expected to occur if all conditions met
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Table 4 Magnitude of adverse effect
Descriptor Examples of descriptions
Minimal
Mild reversible short term adverse health effects to individuals in highly localised area
Highly localised and contained environmental impact, affecting a few (less than ten)
individuals members of communities of flora or fauna,
no discernible ecosystem impact
Local/regional short-term adverse economic effects on small organisations (businesses,
individuals), temporary job losses
No social disruption
Minor
Mild reversible short term adverse health effects to identified and isolated groups
Localised and contained reversible environmental impact, some local plant or animal
communities temporarily damaged, no discernible ecosystem impact or species damage
Regional adverse economic effects on small organisations (businesses ,individuals)
lasting less than six months, temporary job losses
Potential social disruption (community placed on alert)
Moderate
Minor irreversible health effects to individuals and/or reversible medium term adverse
health effects to larger (but surrounding) community (requiring hospitalisation)
Measurable long term damage to local plant and animal communities,
but no obvious spread beyond defined boundaries, medium term
individual ecosystem damage, no species damage
Medium term (one to five years) regional adverse economic effects with
some national implications, medium term job losses
Some social disruption (e.g. people delayed)
Major
Significant irreversible adverse health effects affecting individuals and requiring
hospitalisation and/or reversible adverse health effects reaching beyond the immediate
community
Long term/irreversible damage to localised ecosystem but no species loss
Measurable adverse effect on GDP, some long term (more than five years) job losses
Social disruption to surrounding community, including some evacuations
Massive
Significant irreversible adverse health effects reaching beyond the immediate community
and/or deaths
Extensive irreversible ecosystem damage, including species loss
Significant on-going adverse effect on GDP, long term job losses on a national basis
Major social disruption with entire surrounding area evacuated and impacts on wider
community
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Table 5 Magnitude of beneficial effect
Descriptor Examples of descriptions
Minimal
Mild short term positive health effects to individuals in highly localised area
Highly localised and contained environmental impact, affecting a few (less than ten)
individuals members of communities of flora or fauna, no discernible ecosystem impact
Local/regional short-term beneficial economic effects on small
organisations (businesses, individuals), temporary job creation
No social effect
Minor
Mild short term beneficial health effects to identified and isolated groups
Localised and contained beneficial environmental impact, no discernible ecosystem
impact
Regional beneficial economic effects on small organisations (businesses, individuals)
lasting less than six months, temporary job creation
Minor localised community benefit
Moderate
Minor health benefits to individuals and/or medium term health impacts on larger (but
surrounding) community and health status groups
Measurable benefit to localised plant and animal communities expected to pertain to
medium term
Medium term (one to five years) regional beneficial economic effects with some national
implications, medium term job creation
Local community and some individuals beyond immediate community
receive social benefit
Major Significant beneficial health
Major
Significant beneficial health effects to localised community and specific groups in wider
community
Long term benefit to localised ecosystem(s)
Measurable beneficial effect on GDP, some long term (more than five years) job creation
Substantial social benefit to surrounding community, and individuals in the wider
community.
Massive
Significant long term beneficial health effects to the wider community
Long term, wide spread benefits to species and/or ecosystems
Significant on-going effect beneficial on GDP, long term job creation on a
national basis
Major social benefit affecting wider community
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Appendix C: Classification of Mainman
Unless otherwise noted, all studies were conducted according to Good Laboratory Practice (GLP) and were
fully compliant with all requirements of the standard international test methods used. The classifications for
Mainman are shown in Table 6.
Data quality – overall evaluation
It is acknowledged that there are frequently data gaps in the hazard classification for chemicals which have
been in use internationally for a long time. International programmes such as the OECD High Production
Volume Programme11
, REACH12,
and European Regulation 1107/2009/EC13
are progressively working
towards filling these data gaps. As new information becomes available the staff will update the Hazardous
Substances and New Organisms (HSNO) classifications for those substances.
Table 6 Applicant and staff classifications of the mixture
Hazard Class/Subclass
Mixture classification Method of
classification
Remarks Applicant’s
classification
Staff’s
classification
Mix
ture
data
Read
acro
ss
Mix
ture
rule
s14
Class 1 Explosiveness No No Expert statement
instead of test
Class 2, 3 & 4 Flammability No No
Class 5 Oxidisers/Organic
Peroxides No No
Expert statement
instead of test
Subclass 8.1 Metallic
corrosiveness No ND
Subclass 6.1 Acute toxicity (oral) 6.1E No
The applicant based
this on the data for
the active not for
Mainman
Subclass 6.1Acute toxicity No No
11 OECD (1990) Manual for Investigation of HPV Chemicals. Retrieved on 23 January 2008 at http://www.icca-chem.org/Home/ICCA-
initiatives/High-production-volume-chemicals-initiative-HPV/
12 http://ec.europa.eu/environment/chemicals/reach/reach_intro.htm
13 http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2009:309:0001:0050:EN:PDF
14 Use of mixture rules may not adequately take into account interactions between different components in some circumstances and
must be considered of lower reliability than substance (formulation) data.
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(dermal)
Subclass 6.1 Acute toxicity
(inhalation) No No
Subclass 6.1 Aspiration hazard No
Subclass 6.3/8.2 Skin
irritancy/corrosion No No
Subclass 6.4/8.3 Eye
irritancy/corrosion No 6.4A
The applicant and the
study author did not
consider that the
formulation should be
classified as an eye
irritant, based on
European Union
classification criteria.
However the HSNO
criteria for 6.4A
classification are met
(mean Draize score
≥1, but <3, for corneal
opacity.)
Subclass 6.5A Respiratory
sensitisation No ND
Subclass 6.5B Contact
sensitisation No No
Subclass 6.6 Mutagenicity No ND
Subclass 6.7 Carcinogenicity No ND
Subclass 6.8 Reproductive/
developmental toxicity No ND
Subclass 6.8 Reproductive/
developmental toxicity (via
lactation)
No ND
Subclass 6.9 Target organ
systemic toxicity15
6.9B ND
The applicant
anticipated that
flonicamid would be
classified 6.9B but
staff have not applied
this classification to
flonicamid.
Subclass 9.1 Aquatic ecotoxicity 9.1D based on
biocidal action No
Given the 9.3C
classification there is
15 When appropriate include separate rows to address single as well as repeat dose target organ toxicity, and any of the relevant routes
oral, dermal or inhalation.
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no need for 9.1D
biocidal classification
Subclass 9.2 Soil ecotoxicity No No
Subclass 9.3 Terrestrial
vertebrate ecotoxicity No 9.3C
Flonicamid and
component E
Subclass 9.4 Terrestrial
invertebrate ecotoxicity No No
ND: No Data or poor quality data (according to Klimisch criteria16
) --> There is lack of data. No: Not Classified based on actual relevant data available for the substance --> The data are conclusive and indicate the threshold for classification is not triggered
Physico-chemical properties of the mixture Table 7 Physical and chemical properties of the mixture
Property Results Test method Klimisch Score
(1-4) Reference
Colour brown Visual
observation 1
B. de Ryckel (2005) Shelf-
life stability of IKI-220
50%WG. Walloon
Agricultural Research centre
(CRA-W) Pesticides
Research Department B-
5030 Gembloux- Belgium.
Report number: ISK/FO
20353/Ch.2531/2001/207
Physical state
Free flowing
cylindrical
granules
Visual
observation 1
Odour Slight odour of
ammonia - 1
Oxidizing properties
Does not
contain
oxidizing
components
Expert
Statement 1
U Schmiedel (2001) Expert
statement on the oxidizing
properties of IKI-220 50%
WG. RCC Ltd,
Environmental Chemistry &
Pharmanalytics Division.
CH-4452 Itingen Switzerland
Report no 834107
pH 8.3 (1% in
distilled water) CIPAC MT 75.2 1
B. de Ryckel (2005) Shelf-
life stability of IKI-220
50%WG. Walloon
Agricultural Research centre
(CRA-W) Pesticides
Research Department B-
5030 Gembloux- Belgium.
Report number: ISK/FO
20353/Ch.2531/2001/207
16 Klimisch,, H-J., Andrear, M., & U. Tillmann, 1997. A systematic approach for evaluating the quality of experimental toxicological and
ecotoxicological data. Reg. Toxicol. Pharmacol. 25, 1–5 (1997)
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Explosive properties
Does not
contain
explosive
components
Expert
Statement 1
U Schmiedel (2001) Expert
statement on the explosive
properties of IKI-220 50%
WG. RCC Ltd,
Environmental Chemistry &
Pharmanalytics Division.
CH-4452 Itingen Switzerland
Report no 834096
Tap Density
0.543 g/mL
before
compaction,
0.582 g/mL
after
compaction
CIPAC MT 169 1
B. de Ryckel (2005) Shelf-
life stability of IKI-220
50%WG. Walloon
Agricultural Research centre
(CRA-W) Pesticides
Research Department B-
5030 Gembloux- Belgium.
Report number: ISK/FO
20353/Ch.2531/2001/207
Particle size distribution
90% particles >
250 µm
10% particles <
850 µm
CIPAC MT 170 1
Dustiness
Collected dust
9.2 mg: nearly
dust free
CIPAC MT 171 1
Attrition resistance and
friability of granules 97.1% CIPAC MT 178 1
Flammability
Does not
contain
flammable
components
Evaluation of the
classification of the
components
Mammalian toxicology - Robust study summaries for the mixture
Acute toxicity [6.1]
Acute Oral Toxicity [6.1 (oral)]
In the following study summaries Mainman is referred to as IKI-220 50%WG.
Acute toxicity
Acute toxicity [6.1]
Acute Oral Toxicity [6.1 (oral)]
Type of study Acute oral limit toxicity test in rats
Flag Key study
Test Substance IKI-220 50%WG, Batch No. N88-0012-1
Endpoint LD50
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Value >2000 mg/kg bw (limit dose)
Reference Damme, B., IKI-220 50%WG, Acute oral toxicity study in rats, RCC Ltd,
Toxicology Division, Switzerland, 2001. ISK Document No.2606-822633
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 423, 96/54/EEC, US EPA OPPTS 798.1175
Species Rat
Strain Han Brl: WIST
No/Sex/Group 3M and 3F
Dose Level 2000 mg/kg bw
Exposure Type Oral gavage
Study Summary There were no deaths, no adverse signs of reaction to treatment or any
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion
IKI-220 50%WG should not be classified for 6.1 (oral), based on the lack of
adverse effects at the limit dose used in this study (Section 10.2; User Guide for
Thresholds and Classifications; EPA0109, 2012).
Acute Dermal Toxicity [6.1 (dermal)]
Type of study Acute dermal limit toxicity test in rats
Flag Key study
Test Substance IKI-220 50%WG, Batch No. N88-0012-1
Endpoint LD50
Value >2000 mg/kg bw (limit dose)
Reference Damme, B., IKI-220 50%WG, Acute dermal toxicity study in rats, RCC Ltd,
Toxicology Division, Switzerland, 2002. ISK Document No.2618-842020
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 402, 92/69/EEC, US EPA OPPTS 870.1200
Species Rat
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Strain HanBrl: WIST
No/Sex/Group 5M and 5F
Dose Level 2000 mg/kg bw
Exposure Type Dermal to clipped skin (≥ 10% body surface area) with semi-occlusive dressing for
24 hours
Study Summary
There were no deaths. Slight spots of erythema were noted in 2 females on Days
2 and 3 and a single male on Day 2. There were no other clinical findings or any
macroscopic changes at necropsy
Additional Comments No additional comments
Conclusion
IKI-220 50%WG should not be classified for 6.1 (dermal), based on the lack of
adverse effects at the limit dose used in this study (Section 10.2; User Guide for
Thresholds and Classifications; EPA0109, 2012).
Acute Inhalation Toxicity [6.1 (inhalation)]
Type of study Acute inhalation limit toxicity test in rats (4-hour, nose only)
Flag Key study
Test Substance IKI-220 50%WG, Batch No. N88-0012-1
Endpoint LC50
Value >5.360 mg/l (chemically determined mean concentration of formulated test item in
the test atmosphere)
Reference Decker, U., IKI-220 50%WG, 4-hour acute inhalation toxicity study in rats, RCC
Ltd, Toxicology Division, Switzerland, 2002. ISK Document No.2619-822666
Klimisch Score 1
Amendments/Deviations None of any significance
GLP Y
Test Guideline/s OECD 403, 92/69/EEC, US EPA OPPTS 870.1300
Species Rat
Strain HanBrl: WIST
No/Sex/Group 5M and 5F
Dose Level
5.360 mg/l
mass median aerodynamic diameters and (geometric standard deviations) were
3.09 µm (3.29) and 3.07 µm (3.18) respectively (measured on two occasions)
Exposure Type 4h nose only – dust aerosol
Study summary There were no deaths. Clinical signs consisted of bradypnea, rales (noisy
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breathing), red secretion from the nose, decreased activity, ruffled fur and
hunched posture observed from 1 hour after the end of treatment through to Day1
post exposure. Salivation was noted during the exposure period only. Bradypnea
and particularly rales persisted in both sexes for up to 9 days after exposure
before resolving. Marked transient weight loss was evident in all males and 4
females, the remaining female showing moderate weight loss during the first 3
days after exposure; however, recovery was evident by Day 8. There were no
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion
IKI-220 50%WG should not be classified for 6.1 (inhalation), based on the lack of
adverse effects at the limit concentration used in this study (Section 10.2; User
Guide for Thresholds and Classifications, EPA 0109, 2012).
Skin Irritation [6.3/8.2]
Type of study Dermal irritation/corrosion study in rabbits
Flag Key study
Test Substance IKI-220 50%WG, Batch No. N88-0012-1
Endpoint Skin irritation (mean of Draize scores at 24, 48 and 72 hours)
Value Non-irritant: 0.0 for erythema and oedema
Reference Arcelin, G., IKI-220 50%WG, Primary skin irritation study in rabbits, RCC Ltd,
Toxicology Division, Switzerland, 2001. ISK Document No.2607-822644
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 404, EEC B.4, US EPA OPPTS 870.2500
Species Rabbit
Strain New Zealand White
No/Sex/Group 1M and 2F
Dose Level 0.5 g (moistened with distilled water)
Exposure Type 4h dermal, semi occlusive then rinsed
Study Summary There were no skin reactions in any animal. Scores for erythema/eschar and
oedema were 0 for all observations
Additional Comments No additional comments
Conclusion IKI-220 50%WG should not be classified for 6.3 (skin irritation) based on the
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results from this study (Section 11.2; User Guide for Thresholds and
Classifications; EPA0109, 2012).
Eye Irritation [6.4/8.3]
Type of study Eye irritation/corrosion study in rabbits
Flag Key study
Test Substance IKI-220 50%WG, Batch No. N88-0012-1
Endpoint Eye irritancy (mean of Draize scores at 24, 48 and 72 hours)
Value Irritant: 1.78 conjunctival redness, 1.22 corneal opacity, 1 chemosis, 0 iritis
Reference Arcelin, G., IKI-220 50%WG, Primary eye irritation study in rabbits, RCC Ltd,
Toxicology Division, Switzerland, 2001. ISK Document No.2612-822655
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 405, EEC B.5, US EPA OPPTS 870.2400
Species Rabbit
Strain New Zealand White
No/Sex/Group 1M and 2F
Dose Level 0.1 g undiluted
Exposure Type Ocular
Study Summary
Corneal opacities were seen in all animals from 24 hours after exposure persisting
to Day 10 post exposure in the male, Day 14 in one female and Day 2 in the
remaining female. There was no iritis. Slight to moderate reddening of the
conjunctivae was seen in the male from 1 hour to Day10 after treatment and from
1 hour to Day 7 or Day14 in the two females. Swelling of the conjunctivae (with
partial eversion of the lids or lids half closed) was evident in all animals 1 hour
after treatment. Swelling persisted in 2 animals for up to 72 hours and 24 hours in
the remaining animal. The mean scores were: corneal opacity 1.22, iritis 0,
conjunctivae redness 1.78 and chemosis 1
Additional Comments No additional comments
Conclusion
IKI-220 50%WG should be classified for 6.4 (eye irritation) based on the results
from this study (mean scores for corneal opacity above the threshold for
classification) (Section 12.2; User Guide for Thresholds and Classifications;
EPA0109, 2012).
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Contact Sensitisation [6.5]
Type of study Sensitisation – modified Buehler test in guinea pigs
Flag Key study
Test Substance IKI-220 50%WG, Batch No. N88-0012-1
Endpoint Contact sensitisation on challenge
Value Non sensitiser: <thresholds (negative)
Reference
Arcelin, G., IKI-220 50%WG, Contact hypersensitivity in albino guinea pigs, Buhler
test, RCC Ltd, Toxicology Division, Switzerland, 2002. ISK Document No. 2611-
823882
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 406, EEC B.5, US EPA 870.2600
Species Guinea pig
Strain Ibm: GOHI (Himalayan spotted)
No/Sex/Group 10F control group, 20F treated group
Dose Levels 15% in distilled water, once per week for 3 weeks induction phase. 15% in distilled
water for challenge (maximum non-irritating concentration)
Exposure Type Dermal – 6 hours
Study Summary
No skin reactions were evident after the challenge dose. Comparison was against
concurrent laboratory background data using the positive control 2-
mercaptobenzothiazole
Additional Comments No additional comments
Conclusion
IKI-220 50%WG should not be classified for 6.5B (contact sensitisation) based on
the results from this study (Section 13.2; User Guide for Thresholds and
Classifications; EPA0109, 2012).
General conclusion about acute toxicity classification:
IKI-220 50%WG was shown to have low acute toxicity to rats by the oral, dermal and inhalation routes, and
should be not classified: 6.1 (oral), 6.1 (dermal) or 6.1 (inhalation) (Table 10.1; User Guide for Thresholds
and Classifications; EPA0109, 2012).
IKI-220 50%WG was shown not to induce skin irritation of sufficient severity to require classification for 6.3,
however the scores for eye irritation are slightly over the threshold for classification and therefore IKI-220
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50%WG should be classified 6.4 (Sections 11.2 and 12.2; User Guide for Thresholds and Classifications;
EPA0109, 2012).
IKI-220 50%WG was shown not to have contact sensitisation potential, and should not be classified for 6.5B
(contact sensitisation) (Section 13.2; User Guide for Thresholds and Classifications; EPA0109, 2012).
Dermal absorption
The applicant submitted two dermal absorption studies with IKI-220 50%WG. The first was conducted at a
concentration similar to the lowest field use dilution (0.4 g flonicamid/L), while the second was conducted at
the highest field use dilution (0.07 g flonicamid/L).
Type of study Percutaneous penetration study with human skin in vitro
Flag Key study
Test Substance Field use dilution of [14
C]-IKI-220 50% WG, Lot No. CP-2173
Reference
Roper, C. S., The in vitro percutaneous absorption of radiolabelled
insecticide [14
C]-IKI-220 50% WG at one concentration (worst case
concentration field dilution) of a test preparation through human skin,
Inveresk Research, Scotland, 2005. ISK Document No. 2629
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 428, EEC Sanco/222/rev.6 2004
Test system/Dose levels
[14
C]-IKI-220 was applied in a single preparation (in-use spray dilution, ca
0.4g IKI-220.L-1
) to split-thickness skin membranes mounted in flow
through diffusion cells in-vitro. The dose was 10 µL.cm2 to give a target
application rate of IKI-220 of 0.4 µg.cm2 for 6 hours. Absorption was
assessed over 0 – 48 hours
Study Summary
[14
C]-IKI-220 was applied topically to human skin in vitro at a single
concentration (in-use spray dilution, ca 0.4g IKI-220/L). Most of the applied
dose was washed off the skin at 6 hours post dose with 92.52% of the
dose recovered in the 6h mean skin wash and tissue swab combined. At
6h post dose, the mean absorbed dose was 4.22% (0.18 µg equiv.cm2). At
48 hours post dose the mean absorbed dose and dermal delivery were
7.31% (0.31 µg equiv.cm2) and 7.46% (0.31 µg equiv.cm
2) respectively.
Additional Comments
Reviewers comment: the study report claims that the dilution tested was
the worst case field use dilution. However, a higher dilution is now
proposed for use of IKI-220 50% WG.
Conclusion Penetration of IKI-220 50%WG through human skin membranes was at a
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negligibly low rate and extent at the ‘worst case scenario in use spray
dilution’.
Type of study Percutaneous penetration study with human skin in vitro
Flag Key study
Test Substance Field use dilution of IKI-220 50% WG containing 3H-IKI-220
Reference
Gelis, C., In vitro percutaneous absorption of IKI-220. ADME
BIOANALYSES 75, chemin de Sommières, 30310 VERGEZE, France,
2005. ISK Document No. UL-2632.
Klimisch Score
Amendments/Deviations None considered to impact the integrity of the study
GLP Y
Test Guideline/s OECD 428
Test system/Dose levels
Rat and human split-thickness skin membranes
Highest in use dilution (0.07 g IKI-220/L)
10 mg/cm2 of formulation applied for 6 hours
Four static cells per species with continuous stirring
Study Summary
The diluted formulation was applied to the surface of dermatomed rat and
human skin mounted in static cells. Six hours after application the
formulation remaining on the skin surface was removed. Receptor fluid
was collected 1, 2, 4, 7, 8 and 24 hours after application.
Mean recoveries were 95.68% and 95.43% in humans and rats,
respectively.
The dermal penetration of IKI-220 was calculated based on the sum of
radioactivity measured in the stratum corneum, epidermis, dermis and
receptor fluid. Dermal penetration was 47.34% in rat skin and 12.83% in
human skin.
Additional Comments None
Conclusion Dermal penetration of IKI-220 is 12.83% and 47.34% in human and rat
skin, respectively.
Conclusion on dermal absorption
The dermal penetration of IKI-220 formulated as IKI-220 50% WG in human skin was 7% at the lowest field
use dilution of 0.4 g IKI-220/L. At the highest field use dilution of 0.07 g IKI-220/L, the dermal absorption in
human skin was 13%.
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Ecotoxicity - Robust study summaries for the mixture
In the following summaries Mainman is called IKI-220 50% WG.
Aquatic toxicity
Fish Acute Toxicity (Freshwater Species)
Type of study Full test
Flag Key study
Test Substance Formulation: IKI-220 50% WG
Species Oncorhynchus mykiss (Rainbow trout)
Type of exposure Semi-static for 96 hours. Renewal after 48 hours
Endpoint LC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of IKI-220 50% WG to Rainbow trout
(Oncorhynchus mykiss) in a 96-hour semi-static test. RCC Ltd, Environmental
Chemistry & Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no
823307.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 7
Dose Levels 6.3, 12.5, 25, 50 and 100 mg/L
Analytical measurements
Yes from fresh medium on day 0 and at 48 h and from the old medium at 48 and
96 hours, by HPLC. All concentrations were analysed except 6.3 and 12.5 mg/L
because they were below the NOEC.
Study Summary
The acute toxicity of the test substance to rainbow trout was determined in a 96-hr
semi-static test. The test concentrations were 6.3, 12.5, 25, 50 and 100 mg/L.
The measured concentrations in the test medium were 94-101% of the nominal at
the start and at the end of the test substance so the results are expressed as
nominal concentrations.
The pH values in the test medium and in the control ranged from 7.8 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 7.3
mg/L or above) and the temperature was in the range of 13-15oC.
No mortalities were observed in any group.
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In the control and in the test concentrations up to 25 mg/L, no visible
abnormalities were observed, at 50 mg/L 1 fish was apathetic and at 100 mg/L, all
test fish showed symptoms of toxicity (apathy, tumbling during swimming, fish at
the bottom of the aquarium).
The LC50 is higher than 100 mg/L corresponding to > 51 mg a.i./L.
Conclusion The LC50 is higher than 100 mg/L (corresponding to > 51 mg a.i./L). No
mortality observed at 100 mg/L
Invertebrates Acute Toxicity (Freshwater Species)
Type of study Limit test
Flag Key study
Test Substance Formulation: IKI-220 50% WG
Species Daphnia magna
Type of exposure Static for 48 hours.
Endpoint EC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of IKI-220 50% WG to Daphnia magna in a 48-
immobilisation test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 823320.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202 (1984)
No/Group 2 replicates of 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium on day 0 and at 48 h, by HPLC.
Study Summary
The acute toxicity of the test substance to Daphnia magna was determined in a
48-hr static test. A limit test was performed at 100 mg/L. The measured
concentrations in the test medium were 99-102% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.9 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 8.3
mg/L or above) and the temperature was in the range of 21-22oC.
No immobilised or dead animals were observed in any group.
The EC50 is higher than 100 mg/L corresponding to > 51 mg a.i./L.
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Conclusion The EC50 is higher than 100 mg/L (corresponding to > 51 mg a.i./L). No
immobilisation observed at 100 mg/L
Algae Acute Toxicity (Freshwater Species)
Type of study Full test
Flag Key study
Test Substance IKI-220 50% WG
Species Pseudokirchneriella subcapitata
Type of exposure Static, 72 hours
Endpoint ErC50 and EbC50
Value > 100 mg/L (ErC50)
Reference
A. Peither (2002) Acute toxicity of IKI-220 50% WG to Pseudokirchneriella
subcapitata (formerly Selenastrum capricornutum) in a 72-hour algal growth
inhibition test. RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-
4452 Itingen Switzerland. Report no 823342.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201 (1984)
No/Group 3 replicates for test concentrations and 6 for the control of 104 cells/mL
Dose Levels 4.6, 10, 22, 46 and 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 72 h, by HPLC. The concentrations
below 22 mg/L were not analysed because they are below the NOEC.
Study Summary
The influence of the test substance on the growth of the green algal species
Pseudokirchneriella subcapitata was determined in a 72-hr static test. The
nominal concentrations were 4.6, 10, 22, 46 and 100 mg/L. The measured
concentrations in the test medium were 103-108% of the nominal at the start and
at the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.9 to 8.0 at the
start of the study and from 8.1 to 9.3 at the end, and the temperature was 23oC.
The growth rate in the control was valid according to the guideline (at least a
factor of 16).
The test substance had a statistically significant inhibitory effect on the growth
(biomass and growth rate) at the concentration of 46 mg/L and above. The NOEC
was 22 mg/L (corresponding to 11 mg ai/L) and ErC50 is > 100 mg/L
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(corresponding to > 51 mg ai/L), EbC50 = 85 mg/L (corresponding to 43 mg ai/L).
Conclusion ErC50 is > 100 mg/L (corresponding to > 51 mg ai/L), EbC50 = 85 mg/L
(corresponding to 43 mg ai/L).
Other study
Type of study Activated sludge respiration inhibition
Flag Key study
Test Substance IKI-220 50WG
Species Activated sludge of a wastewater treatment plant treating predominantly domestic
wastewater
Type of exposure 3 hours
Endpoint EC50
Value > 1000 mg/L
Reference
I Gruetzner (2002) Toxicity of IKI-220 50% WG to activated sludge in a respiration
inhibition test. RCC Ltd Environmental Chemistry & Pharmanalytics division CH-
4452 Itingen Switzerland. Report number 834118
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 209 (1984)
No/Group 1 replicates for test concentration and 2 for the control
Dose Levels 12, 32, 100, 320 and 1000 mg/L
Analytical measurements Not required
Study Summary
The inhibitory effect of the test item IKI-220 50% WG on the respiration rate of
aerobic wastewater microorganisms of activated sludge was investigated in a 3-
hour respiration inhibition test
The following nominal concentrations were tested: 12, 32, 100, 320 and 1000
mg/L.
In addition, two controls and three different concentrations of the reference item
3,5-dichlorophenol (5, 16 and 50 mg/L) were tested in parallel. The results of
these treatments confirmed the suitability of the activated sludge and the method
used.
Up to and including the highest concentration of 1000 mg/L, the test substance
had no significant inhibitory effect ( <15%) on the respiration rate of activated
sludge after the incubation period of three hours. Thus, the 3-hour NOEC is 1000
mg/L. EC50 is higher than 1000 mg/L.
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Conclusion EC50 is higher than 1000 mg/L.
General conclusion about aquatic toxicity classification:
Mainman exhibits a very low aquatic toxicity. No classification is triggered except a 9.1D for biocidal activity
which doesn’t apply because Mainman is also classified as 9.3C.
Soil toxicity
Non-Target Plant Toxicity
Study type Seedling emergence
Flag Key study
Test Substance IKI-220 50WG
Species
Cucumber (Cucumis sativus), corn (Zea mays), soybean (Glycine max), oats
(Avena sativa), tomato (Lycopersicon esculentum), radish (Raphanus sativus),
cabbage (Brassica oleracea), perennial ryegrass (Lolium perenne), carrot
(Daucus carotta), lettuce (Lactuca sativa), onion (Alium cepa)
Type of exposure Application to the soil surface
Endpoint Effects on emergence, heights and dry weight of seedling
Value No effects at 150 g ai/ha
Reference
E.R. Stewart (2002) Tier 1 seedling emergence study assessing IKI-220 50% WG
formulation. Stewart Agricultural Research Services Inc. 1893 Highway K
Clarence, Missouri 63437. Study number IB-2002-MG-003-00-00
Klimisch Score 1
Amendments/Deviations None that impacted the test results
GLP Yes
Test Guideline/s OPPTS 850.4000 and OPPTS 850.4100 (drafts, 1996)
No/Group 5 replicates of 10 seeds
Dose Levels 150 g ai/ha
Analytical measurements Not required
Study Summary
Replicates of 10 seeds of 7 dicotyledons species and 4 monocotyledons species
were planted in a sandy loam. Then the test substance was applied on soil
surface at a dose corresponding to 150 g ai/ha. The pots were incubated in
greenhouse conditions. Seedling emergence was assessed at 10, 14 and 21
days after planting and application. Seedlings were also examined for phytotoxic
effects or abnormalities at the same intervals. At the end of the test (21 days),
plant height was measured and dry weight was determined.
No phytotoxic symptoms or abnormalities were detected among treated plots or
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non-treated control. No significant differences were observed on average height
or on average dry weight of any species included in the test.
Conclusion No effects at 150 g ai/ha
Study type Vegetative vigour
Flag Key study
Test Substance IKI-220 50WG
Species
Cucumber (Cucumis sativus), corn (Zea mays), soybean (Glycine max), oats
(Avena sativa), tomato (Lycopersicon esculentum), radish (Raphanus sativus),
cabbage (Brassica oleracea), perennial ryegrass (Lolium perenne), carrot
(Daucus carotta), lettuce (Lactuca sativa), onion (Alium cepa)
Type of exposure Application on seedlings
Endpoint Effects on heights and dry weight of seedling, + phytotoxicity effects
Value No detrimental effects at 150 g ai/ha
Reference
E.R. Stewart (2003) Tier 1 vegetative vigour study assessing IKI-220 50% WG
formulation. Stewart Agricultural Research Services Inc. 1893 Highway K
Clarence, Missouri 63437. Study number IB-2002-MG-004-00-00
Klimisch Score 2: no information on the number of plants per replicates
Amendments/Deviations
The minimum number of plant per replicates (5 for dicots and 10 for monocots)
was not reached for some species and some replicates. No information in the
report about the actual number.
GLP Yes
Test Guideline/s OPPTS 850.4000 and OPPTS 850.4150 (drafts, 1996)
No/Group 5 replicates of an unknown number of plants
Dose Levels 150 g ai/ha
Analytical measurements Not required
Study Summary
7 dicotyledons species and 4 monocotyledons species were planted in a sandy
loam. Then the test substance was applied on seedlings (growth stage tillering for
oat and ryegrass, and 2 to 5-6 leaves for the other species) at a dose
corresponding to 150 g ai/ha. The pots were incubated in greenhouse conditions.
Plant heights were recorded just before application and at 15 days after
application. Plants were also examined for phytotoxic effects or abnormalities at
the 7 and 15 days. At the end of the test (16 days), dry weight was determined.
No phytotoxic symptoms or abnormalities were detected among treated plots or
non-treated control. No significant differences were observed on average height
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of any species included in the test.
Analysis of dry weights indicated a slight increase in ryegrass treated by the test
substance (0.425 g and 0.351 g for treated and controls plants, respectively).
There was no other effect on dry weight on any of the other species.
Conclusion No detrimental effects at 150 g ai/ha
Study type Effects on adjacent crops and succeeding crops
Flag Disregarded study
Test Substance IKI-220 50 WG
Species
Effects on adjacent plants: beet, lettuce, maize, melon, oat, oil seed rape, pea,
soybean, wheat, pepper, eggplant
Succeeding crops: beet, cucumber, eggplant, lettuce, maize, melon, oat, oil seed
rape, pea, pepper, soybean, wheat, turnip, kidney bean, flax, onion, barley
Type of exposure Effects on adjacent plants: foliar spray
Succeeding crops: soil treatment prior sowing
Endpoint Phytotoxicity
Value None observed
Reference
M Morita & M Iwasa (2002) Impact of IKI-220 50 WG (IBE-3894) on adjacent
crops and succeeding crops. Ishihara Sangyo Kaisha Ltd. Central Research
Institute. Agrochemical Research Laboratory, Biology Group.3-1, 2-Chome, Nishi-
shibukawa; Kusatsu-shi, Shiga-ken, 525-0025 Japan. Report no AL-0201-I-08
Klimisch Score
3 not GLP, no guideline followed, visual assessment of phytotoxicity only, no
biomass endpoint. Monocotylodons are under-represented. Very few details in
the report
Amendments/Deviations None
GLP No
Test Guideline/s None followed
No/Group
Not clear from the report. 6 replicates are mentioned for adjacent crops but no
information about the number of plant per replicate.
Succeeding crops: 5 to 20 plants per pot, no mention about the number of pot per
group.
Dose Levels
Effects on adjacent plants: 100 or 200 g ai/ha with volumes of 300, 500 and 1200
L/ha
Succeeding crops: 100 or 300 g ai/ha
Analytical measurements No
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Study Summary
Adjacent plants: plants grown in pots were treated with a spray solution of the
substance at 100 or 200 g ai/ha. All spray solutions were applied at volumes
rates of 300, 500 or 1200 L/ha using spray gun. After application the plants were
kept in a glasshouse. Phytotoxicity was visually assessed on days 7 and 14.
No phytotoxicity was observed in any conditions.
Succeeding crops: soil was treated with the spray solution of the substance at 100
or 300 g ai/ha using a flat spray tip. Seeds were sown in the treated soil; pots
were kept in a glasshouse. Phytotoxicity was visually assessed after 2, 3 and 4
weeks.
No phytotoxicity was observed in any conditions.
Conclusion No phytotoxicity observed after foliar application or soil treatment
Nitrogen Transformation Test
Study type Effects on nitrogen transformation
Flag Key study
Test Substance IKI-220 50 WG
Species Soil microflora
Type of exposure Static for 28 days
Endpoint Soil nitrogen content (nitrate, nitrite, ammonium, mineral nitrogen)
Value No significant long term effect
Reference
K-H Reis (2002) Effects of IKI-220 50% WG on the activity of the soil microflora in
the laboratory. IBACON GmbH Arheilger Weg 17 64380 Rossdorf Germany.
Report no 10722080
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study
GLP Yes
Test Guideline/s OECD 216 (2000)
No/Group 3 replicates
Dose Levels
0.104 mg/kg (corresponding to the worst case of 80 g ai)/ha for 1 application with
50 % foliage interception of IKI-220 50 % WG at 40 g ai)/ha) and 0.274 mg/kg soil
dry weight (corresponding to 3-times 70 g ai)/ha with 50 % foliage interception of
IKI-220 50 % WG at 105 g (ai)/ha).
Analytical measurements None required
Study Summary
The effects of the substance applied at 0.104 mg/kg and 0.274 mg/kg soil dry
weight on the soil nitrogen transformation of the soil micro-flora were tested in the
laboratory at 20 ± 2°C in the dark. At different sampling times (0, 7, 14 and 28
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days), the soil nitrogen content was determined.
The differences between the soil nitrogen content (nitrate-N) of the IKI-220 50%
WG treated soil and control was less than the trigger value of 25% according to
the OECD guideline 216 at days 0, 14 and 28.
At day 28 after application, the differences between the soil nitrate content of both
IKI-220 50% WG treated soils and control were 14.1% and 6.34% respectively.
The differences were statistically significant, but below the 25% threshold.
At day 28; the difference to control in mineral soil nitrogen content was clearly
below the trigger value of 25%.
The deviation of replicate control samples was below 15% at every sampling date
so the validity criteria were met.
Conclusion No long term influence on soil micro-flora when IKI-220 50% WG is applied
up to 0.274 mg/kg soil dry weight.
Carbon Transformation Test
Study type Effects on carbon transformation
Flag Key study
Test Substance IKI-220 50 WG
Species Soil microflora
Type of exposure Static for 28 days
Endpoint CO2 production
Value No significant long term effect
Reference
K-H Reis (2002) Effects of IKI-220 50% WG on the activity of the soil microflora in
the laboratory. IBACON GmbH Arheilger Weg 17 64380 Rossdorf Germany.
Report no 10722080
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study
GLP Yes
Test Guideline/s OECD 217 (2000)
No/Group 3 replicates
Dose Levels
0.104 mg/kg (corresponding to 80 g ai)/ha for 1 application with 50 % foliage
interception of IKI-220 50 % WG at 40 g ai)/ha) and 0.274 mg/kg soil dry weight
(corresponding to 3-times 70 g ai)/ha with 50 % foliage interception of IKI-220 50
% WG at 105 g ai)/ha ).
Analytical measurements None required
Study Summary The effects of the substance applied at 0.104 mg/kg and 0.274 mg/kg soil dry
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weight on the carbon mineralisation of the soil micro-flora were tested in the
laboratory at 20 ± 2°C in the dark. At different sampling times (0, 7, 14 and 28
days), the soil respiration rate was determined.
The soil respiration rates of IKI-220 50% WG treated soils were clearly below the
OECD guideline 217 trigger value of 25 % at every sampling date.
At day 28 after application, the differences between the respiration rates of IKI-
220 50% WG treated soil and control was 0.88% and 7.90% respectively. The
difference to control of the 1st rate group was not statistically significant different,
whereas the difference of the 2nd rate group was.
With exception of day 14, the deviation of replicate control samples was below 15
%.
Conclusion No long term influence on soil micro-flora when IKI-220 50% WG is applied
up to 0.274 mg/kg soil dry weight.
General conclusion about soil toxicity classification:
No information is available on other organisms than plants so mixture rules will be used to classify Mainman.
Toxicity to terrestrial invertebrates
Bees - Laboratory Tests (acute oral and contact)
Study type Acute toxicity
Flag Supporting study
Test Substance IKI-220 50 WG
Species Apis mellifera
Type of exposure Oral or contact, for 48 or 96 hours (oral test)
Endpoint LD50
Value Contact: > 100 µg/bee
Oral: > 104.3 µg/bee
Reference
S Schmitzer (2002) Laboratory testing for toxicity (acute contact and oral) of IKI-
220 50% WG on honey bees (Apis mellifera L). IBACON GmbH Arheilger Weg 17
64380 Rossdorf Germany. Report no 10721036
Klimisch Score
1 for the contact test
3 for the oral test: no dose response relationship is observed in the test,
consequently it is impossible to estimate a LD50. The value reported (> 104
µg/bee) is highly doubtful.
Amendments/Deviations None that impacted the results of the study
GLP Yes
Test Guideline/s EPPO 170 (1992)
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No/Group 3 replicates of 10 for the oral test
5 replicates of 10 for the contact test
Dose Levels Contact: 100 µg/bee
Oral: 6.8, 14.1, 28.0, 57.0, 104.3 µg/bee (actual intake)
Analytical measurements Not required
Study Summary
The acute toxicity of the substance on honey bees was determined in a laboratory
test with oral and contact exposure.
In the contact test, 10% mortality was observed at 100 µg product/bee after 48
hours. Behavioural impairments like discoordinated movement and apathy was
observed during the whole experiment.
In the oral test, the observation period was extended for 48 hours up to 96 hours
because of delayed mortality in all dose groups.
Oral doses of 104.3, 57.0, 28.0, 14.1 and 6.8 µg/bee led to 53.3%, 30.0%, 26.7,
53.3 and 23.3%, respectively. Behavioural abnormalities attributed to the
substance were discoordinated movement and apathy observed during the whole
experiment. The control mortality was 3.3%.
Control mortality was below the validity criteria of the guideline.
Conclusion
LD50 Contact: > 100 µg/bee (corresponding to 51.1 µg ai/bee)
LD50 Oral: > 104.3 µg/bee (corresponding to 53.3 µg ai/bee) – Unreliable
results
Bees - Tunnel Tests
Study type Tunnel test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera
Type of exposure Direct spray + exposure to residues on sprayed flowers
Endpoint Behaviour, mortality, development of colonies
Value Immediate repellent effect and slight increase of mortality the day of treatment.
High mortality due to direct spray
Reference
G Maurin (2000) Tunnel trail assessment of the product IBE 3894 – short-term
toxicity on honey bees. ACTA 149 rue de Bercy Paris, France. Study number
LYT0001
Klimisch Score 2, not GLP no statistical analysis, uneven behaviour of the bees between tunnels
Amendments/Deviations None
GLP No
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February 2015
Test Guideline/s CEB no 129 (1996)
No/Group 1 tunnel per group divided in 4 plots: 2 treated and 2 untreated used as refuge
Dose Levels 140 g IKI 220 50 WG/ha, “soft” reference: phosalone at 600 g ai/ha
Analytical measurements Not required
Study Summary
A tunnel test was conducted with the test substance to evaluate the effects of a
spray at 140 g/ha on foraging activity, bee behaviour, mortality and development
of colony.
3 tunnels were used over an oil seed rape crop in flower. Each tunnel was divided
in 4 plots: 2 were treated and 2 were left untreated as refuge for foraging bees.
Phosalone at 600 g ai/ha was used as soft reference. Control tunnel was sprayed
with water.
Hives were placed in the tunnel 4 days prior treatment. Foraging activity and
behaviour were observed once to 5 times per day, mortality was assessed once a
day and the colony status was determined before treatment and at the end of the
test (6 days after application).
An immediate (at 1 hour after treatment) repellent effect was observed in the
tunnel treated by the test substance. There was a slight increase in mortality due
to the test substance which was much lower than the direct effect of the spray.
Ratio of mortality before and after treatment is 3.27 for the test substance and
1.26 and 2.18 for the control and the reference, respectively.
There was no noticeable effect on colony development due to the test substance.
Comment
Foraging activity was not similar between tunnels (high activity in control tunnel,
very low in reference tunnel), consequently the mortality which is usually related to
the foraging activity was much higher in the control tunnel. However, there is a
clear pattern in the tunnel treated with the test substance with an increase of
mortality due to the direct spray.
Conclusion Immediate repellent effect and slight increase of mortality the day of
treatment. High mortality due to direct spray
Study type Tunnel test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera
Type of exposure Direct exposure to spray, or exposure to residues on flowers
Endpoint Effects on foraging activity, behaviour, mortality colony development.
Value Direct spray of the substance on bees has an effect on mortality and behaviour on
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the first day. No effects when the substance was sprayed when bees were not
foraging.
Dubious effects on colonies exposed to the substance, recovery within 6 weeks
Reference
M. Barnavon (2003) Tunnel test – acute and short-term effects of IBE 3894 (IKI-
220 50%WG), applied on oil seed rape, on honey bees (Apis mellifera). SOLEVI
SARL 12 rue Felix Perrier 26400 Crest France. Study report: S03ISB.F01VO33
Klimisch Score 1
Amendments/Deviations Pollen collect was impossible due to the small size of the pollen balls
GLP Yes
Test Guideline/s CEB no 129 (2003), EPPO 170 (1992)
No/Group 1 tunnel per group divided in 4 plots: 2 treated and 2 untreated used as refuge
Dose Levels 150 g IKI 220 50 WG/ha, toxic reference: dimethoate at 365 g ai/ha
Analytical measurements Not required
Study Summary
A tunnel test was conducted with the test substance to evaluate the effects of a
spray at 150 g product/ha on foraging activity, bee behaviour, mortality and
development of colony.
4 tunnels were used over an oil seed rape crop in flower. Each tunnel was divided
in 4 plots of 16 m2: 2 were treated and 2 were left untreated as refuge for foraging
bees.
Dimethoate at 365 g ai/ha was used as toxic reference. Control tunnel was
sprayed with water. The test substance was either applied in the morning when
bees were foraging, either in the late evening when no bees were foraging.
Hives were placed in the tunnel 2 days prior treatment. Foraging activity and
behaviour were observed once a day except on the treatment day (15 min before
and after treatment and 60 min after treatment), mortality was assessed once a
day and the colony status was determined before treatment and at the end of the
test (7 days after application).
In this study, during the adaptation phase bee mortality was medium. The day
before treatment, mortality was slightly higher in tunnel n°2 (treatment with the test
substance during foraging activity). In the water (control) tunnel, mortality was
stable after treatment except on day 6 after treatment where a peak was noticed
for all the tunnels. This peak was not treatment related but, most probably, due to
the high temperatures (up to 37° in the tunnels) these days. In the dimethoate
tunnel, mortality was very high after treatment and stayed still at a quite high level
in the following days.
In this context, for the test substance, a slight effect was only seen the day after
application when applied during foraging activity. When the substance was
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applied the same day but in the evening, when there was no more foraging
activity, there was no product effect. Differences in the tox index values of test
substance applied in the morning, in the evening and water were mainly due to
differences between colonies in the different tunnels except for the morning
application, 24 hours after application.
Tox Index = (Mt x Ta)/(Ma x Tt) where Mt and Ma are daily mortality in the treated
tunnel after and before treatment respectively and Tt and Ta are daily mortality in
the control tunnel after and before treatment respectively.
Tox index
Treatment 24 h 48 h 72 h 96 h 120 h
Dimethoate 41.7 23.4 18.3 10.8 8.9
Control 1.0 1.0 1.0 1.0 1.0
Morning
application
5.8 1.3 0.9 0.9 1.0
Evening
application
2.4 3.5 2.5 2.1 2.7
Thus, it is considered that the test substance has a weak effect on bee mortality
the day of application when the product is applied during foraging activity and then
this effect disappeared immediately. When it was applied out of foraging activity
period no effect was not seen.
Dimethoate had a strong and long impact on the foraging activity. The substance
had some impact on foraging activity in the hour following the application and then
this effect disappeared 4 hours after application and the following days when
applied during foraging activity. During this transient period, the observations
have shown that bees were presenting symptoms of loss of their equilibrium when
they tried to forage the flowers. This was not observed when the substance was
applied in the evening without foraging activity.
At the end of the exposure period, colonies belonging to the tunnels treated with
the test substance were slightly weakened compared to the colony belonging to
the water tunnel but stiII much lesser than the colony belonging to the dimethoate
tunnel. The queen activity was not disturbed. After return to the apiary, for the
last assessment, 6 weeks after treatment, colonies were recovering and at that
time only the colony having received the dimethoate treatment appeared as weak.
It was difficult to conclude that the effects seen in the substance tunnels were
product related.
Reserves and brood were reduced during the study in the four tunnels, which was
normal for beehives staying within a tunnel. This reduction was however higher in
the dimethoate tunnel. After return to the apiary, for the last assessment, 6 weeks
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after treatment, reserves were much higher in all the tunnels, much probably due
to the excellent weather conditions of the summer. The dimethoate colony was
still weaker than the others. There was no difference anymore between the three
other tunnels even if the colony belonging to the substance treatment out of
foraging activity period had a slight heterogeneous brood.
Conclusion
Direct spray of the substance on bees has an effect on mortality and
behaviour on the first day. No effects when the substance was sprayed
when bees were not foraging.
Dubious effects on colonies exposed to the substance, recovery within 6
weeks
Study type Tunnel test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera
Type of exposure Direct exposure to spray, and exposure to residues on flowers
Endpoint Effects on foraging activity, behaviour, mortality colony development.
Value Weak effect on mortality and on bee behaviour and foraging activity the day of
treatment
Reference
M. Barnavon (2002) Tunnel test – acute and short-term effects of IBE 3894 (IKI-
220 50%WG), applied on white mustard, on honey bees (Apis mellifera). SOLEVI
SARL 12 rue Felix Perrier 26400 Crest France. Study report: S03ISB.F01VO01
Klimisch Score 1
Amendments/Deviations Assessment of beehive after return to apiary not done due to bad weather
conditions
GLP Yes
Test Guideline/s CEB no 129 (1996), EPPO 170 (1992)
No/Group 2 tunnels for the treated group, 1 tunnel for water control divided in 4 plots: 2
treated and 2 untreated used as refuge
Dose Levels 160 g IKI 220 50 WG/ha
Analytical measurements Not required
Study Summary
A tunnel test was conducted with the test substance to evaluate the effects of a
spray at 160 g product /ha on foraging activity, bee behaviour, mortality and
development of colony.
3 tunnels were used over a white mustard crop in flower. Each tunnel was divided
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in 4 plots of 16 m2: 2 were treated and 2 were left untreated as refuge for foraging
bees.
Control tunnel was sprayed with water. The test substance was applied early in
the afternoon when bees were foraging.
Hives were placed in the tunnel 22 days prior treatment. Foraging activity and
behaviour were observed once a day except on the treatment day (60 min before
treatment and 15 min after treatment), mortality was assessed once a day and the
colony status was determined before treatment and at the end of the test (11 days
after application).
In this study, during the adaptation phase bee mortality was quite low. The
adaptation period was long due to the limited evolution of the crop stage (fresh
weather conditions).
The product effect was only seen immediately after application. The natural
mortality was the same for the three tunnels, respectively 41, 46 and 64 bees, at
the time of application and the number of bees foraging at the time of application
was between 191 and 270 bees for 85 m2. 46 and 12 bees died due test
substance in tunnel 2 and in tunnel 3 representing respectively less than 17 and
6.3% of the bees present at the treatment time.
The following days bee mortality was similar for the three tunnels. Thus, it is
considered that the substance has a weak effect on bee mortality the day of
application and then this effect disappeared quickly.
The substance had a very limited and transient impact on foraging activity
immediately after application in tunnel 3 and then this effect disappeared the
following days. This was not the case in tunnel 2.
Substance effects on bee behaviour were seen only the day of application and
nothing was observed later on.
After return to the apiary, no assessment was done due to the bad weather
conditions and the winter time.
Reserves and brood were reduced during the study in the three tunnels which was
normal for beehives staying five weeks within a tunnel. This reduction was similar
for the three tunnels.
Conclusion Weak effect on mortality and on bee behaviour and foraging activity the day
of treatment
Study type Tunnel test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera
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Type of exposure Direct exposure to spray, and exposure to residues on flowers
Endpoint Effects on foraging activity, behaviour, mortality colony development.
Value
Weak effect on mortality and on bee behaviour and foraging activity the day of
treatment.
Transient effect on bee colony
Reference
M. Barnavon (2002) Tunnel test – acute and short-term effects of IBE 3894 (IKI-
220 50%WG), applied on oil seed rape, on honey bees (Apis mellifera). SOLEVI
SARL 12 rue Felix Perrier 26400 Crest France. Study report: S02ISB.F01VO14
Klimisch Score 1
Amendments/Deviations Assessment of mortality was not done every day
GLP Yes
Test Guideline/s CEB no 129 (1996), EPPO 170 (1992)
No/Group 2 tunnels for the treated group, 1 tunnel for water control divided in 4 plots: 2
treated and 2 untreated used as refuge
Dose Levels 160 g IKI 220 50 WG/ha
Analytical measurements Not required
Study Summary
A tunnel test was conducted with the test substance to evaluate the effects of a
spray at 160 g product /ha on foraging activity, bee behaviour, mortality and
development of colony.
2 tunnels were used over oilseed rape crop in flower. Each tunnel was divided in
4 plots of 16 m2: 2 were treated and 2 were left untreated as refuge for foraging
bees.
Control tunnel was sprayed with water. The test substance was applied early in
the afternoon when bees were foraging.
Hives were placed in the tunnel 7 days prior treatment. Foraging activity and
behaviour were observed once a day except on the treatment day (60 min before
treatment and 15 min after treatment), mortality was assessed once a day and the
colony status was determined before treatment and at the end of the test (23 days
after application).
In this study, during the adaptation phase bee mortality was quite low.
The product effect was only seen immediately after application. The natural
mortality was the same for the 2 tunnels, respectively 57 and 41 bees, at the time
of application and the number of bees foraging at the time of application was
between 549 and 604 bees for 85 m2. 120 bees died due test substance
representing respectively less than 20% of the bees present at the treatment time.
The following days bee mortality was similar for the three tunnels. Thus, it is
considered that the substance has a weak effect on bee mortality the day of
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application and then this effect disappeared quickly.
The substance had a very limited and transient impact on foraging activity
immediately after application and then this effect disappeared the following days.
Substance effects on bee behaviour were seen only the day of application and
nothing was observed later on. Nevertheless, queen activity seemed to be
perturbed (laying activity was lower and even stopped). However at the end of the
test, 23 days after application, no difference was observed between control and
treated colonies.
After return to the apiary, no assessment was done due to the bad weather
conditions and the winter time.
Reserves and brood were reduced during the study in the 2 tunnels which was
normal for beehives staying within a tunnel. This reduction was higher for the
treated colony during the study but was not observed at the end of the study.
Conclusion
Weak effect on mortality and on bee behaviour and foraging activity the day
of treatment.
Transient effect on bee colony
Study type Tunnel test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera
Type of exposure Direct exposure to spray, and exposure to residues on flowers
Endpoint Effects on foraging activity, behaviour, mortality, storage of food, area of frames
covered by brood or food.
Value Effects on foraging activity, bee behaviour and number of bees entering and
leaving the hives limited to the day of treatment. No other effects were observed.
Reference
M. Barnavon (2000) Tunnel test – acute and short-term effects of IBE 3894,
applied on white mustard, on honey bees (Apis mellifera). SOLEVI SARL 12 rue
Felix Perrier 26400 Crest France. Study report: S00ISB008IVO55
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s CEB no 129 (1996), EPPO 170 (1992)
No/Group
1 tunnel for the treated group, for water control and non-toxic reference phosalone
(Zolone Flo at 1.2 L/ha) divided in 4 plots: 2 treated and 2 untreated used as
refuge
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Dose Levels 140 g IKI 220 50 WG/ha
Analytical measurements Not required
Study Summary
A tunnel test was conducted with the test substance to evaluate the effects of a
spray at 140 g product /ha on foraging activity, bee behaviour, mortality and
development of colony.
1 tunnel was used for each group over a white mustard crop in flower. Each
tunnel was divided in 4 plots of 16 m2: 2 were treated and 2 were left untreated as
refuge for foraging bees.
Control tunnel was sprayed with water. The test substance was applied early in
the afternoon when bees were foraging.
Hives were placed in the tunnel 6 days prior treatment. Foraging activity and
behaviour were observed twice a day except on the treatment day (15 min before
treatment and 15 min and 1 hour after treatment), mortality was assessed once a
day and the colony status was determined before treatment and 3 days after the
end of the 7-d exposure period.
There was no effect on bee mortality in all 3 tunnels, when pre-treatment and
post-treatment mortalities are compared.
The substance had a very limited and transient impact on foraging activity
immediately after application and then this effect disappeared the following days.
Substance effects on bee behaviour and number of bees entering and leaving the
hives were seen only the day of application and nothing was observed later on.
There was no effect on duration of flower visits.
There was no difference on the condition of the colony when compared with the
non-toxic reference.
Conclusion
Effects on foraging activity, bee behaviour and number of bees entering and
leaving the hives limited to the day of treatment. No other effects were
observed.
Bees - Field Tests
Study type Field test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera carnica
Type of exposure Exposure to dried residues on flower in field conditions
Endpoint Mortality, foraging activity (flight density), sublethal effects, colony assessment
(food stores, brood status, hive population), strength of colony
Value No effect
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February 2015
Reference
S. Schmitzer (2008) Toxicity testing of Teppeki (IKI-220 50% WG) on Honey bees
(Apis mellifera L.) in the field. IBACON GmbH Arheilger Weg 17 64380 Rossdorf
Germany. Report no 36061040
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s EPPO 170 (2000)
No/Group 1 treated plot + 1 untreated with 4 bee colonies
Dose Levels 80 g ai/ha (160 g formulation/ha in 400 L of water/ha)
Study Summary
The aim of the study was to determine the risk to honey bees after contact with
dried residues of the substance applied at 80 g ai/ha, under full field conditions.
Flowering crops (Phacelia tanacetifolia) were sprayed with the substance after
bee flight (evening application). The trial was carried out on 2 different plots of
about 2300 m2 and a distance of approximately 5 km. 4 bee colonies were set up
on each plot 4 days before application. The development of the colonies was
observed until test end on day 21 following application.
The mean number of dead bees per colony per day was 4.1 in the treated plot
after treatment and 24.3 before application. In control it was 13.4.
No statistical difference was observed in flight intensity: mean was 19.4
bees/m2/days on the treated plot and 16.3 in the control plot.
No poisoning symptom or marked reaction to exposure were noted at any time
until the end of the test.
No effects on brood were observed, each colony had a sufficient amount of all
brood stages without any indication of test item related effect.
No substance related influence on the overall strength of the colony were
observed, this was evaluated by counting the number of gaps occupied by bees.
In the treated colonies at days 14 and 21, the number was 93% of the initial value,
and 88 and 78% in the control plot.
Pollen was checked to confirm the exposure to the crop: 47-66% of the collected
pollen came from Phacelia.
Conclusion No effects at 80 g ai/ha in field conditions
Study type field test
Flag Weight of evidence
Test Substance IKI-220 50 WG
Species Apis mellifera carnica
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Type of exposure Exposure to dried residues on flower in field conditions
Endpoint
Mortality, foraging activity (flight density), sublethal effects, colony assessment
(food stores, brood status, hive population), strength of colony, brood
development to adulthood
Value Reduction of development success of eggs from to 3 d-old. No other effects.
Reference
L. Jeker (2008) Teppeki (IKI-220 50% WG) A field study to evaluate side effects
on Honey bees (Apis mellifera L.) following the application on Phacelia
tanacetifolia after daily bee-flight in Germany. . RCC Ltd, Zelgliweg 1 CH-4452
Itingen Switzerland Report no B47520
Klimisch Score 2 due to bias on the assessment of foraging activity and mortality in field (see
comment below)
Amendments/Deviations None that had impacted the results of the study
GLP Yes
Test Guideline/s EPPO 170 (2000)
No/Group 1 treated plot + 1 untreated with 4 bee colonies
Dose Levels 80 g ai/ha (160 g formulation/ha in 300 L of water/ha)
Study Summary
The aim of the study was to determine the effects to honey bees after contact with
dried residues of the substance applied at 80 g ai/ha in 300 L water /ha, under full
field conditions. Flowering crops (Phacelia tanacetifolia) were sprayed with the
substance after bee flight (evening application). The trial was carried out on 2
different plots of about 2000 m2 and a distance of approximately 3 km. 4 bee
colonies were set up on each plot 13 days before application. The development
of the colonies was observed until test end on day 25 following application.
The study is considered as valid: less than 15% of mortality is observed in control
brood stages (observed mortality 7.4%).
A statistically significant reduction of development success of eggs of 1 to 3 days
was observed in the colonies of the treated plot (78.0% vs 89.5% in control). No
effects were observed on other brood stages.
There were no significant effects on mortality of pupae and adult worker bees
assessed in bee traps; and on condition of colonies (% of comb covered by stores
or by life stages or by vacant cells).
No significant effects were observed on foraging activity and mortality in field.
Comment
There were more colonies in the neighbourhood of the control plot than of the
treated plot so more bees were foraging in the control plot. Consequently the
foraging activity and mortality in field assessment are biased in this study.
The report concludes that the effects on eggs are not biologically significant
because there is no overall effect on brood.
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Conclusion Reduction of development success of eggs from to 3 d-old. No other effects
Other non-target arthropods - Laboratory Tests
Study type Tests with glass plates
Flag Supportive study
Test Substance IKI-220 50 WG
Species Aphidius rhopalosiphi, T. pyri and C. septempunctata
Type of exposure Residues on glass plates, for 48 h for parasitic wasps, for 7 days for predatory
mites, till pupation for ladybirds
Endpoint Corrected mortality
Value
Aphidius rhopalosiphi: 22.2% at 80 g ai/ha, 55.5% at 210 g ai/ha
T. pyri: 100% at both application rates
C. septempunctata: 30% at both application rates
Reference
JP Jansen (2001) Screening test with IKI-220 50WG: side effects on Aphidius
rhopalosiphi, T. pyri and C. septempunctata on glass plates in the laboratory.
Agricultural Research Center - Department of Biological Control and Plant Genetic
Resources – Chemin de Liroux 2, B-5030 Gembloux Belgium. Report number
none
Klimisch Score 2 non GLP study but performed according to standard guidelines
Amendments/Deviations None
GLP No
Test Guideline/s
A. rhopalosiphi: Mead-Briggs, 1992;
T. pyri: Overmeer, 1988;
C. septempunctata: Pinsdorf, 1988
No/Group
A. rhopalosiphi test: 3 replicates of 5 males and 5 females for control and treated
groups, 1 replicate for the toxic reference (deltamethrin at 1 g ai/ha).
T. pyri test: 3 replicates of 20 protonymphs for all groups
C. septempunctata test: 20 replicates of 1 larvae, 10 replicates for the toxic
reference
Dose Levels 80 g and 210 g a.i./ha in 200 L water/ha
Study Summary
In order to determine the initial toxicity of IKI-220 50WG to adult of the parasitoid
wasp Aphidius rhopalosiphi, protonymph of the predatory mite Typhlodromus pyri
and larvae of the ladybird Coccinella septempunctata, a screening test was
carried out in the laboratory. Test product was applied at rates of 80 g and 210 g
(ai)/ha on glass plates. Additional glass plates were treated with deltamethrin (1 g
(ai)/ha) as toxic standard.
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A. rhopalosiphi : At the end of exposure, corrected mortality with IKI-220 50WG at
80 g and 210 g (ai)/ha were, respectively, 22.5% and 55.5%. Mortality in units
treated with toxic standard reached 100.0%.
C. septempuncata : Young ladybirds larvae were exposed to test product applied
on glass plates till pupation occurred. Preimaginal corrected mortalities reached
30% for IKI-220 50WG at both application rate and 100.0% for toxic standard.
T. pyri: Mortality of protonymphs exposed to IKI-220 50WG at both doses and to
toxic standard reached 100.0%.
Conclusion
Aphidius rhopalosiphi: 22.2% at 80 g ai/ha, 55.5% at 210 g ai/ha
T. pyri: 100% at both application rates
C. septempunctata: 30% at both application rates
Study type Test with application on inert surface (sand) in laboratory
Flag Key study
Test Substance IKI-220 50 WG
Species Poecilus cupreus
Type of exposure 14 d- exposure to dried residues on sand
Endpoint Corrected mortality
Value 3.3%
Reference
JP Jansen (2002) Side effects of IKI-220 50WG on adults of the carabid beetle
Poecilus cupreus L. (Coleoptera, carabidae) in the laboratory. Agricultural
Research Center - Department of Biological Control and Plant Genetic Resources
– Chemin de Liroux 2, B-5030 Gembloux Belgium. Report number PC.01/2001
Klimisch Score 1
Amendments/Deviations None that had an impact on the results
GLP Yes
Test Guideline/s Heimbach, 1994
No/Group 5 units of 3 males and 3 females for control, treated groups, and for the reference
substance (dimethoate).
Dose Levels
45 g ai/ha in 400 L water /ha (the report mentions that this is equivalent to a worst
case PEC after two applications of 80 g ai)/ha at 21 days interval for a soil-
dwelling species, assuming 50% crop interception)
Study Summary
A laboratory study was carried out to determine the effects of the insecticide IKI-
220 50WG on adults of the carabid beetle Poecilus cupreus. The test product
was applied at a rate of 45 g ai/ha to 5 exposure unit made of plastic boxes filled
65
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February 2015
with sand, each containing six beetles (3 males, 3 females). Additional groups of
5 exposure units were treated with water as control or dimethoate as toxic
standard. Beetles were fed with housefly pupae. Food consumption and
observation of the condition of the beetles were recorded throughout the 14 days
of the test.
After 14 days of exposure, no beetle was found dead in the units treated with
water and one with IKI-220 50WG. This corresponds to an observed and
corrected mortality of 3.3% for IKI-220 50WG. Three beetles were also found
affected in units treated with IKI-220 50WG between day 2 and day 4
assessments, but they recovered a normal behaviour at further assessment.
Mortality in the toxic standard reached 100.0%. No reduction of food consumption
was observed with IKl-220 50WG, compared with control.
These results showed that IKI-220 50WG is harmless (< 25% of effects) to adults
of the carabid beetle P. cupreus on inert surface in the laboratory.
Conclusion Corrected mortality: 3.3%
Other non-target arthropods - Extended Laboratory Tests
In the following studies, reduction of beneficial capacity (E) is calculated as follows:
E (%) = 100- {(100-M) x R}
with M = corrected mortality and R = reproductive ratio
Study type Test with bean leaves in laboratory
Flag Key study
Test Substance IKI-220 50 WG
Species Typhlodromus pyri
Type of exposure 7-d exposure to dried residues on bean leaves
Endpoint Mortality and fertility
Value No effect on fertility, 23.3% corrected mortality
Reference
JP Jansen (2002) Side effects of IKI-220 50WG on the predacious mite
Typhlodromus pyri Scheuten (Acari; Phytoseiidae) on detached bean leaves in the
laboratory. Agricultural Research Center - Department of Biological Control and
Plant Genetic Resources – Chemin de Liroux 2, B-5030 Gembloux Belgium.
Report number TE.01/2002
Klimisch Score 1
Amendments/Deviations None that had an impact on the results
GLP Yes
Test Guideline/s IOBC/WPRS 1988, Hassan, 1994
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February 2015
No/Group 5 units of 20 protonymphs for control and treated groups, 2 for the reference
substance (lambda-cyhalothrin at 1 g ai/ha).
Dose Levels
85 g ai/ha in 200 L water /ha (the report mentions that this is equivalent to 2
applications of 80 g ai/ha at 21 days of interval considering the degradation rate of
the active)
Study Summary
A study was carried out to determine the effects of the insecticide IKI-220 50WG
on protonymphs of the predacious mite T. pyri (Acari; Phytoseiidae) under
extended laboratory test conditions.
Test product was applied at a dose of 85 g (ai)/ha on detached bean leaves.
Additional leaves were treated with water as control and with lambda-cyhalothrin
as toxic standard. Two set of leaves were treated at the same time. First set was
used for initial exposure, second set for assessment of fertility the second week of
experiments.
After treatment, leaves were carefully placed on wetted cotton pad and twenty 2-3
day old protonymphs of T. pyri were confined on each leaf. 5 exposure units were
assembled for control and test product and 2 for toxic standard. After 7 days,
surviving mites were harvested, sexed and transferred to second set of leaves for
fecundity assessment. Mean mortality in each group was corrected with the value
observed in the control. After 7 days, the number of protonymphs and eggs found
in each unit was counted and mean fertility performance calculated. Based on
mortality and reproductive performance, reduction of beneficial capacity was
calculated for each group and products were rated according the IOBC
classification for extended-laboratory test.
At day 7 of exposure, a mean mortality of 14.5%, 34.4% and 100.0% was
observed in control, IKI-220 50WG and toxic standard, respectively. Corrected
mortality for IKI-220 50WG was 23.3%.
Mean egg production was 2.71 eggs/female and 2.87 eggs/female for control and
IKI-220 50WG, respectively. As no females survived in toxic standard units,
fertility assessment was not realised with this product.
Reduction of beneficial capacity was equal to 18.8% compared to control for IKI-
220 50WG and 100.0% for toxic standard. Based on these results, IKI-220 50WG
is considered as harmless (less than 25% of effects) on natural substrate in the
laboratory.
Conclusion No effect on fertility. Corrected mortality: 23.3%
Study type Test with barley seedlings in laboratory
Flag Key study
Test Substance IKI-220 50 WG
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Species Aphidius rhopalosiphi
Type of exposure 48-h exposure to dried residues on barley seedlings
Endpoint Mortality and fertility
Value No effect on fertility, 4.4% corrected mortality
Reference
JP Jansen (2002) Side effects of IKI-220 50WG on adults of the parasitic wasp
Aphidius rhopalosiphi (Hym., Aphidiidae) on plants in the laboratory. Agricultural
Research Center - Department of Biological Control and Plant Genetic Resources
– Chemin de Liroux 2, B-5030 Gembloux Belgium. Report number AE.03/2001
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s Mead-Briggs & Longley (1997), Hassan (1994)
No/Group 5 replicates of 5 wasps for control and treated groups, 2 for the reference
substance (dimethoate at 35 g ai/ha).
Dose Levels
85 g ai/ha in 400 L water /ha (the report mentions that this is equivalent to 2
applications of 80 g ai/ha at 21 days of interval considering the degradation rate of
the active)
Study Summary
A study was carried out to determine the effects of the insecticide IKI-220 50WG
on adults of the aphid parasitoid Aphidius rhopalosiphi (Hym; Aphidiidae) under
extended laboratory test conditions.
Test product was applied at a dose of 85 g ai/ha to barley seedlings. Additional
seedlings were treated with water as control and with dimethoate as toxic
standard. Five mated female wasps were confined on each seedling, using a
perspex cage, for a period of 48h. During exposure, wasps were observed 10
times and their position (treated or untreated surfaces) were noted to detect any
repellent effects. After 48 hours, surviving females were harvested, 14 were
selected randomly in each group - assuming that this number survive and were
assessed for reproductive performance. They were individually released for a
period of 24 hours onto barley seedlings infested with cereal aphids (Sitobion
avenae). 10 to 13 days later, number of parasitized aphids (mummies) was
counted in each seedling and mean mummy production calculated for each test
group. Based on mortality and reproductive performance, reduction of beneficial
capacity was calculated and products rated according to the IOBC classification
for extended-laboratory test.
After 48h of exposure to treated plants, 22/25 parasitoids were collected on plants
treated with IKI-220 50WG, 23/25 in the control and none in the toxic standard.
Corrected mortality for IKI-220 50WG was equal to 4.4 %. A mean production of
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8.14, and 7.36 mummies/female was obtained for control and IKI- 220 50WG. As
no females survived with toxic standard, no fertility assessment was realized with
this product.
Reduction in beneficial capacity with IKI-220 50WG was equal to 13.6%.
According to the IOBC rating, IKI-220 50WG can be considered as harmless (<
25% effects) to the parasitic wasp A. rhopalosiphi under extended laboratory test
conditions.
Conclusion No effect on fertility. Corrected mortality: 4.4%
Study type Test with bean leaves in laboratory
Flag Key study
Test Substance IKI-220 50 WG
Species Chrysoperla carnea
Type of exposure Exposure to dried residues on bean leaves, till pupation
Endpoint Mortality and fertility
Value No effect on fertility, 18.8% corrected mortality
Reference
JP Jansen (2002) Side effects of IKI-220 50WG on larvae of the green lacewing
Chrysoperla carnea (Neuroptera, Chrysopidae) on detached bean leaves treated
in the laboratory. Agricultural Research Center - Department of Biological Control
and Plant Genetic Resources – Chemin de Liroux 2, B-5030 Gembloux Belgium.
Report number CCE.02/2001
Klimisch Score 1
Amendments/Deviations None that had an impact on the results
GLP Yes
Test Guideline/s Bigler 1988
No/Group 40 units of 1 larvae for control and treated groups, 20 for the reference substance
(lambda-cyhalothrin at 1 g ai/ha).
Dose Levels
85 g ai/ha in 200 L water /ha (the report mentions that this is equivalent to 2
applications of 80 g ai/ha at 21 days of interval considering the degradation rate of
the active)
Study Summary
A study was carried out to determine the effects of the insecticide IKI-220 50WG
on larvae of the green lacewing C. carnea (Neuroptera; Chrysopidae) under
extended-lab test conditions.
IKI-220 50WG was applied at a rate of 85 g ai/ha. Additional leaves were treated
with water as control and with lambda-cyhalothrin as toxic standard.
After treatment, one 2-3-day old larvae of C. carnea was released in each unit.
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Larvae were fed 5 times a week with Ephestia Kuhniella sterilised eggs, till
pupation.
Larval mortality was assessed daily. After pupation, pupae were carefully
transferred to plastic petri dishes for adult emergence. After emergence,
observed preimaginal mortalities were calculated and corrected with the value of
the corresponding control. When they emerged, adults were transferred in fertility
assessment units and egg production was followed during a 4-week period. Mean
number of viable eggs/female/day was used as a measure of reproductive
performance.
A preimaginal mortality of 20.0% was obtained for control. With IKI-220 50WG,
observed and corrected preimaginal mortality reached 35.0% and 18.8%
respectively. All larvae were killed by toxic standard. A mean production of 157.1
and 186.3 viable eggs/female was obtained in control and IKI-220 50WG units.
As no females survived, assessment of fertility with toxic standard was not
possible.
According to corrected mortalities and mean viable egg production, reduction of
beneficial capacity with IKI-220 50WG was equal to 3.65%. These results show
that IKI-220 50WG is harmless for larvae of C. carnea under extended laboratory
conditions.
Conclusion No effect on fertility. Corrected mortality: 18.8%
Study type Test with broad bean leaves in laboratory
Flag Supportive study
Test Substance IKI-220 50 WG
Species Hoverfly Episyrphus balteatus
Type of exposure Exposure to dried residues on broad bean leaves, till pupation
Endpoint Mortality and fertility
Value No statistically significant effect on fertility, 2.6% corrected mortality
Reference
JP Jansen (2002) Side effects of IKI-220 50WG on larvae of the hoverfly
Episyrphus balteatus (DeGeer) (Diptera, Syrphidae) on plants in the laboratory.
Agricultural Research Center - Department of Biological Control and Plant Genetic
Resources – Chemin de Liroux 2, B-5030 Gembloux Belgium. Report number
EBE.01/2002
Klimisch Score 2: no existing guideline but the method similar to other non-target arthropods
method
Amendments/Deviations None that had an impact on the results
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February 2015
GLP Yes
Test Guideline/s None existing
No/Group 10 units of 5 larvae for control and treated groups, 5 for the reference substance
(lambda-cyhalothrin at 1 g ai/ha).
Dose Levels
85 g ai/ha in 400 L water /ha (the report mentions that this is equivalent to 2
applications of 80 g ai/ha at 21 days of interval considering the degradation rate of
the active)
Study Summary
A study was carried out to determine the effects of the insecticide IKI-220 50WG
on larvae of hoverfly E. balteatus (Diptera; Syrphidae) under extended-lab test
conditions. As no standardised and validated methods exist, trials were
conducted according to an original method. Test products were applied till run-off
on broad bean seedlings. IKI-220 50WG was applied at a rate of 85 g (ai)/ha, and
additional seedlings were treated with water as control and with lambda-
cyhalothrin as toxic standard. 2 to 3 hours after treatments, E.balteatus larvae
were confined on treated seedling till pupation. After adult emergence, observed
preimaginal mortalities were calculated and corrected with the value of the
corresponding control. When they emerged, adults were transferred in fertility
assessment units and egg production was followed during five 2-day periods.
Mean number of viable eggs/female was used as a measure of reproductive
performance.
A preimaginal mortality of 22.0%, 24.0% and 100.0% was obtained for control, IKI-
220 50WG and toxic standard, respectively. Corrected preimaginal mortality
reached 2.6% for IKI-220 50WG. A mean production of 197.9 and 138.00 viable
eggs/female was obtained in control and IKI-220 50WG units. IKI-220 50WG
female fertility was not statistically different from control. As no females survived,
assessment of fertility with toxic standard was not possible.
According to corrected mortalities and mean viable eggs production, reduction of
beneficial capacity with IKI-220 50WG was equal to 32.1%. If reduction of
beneficial capacity was mostly the consequence of fertility test results, mean
viable egg production of IKI-220 50WG female was not statistically different from
control.
Conclusion No statistically significant effect on fertility, corrected mortality: 2.6%
Study type Test with bean leaves in laboratory
Flag Key study
Test Substance IKI-220 50 WG
Species Coccinella septempunctata
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Type of exposure Exposure to dried residues on bean leaves, till pupation
Endpoint Mortality and fertility
Value No effect on fertility, 6.1% corrected mortality
Reference
JP Jansen (2002) Side effects of IKI-220 50WG on larvae of the ladybird
Coccinella septempunctata L. (Coleoptera, Coccinellidae) on detached bean
leaves in the laboratory. Agricultural Research Center - Department of Biological
Control and Plant Genetic Resources – Chemin de Liroux 2, B-5030 Gembloux
Belgium. Report number CSE.01/2002
Klimisch Score 1
Amendments/Deviations None that had an impact on the results
GLP Yes
Test Guideline/s Bigler (1988)
No/Group 40 units of 1 larvae for control and treated groups, 20 for the reference substance
(lambda-cyhalothrin at 1 g ai/ha).
Dose Levels
85 g ai/ha in 200 L water /ha (the report mentions that this is equivalent to 2
applications of 80 g ai/ha at 21 days of interval considering the degradation rate of
the active)
Study Summary
A study was carried out to determine the effects of the insecticide IKI-220 50WG
on larvae of the ladybird C. septempunctata (Coleoptera; coccinellidae) under
extended-lab test conditions. Trials were conducted with exposure units using
detached bean leaves treated in the laboratory instead of glass plates. IKI-220
50WG was applied at a rate of 85 g (ai)/ha. Additional leaves were treated with
water as control and with lambda-cyhalothrin as toxic standard.
After treatment, one 2-3-day old larvae of C. septempunctata was released in
each unit. 40 units were prepared for control and IKI-220 50WG and 20 for toxic
standard. Larvae were fed 5 times a week with aphids, till pupation. Larval
mortality was assessed daily and pupae were left to emerge. After emergence,
observed preimaginal mortalities were calculated and corrected with the value of
the corresponding control. When they emerged, adults were transferred in fertility
assessment units and egg production was followed during a 4-week period since
first eggs were found. Mean number of viable eggs/female was used as a
measure of reproductive performance.
A preimaginal mortality of 17.5%was obtained for control. With IKI-220 50WG,
observed and corrected preimaginal mortality reached 22.5% and 6.1%,
respectively. All larvae were killed by toxic standard. A mean production of 116.2
and 99.6 viable eggs/female was obtained in control and IKI-220 50WG units. As
no females survived, assessment of fertility with toxic standard was not possible.
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According to corrected mortalities and mean viable eggs production, reduction of
beneficial capacity with IKI-220 50WG was equal to 19.50%.
Conclusion No effect on fertility, corrected mortality: 6.1%
General conclusion about toxicity to terrestrial invertebrates classification:
Tunnel tests and field studies on bees performed at rates of 70 to 80 g ai/ha show that whereas a direct
spray on bees would have a transient impact on mortality and foraging activity, applications when no bees
are foraging (in the evening) do not have any effect on bees.
Laboratory and extended laboratory tests performed with different types of non-target soil-dwelling or foliar
arthropods, show that no adverse effects are expected in the use conditions of Mainman.
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Application for approval to import Mainman for release (APP202145)
February 2015
Appendix D: Active ingredient and metabolites
Identity
As this is the first full Part 5 application considered for this active ingredient, general data about flonicamid
are provided in the following tables.
Table 8 Identification of flonicamid
IUPAC name N-cyanomethyl-4-trifluoromethylnicotinamide
CAS name N-(cyanomethyl)-4-(trifluoromethyl)-3-pyridinecarboxamide
Common name Flonicamid, Code IKI-220
Molecular formula C9H6F3N3O
CAS Number 158062-67-0
Molecular weight 229.16 g/mole
Structural formula
Purity Minimum 97%
Other international classification &
labelling
Not classified in the EU yet but a classification acute cat4 H302 (Xn
R22) has been proposed.
Mode of action
Flonicamid is a selective homopteran feeding blocker in IRAC Group 9C, and is the only compound in this
chemical subgroup. Numerous assays on the mode of action of flonicamid have been performed. These
include: inhibition studies of acetylcholinesterase, nicotinic and muscarinic acetylcholine receptors,
respiration (including Complex I, II, III, and IV inhibitors, phosphorylation inhibitors, uncouplers of oxidative
phosphorylation), GABA-receptor (in vivo and in vitro), glutamate-receptor (agonist and antagonist),
octopamine receptor agonist, nitric oxide synthase, nitric oxide receptor agonist, Na-channel, L-type Ca-
channel, ryanodine Ca-channel, and calcium ATPase (body wall contraction assay). In all of the above
assays, flonicamid does not act on these targets as do commercial standards within that mode of action.
These assays cover most of the known insecticide modes of action.
The rapid onset of toxicity is inconsistent with chitin synthesis inhibition, juvenile hormone, and ecdysone
agonist activity, meaning it cannot be an insect growth regulator. Because flonicamid does not respond in
74
Application for approval to import Mainman for release (APP202145)
February 2015
these assays, it suggests a novel mode of action that is differentiated from other insecticides commonly
used. There is some evidence that flonicamid targets the insect potassium A-type channel.
Hazard classification
Table 9 Applicant and Staff classifications of the active ingredient
Hazard Class/Subclass
Active ingredient
classification
Method of
classification
Remarks
Ap
plican
t’s
cla
ssif
icati
on
Sta
ff’s
cla
ssif
icati
on
Test
resu
lts
Read
acro
ss
Class 1 Explosiveness No No Expert statement
instead of test
Class 2, 3 & 4 Flammability No No
Class 5 Oxidisers/Organic Peroxides No No Expert statement
instead of test
Subclass 8.1 Metallic corrosiveness No ND
Subclass 6.1 Acute toxicity (oral) 6.1E 6.1D
It is not clear why
the applicant
proposed 6.1E: the
LD50 is within the
range for 6.1DE
classification
Subclass 6.1Acute toxicity (dermal) No No
Subclass 6.1 Acute toxicity (inhalation) No No
Subclass 6.1 Aspiration hazard No ND
Subclass 6.3/8.2 Skin irritancy/corrosion No No
Subclass 6.4/8.3 Eye irritancy/corrosion No No
Subclass 6.5A Respiratory sensitisation No ND
Subclass 6.5B Contact sensitisation No No
Subclass 6.6 Mutagenicity No No
Subclass 6.7 Carcinogenicity No No
Subclass 6.8 Reproductive/ developmental
toxicity No No
Subclass 6.8 Reproductive/ developmental
toxicity (via lactation) No No
Subclass 6.9 Target organ systemic toxicity 6.9B No
No significant target
organ toxicity of
relevance for
human observed at
doses in the
guidance value
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Application for approval to import Mainman for release (APP202145)
February 2015
range for
classification
Subclass 9.1 Aquatic ecotoxicity No No
Subclass 9.2 Soil ecotoxicity No No
Subclass 9.3 Terrestrial vertebrate
ecotoxicity No 9.3C Based on bird data
Subclass 9.4 Terrestrial invertebrate
ecotoxicity No No
NA: Not Applicable --> For instance when testing is technically not possible: testing for a specific endpoint may be omitted, if it is
technically not possible to conduct the study as a consequence of the properties of the substance: e.g. very volatile, highly reactive or
unstable substances cannot be used, mixing of the substance with water may cause danger of fire or explosion or the radio-labelling of
the substance required in certain studies may not be possible.
ND: No Data or poor quality data (according to Klimisch criteria17
) --> There is lack of data.
No: Not Classified based on actual relevant data available for the substance --> The data are conclusive and indicate the threshold for
classification is not triggered.
Physico-chemical properties of the active ingredient
Table 10 Physico-chemical properties of flonicamid
Property Result Test
method
Klimisc
h Score
(1-4)
Reference
Colour
Light beige at 21.9°C (lot #9809)
(ref1)
Off-white at 25°C (lot #9803) (ref 2)
OPPTS
830.6302 1
1) JA Pelton (2000) IKI-220
TGAI – Appearance, pH, and
relative density. Ricerca Inc.
Analytical services 7528 Auburn
Road Painesville OH 44077-
1000. Report no 012575-1
2) GG Sweetapple (1999) IKI-
220 – Melting point, relative
density, physical state, colour
and door. Ricerca Inc. Analytical
services 7528 Auburn Road
Painesville OH 44077-1000.
Report no 010153-1
Physical state
Solid powder at 24.9°C (lot #9809)
(ref1)
Solid powder at 25°C (lot #9803) (ref
2)
OPPTS
830.6303 1
Odour Odourless at 24.9°C (lot #9809) (ref1)
Odourless at 25°C (lot #9803) (ref 2)
OPPTS
830.6304 1
Oxidizing
properties
The substance is not supposed to be
oxidizing as the oxygen and fluorine
in the molecule are bonded only to
UN
recommen
dations on
transport of
1
U Schmiedel (2001) Expert
Statement on the oxidizing
properties of IKI-220 technical.
RCC Ltd , Environmental
17 Klimisch,, H-J., Andrear, M., & U. Tillmann, 1997. A systematic approach for evaluating the quality of experimental
toxicological and ecotoxicological data. Reg. Toxicol. Pharmacol. 25, 1–5 (1997)
76
Application for approval to import Mainman for release (APP202145)
February 2015
carbon and hydrogen. Dangerous
Goods,
Annex 6 3rd
edition,
1999
Chemistry & Pharmanalytics
Division. CH-4452 Itingen
Switzerland. Report no 834030
pH 4.5 at 25 o
C (1% aqueous solution) OPPTS
830.7000 1
JA Pelton (2000) IKI-220 TGAI
– Appearance, pH, and relative
density. Ricerca Inc. Analytical
services 7528 Auburn Road
Painesville OH 44077-1000.
Report no 012575-1
Explosive
properties
Not explosive based on absence of
structural features and decomposition
energy (measured by differential
scanning calorimetry) which is below
the UN limit of concern of 500 J/g (it
was 374 J/g, the exothermic peak
was at 280oC.)
UN
recommen
dations on
transport of
Dangerous
Goods,
Annex 6 3rd
edition,
1999
1
U. Schmiedel (2001) Expert
statement on the explosive
properties of IKI-220 technical.
RCC Ltd , Environmental
Chemistry & Pharmanalytics
Division. CH-4452 Itingen
Switzerland. Report no 834028
Henry’s Law
constant 4.2 x 10
-8 Pa-m
3/mole at 20
oC
Estimation
based on
vapour
pressure
and water
solubility
1
JE Schetter (1999) IKI-220 –
Vapour pressure. Ricerca Inc.
Analytical services 7528 Auburn
Road Painesville OH 44077-
1000. Report no 010341-1
Melting point 157.5 oC
OPPTS
830.7200 1
GG Sweetapple (1999) IKI-220
– Melting point, relative density,
physical state, colour and door.
Ricerca Inc. Analytical services
7528 Auburn Road Painesville
OH 44077-1000. Report no
010153-1
Boiling point Decomposition before boiling
OECD 103
(1995)
JMAFF 12
Nohsan
8147
1
A.Tognucci (2002)
Determination of the boiling
point / boiling range of IKI-220
PAI. RCC Ltd Environmental
Chemistry & Pharmanalytics
Division CH-4452 Itingen
Switzerland. Study report
842001
Vapour pressure
(Pa)
At 20.0 oC: 9.43 x 10
-7
At 25.0 oC: 2.55 x 10
-6
OPPTS
830.7950
(gas
saturation
method)
1
JE Schetter (1999) IKI-220 –
Vapour pressure. Ricerca Inc.
Analytical services 7528 Auburn
Road Painesville OH 44077-
1000. Report no 010341-1
77
Application for approval to import Mainman for release (APP202145)
February 2015
Relative Density 1.531 (20
oC/20
oC) lot #9809
1.54 (20 o
C/20 o
C) lot #9803
OPPTS
830.7300 1
JA Pelton (2000) IKI-220 TGAI
– Appearance, pH, and relative
density. Ricerca Inc. Analytical
services 7528 Auburn Road
Painesville OH 44077-1000.
Report no 012575-1
GG Sweetapple (1999) IKI-220
– Melting point, relative density,
physical state, colour and door.
Ricerca Inc. Analytical services
7528 Auburn Road Painesville
OH 44077-1000. Report no
010153-1
Surface Tension
47.3 mN/m at 25 o
C±1
47.0 mN/m at 40 o
C±1
The substance is surface active
however the test was performed with
concentration > 1g/L (around 10 g/L)
Mettens
(equivalent
to EEC A5)
2
B de Ryckel (2002) Relative
self-ignition temperature,
flammability and surface tension
of IKI-220 TGAI. Centre de
recherches Agronomiques de
Gembloux. Departement de
Phytopharmacie 5030
Gembloux Belgique. Report no
20334.
Water Solubility 5.2 g/L at 20 o
C
OPPTS
830.7840
(shake
flask
method)
1
RT O’Donnell (1999) IKI-220,
PAI (Lot #9803) – Water
solubility. Ricerca Inc. Analytical
services 7528 Auburn Road
Painesville OH 44077-1000.
Report no 010251-1
Solvent
Solubility (20°C)
(g/100 mL
solvent)
Solvent Lot#9
809
lot
#980
3
Acetone 17.32 18.67
Ethylacetat
e
3.57 3.39
Methanol 9.76 11.06
Dichlorome
thane
0.40 0.45
Toluene 0.03 0.055
Hexane 0.000
03
0.000
02
n-octanol 0.26 0.30
Acetonitrile 12.04 14.61
Isopropyl
alcohol
1.46 1.87
OPPTS
830.7840 1
LP Dudones (1999) IKI-220, PAI
(lot #9809) – Organic solvent
solubility. Ricerca Inc. Analytical
services 7528 Auburn Road
Painesville OH 44077-1000.
Report no 011201-1
RT O’Donnell (1999) IKI-220,
PAI (lot #9803) – Organic
solvent solubility. Ricerca Inc.
Analytical services 7528 Auburn
Road Painesville OH 44077-
1000. Report no 010250-1
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Log Kow Kow = 1.9
Log Kow = 0.3 at 29.8 o
C
OPPTS
830.7570
(HPLC)
1
LP Dudones (1999) IKI-220, PAI
(lot #9803) – Octanol/water
partition coefficient. Ricerca Inc.
Analytical services 7528 Auburn
Road Painesville OH 44077-
1000. Report no 010252-1
Flammability
Not flammable. Substance melts on
contact with flame or become liquid
but it does not ignite.
EEC A10 B de Ryckel (2002) Relative
self-ignition temperature,
flammability and surface tension
of IKI-220 TGAI. Centre de
recherches Agronomiques de
Gembloux. Departement de
Phytopharmacie 5030
Gembloux Belgique. Report no
20334.
Auto
flammability
No relative self-ignition temperature
up to 400 o
C
- E
E
C
A
1
6
-
Dissociation
constant pKa = 11.60 ± 0.03 at 20 ± 1
oC
OPPTS
830.7370 1
RC Beckwith (1999) IKI-220 (lot
#9803) – dissociation constant.
Ricerca Inc. Analytical services
7528 Auburn Road Painesville
OH 44077-1000. Report no
010141-1
Thermal stability
in air
No relevant endothermic or
exothermic reaction up to 150 oC.
OECD 113
(by DSC) 1
A Tognucci (2002) Screening of
the thermal stability in air of IKI-
220 PAI. RCC Ltd ,
Environmental Chemistry &
Pharmanalytics Division. CH-
4452 Itingen Switzerland.
Report no 842000
Particle size
distribution
% of
particles
Size of
particles
(mm)
10.34 > 1.190
35.13 1.190 –
0.500
47.21 0.500 -
0.250
6.44 0.250 -
0.106
0.76 < 0.106
OECD 110
(1981)
CIPACT
MT 59
1
L.F. Piasentini de Campos
(2002) Particle size distribution
of Flonicamid tecnico. Bioagri
Laboratorios Ltda Piracicaba
S.P. Brazil. Study number RF-
1213.018.080.02
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Mammalian toxicology - Robust study summaries for the active ingredient and
metabolites
In the following study summaries the active ingredient flonicamid is referred to by the development code IKI-
220.
Acute toxicity [6.1]
Acute Oral Toxicity [6.1 (oral)]
Type of study Acute oral toxicity test in rats
Flag Key study
Test Substance IKI-220 technical, Lot No.9809
Endpoint LD50
Value 884 mg/kg bw males, 1768 mg/kg bw females
Reference
Ridder, W. E., Yoshida, M., Watson, M., Acute oral toxicity study in rats
with IKI-220 technical, RCC Ltd, Toxicology and Metabolism, USA, 2001.
ISK Document No.2008 010276-1-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 401, US EPA OPPTS 870.1100
Species Rat
Strain Sprague Dawley Crl:CD®(SD)BR(IGS)
No/ Group 5M and 5F
Dose Levels 625, 1250, 2500, 5000 mg/kg bw
Exposure Type Oral gavage
Study Summary
Treatment related deaths were 5/5 males and 5/5 females at 5000
mg/kg/bw, 5/5 males and 4/5 females at 2500 mg/kg bw and 5/5 males
and 1/5 females at 1250 mg/kg bw. There were no deaths at 625 mg/kg
bw. Deaths were generally within 4 hours of dosing but also occurred up to
48 hours post dosing. The principal clinical signs in decedents included
decreased activity, loss of mobility, laboured breathing, prostration,
convulsion, cold to touch and sensitive to noise. Ataxia was common
amongst females. Additional signs amongst surviving females were
laboured breathing, rales, prostration, cold to touch, tremors, anogenital
staining and few or no faeces. Initial weight loss was evident at all doses
with subsequent recovery in those animals that survived to Day 14.
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Necropsy revealed dark red-black spots on the serosal surface of the
stomach in 1 male at 1250 mg/kg bw and 2 females each at 2500 and
5000 mg/kg bw. All were decedents within 48 hours of treatment
Additional Comments No additional comments
Conclusion
IKI-220 technical should be classified as 6.1D (oral), based on the male LD
50 value of 884 mg/kg bw in males and 1768 mg/kg bw in females (Section
10.2; User Guide for Thresholds and Classifications; EPA0109, 2012).
Acute Dermal Toxicity [6.1 (dermal)]
Type of study Acute dermal limit toxicity test in rats
Flag Key study
Test Substance IKI-220 technical, Lot No. 9809
Endpoint LD50
Value >5000 mg/kg bw (limit dose)
Reference
Ridder, W. E., Yoshida, M., Watson, M., Acute dermal toxicity (LD50) study
in rats with IKI-220 technical, RCC Ltd, Toxicology and Metabolism, USA,
2002. ISK Document No.2009 010278-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 402, US EPA OPPTS 870.1200
Species Rat
Strain Sprague Dawley Crl:CD®(SD)BR(IGS)
No/ Group 5M and 5F
Dose Level 5000 mg/kg bw undiluted – moistened
Exposure Type Dermal, 24h occluded
Study Summary
There were no deaths. Principal clinical signs were coloured material
around the nose and eye(s) and anogenital staining observed in most
animals from 1 hour to 3 days post dosing but resolving by Day 4. There
were no treatment related effects on bodyweight gain or any macroscopic
changes evident at necropsy.
Additional Comments No additional comments
Conclusion IKI-220 technical should not be classified for 6.1 (dermal), based on the
lack of adverse effects at the limit dose used in this study (Section 10.2;
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User Guide for Thresholds and Classifications; EPA0109, 2012).
Acute Inhalation Toxicity [6.1 (inhalation)]
Type of study Acute inhalation limit toxicity test in rats (4-hour, nose only)
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint LC50
Value >4.90 mg/l
Reference
Paul, G. R., Bowden, A.M., Coombs, D.W., IKI-220 Technical Acute (four-
hour) inhalation in rats, Huntingdon Life Sciences Ltd, UK, 2000. ISK
Document No.2010-994326
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 403, 92/69/EEC, US EPA OPPTS 870.1300
Species Rat
Strain Sprague Dawley
No/Sex/Group 5M and 5F
Dose Levels
4.90 mg/l
mass median aerodynamic diameter and geometric standard deviation
were 4.8 µm and 2.37 respectively
Exposure Type 4h nose only – dust aerosol
Study summary
There were no deaths. Clinical signs were confined to exaggerated
breathing in some rats from 30 minutes of exposure and all rats from 3
hours into the exposure period, persisting up to Day 2 post dosing. Brown
staining around the snout and jaws was seen in all treated animals after
exposure, persisting up to Day 2 in females. All signs had resolved by Day
3 post dosing. There were no treatment related effects on bodyweight gain,
food or water intake, macroscopic pathology or organ weights.
Additional Comments
The mass median aerodynamic diameter of of the particulate atmosphere
was 4.8 µm and was marginally higher than the acceptable range (1 µm to
4 µm) for an acute inhalation study. This was reported to be attributable to
the nature of the test material. The authors note that approximately 66% of
the particulate were less than 7 µm in aerodynamic diameter and are
therefore considered of a respirable size.
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Conclusion
IKI-220 technical should not be classified for 6.1 (inhalation), based on the
lack of adverse effects at the limit concentration used in this study (Section
10.2; User Guide for Thresholds and Classifications; EPA0109, 2012).
Skin Irritation [6.3/8.2]
Type of study Dermal irritation/corrosion study in rabbits
Flag Key study
Test Substance IKI-220 Technical, Lot No. 9809
Endpoint Dermal irritancy (mean of Draize scores at 24, 48 and 72 hours)
Value Non irritant: 0.0 for erythema and oedema
Reference
Ridder, W. E., Yoshida, M., Watson, M., Acute dermal study in albino
rabbits with IKI-220 technical, RCC Ltd, Toxicology and Metabolism, USA,
2000. ISK Document No.2011 010280
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 404, US EPA OPPTS 870.2500
Species Rabbit
Strain New Zealand White
No/ Group 6M
Dose Levels 0.5g undiluted - moistened
Exposure Type 4h dermal, semi occlusive
Study Summary There were no skin reactions in any animal. Scores for erythema/eschar
and oedema were 0 for all observations
Additional Comments No additional comments
Conclusion
IKI-220 technical should not be classified for 6.3 (skin irritation) based on
the results from this study (Section 11.2; User Guide for Thresholds and
Classifications; EPA0109, 2012).
Eye Irritation [6.4/8.3]
Type of study Eye irritation/corrosion study in rabbits
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
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Endpoint Eye irritancy (mean of Draize scores at 24, 48 and 72 hours)
Value Non irritant: 0.4 for conjunctival redness, 0.06 for chemosis, 0.0 for corneal
opacity and iritis
Reference
Ridder, W. E., Yoshida, M., Watson, M., Acute eye irritation study in albino
rabbits with IKI-220 technical, RCC Ltd, Toxicology and Metabolism, USA,
2000. ISK Document No.2012 010281-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 405, US EPA OPPTS 870.2400
Species Rabbit
Strain New Zealand White
No/ Group 6M
Dose Levels 0.1 ml (approx.70 mg)
Exposure Type Ocular
Study Summary
There were no corneal or iridal effects. Conjunctival redness, chemosis
and discharge were seen in all animals 1 hour post application. Redness
became less severe by 24 hours and resolved by 72 hours. Chemosis
resolved in most animals by 24 hours and discharge in all animals resolved
by 24 hours. Draize scores were cornea 0, iris 0, conjunctival redness 0.4,
chemosis 0.06. There were no other treatment related effects.
Additional Comments No additional comments
Conclusion
IKI-220 technical should not be classified for 6.4 (eye irritation) based on
the results from this study (Section 12.2; User Guide for Thresholds and
Classifications; EPA0109, 2012).
Contact Sensitisation [6.5]
Type of study Sensitisation study – Magnusson and Kligman Maximisation test in guinea
pigs
Flag Key
Test Substance IKI-220 Technical, Lot No. 9809
Endpoint Contact sensitisation on challenge
Value Non sensitiser: <thresholds (negative)
Reference Ridder, W. E., Watson, M., Dermal sensitisation study (maximisation
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design) in guinea pigs with IKI-technical, RCC Ltd, Toxicology and
Metabolism, USA, 2000. ISK Document No.2013 010282-1-2
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 406, US EPA 870.2600
Species Guinea pig
Strain Hartley albino
No/Sex/Group 10F control group, 20F treated group
Dose Levels
10% in mineral oil distilled water, once per week for 2 weeks induction
phase. 10% in mineral oil for challenge (maximum non irritating
concentration)
Positive control – dinitrochlorobenzene 0.8% in mineral oil
Exposure Type Dermal -24 hours occluded
Study Summary Only a slight response was seen on challenge equating to mild irritation
and thus assessed as negligible rather than sensitisation.
Additional Comments No additional comments
Conclusion
IKI-220 technical should not be classified for 6.5B (contact sensitisation)
based on the results from this study (Section 13.2; User Guide for
Thresholds and Classifications; EPA0109, 2012).
General conclusion about acute toxicity classification:
IKI-220 technical demonstrated acute oral toxicity albeit at a relatively low classification of 6.1D
(oral). No classifications are required for acute dermal or inhalation toxicity (6.1 dermal or
inhalation).
IKI-220 technical did not demonstrate skin or eye irritation and does not require classification for
6.3 and 6.4.
IKI-220 technical did not show contact sensitisation potential and should not be classified for
contact sensitisation (6.5B).
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Genotoxicity [6.6]
In Vitro Studies
Study type/Test Guideline Result Reference
Ames Reverse mutation test.
OECD 471, US EPA OPPTS
870.5100
Non mutagenic,
negative with or
without metabolic
activation
Matsumoto, K., IKI-220 technical: reverse
mutation test, Institute of Environmental
Toxicology, Japan, 2002. ISK Document No.
2018 IET 00-0147
In vitro cytogenetics test.
OECD 473, US EPA OPPTS 870.5375
Non clastogenic,
negative with or
without metabolic
activation
Matsumoto, K., IKI-220 technical: in vitro
cytogenetics test, Institute of Environmental
Toxicology, Japan, 2002. ISK No. 2018 IET 00-
0149
In vitro mouse lymphoma gene mutation test. OECD 476, US EPA OPPTS 870.5300
Non mutagenic,
negative with or
without metabolic
activation
Matsumoto, K., IKI-220 technical: in vitro
mouse lymphoma gene mutation test, Institute
of Environmental Toxicology, Japan, 2002. ISK
Document No. 2018 IET 00-0150
Comet assay in mouse colon, liver and lung. Tice et al., 2000 International Workshop on Genotoxicity Test Procedures, 1999
Non genotoxic in
tested tissues.
Sasaki, Y. F., A novel insecticide: the comet
assay with mouse colon, liver and lung.
Hachinohe National College of Technology,
Japan, 2002. ISK Document No.2412 AN-2106
Conclusion Results from the genotoxicity assays were
negative
In Vivo Studies
Type of study Mouse micronucleus test
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint Clastogenicity
Value Non clastogenic
Reference Matsumoto, K., IKI-220 technical: micronucleus test in mice, Institute of
Environmental Toxicology, Japan, 2002. ISK Document No. 2018 IET 00-0148
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 474, US EPA OPPTS 870.5395
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Species Mouse
Strain Crj:CD-1
No/Sex/Group 5M and 5F
Dose Levels
250, 500, 1000 mg/kg bw males; 125, 250, 500 mg/kg bw females - by oral
gavage
Positive control – mitomycin C
Study Summary
There were no deaths or clinical signs. There was no increase in the frequency of
micronucleated polychromatic erythrocytes or any decrease in the polychromatic
erythrocyte ratio. Treatment with IKI-220 technical did not induce micronuclei in
the bone marrow of mice.
Additional Comments No additional comments
Conclusion The test substance did not demonstrate any clastogenic activity
Type of study DNA repair test (Unscheduled DNA Synthesis) in rats
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint DNA repair synthesis
Value Negative
Reference
Mehmood, Z., IKI-220 technical: In vivo DNA repair (UDS) test using rat
hepatocytes, Huntingdon Life Sciences Ltd., UK, 2003. ISK Document No. 269-
032007
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 486, 2000/32/EC Annex 4G-B.39, US EPA OPPTS 870.5550
Species Rat
Strain Sprague-Dawley
No/Sex/Group 5M
Dose Levels 600, 2000 mg/kg bw – oral gavage
Study Summary
There were no deaths. Clinical signs seen at both dosages were under activity,
flattened posture, abnormal gait, irregular respiration, reduced body tone and
reddened skin colour with the additional signs at 2000 mg/kg bw of prominent
eyes and partially closed eyelids. Treated animals did not show any increase in
the gross or net nuclear grain count compared with controls. Significant increases
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were seen in the positive control groups treated with dimethylnitrosamine or 2-
acetylaminofluorene.
Additional Comments No additional comments
Conclusion IKI-220 technical was negative in the assay and did not cause any unscheduled
DNA synthesis
General conclusion about genotoxicity classification:
IKI-220 technical was not mutagenic or clastogenic. The combined in vitro and in vivo data indicate that no
classification for genotoxicity (6.6) is required.
Carcinogenicity [6.7]
Type of study Carcinogenicity/Toxicity in rats
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoints
Non-Neoplastic Effects
LOAEL: 1000 ppm (36.5 mg/kg bw/day) males, 5000
ppm (219 mg/kg bw/day)females – altered kidney
micropathology in males, liver and kidney in females,
decreased body weight and body weight gain
NOAEL: 200 (7.32 mg/kg bw/day) ppm males, 1000
ppm (44.1 mg/kg bw/day) females
Neoplastic Effects
LOAEL: N/A
NOAEL: 1000 ppm (36.5 mg/kg bw/day) males,
5000 ppm (219 mg/kg bw/day) females
Tumours N/A
Malignant/Benign N/A
Background Incidence N/A
Time of Onset N/A
Survival N/A
Reference
Kuwahara, M., IKI-220 technical: Combined chronic toxicity and carcinogenicity
study in rats, The Institute of Environmental Toxicology, Japan, 2002. ISK
Document No. 2036 IET 98-0142
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s 87/302/EEC 1987, US EPA OPPTS 870.4300
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Species Rat
Strain Wistar Jcl:Wistar
No/Sex/Group 76M and 76F (14 and 10 per sex per group sacrificed after 26 and 52 weeks
respectively for toxicity assessments)
Dose Levels
0, 50 (males only), 100 (males only), 200, 1000, 2000 (females only) ppm –
equivalent to 0, 1.84, 3.68, 7.32, 36.5 mg/kg bw/day males and 0, 8.92, 44.1, 219
mg/kg bw/day females respectively
Exposure Type Admixture with the diet
Study Summary
There was no evidence of carcinogenic activity at any of the dosages investigated.
Females receiving 5000 ppm showed lower food intake and food efficiency with
concomitant lower weight gain during the first 52 weeks of treatment with
bodyweight at termination remaining lower than concurrent controls. Some
association with palatability cannot be discounted. There was a significant
decrease in rearing behaviour during weeks 5 to 12. Fluctuations in haematocrit,
haemoglobin concentration, erythrocyte count and/or mean corpuscular
haemoglobin concentration indicated mild anaemia, particularly in the later period
of treatment. Blood chemistry revealed increased ү-glutamyl transpeptidase and
cholesterol with decreased triglycerides. Significant increases in absolute and/or
relative liver (from Week 26) and kidney (from Week 52) weights were noted.
Necropsy revealed increased incidences of accentuated lobular pattern and dark
colour of the liver. Histopathology revealed hepatocellular hypertrophy of the liver
and cytoplasmic vacuolation of the proximal tubular cells of the kidney. Additional
histopathological findings noted in the later stages of treatment included increased
incidences of atrophy of striated muscle fibres (m.triceps surae – calf muscle),
cataract, retinal atrophy, hepatic foci of cellular alterations (eosinophilic cell foci),
chronic nephropathy and brown pigment deposition in renal proximal tubular cells.
At 1000 ppm a significant decrease in rearing behaviour was noted amongst
males during weeks 10 to 30, decreased body weight and body weight gain was
also observed although some association with palatability cannot be discounted.
Increased liver weight was evident from Week 26 and increased kidney weight
from Week 52. Histopathology in males revealed increased incidences of hyaline
droplet deposition and tubular basophilic changes and granular casts of the kidney
during the early phase of treatment. These changes during the later stages of
treatment occurred at an incidence comparable with controls whereas the
incidence of chronic nephropathy was significantly increased. There were no in life
or pathological changes seen in treated females
At 200 ppm although there was a significant increase in the incidence of hyaline
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droplet deposition in kidney proximal tubular cells in males the finding was
considered not to be adverse as it was noted only at 26 weeks without any
increase in degenerative kidney lesions seen throughout the whole treatment
period. Liver weight was slightly increased from Week 26. There were no in life or
pathological changes seen in treated females.
At 100 and 500 ppm (males only) there were no treatment related changes.
Additional Comments
A short term GLP study1 revealed that IKI-technical had the potential of binding to
a male specific protein ‘α2µ-globulin’ (identified imunohistochemically in hyaline
droplets) and the protein-test substance complex deposited in the proximal tubules
caused degenerative changes in the kidney. In the present study similarly hyaline
droplets deposited in the proximal tubular cells were seen in males at 200 ppm or
higher in the early phases of treatment. Deposition of α2µ-globulin causes
degenerative changes consisting of tubular basophilic change and granular casts
in dilated tubules. Although seen early in the treatment period compared with
controls the incidences at termination were comparable, thus the absence of
degenerative renal lesions including tubular basophilic change and chronic
nephropathy in these males indicated the hyaline droplet deposition itself was
treatment related however it was not an adverse effect.
Conversely, the increased incidence of cytoplasmic vacuolation of proximal tubular
cells in the kidney of females at 5000 ppm at all examinations was considered to
be treatment related and adverse with a higher incidence of chronic nephropathy.
1 Kuwahara, M., IKI-220 technical: 28 day dose range finding study in rats, The
Institute of Environmental Toxicology, Japan, 2002. ISK Document No. 2025 IET
98-0140
Conclusion IKI-220 technical did not demonstrate any carcinogenic potential. Target organs
identified were the kidney and liver.
Type of study Carcinogenicity study in mice
Flag Supporting
Test Substance IKI-220 technical, Lot No. 9809
Endpoints
Non-Neoplastic Effects
LOAEL: 250 ppm – 29/38 mg/kg bw/day
males/females respectively – altered lung pathology
NOAEL: <250 ppm - 29/38 mg/kg bw/day
males/females respectively
Neoplastic Effects
LOAEL: 250 ppm - 29/38 mg/kg bw/day
males/females respectively – alveolar/bronchiolar
adenoma
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NOAEL: <250 ppm - 29/38 mg/kg bw/day
males/females respectively
Tumours Lung tumours – alveolar/bronchiolar
Malignant/Benign
Increased incidence of carcinoma in males at 750 ppm and 2250 ppm and
females at 2250 ppm. Increased incidences of adenoma in all dose groups of both
sexes. Combined incidences 45-60% males, 37-53% females, across all treated
groups
Background Incidence 17% males, 15% females
Time of Onset Terminal
Survival There was no adverse effect of treatment on mortality
Reference
Ridder, W. E., Watson, M., An oncogenicity study in mice with IKI-220 technical,
Ricerca Biosciences Ltd, Toxicology and Pharmacology, USA, 2003. ISK
Document No. 2002 011885-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 451, US EPA OPPTS 870.4200
Species Mouse
Strain Crl:CD-1® (ICR) BR VAF/plus
No/Sex/Group 80M and 80 F (control and 2250 ppm groups – 10M and 10F sacrificed at 26 and
52 weeks for interim evaluations), 60M and 60F at 250 and 750 ppm
Dose Levels 0, 250, 750, 2250 ppm equivalent to 0, 29, 88, 261 mg/kg bw/day males and 0, 38,
112, 334 mg/kg bw/day females respectively
Exposure Type Admixture with the diet
Study Summary
There were no treatment related effects on mortality, clinical signs, bodyweight
gain, food consumption or leucocyte differentials at any dose. Treatment related
increases in liver weight were evident in both sexes at all dosages at 26 and 52
weeks and termination. At termination necropsy revealed an increased number of
animals with lung masses or nodules in treated groups which correlated with an
increase in the incidence of primary lung tumours (alveolar/bronchiolar) in both
sexes at all doses. The incidence in males ranged from 45 – 60% and 37 – 53% in
females compared with 17% and 15% in male and female concurrent control
groups. Additional histopathological findings consisted of an increased incidence
of alveolar/bronchiolar hyperplasia in treated mice, an increased incidence of
hepatocellular hypertrophy in the liver of males at all dosages and females at 2250
ppm and an increased incidence of extramedullary haemopoiesis in the spleen of
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males at all dosages. Decreased cellularity (hypocellularity) was noted in both
sexes at 2250 ppm. The incidence of this finding was increased in males at 750
and 2250 ppm and all treated groups of females.
Additional Comments No no effect level for tumour incidence or toxicity was established in this study
Conclusion
IKI-220 technical demonstrated carcinogenic potential in the lung manifest as
increased incidences principally of bronchiolar/alveolar epithelial adenoma. No
NOAEL was demonstrated in this study. The liver and kidney were identified as
target organs
Type of study Carcinogenicity study in mice
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoints
Non-Neoplastic Effects
LOAEL: 250 ppm 30.3/36.3 mg/kg bw/day
males/females respectively – altered lung pathology
NOAEL: 80 ppm 10.0/11.8 mg/kg bw/day
males/females respectively
Neoplastic Effects
LOAEL: 250 ppm males (30.3 mg/kg bw/day),
increased incidence of lung adenoma, >250 ppm
females (36.3 mg/kg bw/day)
NOAEL: 80 ppm males, 250 ppm females (10.0/36.3
mg/kg bw/day males/females)
Tumours Lung adenoma – alveolar/bronchiolar, males only 42%
Malignant/Benign Benign
Background Incidence Males 16%
Time of Onset Termination
Survival No treatment related effects
Reference
Nagaoka, T., Dietary carcinogenicity study of IKI-220 technical in mice, Shin
Nippon Biomedical Laboratories Ltd, Japan, 2004. ISK Document No. 2053 SBL
40-50
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 451, US EPA OPPTS 870.4200
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Species Mouse
Strain Crj:CD-1 (ICR)
No/Sex/Group 50M and 50F
Dose Levels 0, 10, 25, 80 250 ppm equivalent to 0, 1.20, 3.14, 10.0, 30.3 and 0, 1.42, 3.67,
11.8, 36.3 mg/kg bw/day males/females respectively
Exposure Type Admixture with the diet
Study Summary
There was no adverse treatment related effect on survival or any treatment related
effects on clinical signs, food consumption, bodyweight gain or organ weights at
any of the dosages investigated. Necropsy revealed in males only receiving 250
ppm to have an increased incidence of lung masses, histopathologically these
were identified as pulmonary alveolar/bronchiolar adenoma. In both sexes at 250
ppm there was an increase in the non neoplastic finding of
hyperplasia/hypertrophy of the epithelial lining of terminal bronchioles albeit slight
in all affected animals.
Additional Comments No additional comments
Conclusion
IKI-220 technical demonstrated carcinogenic potential in the lung manifest as
increased incidences principally of bronchiolar/alveolar epithelial adenoma in
males at the highest dose investigated. The lung was also identified as a target
organ.
Type of study Lung cell cycle analysis in the mouse
Flag Supporting study
Test Substance IKI-220, Lot No. 9809
Reference
Nomura, M., IKI-220: Cell cycle analysis using BRDU in the mouse lung by
dietary admixture administration for three days, Safety Sciences Group,
Safety Science Research Laboratory, Ishihara Sangyo Kaisha, Japan,
2003. ISK Document No. 2047 AN-2110
Klimisch Score
2 – reliable with restrictions - on the basis that the study was conducted
‘in-house’ by the Applicant Ishihara Sangyo Kaisha. The Applicants’ own
Compliance Statement includes the comment ‘the study was not conducted
or reported in compliance with Good Laboratory Practice (GLP)
Regulations. However, we believe that the results of this study are
scientifically valid and would not have been different if the study had been
conducted under GLP regulations’
Amendments/Deviations None
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GLP Conducted in accordance with GLP principles
Test Guideline/s Not applicable
Species Mouse
Strain Crj: CD-1 (ICR)
No/Sex/Group 5M
Dose Levels
0, 80, 250, 750, 2250 ppm – equivalent to 0, 12.3, 40.9, 129.6, 339.3
mg/kg bw/day – admixture with the diet. Bromodeoxyuridine (Brdu – a cell
cycle marker) was injected 14 and 2 hours prior to sacrifice
Study Summary
There were no deaths or any adverse treatment related clinical signs.
There were no macropathological changes. Histopathological lung sections
were stained with anti-Brdu monoclonal antibody then examined
immunohistochemically. The terminal bronchiole was targeted for
assessment. The Brdu Index was higher at 250, 750 and 2250 ppm
compared with concurrent controls. At 80 ppm values were comparable
with the control indicating a threshold between 80 ppm and 250 ppm for
effects in the terminal bronchiolar epithelia of the lung – an increase in the
number of cells in the synthesis phase of the cell cycle. The results
correlate with the carcinogenicity study in which increased numbers of
bronchioloalveolar tumours were seen at 250 ppm or higher.
Additional Comments No additional comments
Conclusion IKI-220 demonstrated effects in the terminal bronchioles of the lung
causing increased numbers of cells in the synthesis phase.
Type of study Toxicity/reversibility study in the mouse
Flag Supporting study
Test Substance IKI-220 technical, Lot No. 9809
Reference
Nomura, M., IKI-220: Toxicological effect on the mouse lung and its
reversibility by dietary administration for 28 days followed by a 28-day
recovery period, Safety Sciences Group, Safety Science Research
Laboratory, Ishihara Sangyo Kaisha, Japan, 2003. ISK Document No. 2049
AN-2140
Klimisch Score
2 – reliable with restrictions - on the basis that the study was conducted
‘in-house’ by the Applicant Ishihara Sangyo Kaisha. The Applicants’ own
Compliance Statement includes the comment ‘the study was not conducted
or reported in compliance with Good Laboratory Practice (GLP)
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Regulations. However, we believe that the results of this study are
scientifically valid and would not have been different if the study had been
conducted under GLP regulations’
Amendments/Deviations None
GLP Conducted in accordance with GLP principles
Test Guideline/s Not applicable
Species Mouse
Strain Crj:CD-1 (ICR)
No/Sex/Group
5F with 5 per group sacrificed at the end of 28 days of treatment then 5 per
group sacrificed after 7, 14 or 28 days recovery – admixture with the diet.
Bromodeoxyuridine (Brdu – a cell cycle marker) was injected 2 hours prior
to sacrifice.
Dose Levels 0, 2250 ppm equivalent to 0, 295 – 303 mg/kg bw/day
Study Summary
The reversibility of IKI-220 induced effects in the terminal bronchiolar
epithelial cells of mice was assessed by measuring the uptake of Brdu (a
measure of the number of cells in the synthesis (S) phase of the cell cycle.
The number indicates cells preparing to divide or have passed the
synthesis phase.
There were no deaths or any clinical signs considered attributable to
treatment. At necropsy slight enlargement of the liver noted at the end of
the treatment period however this was not evident in any of the recovery
groups.
The terminal bronchioles were examined for counting of Brdu positive cells.
Clara cells in the same tissues were also examined. The Brdu Index was
significantly increased in treated animals at the end of 28 days treatment
compared with concurrent controls, however, there was no difference
between the treated and control group after 7, 14 or 28 days of recovery.
There were no histopathological findings in any of the tissues. Anti-Clara
cell antibodies also increased in the terminal bronchioles at the end of the
treatment period. Morphologically altered Clara cells returned to normal
within 7 days of recovery. The altered morphology consisted of greater
length, elongation and protruded cytoplasm with enlarged and increased
numbers of secretory granules in the cytoplasm. These findings were
consistent with those seen with styrene toxicity with the difference that IKI-
220 was not cytotoxic to the terminal bronchioles and did not induce
necrosis. Whereas a metabolite of styrene has been shown to be the
cause of effects on Clara cells related metabolites of IKI-220 did not show
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any cytotoxic or mitogenic activity on the target tissue. Thus, the effects of
IKI-220 on the terminal bronchioles of mouse lung were readily reversible
on withdrawal of exposure, returning to normal within 7 days and without
adverse findings in the lung.
Additional Comments No additional comments
Conclusion
IKI-220 demonstrated activity affecting the Clara cells in terminal
bronchioles, however, the effect was completely reversible with a few days
of withdrawal of exposure to the test substance.
Type of study Comparative study of cell cycle analysis between 3 strains of mice
Flag Supporting study
Test Substance IKI-220 technical, Lot No. 9809
Reference
Nomura, M., IKI-220: A comparative study among three mouse strains on
cell cycle analysis in the lung by dietary administration of IKI-220 or
Isoniazid for three days, Safety Sciences Group, Safety Science Research
Laboratory, Ishihara Sangyo Kaisha, Japan, 2003. ISK Document No. 2051
AN-2200
Klimisch Score
2 – reliable with restrictions - on the basis that the study was conducted
‘in-house’ by the Applicant Ishihara Sangyo Kaisha. The Applicants’ own
Compliance Statement includes the comment ‘the study was not conducted
or reported in compliance with Good Laboratory Practice (GLP)
Regulations. However, we believe that the results of this study are
scientifically valid and would not have been different if the study had been
conducted under GLP regulations’
Amendments/Deviations None
GLP Conducted in accordance with GLP principles
Test Guideline/s Not applicable
Species Mouse
Strain Crj: CD-1 (ICR), Jcl: B6C3F1, Jcl: C57/6J
No/Sex/Group 5M
Dose Levels
0, 2250 ppm IKI-220 technical, 2250 ppm Isoniazid (positive control)
equivalent to:
IKI-220 mg/kg bw/day Isoniazid mg/kg bw/day
CD-1 299 298
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B6C3F1 316 325
C57 306 290
Admixture with the diet
Study Summary
There were no deaths or any adverse treatment related clinical signs.
Macropathology did not reveal any treatment related changes. Histology
sections were then examined immunohistochemically after staining with
anti-Brdu monoclonal and anti-Clara cell antibody and CC-10 polyclonal
antibodies. The terminal bronchioles were then examined.
The Brdu index in the CD-1 strain was significantly increased compared
with the concurrent controls, however, the values for the B6C3F1 and C57
strains were comparable with the concurrent control group indicating an
overt difference between strains for IKI-220 induced mitogenic activity in
target cells of the lung terminal bronchioles. The CD-1 was the most
sensitive strain and was the strain used for the carcinogenicity
investigation. Isoniazid induced a more prominent effect than did IKI-220,
affecting all three strains, including the most resistant C57 strain. Isoniazid
increased cell turnover in the terminal bronchiolar cells of the lung of all
three strains whereas IKI-220 was only effective in the CD-1 strain.
CC-10 reaction percentages were similar in all three strains indicating
approximately 80% of the cells in the terminal bronchioles were Clara cells.
Although there were no quantitative differences between the cells
qualitative differences such as cell turnover labelled with Brdu did exist
between the strains. Thus, the Clara cells in the sensitive CD-1 strain
(higher background of spontaneous lung tumours) seemed to be
congenitally predisposed to divide more readily in response to mitogenic
stimuli. However, the Clara cells in the more resistant strains may not be
similarly predisposed. Consequently a unique adenoma related gene
named ‘pas: pulmonary adenoma susceptibility’ has been reported to
predispose some inbred mouse strains to the development of lung
tumours. Such a gene may in part be responsible for the effects caused by
administration on IKI-220 in mice.
Additional Comments No additional comments
Conclusion
IKI-220 demonstrated effects in the terminal bronchioles (Clara cells) in
CD-1 mice but was not clearly apparent in B6C3F1 and even less so in the
C57 strain, values being comparable with controls. Thus, there was a clear
differentiation in strain sensitivity of mice to IKI-220.
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Type of study Comparative rat and mouse cell cycle analysis in the lung
Flag Supporting
Test Substance IKI-220 technical, Lot No. 9809
Reference
Nomura, M., IKI-220: A comparative study between mice and rats on cell
cycle analysis in the lung by dietary admixture for three and seven days,
Safety Sciences Group, Safety Science Research Laboratory, Ishihara
Sangyo Kaisha, Japan, 2003. ISK Document No. 2048 AN-2130
Klimisch Score
2 – reliable with restrictions - on the basis that the study was conducted
‘in-house’ by the Applicant Ishihara Sangyo Kaisha. The Applicants’ own
Compliance Statement includes the comment ‘the study was not conducted
or reported in compliance with Good Laboratory Practice (GLP)
Regulations. However, we believe that the results of this study are
scientifically valid and would not have been different if the study had been
conducted under GLP regulations’
Amendments/Deviations None
GLP Conducted in accordance with GLP principles
Test Guideline/s Not applicable
Species Mouse. rat
Strain Crj: CD-1 (ICR) mouse, Jcl: Wistar rat
No/Sex/Group 5F
Dose Levels
0, 2250 ppm mice (equivalent to 374 – 286 mg/kg bw/day), 5000 ppm rats
(equivalent to 392 – 403 mg/kg bw/day). Admixture with the diet.
Bromodeoxyuridine (Brdu – a cell cycle marker) was administered 2 hours
prior to sacrifice.
Study Summary
There were no deaths or any adverse clinical signs of reaction. There were
no macropathological findings. Histopathological lung samples were
stained with Brdu monoclonal antibody then examined
immunohistochemically counting positive nuclei in terminal bronchioles.
The Brdu Index was significantly higher in the treated mice than concurrent
controls after 3 and 7 days indicating an increase in the number and
incidence of cells in the DNA synthesis (S) phase of cell cycle preparing for
cell division or past the S phase. Values for treated rat were comparable
with concurrent controls on both occasions indicating no such activity as
seen in the mouse. These results indicated IKI-220 had mitogenic activity
in the terminal bronchioles of the mouse but not the rat. The species
difference response was similar to that seen with styrene toxicity.
A much higher incidence of spontaneous bronchioloalveolar tumours in
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aged mice than aged rats coupled with some toxic effect of IKI-220 that
stimulates cell turnover allowing expression of additional background
spontaneous tumours might be associated with this higher level of induced
increase of cell cycle marker of Brdu in terminal bronchioles. The results of
this study suggest the increased incidence of lung tumours in CD-1 mice
was due to an increased number of cells in the DNA synthesis phase of the
cell cycle. There was no similar response to IKI-220 in rats. Information in
the literature indicates that humans respond to similar chemicals (such as
Isoniazid) more like rats than mice and thus are unlikely to develop lung
tumours in response to exposure to IKI-220. Findings in the mouse are not
likely to translate to humans.
Additional Comments No additional comments
Conclusion IKI-220 demonstrated mitogenic effects in the terminal bronchioles of mice
but not rats indicating species specificity
Type of study Lung cell cycle analysis in the mouse
Flag Supporting study
Test Substance IKI-220 Lot No. 9809 , TFNG Lot No. 0006, TFNA Lot No.0006, TFNA-AM
Lot No. 0006
Reference
Nomura, M., Cell cycle analysis using Brdu in the mouse lung following
short term dietary administration of IKI-220 and its metabolites: TFNG,
TFNA and TFNA-AM, Safety Sciences Group, Safety Science Research
Laboratory, Ishihara Sangyo Kaisha, Japan, 2003. ISK Document No. 2050
AN-2163
Klimisch Score
2 – reliable with restrictions - on the basis that the study was conducted
‘in-house’ by the Applicant Ishihara Sangyo Kaisha. The Applicants’ own
Compliance Statement includes the comment ‘the study was not conducted
or reported in compliance with Good Laboratory Practice (GLP)
Regulations. However, we believe that the results of this study are
scientifically valid and would not have been different if the study had been
conducted under GLP regulations’
Amendments/Deviations None
GLP Conducted in accordance with GLP principles
Test Guideline/s Not applicable
Species Mouse
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Strain Crj: CD-1 (ICR)
No/Sex/Group 5M
Dose Levels
0, 2250 ppm for 3 or 7 days equivalent to 389/330 (IKI-220), 402/318
(TFNG), 364/336 (TFNA), 385/332 (TFNA-AM) mg/kg bw/day for 3/7 days
respectively. – admixture with the diet. Bromodeoxyuridine (Brdu – a cell
cycle marker) was injected 3 hours prior to sacrifice
Study Summary
There were no deaths or any adverse treatment related clinical signs.
There were no macropathological changes. Histopathological lung samples
were treated with anti-Brdu monoclonal antibodies then examined
immunhistochemically.
The Brdu Index in the terminal bronchiole was significantly increased in the
groups treated with IKI-220, being higher after 3 days than 7 days. There
were no differences in the Brdu Index for groups receiving any of the
metabolites compared with concurrent controls. Thus, only IKI-220 induced
an increase of Brdu positive cells in the terminal bronchiolar epithelia of
mouse lung.
Additional Comments No additional comments
Conclusion Comparison of potential mitogenic activity of IKI-220 and its metabolites
showed only the parent material to have any effects.
Type of study Applicants review of lung findings in the mouse oncogenicity studies
Flag Weight of evidence
Test Substance IKI-220 technical
Reference
Nomura, M., IKI-220: Discussion on lung finding observed in mouse
oncogenicity, Safety Sciences Group, Safety Science Research
Laboratory, Ishihara Sangyo Kaisha, Japan, 2003. ISK Document No.
2052A AN-2052
Study Summary
In summary:
Only the mouse showed significant increases in bronchioloalveolar
tumours in response to dietary administration of IKI-220 (Flonicamid)
technical. The tumours were not life threatening as nearly all were detected
at the terminal sacrifice.
There were no treatment related tumours seen in rats.
The increase in bronchioloalveolar tumours in mice was associated with an
increase in alveolar/bronchiolar hyperplasia/hypertrophy.
Short term investigations in mice with dietary administration of IKI-220
resulted in increased cell turnover of Clara cells of the terminal bronchioles
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as measured by Brdu staining.
These short term studies did not result in any cytotoxic effects of IKI-220 in
the lung bronchioles, suggesting the effect of IKI-220 was a mitogenic
effect stimulating cell turnover.
IKI-220 is not mutagenic.
There is a threshold level for mitogenic activity between 80 and 250 ppm
administration.
There is a clear species difference between mice and rats as assessed by
lung tumour incidence and Brdu Index analysis.
IKI-220 induced a unique mitogenic activity in the terminal bronchiolar
epithelial cells (non-ciliated Clara cells) in mice but its three animal
metabolites had no similar effect.
The increased Brdu Index in Clara cells in mice and activated Clara cells in
target tissue were clearly reversible within a week of withdrawal from IKI-
220, with no damage to the lung.
The IKI-200 induced increase in the Brdu Index appears to be related to
the different sensitivity of strains of mice to development of spontaneous
tumours in the lungs as seen in CD-1 mice but not B6C3F1 (intermediate
sensitivity) and C57 (least sensitivity) mice.
Dietary administration of Isoniazid which is structurally similar to IKI-220
and which has also been shown to increase levels of spontaneous lung
tumours in mice but not in rats caused an increase in Brdu Index in all
three strains of mice. The effect of IKI-220 on the target cell of mice lungs
is clearly weaker than that of Isoniazid.
Thus, it was concluded that IKI-220 induction of increased
bronchioloalveolar tumours is unique to the mouse, especially to sensitive
strains such as the CD-1 strain which have incidences of spontaneous
tumours. A threshold between 80 and 250 ppm has been demonstrated for
the pre-neoplastic event in the CD-1 strain. This promoter effect is readily
reversible and lung tissue returns to normal within 1 week of cessation of
exposure to IKI-220. The absence of similar effects in the lung tissue of
rats suggest the effects seen in mice may not translate to other species
including humans.
Isoniazid, used to treat tuberculosis patients, has also been shown to
induce a high incidence of bronchioloalveolar tumours in mice but not in
rats. Comparison of IKI-220 and Isoniazid showed that Isoniazid was more
potent in causing cell turnover in mouse lungs than IKI-220. The IARC has
classified Isoniazid in Group 3 – cannot be classified as to its
carcinogenicity to humans. The overall conclusion is that a non-linear risk
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assessment should be used to assess human risk for the effects observed
in mouse lungs treated with IKI-220 based on demonstration of a threshold
for effect, confirmation that IKI-220 is not mutagenic, there is rapid
reversibility without tissue damage and specificity of the effects to the CD-1
mouse strain with no effects in other mouse strains or the rat.
Additional Comments No additional comments
Conclusion
The mechanism of tumour induction is specific to the CD-1 mouse
exacerbated by its higher sensitivity due to the much higher numbers of the
target Clara cells in the bronchiolar/alveolar epithelium compared with
other mouse strains. No similar effect was demonstrated on other mouse
strains or in the rat. A threshold dose was demonstrated, there was no
dose relationship and the occurrence of tumours did not affect survival as
occurrence was observed at terminal sacrifice, primarily being benign
adenomas. Human lung has a much lower number of Clara cells in the
target tissue and thus classification (6.7) is considered not to be
appropriate.
General conclusion about carcinogenicity classification:
IKI-220 technical was not carcinogenic in the rat.
In the mouse exposure to IKI-220 technical resulted in increased incidences of primary lung tumours
(alveolar/bronchiolar adenomas and carcinomas) in both sexes. At dosages that did not induce tumours pre
neoplastic changes were noted in the lungs. Investigations to elucidate the aetiology of tumour formation in
the CD-1 strain demonstrated a dose related increase in cell proliferation in epithelial cells of the
alveolar/bronchiolar region. The induction of cell proliferation was shown to be readily and fully reversible
within 7 days of the withdrawal of treatment. The Clara cell was shown to be a specific target of IKI-220
undergoing reversible hypertrophy/hyperplasia and morphological elongation in response to the test
substance. Furthermore there was no evidence of secondary tissue responses in the areas surrounding the
activated Clara cells. Thus, the pattern of effects suggested a mitogenic rather than cytotoxic effect of IKI-
220 technical. These effects were seen only with the parent molecule, not with any of its metabolites.
Further investigation showed the effects to be species specific and strain sensitive. The proportion of Clara
cells in the lung of the mouse strains examined was approximately 80%, whereas in the rat this is only about
35%. The CD-1 mouse has the highest incidence of spontaneous bronchiola/alveolar tumours. This
combined with the proportion of immature Clara cells in the lung epithelium and higher sensitivity to
stimulated cell division would account for the high incidence of lung tumours seen.
Investigations demonstrated a threshold level for the mitogenic response, and IKI-220 technical did not
demonstrate any mutagenic or genotoxic effect at the DNA, gene and chromosome levels of organisation.
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Thus, there is sound rationale to support the contention that IKI-220 induced bronchioalveolar tumours is
unique to the mouse and specifically to the CD-1 strain.
In the human lung the Clara cell complement is very low in respiratory and terminal bronchioles, 22% and
11% respectively. Epidemiological studies on Isoniazid, a structurally similar molecule, revealed no
relationship between its therapeutic use and the occurrence of human lung tumours. Thus, it is concluded
that a non-linear risk assessment is appropriate for IKI-220 technical based on the absence of genotoxicity,
the species specific nature of the lung effect, its ready reversibility without tissue damage and the
demonstration of consistent NOEL doses for both the mechanism and tumour formation. Consequently, it is
considered classification (6.7) is not required.
Reproductive/Developmental Toxicity [6.8]
Developmental Toxicity
Type of study Rat embryofoetal development study
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoints
Parental Toxicity
LOAEL: 500 mg/kg bw/day altered liver and kidney
micropathology
NOAEL: 100 mg/kg bw/day
Foetal Toxicity
LOAEL: 500 mg/kg bw/day – increased incidence of
skeletal anomalies
NOAEL: 100 mg/kg bw/day
Reference Hojo, H., IKI-220 technical: A teratogenicity study in rats, The Institute of
Environmental Toxicology, Japan, 2002. ISK Document No. 2031 IET 00-0023
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 414, US EPA OPPTS 870.3700
Species Rat
Strain Jcl:Wistar
No/ Group 24F
Dose Levels 0, 20, 100. 500 mg/kg bw/day
Exposure Type Oral gavage, Days 6 – 19 of gestation
Study Summary
The only treatment related findings amongst maternal rats were confined to 500
mg/kg bw/day. These consisted of slightly lower food intake during the dosing
period, a significant increase in liver weight, hypertrophy of centrilobular
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hepatocytes in the liver (13/14) and vacuolation of proximal tubular cells in the
kidney (24/24). At necropsy there was an increase in placental weights whilst
examination of foetuses revealed an increase in skeletal anomalies (specifically
an increased incidence of cervical (extra) rib) considered to reflect maternal
toxicity.
There were no treatment related effects in maternal rats or foetuses at 20 or 100
mg/kg bw/day.
Additional Comments No additional comments
Conclusion
Maternal toxicity was confined to slightly lower food intake and changes to the
liver with a correlated finding of a slight increase in skeletal (extra rib) anomalies,
a foetal finding commonly considered an indicator of maternal toxicity.
Type of study Rabbit embryofoetal development study
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoints
Parental Toxicity LOAEL: 25 mg/kg bw/day – impaired weight gain
NOAEL: 7.5 mg/kg bw/day
Foetal Toxicity LOAEL: >25 mg/kg bw/day
NOAEL: 25 mg/kg bw/day
Reference Takahashi. K., IKI-220 technical: Teratogenicity study in rabbits, The Institute of
Environmental Toxicology, Japan, 2002. ISK Document No. 2032A IET 00-0025
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 414, US EPA OPPTS 870.3700
Species Rabbit
Strain Japanese white Kbl:JW
No/ Group 25F
Dose Levels 0, 2.5, 7.5, 25 mg/kg bw/day
Exposure Type Oral gavage, Days 6 to 27 of gestation
Study Summary
The only treatment related findings were significantly lower maternal bodyweight
gain and food intake during the dosing period seen at 25 mg/kg bw/day. There
were no adverse maternal findings at 2.5 or 7.5 mg/kg bw/day. There was no
adverse treatment related effects on embryofoetal development at any of the
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dosages investigated.
External, visceral, and skeletal examinations of live fetuses revealed several types
of malformations and variations in all groups including the control. However, these
findings were not considered to be adverse for a number of reasons including that
many of the malformations were within the historical control range, and the
frequency of malformations did not significantly increase with dose.
Additional Comments No additional comments
Conclusion Maternal toxicity was limited to lower weight gain and food intake, however, there
were no adverse effects on embryofoetal development.
Reproductive toxicity
A preliminary study was performed but details are not reported here:
Takahashi, K., IKI-220 technical: Reproductive toxicity study in rats preliminary study, Institute of
Environmental Toxicology, Japan, 2002. ISK Docment No. 2046 IET 99-0084
Type of study Rat 2 generation reproduction study
Flag Key
Test Substance IKI-220 technical
Endpoints
Parental Toxicity –
F0 and F1 generation
LOAEL: 1800 ppm – 109.1/124.8 (F0),
163.8/176.8 (F1) males/females – altered kidney
pathology
NOAEL: 300 ppm 22.3/26.5 mg/kg bw/day
males/females (The change in luteinizing hormone
seen in F1 females at 50 ppm was considered to
be a compensatory mechanism rather than a toxic
effect)
Reproductive Toxicity
LOAEL: Not identified
NOAEL: 1800 ppm 109.1/124.8 mg/kg bw/day
males/females F0 generation, 163.8/176.8 mg/kg
bw/day males/females F1 generation
Developmental
toxicity F1 and F2
offspring:
LOAEL: Not identified (males), 1800 ppm females
- 153.4 mg/kg bw/day – delayed vaginal opening in
F1 generation
NOAEL: 1800/300 ppm 163.8/30.5 mg/kg b/day
males/females F1 generation
NOAEL: 1800 ppm (females only F2 generation) –
176.8 mg/kg bw/day
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Reference
Takahashi, K., IKI-220 technical: reproductive toxicity study in rats, The
Institute of Environmental Toxicology, Japan, 2002. ISK Document 2037
No. IET 99-0085
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s US EPA OPPTS 870.3800, OECD 416
Species Rat
Strain Jcl:Wistar
No/Sex/Group 24 males and 24 females
Dose Levels
0, 50, 300, 1800 ppm equivalent to 0, 3.07, 18.3, 109.1 and 0, 3.39, 20.7,
124.8 mg/kg bw/day males/females F0 generation and 0, 4.67, 28.2, 163.8
and 0, 4.95, 30.5, 176.8 mg/kg bw/day males/females F1 generation
respectively respectively
Exposure Type Admixture with the diet
Study Summary
There was no adverse effect of treatment on reproductive capacity at any
of the dosages investigated in either generation.
1800 ppm resulted in increased absolute and relative kidney weights in
adult males of both generations, often appearing pale at necropsy.
Micropathologically there were increased incidences of tubular basophilic
change and granular casts in dilated tubules in both generations.
Immunohistochemical examination of the hyaline droplets revealed the
presence of α2µ-globulin. Incidences of vacuolation of proximal tubular
cells in the kidney were increased in adult females of both generations.
Delayed vaginal opening was seen in F1 females at 1800 ppm (but not F2
generation females).
Findings at 300 ppm were confined to increased relative kidney weight in
F1 males with an increased incidence of hyaline droplet deposition in the
proximal tubule cells of males of both generations. Because of the absence
of any tubular basophilic changes or granular casts this change was
considered not to be adverse.
There were no treatment related findings at 50 ppm in either sex or
generation.
Additional Comments No additional comments
Conclusion Treatment with IKI-220 did not result in any in life signs of toxicity in either
generation. Reproductive capacity was unaffected in both generations.
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Toxicity was confined to kidney changes consistent with findings in
subchronic and chronic toxicity studies. Developmental toxicity was
confined to F1 females at the highest dose where there was a delay in
sexual maturation manifest as delayed vaginal opening, however,
reproductive capacity was unaffected.
Type of study IKI-220 oestrogen receptor binding affinity study in rats
Flag Supporting study
Test Substance IKI-220 technical
Endpoint Receptor binding affinity – very low binding affinity for both α- and β-
receptors
Reference
Takahasi, K., IKI-220 technical: reproductive toxicity study in rats additional
observations, The Institute of Environmental Toxicology, Japan, 2002. ISK
Document No. 2040 IET 01-8008
Klimisch Score 2 – reliable with restrictions
Amendments/Deviations None
GLP Not specified
Test Guideline/s Not applicable
Species Rat
Strain Jcl:Wistar
No/Sex/Group 8M and 8F
Dose Levels 0, 50, 300, 1800 ppm equivalent to 0, 3.7, 22.3, 132.9 and 0, 4.4, 26.5,
153.4 mg/kg bw/day males/females respectively
Exposure Type Admixture with the diet
Study Summary
The serum samples used for this investigation were those taken from F1
adults of the previously cited rat reproduction study.
Measurement of serum hormones revealed a significant increase in LH in
females at 300 ppm and a slight decrease in 17β- oestradiol in females at
1800 ppm. No changes were seen in the hormone levels of males.
Oestrogen receptor binding affinity was very low indicating this not to be
the mechanism for IKI-220 anti-oestrogen effects. IKI-220 may cause an
increase in gonadotrophin levels as a result of slight inhibition of 17β-
oestradiol production rather than an acceleration of gonadotrophin
production in females since no change were noted in treated males. Thus,
it was concluded that IKI-220 technical causes a decrease in blood 17β-
oestradiol levels of females at 1800 ppm resulting in decreases in ovary
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and uterine weights and an increase in the day of age of vaginal opening.
Additional Comments
The study report does not contain an author signature page or any
reference to conduct in compliance with GLP principles, however, the data
are considered reliable.
Conclusion
The slightly later sexual maturation of F1 females at 1800 ppm in the
reproductive study was attributable to IKI-220 technical lowering 17β-
oestradiol blood levels.
General conclusion about reproductive/developmental toxicity classification:
IKI-220 technical did not adversely affect fertility or reproductive capacity. There were indications suggestive
of interference with sexual maturation of F1 offspring manifest principally as slightly delayed vaginal opening
in F1 offspring however this was not repeated for F2 offspring. Micropathological changes in the kidneys
were seen in adult males and females of the F0 and F1 generations, a finding consistent with the sub chronic
toxicity studies.
IKI-220 did not exert any adverse effect on embryofoetal development. Although an increased incidence of
extra (cervical) rib was noted in the rat at the highest dose investigated this is commonly associated with
maternal toxicity rather than a direct effect of treatment and thus is not relevant to human risk. (Preliminary
investigations in the rat up to 1000 mg/kg bw/day resulted in deaths and liver and kidney changes indicating
the highest dose in the main study (500 mg/kg bw/day) was on the margins of anticipated maternal toxicity).
Reviewers comment: It should be noted that the European Food Safety Authority (EFSA) and subsequent
European Chemical Agency (ECHA) reviews differed in their respective conclusions regarding embryofoetal
development. The EFSA concluded that there was evidence of adverse embryofoetal developmental effects
in the rat and the rabbit at (essentially) non-maternally toxic doses (500 and 7.5 mg/kg bw/day respectively)
consisting of an increased incidence of cervical ribs in rats and an increase in the incidence particularly of
visceral malformations in rabbits. In the rat study there was evidence of treatment related adverse maternal
effects at 500 mg/kg bw/day and thus a ‘disturbance’ in the number of foetuses with extra ribs (which was
within the historical background control range) is consistent with a ‘stress’ response rather than any direct
effect on development, if indeed it is an ‘effect’ at all. With regard to the rabbit the incidences of
malformations were in fact within the historical control range, were not consistent nor were they dose related.
Also, no increase in extra ribs was noted which, given the argument that this was an effect in the rat might
reasonably be expected to be seen in the rabbit if it were truly a treatment related direct effect on
embryofotal development. Thus, the Reviewer concurs with the conclusions of the ECHA that there is no
direct adverse effect of Flonicamid on embryofoetal development.
Given the marginal nature of the findings in the reproductive study, confined to delayed vaginal opening in
the high dose F1 females only but without any adverse effect on reproductive capacity no classification
(6.8B) is considered necessary. No classification for 6.8C is required.
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Target Organ Systemic Toxicity [6.9]
Preliminary dose range finding studies were conducted for the rat and the dog but the details are not presented:
Ridder., W.E., Yoshida, M., Watson, M., A 28-day oral toxicity study in dogs with IKI-220 technical, Ricerca LLC,
USA, 2001. Document No. 2014 010871-1
Kuwahara, M., IKI-220 technical: 28-day dose range finding study in rats, Institute of Environmental Toxicology,
Japan, 2002. ISK Document No. 2025 IET 98-0140
Type of study 28 day dermal toxicity study in rats
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint
LOAEL: Not identified >1000 mg/kg bw/day
NOAEL: 1000 mg/kg bw/day
Reference
Ridder, W., A 28 day repeated dose dermal toxicity study in rats with IKI-220,
Toxicology and Pharmacology, Ricerca, LLC, USA, 2001. ISK Document No. 2017
012074-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 410, US EPA OPPTS 870.3200
Species Rat
Strain Crl:CD® (SD) (IGS) BR
No/Sex/Group 10M and 10F
Dose Levels 0, 20, 150, 1000 mg/kg bw/day, 6h per day, occluded
Study Summary There were no treatment related effects on any of the parameters recorded at any
of the dosages investigated.
Additional Comments No additional comments
Conclusion No classification required
Type of study Mouse 90 day toxicity study
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint LOAEL: 1000 ppm, equivalent to 153.9/191.5 mg/kg bw/day males/females
respectively – altered spleen micropathology in both sexes and liver in males
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NOAEL: 100 ppm equivalent to 15.25/20.1 mg/kg bw/day males/females
respectively
Reference Ridder, W., A 13 week feeding study in mice with IKI-220, Toxicology and
Pharmacology, Ricerca, LLC, USA, 2001. ISK Document No. 2020 8090-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 408, US EPA OPPTS 870.3100
Species Mouse
Strain Crl:CD-1® (ICR) BR
No/Sex/Group 10M and 10F
Dose Levels 0, 100, 1000, 7000 ppm equivalent to 0, 15.25, 153.9, 1069 and 0, 20.10, 191.5,
1248 mg/kg bw/day males/females respectively. By admixture with the diet
Study Summary
There were no deaths. There were no clear treatment related clinical signs seen at
any dosage. Lower weight gain in both sexes at 7000 ppm generally during the
first 5 weeks of treatment combined with lower food intake was considered to be
attributable to palatability rather than a toxic effect of treatment. Although
subsequent weight gain was evident comparability with controls was not regained
by termination. Haematological investigations revealed significantly lower numbers
of red blood cells, decreased haemoglobin, haematocrit, mean cell volume and
mean corpuscular haemoglobin in both sexes receiving 7000 ppm suggesting a
treatment related anaemia. There were no haematological changes at 100 or 1000
ppm. Blood chemistry revealed significantly increased creatinine, bilirubin, sodium
and chloride with decreased potassium in males at 7000 ppm whilst females at
this dose revealed increased blood glucose. The increased total bilirubin could be
due to the decreased red blood cell count reflecting a treatment related red blood
cell destruction whilst the increased blood sugar could reflect liver changes. At
necropsy absolute and relative liver and spleen weights were increased in both
sexes at 7000 ppm compared with controls, the spleen change possibly being
related to anaemia. Histopathological investigation revealed centrilobular
hepatocellular hypertrophy in the liver of both sexes at 7000 ppm and males at
1000 ppm, increased extramedullary haemopoiesis and pigment deposition in the
spleen of both sexes at 7000 ppm and 1000 ppm and increased pigment
deposition in the bone marrow in both sexes at 7000 ppm. The spleen and marrow
changes were considered to reflect the anaemia whilst the liver changes were
considered to reflect possible enzyme induction.
Additional Comments No additional comments
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Conclusion The liver, spleen and bone marrow were identified as target organs.
Type of study Rat 90 day toxicity study
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint
LOAEL: 1000/5000 ppm males/females equivalent to 60.0/340.1 mg/kg bw/day
males/females respectively – altered kidney micropathology both sexes, liver in
females
NOAEL: 200/1000 ppm males/females equivalent to 12.11/72.3 mg/kg bw/day
males/females respectively
Reference
Kuwahara, M., IKI-220 technical: 90-day subchronic oral toxicity study in rats, The
Institute of Environmental Toxicology, Japan, 2002. ISK Document No. 2015 IET
98-0141
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 148, US EPA OPPTS 870.3100
Species Rat
Strain Jcl:Wistar
No/Sex/Group 12M and 12F
Dose Levels
0, 50 (males), 200, 1000, 2000 (males), 5000 (females) ppm equivalent to 0,
3.079, 12.11, 60.0, 119.4 and 0, 14.52, 73.3, 340.1 mg/kg bw/day males/females
respectively. By admixture with the diet
Study Summary
Treatment at 5000 ppm, assessed in females only resulted in significantly lower
food intake compared with controls. Haematocrit was significantly decreased
whilst mean corpuscular haemoglobin concentration was increased. Triglyceride
was decreased. Absolute and relative liver and kidney weights were increased
with histopathology revealing increased incidences of centrilobular hypertrophy in
the liver and cytoplasmic vacuolation of proximal tubular cells in the kidney when
compared with concurrent controls
At 2000 ppm, assessed in males only, findings were confined to post sacrifice
investigations and consisted of increased absolute and relative kidney weight with
kidneys noted at necropsy to be pale. Histopathology revealed deposition of
hyaline droplets containing α2µ-globulin in proximal tubules of the kidney of all
animals. The incidences of granular casts in dilated tubules and basophilic change
of tubular cell, considered to be degenerative change due to α2µ-globulin
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deposition were significantly increased. There was an increased incidence of
centrilobular hepatocellular hypertrophy in the liver.
At 1000 ppm findings were confined to males only and consisted of increased
absolute and relative kidney weight with hyaline droplet deposition in proximal
tubular cells, basophilic changes in tubular cells and granulated casts in dilated
tubules of the kidney. There were no treatment related effects in females.
At 200 ppm the only treatment related change in either sex was confined to 8/12
males showing hyaline droplet deposition in proximal tubular cells of the kidney,
however this was considered not to be an adverse effect.
There were no treatment related changes in any parameters at 50 ppm, assessed
in males only.
Additional Comments
Reviewers comment: the findings relating to alpha-2u-globulin-mediated
nephropathy are not considered relevant to humans. Therefore for human
health risk assessment purposes the LOAEL in males could be
considered to be 2000 ppm (equivalent to 119.4 mg/kg bw/day and the
NOAEL 1000 ppm (equivalent to 60 mg/kg bw/day).
Conclusion The liver and kidney were identified as target organs
Type of study Dog 90 day toxicity study
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint
LOAEL: 20 mg/kg bw/day – acute vomiting episodes post dosing
NOAEL: 8 mg/kg bw/day
Reference Ridder, W. E., A 90-day toxicity study in dogs with IKI-220 technical, Toxicology
and Pharmacology, Ricerca LLC, USA, 2001. ISK Document No. 2016 011509-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 409, US EPA OPPTS 870.3150
Species Dog
Strain Beagle
No/Sex/Group 4M and 4F
Dose Levels 0, 3, 8, 20, 50 (females only) mg/kg bw/day. Oral capsule
Study Summary After the first dose, clinical signs of an acute response to the test substance –
vomiting, collapse, prostration, convulsions – were seen in 2 females receiving 50
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mg/kg bw/day whilst vomiting was seen in 2 males receiving 20 mg/kg bw/day.
Clinical signs persisted amongst the females at 50 mg/kg bw/day with associated
decreased food intake, inappetance, anorexia and weight loss resulting in 1
female being sacrificed in Week 4 whilst another was removed from treatment in
Week 9 after which there was complete reversal of the acute signs. Vomiting post
dosing was a persistent sign amongst males at 20 mg/kg bw/day. Clinical
pathology revealed changes in red blood cell and reticulocyte counts in females at
50 mg/kg bw/day in Week 7 but which were not evident at termination. Increased
total protein in blood chemistry was evident in males at 20 mg/kg bw/day in Week
7 but was not evident at termination. No treatment related changes were evident
in urinalysis at any dosage. Histological examinations of tissues from the female
that was sacrificed at 50 mg/kg bw/day revealed mild oedema of the pancreas,
mild evolution of the thymus and vacuolation of the tubules in the inner cortex of
the kidney, the latter finding also seen in another female at this dose. A third
female at 50 mg/kg bw/day had mild haemorrhage at the ileo-colic junction. There
were no other treatment related changes in any animal at any dose.
Additional Comments
No additional comments.
Reviewer comment - The report author indicated a suggested treatment related
effect on thymus weight (decreased) in males at 20 mg/kg bw/day however did not
indicate this in the report summary. There were no histopathological changes
reported for the thymus.
Conclusion
Clinical signs were considered principally attributable to an immediate acute
response to the test substance rather than systemic. Systemic toxicity was evident
with decreased food intake and weight loss in females. No obvious target organs
were identified.
Reviewer comment – The changes in kidney micropathology seen in 2 females at
50 mg/kg bw/day are consistent with renal changes seen in female rats.
Type of study Dog 52 week toxicity
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint
LOAEL: 20 mg/kg bw/day – acute vomiting episodes post dosing and
haematological changes in both sexes, lower weight gain in females
NOAEL: 8 mg/kg bw/day
Reference
Ridder, W. E., Watson, M., A 52 week oral toxicity study in dogs with IKI-220
technical, Toxicology and Pharmacology, Ricerca, LLC, USA, 2002. ISK
Document No. 2044 012075-1-1
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Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 409, US EPA OPPTS 870.3150
Species Dog
Strain Beagle
No/Sex/Group 6M and 6F
Dose Levels 0, 3, 8, 20 mg/kg bw/day. Oral capsule
Study Summary
There were no treatment related deaths at any dose. All males and 4/6 females at
20 mg/kg bw/day vomited after the first dose with one male and one female
showing decreased activity and ataxia. Incidences of vomiting continued post
dosing but eventually resolved as tolerance improved. These findings are an acute
response to the test substance rather than any systemic response. There were no
treatment related clinical signs at any other dosage. Lower bodyweight gain was
evident in females receiving 20 mg/kg bw/day. There were no treatment related
changes in clinical chemistry parameters at any dosage. The only notable
haematological change was a significantly increased reticulocyte count in both
sexes at 20 mg/kg bw/day at termination, a finding consistent with that seen in the
earlier 90 day study suggesting a pattern affecting red blood cells. There were no
treatment related macroscopic changes noted at necropsy or any
histopathological changes in any tissues at any dosage.
Additional Comments No additional comments
Conclusion
Initial clinical signs post dose were considered to be an acute response to the test
substance which were eventually tolerated. Indications of systemic toxicity were
confined to lower weight gain in females and haematological changes in both
sexes, however, no target organs were identified.
Type of study Acute neurotoxicity study in rats
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint
LOAEL: Neurotoxicity: >600 (males), >1000 (females) mg/kg bw; systemic toxicity:
1000 mg/kg bw NB – neurotoxicity/pathology was evaluated only in males treated
at 600 mg/kg bw and females at 1000 mg/kg bw
NOAEL: Neurotoxicity: 600 (males), 1000 (females) mg/kg bw; systemic toxicity:
600 (males), 300 (females) mg/kg bw
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Reference
Ridder, W. E., Watson, M., An acute neurotoxicity study in rats with IKI-220
technical, Toxicology and Pharmacology, Ricerca Biosciences LLC, USA, 2002.
ISK Document No. 2043 012076-1-2
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 424, US EPA 870.6200
Species Rat
Strain Sprague Dawley Crl:CD® (SD) IGS BR
No/Sex/Group 10M and 10F
Dose Levels 0, 100, 300, 600 (males only), 1000 (5M and 10F) mg/kg bw/day. Oral gavage
Study Summary
A single male at 1000 mg/kg bw exhibited impaired locomotion, decreased activity
and tremors at 30 and 60 minutes post dosing and was found dead the following
day. A single female at 1000 mg/kg bw exhibited slightly impaired mobility,
tremors and abnormal gait at 30 and 60 minutes post dosing but subsequently
recovered. These events, at a dosage higher than the LD50 value in males were
considered to be attributable to acute systemic toxicity rather than neurotoxicity.
There were no other clinical signs considered attributable to treatment at any
dosage. There were no treatment related neurotoxic or neuropathological effects
at the highest dosage investigated in each sex – 600 mg/kg bw in males and 1000
mg/kg bw in females, thus IKI-220 is not considered a neurotoxin.
Additional Comments No additional comments
Conclusion There was no evidence of neurotoxicity
Type of study 90 day repeat dose neurotoxicity study in rats
Flag Key
Test Substance IKI-220 technical, Lot No. 9809
Endpoint
LOAEL: Neurotoxicity: >10000 ppm (625/722 mg/kg bw/day males/females).
Systemic toxicity: 10000 ppm (625/722 mg/kg bw/day males/females).
NOAEL: Neurotoxicity: 10000 ppm (625/722 mg/kg bw/day males/females).
Systemic toxicity: 1000 ppm (67/81 mg/kg bw/day males/females)
Reference
Schaefer, G. J., A dietary subchronic (90-day) neurotoxicity study on IKI-220
technical in rats, WIL Research Laboratories, USA, 2003. ISK Document No. 2039
449002
Klimisch Score 1
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Amendments/Deviations None
GLP Y
Test Guideline/s OECD 424, US EPA OPPTS 870.6200
Species Rat
Strain Crl:CD® (SD) IGS BR
No/Sex/Group 10M and 10F
Dose Levels 0, 200, 1000, 10000 ppm equivalent to 0, 13, 67, 625 and 0, 16, 81, 772 mg/kg
bw/day males/females respectively – admixture with the diet
Study Summary
There were no deaths or any treatment related clinical signs at any dosage. Lower
weight gain was evident in both sexes at 10,000 ppm throughout the treatment
period, being 32% and 12% for males and females respectively compared with
concurrent controls by termination. Concomitant lower food intake was seen in
both sexes at 10,000 ppm. There were no signs indicative of neurotoxicity as
assessed by locomotor activity assessments at any dose nor were there any
treatment related macroscopic or neuromicropathological changes at any dose.
Additional Comments No additional comments
Conclusion There was no evidence of neurotoxicity
1. General conclusion about target organ systemic toxicity:
In the rat the liver and kidneys were identified as target organs in both sexes although there was a clear
difference between the sexes for morphological and sensitivity for renal changes. These can be separated
as a treatment related albeit not adverse effect in the kidney of males only resulting from hyaline deposition
resulting from IKI-220 binding to α2µ-globulin and a more chronic, degenerative renal change seen in both
sexes in the rat and to a lesser degree the dog. There was an indication of mild anaemia in the dog.
It should be noted that α2µ-microglobulin is a male rat specific protein and thus effects mediated by this
mechanism have no relevance to humans. The degerative renal changes in rats occurred with a LOAEL
greater than those within the guidance range values for classification.
In the mouse the principal target organs were the liver and haemopoietic system and lung. There was only
slight evidence of kidney dysfunction.
The liver changes seen in rats and mice were consistent irrespective of the duration of exposure, being
increased incidences of hepatocellular hypertrophy. There was no evidence of this change developing any
further and hepatocellular hypertrophy is consistent with a change associated with enzyme induction. Thus,
in the absence of any degenerative change this is considered not of any significant human relevance.
Dermal exposure is well tolerated.
IKI-220 technical does not induce any neurotoxic effects.
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It is therefore concluded that no classification (6.9) is required.
2. Metabolites
Investigations were conducted on five metabolites, TFNA, TFNA-AM, TFNG, TFNG-AM and TFNA-OH
consisting of acute oral LD50 and bacterial reverse mutation assessments. 90 day toxicity studies were also
conducted with TFNA and TFNG.
Type of study Acute oral limit toxicity study in rats
Flag Supporting study
Test Substance TFNA, Batch No. 0006
Endpoint LD50
Value >2000 mg/kg bw (limit dose)
Reference Damme, B., TFNA: Acute oral toxicity study in rats, RCC Ltd, Toxicology Division,
Switzerland, 2002. ISK Document No.2101 834142
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 423, 96/54/EEC, US EPA OPPTS 870.1000
Species Rat
Strain Han Brl: WIST
No/Sex/Group 3M and 3F
Dose Level 2000 mg/kg bw
Exposure Type Oral gavage
Study Summary There were no deaths, no adverse signs of reaction to treatment or any
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion No classification required
Type of study 90 day toxicity study in rats
Flag Supporting study
Test Substance TFNA, Lot No. 9709
Endpoint
LOAEL: >2000 ppm males (136 mg/kg bw/day), >5000 ppm females (409 mg/kg
bw/day)
NOAEL: 2000 ppm males (136 mg/kg bw/day), 5000 ppm females (409 mg/kg
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bw/day)
Reference Nagaike, M., TFNA: 90-day subchronic oral toxicity study in rats, Ishihara Sangyo
Kaisha, Japan, 2003. ISK Document No. 2112 AN-1992
Klimisch Score 2 – reliable with restrictions
Amendments/Deviations None
GLP N
Test Guideline/s Not applicable
Species Rat
Strain Jcl:Wistar
No/Sex/Group 5M and 5F
Dose Levels
0, 50 (males only), 200 (females only), 2000 (males only), 5000 (females only ppm
equivalent to 0, 3.42, 15.9, 136, 409 mg/kg bw/day respectively. Admixture with
the diet
Study Summary There were no treatment related changes in any of the parameters assessed at
any dosage.
Additional Comments No additional comments
Conclusion There was no evidence of toxicity
Type of study Acute oral limit toxicity in rats
Flag Supporting study
Test Substance TFNA-AM. Batch No. 0006
Endpoint LD50
Value >2000 mg/kg bw (limit dose)
Reference Damme, B., TFNA-AM: Acute oral toxicity study in rats, RCC Ltd, Toxicology
Division, Switzerland, 2002. ISK Document No.2102 834750
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 423, 96/54/EEC, US EPA OPPTS 870.1000
Species Rat
Strain Han Brl: WIST
No/Sex/Group 3M and 3F
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Dose Level 2000 mg/kg bw
Exposure Type Oral gavage
Study Summary There were no deaths, no adverse signs of reaction to treatment or any
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion No classification required
Type of study Acute oral limit toxicity study in rats
Flag Supporting study
Test Substance TFNG, Batch No. 0006
Endpoint LD50
Value >2000 mg/kg bw (limit dose)
Reference Damme, B., TFNG: Acute oral toxicity study in rats, RCC Ltd, Toxicology Division,
Switzerland, 2002. ISK Document No.2103 834761
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 423, 96/54/EEC, US EPA OPPTS 870.1000
Species Rat
Strain Han Brl: WIST
No/Sex/Group 3M and 3F
Dose Level 2000 mg/kg bw
Exposure Type Oral gavage
Study Summary There were no deaths, no adverse signs of reaction to treatment or any
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion No classification required
Type of study 90 day toxicity study in rats
Flag Supporting study
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Test Substance TFNG, Lot No. 9709
Endpoint
LOAEL: >2000 ppm males (135 mg/kg bw/day), >5000 ppm females (411 mg/kg
bw/day)
NOAEL: 2000 ppm males (135 mg/kg bw/day), 5000 ppm females (411 mg/kg
bw/day)
Reference Nagaike, M., TFNG: 90-day subchronic oral toxicity study in rats, Ishihara Sangyo
Kaisha, Japan, 2003. ISK Document No. 2112 AN-1993
Klimisch Score 2 – reliable with restrictions
Amendments/Deviations None
GLP No
Test Guideline/s Not applicable
Species Rat
Strain Jcl:Wistar
No/Sex/Group 5M and 5F
Dose Levels
0, 50 (males only), 200 (females only), 2000 (males only), 5000 (females only ppm
equivalent to 0, 3.56, 16.5, 135, 411 mg/kg bw/day respectively. Admixture with
the diet
Study Summary There were no treatment related changes in any of the parameters assessed at
any dosage.
Additional Comments No additional comments
Conclusion There was no evidence of toxicity
Type of study Acute oral limit toxicity study in rats
Flag Supporting study
Test Substance TFNG-AM, Batch No. 0006
Endpoint LD50
Value >2000 mg/kg bw (limit dose)
Reference Damme, B., TFNG-AM: Acute oral toxicity study in rats, RCC Ltd, Toxicology
Division, Switzerland, 2002. ISK Document No.2104 834772
Klimisch Score 1
Amendments/Deviations None
GLP Y
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Test Guideline/s OECD 423, 96/54/EEC, US EPA OPPTS 870.1000
Species Rat
Strain Han Brl: WIST
No/Sex/Group 3M and 3F
Dose Level 2000 mg/kg bw
Exposure Type Oral gavage
Study Summary There were no deaths, no adverse signs of reaction to treatment or any
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion No classification required
Type of study Acute oral limit toxicity study in rats
Flag Supporting study
Test Substance TFNA-OH, Batch No. 0010
Endpoint LD50
Value >2000 mg/kg bw (limit dose)
Reference Damme, B., TFNA-OH Acute oral toxicity study in rats, RCC Ltd, Toxicology
Division, Switzerland, 2002. ISK Document No.2105 834783
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 423, 96/54/EEC, US EPA OPPTS 870.1000
Species Rat
Strain Han Brl: WIST
No/Sex/Group 3M and 3F
Dose Level 2000 mg/kg bw
Exposure Type Oral gavage
Study Summary There were no deaths, no adverse signs of reaction to treatment or any
macroscopic changes evident at necropsy.
Additional Comments No additional comments
Conclusion No classification required
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Study type/Test Guideline Result Reference
Ames Reverse mutation test.
OECD 471, US EPA OPPTS 870.5100, Commission Directive 2000/32/EEC, L1362000, Annexe 4D
Non mutagenic,
negative with or
without metabolic
activation
Wollny, H-E., Salmonella typhimurium and
escherichia coli reverse mutation assay with
TFNA, RCC Cytotest Cell Research GmbH,
Germany, 2002. ISK Document No. 2106
716901
Ames Reverse mutation test. OECD 471, US EPA OPPTS 870.5100 Commission Directive 2000/32/EEC, L1362000, Annexe 4D
Non mutagenic,
negative with or
without metabolic
activation
Wollny, H-E., Salmonella typhimurium and
escherichia coli reverse mutation assay with
TFNA-AM, RCC Cytotest Cell Research
GmbH, Germany, 2002. ISK Document No.
2106 716902
Ames Reverse mutation test. OECD 471, US EPA OPPTS 870.5100, Commission Directive 2000/32/EEC, L1362000, Annexe 4D
Non mutagenic,
negative with or
without metabolic
activation
Wollny, H-E., Salmonella typhimurium and
escherichia coli reverse mutation assay with
TFNG-AM, RCC Cytotest Cell Research
GmbH, Germany, 2002. ISK Document No.
2108 716904
Ames Reverse mutation test. OECD 471, US EPA OPPTS 870.5100, Commission Directive 2000/32/EEC, L1362000, Annexe 4D
Non mutagenic,
negative with or
without metabolic
activation
May, K., TFNG Bacterial reverse mutation test,
Huntingdon Life Sciences Ltd, UK, 2002. ISK
Document No. 2110 ISK 268-023923
Ames Reverse mutation test. OECD 471, US EPA OPPTS 870.5100, Commission Directive 2000/32/EEC, L1362000, Annexe 4D
Non mutagenic,
negative with or
without metabolic
activation
Wollny, H-E., Salmonella typhimurium and
escherichia coli reverse mutation assay with
TFNA-OH, RCC Cytotest Cell Research
GmbH, Germany, 2002. ISK Document No.
2109 716905
Conclusion None of the metabolites investigated
demonstrated mutagenic potential
General conclusion about mammalian toxicology of active ingredient and metabolites
In contrast to the parent material, IKI-220 technical, the metabolites TFNA and TFNG did not elicit adverse
effects in the liver or kidneys in 90-day studies in rats indicating them to be a lower order of toxicity. Similarly,
acute studies with the metabolites TFNA, TFNG, TFNA-AM, TFNG-AM and TFNA-OH indicated all to be a
lesser order of toxicity than the parent material IKI-220 technical.
The active ingredient IKI-220 should be classified 6.1D for acute oral classifitcation. No classification is
required for any of the metabolites to the extent to which they were investigated.
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Table 11 Summary of studies for the active ingredient with NOAEL and LOAEL values and key
effects.
Study type NOAEL LOAEL Key effect
Rat 28 day dermal toxicity 1000 mg/kg bw/day No effects No adverse findings
Mouse 90 day toxicity
100 ppm (15.25/20.1
mg/kg bw/day
males/females
respectively
1000 ppm (153.9/191.5
mg/kg bw/day
males/females
respectively)
Altered spleen
micropathology both
sexes and liver
micropathology in
males
Rat 90 day toxicity*
200/1000 ppm
males/females
(12.11/72.3 mg/kg
bw/day males/females
respectively)
1000/5000 ppm
males/females
(119.4/340 mg/kg
bw/day males/females
respectively)
Altered kidney
micropathology in
males, liver
micropathology in
females
Dog 90 day toxicity 8 mg/kg bw/day 20 mg/kg bw/day
Micropathological
changes in the kidney
in females
Dog 52 week toxicity 8 mg/kg bw/day 20 mg/kg bw/day Mild anaemia.
Rat toxicity/carcinogenicity
study
Non-neoplastic:
200/1000 ppm
males/females
(7.32/44.1 mg/kg
bw/day males/females
respectively)
Non-neoplastic:
1000/5000 ppm
males/females
(36.5/219 mg/kg bw/day
males/females
respectively)
Non-neoplastic: Altered
kidney micropathology
both sexes, liver in
females, reduced body
weight/body weight
gain
Neoplastic: 1000/5000
ppm males/females
(36.5/219 mg/kg bw/day
males/females
respectively)
Neoplastic:
No effects
Neoplastic: no
carcinogenic effects
Mouse carcinogenicity study
(2 studies assessed)
Non-neoplastic: 80 ppm
(10/11.8 mg/kg bw/day
males/females
respectively
Neoplastic: 80/250 ppm
males/females
(11.8/36.3 mg/kg
bw/day males/females
respectively)
Non-neoplastic: 250
ppm (30.3/36.3 mg/kg
bw/day males/females
respectively
Neoplastic: 250 ppm
males (30.3/38 mg/kg
bw/day).
Non-neoplastic: Altered
lung and liver pathology
Neoplastic: Increased
incidence
alveolar/bronchiolar
epithelial adenoma
Rat 2 generation study
Reproductive capacity:
1800 ppm, (109.1/163.9
mg/kg males/females
respectively)
Reproductive capacity:
No effects
Reproductive capacity:
No effects
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Parental toxicity:
300 ppm, (18.3/20.7
(F0), 28.2/30.5 (F1)
mg/kg bw/day
males/females
respectively)
Parental toxicity:
1800 ppm – 109.1/124.8
(F0), 163.8/176.8 (F1)
males/females
Parental toxicity:
Altered kidney
micropathology
Developmental toxicity:
1800/300 ppm
(163.8/30.5 mg/kg b/day
males/females F1
generation)
Developmental toxicity:
No effects (males), 1800
ppm (females) -153.4
mg/kg bw/day
Developmental toxicity:
Delayed vaginal
opening
Rat developmental
study
Maternal toxicity:
100 mg/kg bw/day
Maternal toxicity:
500 mg/kg bw/day
Maternal toxicity:
Altered liver and kidney
micropathology
Embryofoetal toxicity:
100 mg/kg bw/day
Embryofoetal toxicity:
500 mg/kg/bw day
Embryofoetal toxicity:
Increased incidence of
skeletal (extra rib)
anomalies
Rabbit developmental toxicity
Maternal toxicity: 7.5
mg/kg bw/day
Embryofoetal toxicity: 25
mg/kg bw/day
Maternal toxicity: 25
mg/kg bw/day
Embryofoetal toxicity:
No effects
Maternal toxicity:
Impaired weight gain
Embryofoetal toxicity:
No findings
* Note that for human health risk assessment purposes the LOAEL in males should be considered to be 2000 ppm
(equivalent to 119.4 mg/kg bw/day and the NOAEL 1000 ppm (equivalent to 60 mg/kg bw/day), based on the lack of
human relevance of the the findings relating to alpha-2u-globulin-mediated nephropathy
Toxicokinetics
Type of study Pharmacokinetics in rats – single dose
Flag Key
Test Substance [14
C]IKI-220, Lot No. CP-2173
Reference
Neal, T. R., Savides, M. C., Pharmacokinetics of an oral dose of [14
C]IKI-
220 in Sprague-Dawley rats, Ricerca, LLL, USA, 2001. ISK Document
No.2005 100002-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 417, US EPA OPPTS 870.7485
Species Rat
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Strain Sprague-Dawley Crl:CD® (SD)IGS BR
No/Sex/Group 5M and 5F test groups, 1M and 1F control group
Dose Levels O, 2, 400 mg/kg bw. Oral gavage
Study Summary
Following administration the radiolabelled material was rapidly absorbed,
quickly achieved peak plasma radio-concentrations and was rapidly
eliminated at 2 mg/kg bw and in females at 400 mg/kg bw.. The
pharmacokinetic profile was similar between the sexes sat the low dose but
different at the high dose. In females at 400 mg/kg bw the half life was
about 6.8 hours. This was similar to the half life of about 4.5 hours at 2
mg/kg bw. The average half life for males at 2 mg/kg bw was similar to that
seen in females. At 400 mg/kg bw however the plasma concentrations of
the males reached a plateau that lasted for several hours and the average
half life increased to about 11.6 hours. This was significantly different from
the half life of females at 400 mg/kg bw and males at 2 mg/kg bw. The
AUC approximated being proportional to the dose level. The data
suggested that higher doses lead to greater bioavailability. There were no
significant differences in AUC between males and females at any given
dose level. The maximum plasma concentrations (Cmax) also approximated
being proportional to dose level.
Additional Comments No additional comments
Conclusion
Following a single dose the radiolabelled material was rapidly absorbed
and peak plasma concentrations rapidly achieved in both sexes at both
dosages.
Type of study Absorption, distribution, metabolism and excretion study in rats - single
dose
Flag Key study
Test Substance [14
C]IKI-220, Lot No. CP-2173
Reference
Neal, T. R., Savides, M. C., Dow, P., Study of the elimination and
distribution of radiolabel following single oral administration of [14
C]IKI-220
to Sprague-Dawley rats, Ricerca, LLL, USA, 2002. ISK Document No.
2006 10005-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 417, US EPA 870.7485
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Species Rat
Strain Sprague-Dawley Crl:CD® (SD)IGS BR
No/Sex/Group 14M and 14F test groups, 1M and 1F control group.
Dose Levels
0, 2, 400 mg/kg bw. Oral gavage
Both sexes at 2 mg/kg bw were sacrificed after 0.5, 6, 24 and 168 hours
post dosing.
Males at 400 mg/kg bw were sacrificed after 3, 14.5, 24 and 168 hours
post dosing
Females at 400 mg/kg bw were sacrificed after 1, 8, 24 and 168 hours after
dosing
Study Summary
Following oral administration radioactivity was readily absorbed and
excreted at both doses. Over 168 hours approximately 90% of
administered radioactivity was contained in urine, the majority occurring
within the first 24 hours. Faecal elimination accounted for only about 5% of
the administered dose.
Tissue levels of radioactivity rose rapidly with maximum concentrations
mirroring those of blood. Radioactivity was detected in all tissues for the
early time points although by 168 hours, where detectable, levels had
decreased generally by 50 to 100 fold. At 168 hours post dose less than
2% of the administered radioactivity was contained in the carcass. Tissues
contained negligible levels of radioactivity, the liver showing the highest
amount (<0.15%).
Increasing the dose level had little effect on the disposition of radioactivity
and there was no accumulation of radioactivity in tissues or the residual
carcass. No sex related differences were observed in any of the
parameters measured.
Additional Comments No additional comments
Conclusion
Following administration the radiolabelled dose was rapidly absorbed and
excreted in both sexes in the urine. Increasing the dose had little effect of
disposition of radioactivity and was without accumulation in tissues or
residual carcass nor were there any sex-related differences
Type of study Absorption, distribution, metabolism and excretion study in rats - multiple
dose
Flag Key study
Test Substance [14
C]IKI-220, Lot No. CP-2173
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Reference
Neal, T. R., Savides, M. C., Dow, P., Study of the elimination and
distribution of radiolabel following multiple oral administrations of [14
C]IKI-
220 to Sprague-Dawley rats, Ricerca, LLL, USA, 2002. ISK Document No.
2006 10007-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 417, US EPA 870.7485
Species Rat
Strain Sprague-Dawley Crl:CD® (SD)IGS BR
No/Sex/Group 14M and 14F test groups, 1M and 1F control
Dose Levels 2 mg/kg bw/day [
12C] for 14 days followed by a single dose of [
14C]
Animals were sacrificed 0.5, 6, 24 and 168 hours after the final dose.
Study Summary
Following oral administration radioactivity was readily absorbed and
excreted at both doses. Over 168 hours approximately 90% of
administered radioactivity was contained in urine, the majority occurring
within the first 24 hours. Faecal elimination accounted for only about 7% of
the administered dose.
Tissue levels of radioactivity rose rapidly with maximum concentrations
mirroring those of blood. Radioactivity was detected in all tissues for the
early time points although by 168 hours, where detectable, levels had
decreased generally by 50 to 100 fold. At 168 hours post dose less than
2% of the administered radioactivity was contained in the carcass. Tissues
contained negligible levels of radioactivity, the liver showing the highest
amount (<0.12%).
There was no accumulation of radioactivity in tissues or residual carcass.
No sex related differences were observed in any of the parameters
measured. Repeat dosing of IKI-220 to the rat had no effect on the
deposition of radioactivity compared with a single dose at the same dose
level.
Additional Comments No additional comments
Conclusion
Repeat administration of radiolabelled dose was rapidly absorbed and
excreted, predominantly in urine. There was no accumulation in tissues or
residual carcass nor were there any sex-related differences. Repeat dosing
had no effect on disposition of radioactivity compared with single dosing.
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Type of study Biliary excretion study in rats
Flag Key
Test Substance [14
C]IKI-220, Lot No. CP-2173
Reference
Dow, P., Study of the biliary elimination of radiolabel following oral
administration of [14
C]IKI-220 to Sprague-Dawley rats, Ricerca LLC, USA,
2002. ISK Document No. 2034 13364-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s OECD 417, US EPA OPPTS 870.7485
Species Rat
Strain Sprague-Dawley Crl:CD® (SD) IGS BR
No/Sex/Group 4m and 4 F. Control 1M and 1 F
Dose Levels 0, 2, 400 mg/kg bw. Single dose
Study Summary
Radioactivity was rapidly absorbed and excreted, predominantly in the
urine (80 – 85% of radioactivity at both doses). Biliary excretion was not a
principal route of elimination of radioactivity – accounting for only about 5%
of the administered doses. Increasing the dose had little effect on the
deposition of radioactivity and there was no accumulation of radioactivity in
the residual carcass. No sex related differences were observed in any of
the parameters measured.
Additional Comments No additional comments
Conclusion
Orally administered radiolabelled dose was rapidly absorbed and excreted,
predominantly in urine. Biliary excretion was not a significant route of
elimination of radioactivity. Increasing the dose had little effect on
radioactive disposition or accumulation in the residual carcass. There were
no sex-related differences in parameters assessed.
Type of study Metabolic pathway in rats
Flag Key study
Test Substance [14
C]IKI-220, Lot No. CP-2173
Reference Gupta, K. S., Shah, J. F., McClanahan, R. H., Metabolism of [14
C]IKI-220 in
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rats, Ricerca, LLC, USA, 2002. ISK Document No. 2033 010052-1
Klimisch Score 1
Amendments/Deviations None
GLP Y
Test Guideline/s US EPA 870.7485
Species Rat
Dose Levels
Analyses were conducted using biological samples obtained from earlier
cited studies assessing biliary excretion and absorption, metabolism
distribution and elimination of radiolabelled IKI-220 after single and multiple
administrations.
Study Summary
Excretion of IKI-220 and its metabolites occurred primarily in the urine and
to a much lesser extent the faeces. IKI-220 was metabolised by several
routes including nitrile hydrolysis, amide hydrolysis, N-oxidation and
hydroxylation of pyridine ring. Combinations of pathways occurred leading
to the formation of multiple metabolites.
The principal urinary metabolites were IKI-220 (unchanged) and TFNA-AM
with much lower detection of TFNA oxide, TFNG-AM, TFNA conjugate. A
range of other metabolites were detected at low levels
Biliary metabolites were confined to IKI-220 (unchanged) and TFNA-AM.
Faecal metabolites consisted of IKI-220 (unchanged), TFNA-AM, TFNA,
mix of TFNA-AM, N-oxide conjugate and TFNA conjugate.
The principal liver metabolites were IKI-220 (unchanged), TFNG, TFNG-
AM, and TFNA-AM.
Additional Comments No additional comments
Conclusion
Comparison of single and multiple doses studies revealed similar
metabolites observed in the urine indicating no induction of metabolism
had occurred. Distribution of residues was also similar between single and
multiple dose studies.
Summary of toxicokinetic studies
IKI-200 is rapidly absorbed following oral administration and is widely distributed. Excretion is primarily via
the urine, with a small proportion excreted via faeces and bile. Repeated dosing or increasing dose did not
influence the tissue distribution. There were no significant differences between sexes in the parameters
assessed. Metabolism is not extensive with the majority of IKI-220 being excreted via the urine, feces and
bile as the parent compound. A major urinary metabolite is TFNA-AM.
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Environmental fate - Robust study summaries for the active ingredient and
metabolites
List of metabolites mentioned in the following studies, their structure can be found in Figure 1, on page 134.
TFNG-AM: N-(4-trifluoromethylnicotinoyl)-glycinamide
TFNG: N-(4-trifluoromethyl-nicotinonyl)glycine
TFNA: 4- trifluoromethylnicotinic acid
TFNA-AM: 4-trifluoromethylnicotinamide
TFNA-OH: 6-hydroxy-4-trifluoromethylnicotinic acid
Water compartment
Ready Biodegradation
Type of study Ready biodegradation (respirometry)
Flag Supporting study as a higher tier study is available (water/sediment degradation
test)
Test Substance IKI-220
Species Domestic sewage
Endpoint Degradation rate measured by %CO2 evolved
Value 6.6% at 28 days
Reference
P. M. Bidinotto (2002) Immediate biodegradability of flonicamid tecnico, using the
open system. Bioagri Laboratorios Ltda Piracicaba/SP Brazil. Report no RF-
1213.211.039.02
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 301B (1992)
Dose Levels 10 mg carbon/L
Analytical measurements CO2 evolved
Study Summary
The biodegradation of the test substance was studied with the respirometry
method over 28 days at 21 to 24oC.
The following conditions were included in the test (duplicates):
- Blank: mineral medium + inoculum - Reference substance: mineral medium + inoculum + sodium benzoate - Test substance: mineral medium + inoculum + IKI-220 - Inhibition control: mineral medium + inoculum + sodium benzoate + IKI-
220 - Abiotic control: IKI-220 + mercury chloride
More than 60% of degradation was reached in the reference substance after the
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10 day interval, and 72% at 28 days. More than 25% of degradation was observed
at 14 day in the inhibition treatment and 48.6% at 28 days.
No abiotic degradation was observed in the corresponding flasks.
The biodegradation of flonicamid was 6.6% at 28 days, so the substance is not
readily biodegradable.
Conclusion Not readily biodegradable
Hydrolysis
Study type Hydrolysis in function of pH and temperature
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Hydrolysis rate at pH 5, 7 and 9 at 25 and 50oC
Value Stable at pH below 7 and 25 oC. At pH9 and 25
oC, Half-life = 204 d
Reference
KJ Walsh & MD Murray (2002) A hydrolysis study of 14
C-IKI-220 in water. Ricerca
LLC. Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio,
44077-1000. Report no 008076-2
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 111 (1981)
Dose Levels ca 1 µg/mL
Analytical measurements HPLC with radiochemical flow detection
Study Summary
The rate and route of hydrolysis of the substance labelled with 14
C at the C-3
carbon in the pyridyl ring, was determined in sterile buffer solutions as a function
of pH and temperature.
Buffer solutions prepared at pH 5, 7 and 9 were incubated for 30 days (120 days
for pH 9 samples) at 25 ± 1 o
C and for 120 days at 50 ± 1 o
C in darkness; treated
pH 4 buffer samples were also included in the experiment at 50 o
C.
No hydrolysis was observed below pH 7 and only at the elevated temperature at
this pH. The half-life was 578 days in these conditions.
The half-life at pH 9 and 25 oC was 204 days. The major hydrolytic product TFNG-
AM, rose to 30.5% by the last sampling point. A second product TFNG, was
initially observed at day 77 at 1.0%, increasing only to 2.0% by day 120.
IKI-220 is hydrolysed initially at the nitrile group to form the amide (TFNG-AM)
which can then be hydrolysed to the glycine (acid) derivative TFNG. The initial
step is observed under alkaline conditions.
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Conclusion Hydrolysis would not contribute significantly to the degradation of the
substance in use conditions.
Study type Hydrolysis in function of pH and temperature
Flag Key study
Test Substance Metabolite TFNA with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Hydrolysis rate at pH 5, 7 and 9 at 25 and 50oC
Value Stable
Reference
MD Murray & KJ Walsh (2000) A hydrolysis study of 14
C-TFNA in water. Ricerca
LLC. Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio,
44077-1000. Report no 008077-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 111 (1981)
Dose Levels ca 1 µg/mL
Analytical measurements HPLC with radiochemical flow detection
Study Summary
The rate and route of hydrolysis of the substance labelled with 14
C at the C-3
carbon in the pyridyl ring, was determined in sterile buffer solutions as a function
of pH and temperature.
Buffer solutions prepared at pH 5, 7 and 9 were incubated for 30 days (120 days
for pH 9 samples) at 25 ± 1 o
C and for 120 days at 50 ± 1 o
C in darkness; treated
pH 4 buffer samples were also included in the experiment at 50 o
C.
No hydrolysis was observed in any samples at 25 o
C. Hydrolysis of TFNA
incubated at 50 o
C was minimal during the course of the study (120 days). The
radiopurity did not fall below 99% at the study termination.
Conclusion Hydrolysis would not contribute significantly to the degradation of the
substance in use conditions.
Aqueous Photolysis
Study type Aqueous photolysis
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Photolysis half-life
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Value 267 days in continuous radiation.
Reference
KJ Walsh (2002) A photolysis study of [14
C]-IKI-220 in water. Ricerca LLC.
Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio, 44077-
1000. Report no 011050-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OPPTS 835.2210
Dose Levels ca 1 µg/mL
Analytical measurements HPLC with radiochemical flow detection
Study Summary
The substance at a concentration of ca. 1 µg/mL in pH 7.00 ± 0.05 phosphate
buffer was continuously exposed to simulated sunlight (xenon arc lamp with filters)
for 15 days at a temperature of 23 ± 2 oC. At days 0, 1, 3, 7, 10 and 15, light-
exposed and dark control samples were analysed directly by HPLC.
In the dark control samples, the substance did not degrade over the 15 days
period.
The substance degraded slightly after 15 days of continuous irradiation. None of
the degradates comprises more than 5% of the applied radioactivity. The major
degradate TFNA-AM was 2.4% at t0 and increased to 2.9% by day 15. The
calculated half-life was 267 days with continuous radiation. The quantum yield is
0.0319%.
Conclusion Photolysis is not a significant degradation pathway for the substance.
Water/sediment aerobic biodegradation
Study type Aerobic degradation in 2 water/sediment systems at 20oC, in the dark
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Identification of metabolites and DT50
Value Whole system: 43.6 and 35.7 days
Reference
M. D. Murray & K J. Sandacz Herczog (2002) Aerobic Aquatic Sediment
Metabolism of [14
C]IKI-220. Environmental and metabolic fate. 7528 Auburn Road
Painesville Ohio, 44077-1000. Report no 011052-1
Klimisch Score 1
Amendments/Deviations None that had an effect on the study results.
GLP Yes
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Test Guideline/s SETAC-Europe (March 1995), U.S. EPA Pesticide Assessment Guidelines
Subdivision N Chemistry: Environmental Fate, Series 162-4.
Dose Levels 0.03 mg/L
Study Summary
The fate of [14
C]IKI-220, [pyridyl-14
C]-N-cyanomethyl-4-
trifluoromethylnicotinamide, in aerobic water/sediment systems was studied in
order to determine the rate of degradation of the test compound as well as the
pattern of metabolite formation.
IKI-220, labelled in the 3 position of the pyridyl ring, was applied to individual
samples from each of two different sediment/water test systems at an application
rate of 0.03 ppm, based on the total volume of water.
A system from river Rhine (called EFS 163) and another one from a pond from
Switzerland (EFS 164) were used in this study. They have the following
characteristics: silty clay loam with pH 7.11, 10.23% of organic carbon (EFS 164)
and sandy loam with pH 8.10 and 0.74% of organic carbon.
The test systems were sampled at intervals over a 145-day (EFS-163) and 136-
day (EFS-164) period under aerobic conditions. At each sampling interval the
water and sediment phases of the sample were separated and analysed for IKI-
220 and degradates. In addition, the release of radioactivity as 14
CO2 and
radiolabeled volatiles was monitored for the duration of the study. The mass
balance of the applied radioactivity (i.e., based on the sum of the water phase,
sediment extracts, volatile and bound radioactivity) was greater than 90% in both
water/sediment systems at all sampling points, averaging 99.1% in the EFS-163
system samples and 103.1% in the EFS-164 system samples.
Under aerobic aquatic conditions IKI-220 was extensively degraded over the
duration of the study. The DT50 and DT90 values for the EFS-163 sediment/water
system were 37.3 and 123.8 days, respectively, in the water phase and 43.6 and
144.8 days, respectively, in the entire system. In the EFS- 164 system, the DT50
and DT90 values were 30.3 and 100.5 days, respectively, in the water phase and
35.7 and 118.7 days, respectively, in the entire system.
In each test system, 14
C levels in the water phase and IKI-220 levels in the entire
system declined steadily over the sampling period, while radiolabel in bound
residues and the release of 14
CO2, increased over time. By Day 145 of the EFS-
163 test system, the bound residues exceeded 38% of the applied dose; while in
the EFS-164 test system, the bound residues increased to greater than 75% of
the applied dose by Day 136. The accumulated radioactivity in the NaOH traps
increased throughout the study to 59.1% (Day 145, EFS-163) and 15.6% (Day
136, EFS-164). The majority of the radioactivity trapped in the NaOH was
attributed to 14
CO2. Release of radiolabeled organic volatiles was minimal
throughout the study in both systems.
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The metabolic pathway for IKI-220 under aerobic aquatic conditions involved
hydrolysis of the cyano and amide bonds in the side chain to produce TFNG,
TFNA-AM and TFNA. Further hydroxylation of the 4- trifluoromethylnicotinic acid
(TFNA) produced TFNA-OH. TFNA-AM and TFNG were generated as degradate
intermediates in both soil sets. Further degradation of the radioactive residues
resulted in incorporation into the soil organic matter and mineralization to 14
CO2.
TFNA-OH and TFNA were the major degradation products in the EFS-164
sediment/water systems exceeding 10% of the applied dose, with levels rising
steadily over the first 6 weeks of the sampling period before declining TFNA-OH
and TFNA were present in the EFS-163 test system at levels not exceeding 2% of
the applied dose. TFNG and TFNA-AM were minor degradates in both test
systems (see Figure 1).
Comment
The European Food and Safety Authority Draft Assessment Report on Flonicamid
(Volume 3, Annex B, B8, July 2005) mentions DT50 for the 2 major metabolites: 60
days in water and 59 days in sediment for TFNA, and 49 days in water for TFNA-
OH, calculated from this study.
Conclusion DT50 whole system are 43.6 and 35.7 days, major metabolites are TFNA-OH
and TFNA.
Figure 1: Proposed metabolic pathway of IKI-220 in aerobic water/sediment systems
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Water/sediment anaerobic biodegradation
No data available.
Bioaccumulation potential
The log Kow is 0.3, indicating a low potential of bioaccumulation. However, the substance displays active
surface properties consequently the log Kow is not a proper indicator of the bioaccumulation potential. A
bioaccumulation study should have been conducted.
Soil compartment
Aerobic Degradation in Soil (routes and rates) – Laboratory studies
Study type Aerobic degradation on soil
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Rate (DT50) and route of degradation
Value DT50 = 1.0 day
Reference
C. J. Hatzenbeler & K. J. S. Herczog (2002) An Aerobic Soil Metabolism Study with
[14
C]IKI-220. Ricerca LLC 7528 Auburn Road Painesville OH 44077-1000 USA.
Study number 6933-96-0186-EF-001-001.
Klimisch Score 2 study is valid but only one soil is tested
Amendments/Deviations None
GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.1 ppm corresponding to 100 g ai/ha
Analytical measurements Duplicate samples were analysed at 0, 0.5, 1, 2, 3, 7, 14 and 30 days after
treatment
Study Summary
A typical U.S. loamy sand agricultural soil was used for this study. The test soil was
treated with [14
C]IKI-220 at a rate of approximately 0.1 ppm. This concentration is
equivalent to the use rate of the compound (100 g a.i./ha) incorporated to a depth of
7 cm. The soil had an organic carbon content of 0.6% and pH of 7.2.
The test soil was acclimated for 43 days, at 20 ± 1 °C, in darkness, at 45% of its
maximum water holding capacity, prior to treatment with an aqueous solution of
[14
C]IKI-220. Duplicate samples were analysed at 0, 0.5, 1, 2, 3, 7, 14 and 30 days
after treatment. The average recovery of applied radioactivity over the 30-day
course of the study was 86.3%. The recovery of radiocarbon was low in the
definitive experiment for sampling times days 3, 7, 14 and 30 days due to very rapid
extensive metabolism and mineralisation which formed 14
CO2. A second set of soil
samples (mass balance experiment) was dosed and sampled at days 3, 7, 14 and
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30 to correct for mass balance and volatiles. The average recovery of applied
radioactivity for the mass balance experiment was 93.3% which is within the
acceptable limits of 90 to 110%.
Extractable radiolabel decreased from 101.4% on Day 0 to 13.7% after 30 days
incubation. Unextractable soil residues increased steadily from 0.7% on Day 0 to
35.2% on Day 30. Evolution of 14
CO2 increased throughout the study, reaching a
maximum of 47.0% of the applied radioactivity after thirty days in this experiment.
TFNA was the major degradate in the soil over the course of the study. TFNA
formed rapidly reaching a peak of 36.4% of the applied radioactivity 3 days after
treatment. Further metabolism of TFNA occurred readily, leading to residue levels of
less than 2% of the applied radioactivity after 14 days. Hydroxylation of TFNA led to
formation of TFNA-OH. This hydroxylated TFNA formed rapidly over a one-week
period, reaching 20.2% of the applied radioactivity then declined to 0.5% after thirty
days.
IKI-220 degraded easily in the loamy sand soil, principally via hydrolysis of the
cyano and amide functionalities on the aliphatic side chain of the molecule. These
reactions led to the formation of TFNA. TFNA was further metabolized to form a
hydroxy analog, TFNA-OH. The DT50 and DT90 values for IKI-220 in this loamy sand
soil, based on a first-order exponential decay, were 1.0 and 3.4 days respectively.
TFNG, TFNG-AM and TFNA-AM were formed as degradate intermediates.
Comment
R software has been used to recalculate the DT50 and DT90 according to the best fit:
best fit is 1st order with a DT50 of 1.04 and a DT90 of 2.04 days
The difference is not significant so the DT50 of the report is used.
Conclusion
DT50 = 1.0 day at 20oC, the proposed pathway is shown in
Figure 2.
Study type Aerobic degradation on soil
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Rate (DT50) and route of degradation
Value DT50 from 0.7 to 1.8 days at 20oC
Reference N.R. Lentz (2002) Rate of degradation of [
14C]IKI-220 in soil. Ricerca LLC 7528
Auburn Road Painesville OH 44077-1000 USA. Study number 013066-1.
Klimisch Score 1
Amendments/Deviations None
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GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.1 ppm
Analytical measurements 0, 0.33, 0.67, 1, 2, 3, 7, 14 and 30 days after treatment
Study Summary
The degradation rate of IKI-220 under aerobic soil conditions was studied in three
European soils. 2 soils were obtained from the United Kingdom, a third standard soil
(2.1) was obtained from Germany.
Samples of each treated soil type were incubated at 50% maximum water-holding
capacity at 20oC in darkness. An additional soil set (U.K.#21) was also treated as
above but incubated at 10°C. The soils were extracted and analyzed by HPLC at
days 0, 0.33, 0.67, 1, 2, 3, 7, 14 and 30.
Recovery of applied radioactivity over the course of the study for the four sets
ranged from an average of 89.4 to 102.8%. Extractable radiolabel decreased to less
than 8.7% of the applied dose in all soils by the end of the sampling period.
Unextractable residues in all soils ranged from 29.6 to 43.3% by the end of
sampling. Evolution of 14
CO2 ranged from 49.3 to 56.6% of the radiolabel in all soil
sets by the end of the respective sampling periods.
IKI-220 degraded rapidly in all soils, principally via hydrolysis of the amide and
cyano functionalities on the aliphatic side chain to form TFNA and TFNG-AM,
respectively. Subsequent hydroxylation of the aromatic ring led to the formation of
TFNA-OH. Minor intermediates TFNG and TFNA-AM were also detected in all of
the soils. TFNA and TFNA-OH were major degradates detected in all of the soils
ranging from 12.2 to 30.6% and 12.1 to 32.7% at concentration maxima for all of the
soils, respectively. Degradate TFNG-AM ranged from 7.8 to 10.2% of applied dose.
Minor degradates TFNG and TFNA-AM were detected at less than 7. 7% of the
applied dose at all sampling points over the course of the study. All of the
degradates were metabolised and mineralised to carbon dioxide and immobilised as
soil-bound residue.
The DT50 and DT90 values ranged from 0.7 to 1.8 days and 2.3 to 6.0 days for the
soils incubated at 20 °C, respectively. The DT50 value was 2.4 days and the DT90
value was 7.9 days for the soil incubated at 10°C. A first-order kinetics model was
used to approximate parent IKI-220 degradation processes in all of the soil sets.
Bedfordshire
soil (UK21)
Birmingham
soil (UK39)
German soil
2.1
Type Loamy sand Loamy sand Sand
pH 6.9 7.0 6.2
% organic matter 2.4 4.6 0.9
Cation exchange capacity 8.5 16.9 3.3
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(meq/100g)
Max water holding capacity
(%)
41.0 64.8 29.5
1st order DT50 (days) 2.4 at 10
oC
(r2=0.9721)
0.7 at 20oC
(r2=0.9898)
0.7
(r2=0.989)
1.8
(r2= 0.9989)
1st order DT90 (days) 7.9 at 10
oC
2.3 at 20oC
2.4 6.0
Comment
R software has been used to recalculate the DT50 and DT90 at 20oC according to the
best fit:
UK21 soil: best fit is 1st order with a DT50 of 0.635 and a DT90 of 2.11 days
UK39 soil: best fit is indeterminate order rate with a DT50 of 0.703 (= DT90/3
according to NAFTA guidelines) and a DT90 of 2.34 days.
German soil: best fit is indeterminate order rate with a DT50 of 1.84 (= DT90/3
according to NAFTA guidelines) and a DT90 of 6.1 days.
These values are very similar to the initial ones so the initial ones of the report will
be used.
Conclusion DT50 from 0.7 to 1.8 days at 20oC.
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Figure 2: Degradation pathway of flonicamid in soils
Study type Aerobic degradation on soil study with metabolites
Flag Key study
Test Substance TFNA metabolite with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint DT50
Value DT50 from 0.3 to 0.5 days at 20oC
Reference
C.J. Hatzenbeler & N.R. Lentz (2002) Rate of degradation of [14
C]TFNA in soil.
Ricerca LLC 7528 Auburn Road Painesville OH 44077-1000 USA. Study number
012064-1
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.1 ppm
Analytical measurements At 0, 0.5, 1, 2, 3, 7, 14 and 30 days after treatment
Study Summary
The degradation rate of TFNA under aerobic soil conditions was studied in three
European soils. 2 soils were obtained from the United Kingdom, a third standard
soil (2.1) was obtained from Germany.
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Samples of each treated soil type were incubated at 50% maximum water-holding
capacity at 20oC in darkness. An additional soil set (U.K.#21) was also treated as
above but incubated at 10°C. The soils were extracted and analyzed by HPLC at
days 0, 0.5, 1, 2, 3, 7, 14 and 30.
Recovery of applied radioactivity over the course of the study for the four sets
ranged from 91.0 to 100.1%. Extractable radiolabel decreased to less than 16.2%
of the applied dose in all soils by the end of the sampling period.
Unextractable residues in all soils ranged from 21.0 to 42.4% by the end of
sampling. Evolution of 14
CO2 ranged from 37.7 to 54.6% of the radiolabel in all soil
sets by the end of the respective sampling periods.
TFNA degraded rapidly in all soils, principally via hydroxylation of the aromatic
ring to 6-hydroxy-4-trifluoromethylnicotinic acid (TFNA-OH). TFNA-OH was the
major degradate in all soils over the course of the study, ranging from 41.6 to
54.9% of the applied dose. TFNA-OH was metabolized and mineralized to carbon
dioxide and immobilized as soil-bound residue.
The DT50 and DT90 values ranged from 0.3 to 0.5 days and 1.0 to 1.5 days for the
soils incubated at 20 °C, respectively. The DT50 value was 1.0 days and the DT90
value was 3.3 days for the soil incubated at 10°C. A first-order kinetics model was
used to approximate parent TFNA degradation processes in all of the soil sets.
Bedfordshire
soil (UK21)
Birmingham
soil (UK39)
German soil
2.1
Type Loamy sand Loamy sand Sand
pH 6.8 6.4 5.7
% organic matter 1.25 5.49 1.03
Cation exchange capacity
(meq/100g)
4.97 13.01 2.02
Max water holding capacity
(%)
21.23 39.88 18.41
1st order DT50 (days) 1.0 at 10
oC
(r2=0.9864)
0.5 at 20oC
(r2=0.9687)
0.3
(r2=0.9899)
0.5
(r2= 0.9912)
1st order DT90 (days) 3.3 at 10
oC
1.5 at 20oC
1.0 1.5
Comment
R software has been used to recalculate the DT50 and DT90 at 20oC according to
the best fit:
UK21 soil: best fit is indeterminate order rate with a DT50 of 0.539 (= DT90/3
according to NAFTA guidelines) and a DT90 of 1.79 days
UK39 soil: best is 1st order with DT50 of 0.297 and a DT90 of 0.987 days.
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German soil: best fit is indeterminate order rate with a DT50 of 0.487 (= DT90/3
according to NAFTA guidelines) and a DT90 of 1.62 days.
These values are very similar to the initial ones so the initial ones of the report will
be used.
Conclusion DT50 from 0.3 to 0.5 days at 20oC.
Study type Aerobic degradation on soil study with metabolites
Flag Key study
Test Substance TFNA-AM metabolite with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint DT50
Value DT50 from 1.0 to 3.82 days at 20oC
Reference N.R. Lentz (2002) Rate of degradation of [
14C]TFNA-AM in soil. Ricerca LLC 7528
Auburn Road Painesville OH 44077-1000 USA. Study number 012696-1
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.01 ppm
Analytical measurements At 0, 0.04, 0.125, 0.25, 0.5, 1, 2, 3, 7, 14 and 30 days after treatment
Study Summary
The degradation rate of TFNA-AM under aerobic soil conditions was studied in
three European soils. 2 soils were obtained from the United Kingdom, a third
standard soil (2.1) was obtained from Germany.
Samples of each treated soil type were incubated at 50% maximum water-holding
capacity at 20oC in darkness. An additional soil set (U.K.#21) was also treated as
above but incubated at 10°C. The soils were extracted and analyzed by HPLC at
days 0, 0.04, 0.125, 0.25, 0.5, 1, 2, 3, 7, 14 and 30.
Recovery of applied radioactivity over the course of the study for the four sets
ranged from 90.0 to 105%. Extractable radiolabel decreased to less than 13.3% of
the applied dose in all soils by the end of the sampling period.
Unextractable residues in all soils ranged from 26.9 to 37.7% by the end of
sampling. Evolution of 14
CO2 ranged from 49.7 to 63.9% of the radiolabel in all soil
sets by the end of the respective sampling periods.
TFNA-AM was metabolized and mineralized to carbon dioxide and immobilized as
soil-bound residue. 14
CO2 was the major metabolite in all soils over the course of
the study.
TFNA-AM degraded rapidly in all soils. The DT50 and DT90 values ranged from 1.0
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to 2.6 days and 3.4 to 8.5 days for the soils incubated at 20 °C, respectively. The
DT50 value was 4.5 days and the DT90 value was 14.8 days for the soil incubated
at 10°C. A first-order kinetics model was used to approximate parent TFNA-AM
degradation processes in all of the soil sets.
Bedfordshire
soil (UK21)
Birmingham
soil (UK39)
German soil
2.1
Type Loamy sand Loamy sand Sand
pH 6.9 7.0 6.2
% organic matter 2.4 4.6 0.9
Cation exchange capacity
(meq/100g)
8.5 16.9 3.3
Max water holding capacity
(%)
41.0 64.8 29.5
1st order DT50 (days) 4.5 at 10
oC
(r2=0.9944)
1.2 at 20oC
(r2=0.993)
1.0
(r2=0.9892)
2.6
(r2= 0.9571)
1st order DT90 (days) 14.8 at 10
oC
3.8 at 20oC
3.4 8.5
Comment
R software has been used to recalculate the DT50 and DT90 at 20oC according to
the best fit:
UK21 soil: best fit is indeterminate order rate with a DT50 of 1.16 (= DT90/3
according to NAFTA guidelines) and a DT90 of 3.84 days
UK39 soil: best fit is indeterminate order rate with a DT50 of 0.99 (= DT90/3
according to NAFTA guidelines) and a DT90 of 3.29 days.
German soil: best fit is 1st order with a DT50 of 3.82 and a DT90 of 12.7 days.
These values are very similar to the initial ones so the initial ones of the report will
be used except for the German soil were the r2 is low.
Conclusion DT50 from 1.0 to 3.82 days at 20oC.
Study type Aerobic degradation on soil study with metabolites
Flag Key study
Test Substance TFNA-OH metabolite with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint DT50
Value DT50 from 1.0 to 3.4 days at 20oC
Reference D.C. Findak & N.R. Lentz (2002) Rate of degradation of [
14C]TFNA-OH in soil.
Ricerca LLC 7528 Auburn Road Painesville OH 44077-1000 USA. Study number
143
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012066-1
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.1 ppm
Analytical measurements At 0, 0.5, 1, 2, 3, 7, 14 and 30 days after treatment
Study Summary
The degradation rate of TFNA-OH under aerobic soil conditions was studied in
three European soils. 2 soils were obtained from the United Kingdom, a third
standard soil (2.1) was obtained from Germany.
Samples of each treated soil type were incubated at 50% maximum water-holding
capacity at 20oC in darkness. An additional soil set (U.K.#21) was also treated as
above but incubated at 10°C. The soils were extracted and analyzed by HPLC at
days 0, 0.5, 1, 2, 3, 7, 14 and 30.
Recovery of applied radioactivity over the course of the study for the four sets
ranged from 91.0 to 104.9%. Extractable radiolabel decreased to less than 13% of
the applied dose in all soils by the end of the sampling period.
Unextractable residues in all soils ranged from 26.2 to 44.4% by the end of
sampling. Evolution of 14
CO2 ranged from 41.5 to 60.0% of the radiolabel in all soil
sets by the end of the respective sampling periods.
TFNA-OH was metabolized and mineralized to carbon dioxide and immobilized as
soil-bound residue. 14
CO2 was the major metabolite in all soils over the course of
the study.
TFNA-OH degraded rapidly in all soils. The DT50 and DT90 values ranged from 1.0
to 2.6 days and 3.4 to 8.7 days for the soils incubated at 20 °C, respectively. The
DT50 value was 4.5 days and the DT90 value was 15 days for the soil incubated at
10°C. A first-order kinetics model was used to approximate parent TFNA-OH
degradation processes in all of the soil sets.
Bedfordshire
soil (UK21)
Birmingham
soil (UK39)
German soil
2.1
Type Loamy sand Loamy sand Sand
pH 6.8 6.4 5.7
% organic matter 1.25 5.49 1.03
Cation exchange capacity
(meq/100g)
4.97 13.01 2.02
Max water holding capacity
(%)
21.23 39.88 18.41
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1st order DT50 (days) 4.5 at 10
oC
(r2=0.9915)
1.0 at 20oC
(r2=0.9529)
1.3
(r2=0.9559)
2.6
(r2= 0.9521)
1st order DT90 (days) 15.0 at 10
oC
3.4 at 20oC
4.2 8.7
Comment
R software has been used to recalculate the DT50 and DT90 at 20oC according to
the best fit:
UK21 soil: best fit is indeterminate order rate with a DT50 of 1.48 (= DT90/3
according to NAFTA guidelines) and a DT90 of 4.9 days
UK39 soil: best fit is 1st order with a DT50 of 1.51 and a DT90 of 5.03 days.
German soil: best fit is indeterminate order rate with a DT50 of 3.4 (= DT90/3
according to NAFTA guidelines) and a DT90 of 11.3 days.
These values are very similar to the initial ones so the initial ones of the report will
be used except for the German soil where the r2 is low.
Conclusion DT50 from 1.0 to 3.4 days at 20oC.
Study type Aerobic degradation on soil study with metabolites
Flag Key study
Test Substance TFNG metabolite with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint DT50
Value DT50 from 0.1 to 1.1 days at 20oC
Reference N.R. Lentz (2002) Rate of degradation of [
14C]TFNG in soil. Ricerca LLC 7528
Auburn Road Painesville OH 44077-1000 USA. Study number 012065-1
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.1 ppm
Analytical measurements At 0, 0.04, 0.125, 0.25, 0.5, 1, 2, 3, 7, 14 and 30 days after treatment
Study Summary
The degradation rate of TFNG under aerobic soil conditions was studied in three
European soils. 2 soils were obtained from the United Kingdom, a third standard
soil (2.1) was obtained from Germany.
Samples of each treated soil type were incubated at 50% maximum water-holding
capacity at 20oC in darkness. An additional soil set (U.K.#21) was also treated as
above but incubated at 10°C. The soils were extracted and analyzed by HPLC at
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days 0, 0.04, 0.125, 0.25, 0.5, 1, 2, 3, 7, 14 and 30.
Recovery of applied radioactivity over the course of the study for the four sets
ranged from 89.5 to 108.8%. Extractable radiolabel decreased to less than 14.7%
of the applied dose in all soils by the end of the sampling period.
Unextractable residues in all soils ranged from 25.0 to 44.6% by the end of
sampling. Evolution of 14
CO2 ranged from 36.0 to 50.3% of the radiolabel in all soil
sets by the end of the respective sampling periods.
TFNG degraded rapidly in all soils via hydrolysis of the amide functionality on the
aliphatic side chain to form 4-trifluoromethylnicotinic acid (TFNA) and subsequent
hydroxylation of the aromatic ring to form 6-hydroxy-4- trifluoromethyl-nicotinic
acid (TFNA-OH). 4-trifluoromethylnicotinamide (TFNA-AM) was generated as a
minor degradative intermediate and was detected at a maximum of 8.3% of the
applied radioactivity. TFNA and TFNA-OH were the major degradates in all soils
over the course of the study. TFNA ranged from 24.5 to 56.7% of the applied dose
and TFNA-OH ranged from 31.5 to 53.5% of the applied dose at concentration
maxima. Both degradates were metabolized and mineralized to carbon dioxide
and immobilized as soil-bound residue.
The DT50 and DT90 values ranged from 0.1 to 1.1 days and 0.4 to 3.5 days for the
soils incubated at 20 °C, respectively. The DT50 value was 0.3 days and the DT90
value was 0.9 days for the soil incubated at 10°C. A first-order kinetics model was
used to approximate parent TFNG degradation processes in all of the soil sets.
Bedfordshire
soil (UK21)
Birmingham
soil (UK39)
German soil
2.1
Type Loamy sand Loamy sand Sand
pH 6.8 6.4 5.7
% organic matter 1.25 5.49 1.03
Cation exchange capacity
(meq/100g)
4.97 13.01 2.02
Max water holding capacity
(%)
21.23 39.88 18.41
1st order DT50 (days) 0.3 at 10
oC
(r2=0.9794)
0.1 at 20oC
(r2=0.9477)
0.2
(r2=0.9577)
1.1
(r2= 0.9739)
1st order DT90 (days) 0.9 at 10
oC
0.4 at 20oC
0.7 3.5
Comment
R software has been used to recalculate the DT50 and DT90 at 20oC according to
the best fit:
UK21 soil: best fit is indeterminate order rate with a DT50 of 0.136 (= DT90/3
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according to NAFTA guidelines) and a DT90 of 0.453 days
UK39 soil: best fit is indeterminate order rate with a DT50 of 0.236 (= DT90/3
according to NAFTA guidelines) and a DT90 of 0.784 days.
German soil: best fit is indeterminate order rate with a DT50 of 1.11 (= DT90/3
according to NAFTA guidelines) and a DT90 of 3.68 days.
These values are very similar to the initial ones so the initial ones of the report will
be used.
Conclusion DT50 from 0.1 to 1.1 days at 20oC.
Study type Aerobic degradation on soil study with metabolites
Flag Key study
Test Substance TFNG-AM metabolite with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint DT50
Value DT50 from 0.2 to 1.0 days at 20oC
Reference N.R. Lentz (2002) Rate of degradation of [
14C]TFNG-AM in soil. Ricerca LLC 7528
Auburn Road Painesville OH 44077-1000 USA. Study number 012697-1
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s SETAC 1995
Dose Levels 0.02 ppm
Analytical measurements At 0, 0.04, 0.125, 0.25, 0.5, 1, 2, 3, 7, 14 and 30 days after treatment
Study Summary
The degradation rate of TFNG-AM under aerobic soil conditions was studied in three
European soils. 2 soils were obtained from the United Kingdom, a third standard soil
(2.1) was obtained from Germany.
Samples of each treated soil type were incubated at 50% maximum water-holding
capacity at 20oC in darkness. An additional soil set (U.K.#21) was also treated as
above but incubated at 10°C. The soils were extracted and analyzed by HPLC at
days 0, 0.04, 0.125, 0.25, 0.5, 1, 2, 3, 7, 14 and 30.
Recovery of applied radioactivity over the course of the study for the four sets ranged
from 90.4 to 104.2%. Extractable radiolabel decreased to less than 4.2% of the
applied dose in all soils by the end of the sampling period.
Unextractable residues in all soils ranged from 31.0 to 41.1% by the end of sampling.
Evolution of 14
CO2 ranged from 47.4 to 71.2% of the radiolabel in all soil sets by the
end of the respective sampling periods.
TFNG-AM degraded rapidly in all soils to form TFNG (N-( 4-
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trifluoromethylnicotinoyl)glycine). TFNG degraded rapidly via hydrolysis of the amide
functionality on the aliphatic side chain to form 4-trifluoromethylnicotinic acid (TFNA)
and subsequent hydroxylation of the aromatic ring to form 6- hydroxy-4-
trifluoromethyl-nicotinic acid (TFNA-OH). TFNG, TFNA and TFNA-OH were the major
degradates in all soils over the course of the study. TFNG ranged from 6.2 to 30.1%
of the applied dose. TFNA ranged from 29.4 to 63.8% of the applied dose and TFNA-
OH ranged from 12.4 to 25.3% of the applied dose. All of the degradates were
metabolized and mineralized to carbon dioxide and immobilized as soil-bound
residue.
The DT50 and DT90 values ranged from 0.2 to 1.0 days and 0.6 to 3.3 days for the
soils incubated at 20 °C, respectively. The DT50 value was 0.7 days and the DT90
value was 2.3 days for the soil incubated at 10°C. A first-order kinetics model was
used to approximate parent TFNG-AM degradation processes in all of the soil sets.
Bedfordshire
soil (UK21)
Birmingham
soil (UK39)
German soil
2.1
Type Loamy sand Loamy sand Sand
pH 6.9 7.0 6.2
% organic matter 2.4 4.6 0.9
Cation exchange capacity
(meq/100g)
8.5 16.9 3.3
Max water holding capacity
(%)
41.0 64.8 29.5
1st order DT50 (days) 0.7 at 10
oC
(r2=0.9933)
0.2 at 20oC
(r2=0.9933)
0.3
(r2=0.9669)
1.0
(r2= 0.9959)
1st order DT90 (days) 2.3 at 10
oC
0.6 at 20oC
1.0 3.3
Comment
R software has been used to recalculate the DT50 and DT90 at 20oC according to the
best fit:
UK21 soil: best fit is indeterminate order rate with a DT50 of 0.159 (= DT90/3 according
to NAFTA guidelines) and a DT90 of 0.527 days
UK39 soil: best fit is simple 1st order with a DT50 of 0.3 and a DT90 of 0.998 days.
German soil: best fit is simple 1st order with a DT50 of 1.02 and a DT90 of 3.39 days.
These values are very similar to the initial ones so the initial ones of the report will be
used.
Conclusion DT50 from 0.2 to 1.0 days at 20oC.
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Anaerobic Degradation in Soil (routes and rates) – Laboratory studies
Study type Anaerobic degradation on soil
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Rate (DT50) and route of degradation
Value DT50 120 and 188 days in the whole system at 20oC
Reference N.R. Lentz (2002) Rate of degradation of [
14C]IKI-220 in soil. Ricerca LLC 7528
Auburn Road Painesville OH 44077-1000 USA. Study number 013066-1.
Klimisch Score 1
Amendments/Deviations None that had an impact on the results
GLP Yes
Test Guideline/s US EPA Assessment guidelines, subdivision N Chemistry: environment fate
section 162-3.
Dose Levels 0.05 ppm applied into the aqueous layer, corresponding to 0.1 ppm in soil
Analytical measurements 0, 1,3, 7, 14, 30, 45, 60, 91, 120, 179, 270 and 365 days after treatment
Study Summary
The degradation rate of IKI-220 in flooded soil under anaerobic soil conditions was
studied in one American soil, incubated at 20oC in dark. Soil properties
were:sandy loam with a pH of 5.4, cation exchange capacity 8.11 meq/100g,
3.85% of organic matter, 37.69% of maximum water holding capacity.
The test systems were sampled at intervals over a 365-day period under
anaerobic conditions. At each sampling interval the water and soil phases of the
sample were separated and analyzed for [14
C]IKI-220 and degradates. In addition,
the release of radioactivity as 14
CO2 and radiolabeled volatiles was monitored for
the duration of the study. The mass balance of the applied radioactivity (i.e.,
based on the sum of the water phase, soil extracts, volatile and bound
radioactivity) remained between 90% and 110% at all sampling points.
Under anaerobic aquatic conditions, the rate of degradation of [14
C]IKI-220 was
moderately rapid. Throughout the study the amount of radioactivity in the soil and
water phases of the system remained steady. The degradation of IKI-220 to TFNA
was prevalent in the system and TFNA increased steadily throughout the study. A
maximum mean TFNA level of 75.7 % of the applied radioactivity was obtained at
365 days. There was no other significant degradation product in the system. The
levels of TFNG, TFNG-AM, TFNA-AM and TFNA-OH did not reach significant
levels and all were <0.5% of the applied radioactivity at any given time in the total
system. Only small amounts of 14
CO2 were produced and reached 0.2% of the
applied radioactivity by 365 days. The amount of bound nonextractable
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radioactivity (PES) slowly increased during the study and reached a mean level of
3.6% of the applied radioactivity by 270 days. There were no volatile organic
compounds evolved in the flooded soil system.
The DT50 and DT90 values of IKI-220 in flooded anaerobic soil were 88 days and
490 days, respectively, in the water phase and 121 days and 555 days,
respectively, in the entire system.
Under anaerobic aquatic conditions, IKI-220 was metabolized at a moderately
rapid rate. The main metabolism process involved the formation of TFNA, most
likely, through the hydrolytic cleavage of the amide bonds in the side chain.
Water phase Total system
1st order DT50 (days) 88 (r
2=0.9847) 121 (r
2=0.9955)
1st order DT90 (days) 490 555
Comment
R software has been used to recalculate the DT50 and DT90 according to the best
fit:
Water phase: best fit is double 1st order with a DT50 of 98.1 days and a DT50 for
the slow phase of 174 days and a DT90 of 501 days.
Whole system: best fit double 1st order with a DT50 of 120 days and a DT50 for the
slow phase of 188 days and a DT90 of 1030 days.
These values are very different to the initial ones so these re-calculated values will
be used.
Conclusion DT50 120 and 188 days in the whole system at 20oC.
Soil photolysis
Study type Soil photolysis
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Photolysis half-life
Value 22 days in continuous illumination, 53 days in dark.
Reference
KJ Walsh (2002) A photochemical degradation of [14
C]-IKI-220 in soil. Ricerca
LLC. Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio,
44077-1000. Report no 011298-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OPPTS 835.2410
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Dose Levels ca 0.1 ppm dry soil
Analytical measurements HPLC with radiochemical flow detection
Study Summary
The photolysis of the substance was studied at a concentration of ca 0.1 ppm on
dried soil which was continuously exposed to simulated sunlight (xenon arc lamp
with filters) for 15 days at a temperature of 20 o
C ± 1. At days 0, 1, 3, 7, 9, 11 and
15, sample extracts were analysed by HPLC.
A typical US loamy sand agricultural soil was used for the study and was dried to
minimize the effects of microbial degradation.
The substance degraded slowly in both dark and light exposed samples. TFNG-
AM was the major metabolite in both dark and light-exposed samples at 13.8%
and 29.5%, respectively. TFNA-AM and TFNG were detected as minor
metabolites (2-5 %). The half-life in dark conditions is 53 days and is 22 days in
continuous illumination.
Conclusion Photolysis is not a significant pathway for the degradation of the substance
in use conditions.
Adsorption/ desorption on soil particles
Study type Batch equilibrium test
Flag Key study
Test Substance IKI-220 with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Koc
Value 8 to 42
Reference
KJ Sandacz (2000) Adsorption and desorption of [14
C]-IKI-220 in soils. Ricerca
Inc. Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio,
44077-1000. Report no 6934-96-0185-EF-001
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 106 (version not stated)
Dose Levels ca 1 and 5 µg/mL in 0.01 M calcium chloride solution
Analytical measurements HPLC
Study Summary
Screening test:
The adsorptive properties of the test substance were studied in 4 soils (3
European and 1 American soil): 2 loamy sands and 2 sandy loams with organic
matter content from 0.7 to 3.0% and pH from 6.5 to 7.6.
The percent of the test substance adsorbed to the soil using a 1:5 soil:test
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substance solution (ca 1 µg/mL ) ratio ranged from 2.8 to 8.3%.
In the run with 1:2 soil:test substance solution (ca 5 µg/mL), the percent of
adsorption remain below 15%.
The concentration of the test substance in the aqueous phase after adsorption
was used to estimate the adsorption coefficients.
EFS-097 EFS-099 EFS-101 EFS-072
Soil type Loamy sand Sandy loam Sandy loam Loamy sand
Origin UK UK Germany Ohio, USA
% organic
carbon 1.2 3.0 1.3 0.7
pH 7.6 6.9 6.8 6.5
% moisture 6.29 16.39 8.58 8.50
Koc for 1:5
ratio and no
HgCl2
12.1 15.1 22.7 34.9
Koc for 1:5
ratio and
0.01% HgCl2
16.2 13.3 18.8 42.1
Koc for 1:2
ratio and
0.01% HgCl2
7.9 11.2 13.4 20.7
The substance is highly mobile to mobile in soils according to the FAO
classification.
Conclusion Koc from 8 to 42. The substance is highly mobile to mobile in soils
Study type Batch equilibrium test
Flag Key study
Test Substance Metabolite TFNA with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Koc
Value 0.00 to 3.05
Reference
GJ Misich & KJ Sandacz Herczog (2002) Adsorption and desorption of [14
C]-TFNA
in soil. Ricerca LLC. Environmental and metabolic fate. 7528 Auburn Road
Painesville Ohio, 44077-1000. Report no 012061-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
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Test Guideline/s OECD 106 (version not stated)
Dose Levels ca 5 µg/mL in 0.01 M calcium chloride solution
Analytical measurements HPLC
Study Summary
Screening test:
The adsorptive properties of the test substance were studied in 4 soils (3
European and 1 American soil): 2 loamy sands and 1 sandy loam and 1 sand with
organic matter content from 0.93 to 5.49% and pH from 5.7 to 7.2.
The percent of the test substance adsorbed to the soil using a 1:1 soil:test
substance solution (ca 5 µg/mL ) ratio ranged from 0.0 to 1.40%.
The concentration of the test substance in the aqueous phase after adsorption
was used to estimate the adsorption coefficients.
EFS-165 EFS-166 EFS-167 EFS-072
Soil type Loamy sand Sandy loam Sandy loam Loamy sand
Origin UK UK Germany Ohio, USA
% organic
carbon 0.727 3.19 0.599 0.541
pH 6.8 6.4 5.7 7.2
Koc 0.35 0.00 3.05 2.67
The substance is highly mobile in soils according to the FAO classification.
Conclusion Koc from 0.00 to 3.05. The substance is highly mobile in soils
Study type Batch equilibrium test
Flag Key study
Test Substance Metabolite TFNA-AM with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Koc
Value 4.53 to 12.11
Reference
KJ Sandacz Herczog et al (2002) Adsorption and desorption of [14
C]-TFNA-AM in
soils. Ricerca LLC. Environmental and metabolic fate. 7528 Auburn Road
Painesville Ohio, 44077-1000. Report no 012694-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 106 (version not stated)
Dose Levels ca 5 µg/mL in 0.01 M calcium chloride solution
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Analytical measurements HPLC
Study Summary
Screening test:
The adsorptive properties of the test substance were studied in 4 soils (3
European and 1 American soil): 2 loamy sands and 1 sandy loam and 1 sand with
organic matter content from 0.98 to 5.49% and pH from 5.7 to 7.2.
The percent of the test substance adsorbed to the soil using a 1:1 soil:test
substance solution (ca 5 µg/mL ) ratio ranged from 1.97 to 5.47%.
The concentration of the test substance in the aqueous phase after adsorption
was used to estimate the adsorption coefficients.
EFS-165 EFS-166 EFS-167 EFS-072
Soil type Loamy sand Sandy loam Sandy loam Loamy sand
Origin UK UK Germany Ohio, USA
% organic
carbon 0.727 3.19 0.599 0.570
pH 6.8 6.4 5.7 7.2
Koc 5.52 2.76 5.16 4.80
Adsorption equilibrium test
The adsorption equilibrium time was also investigated in four US soils and one
German soil.
EFS-116 EFS-150 EFS-167 EFS-170 EFS-200
Soil type Sandy
loam
Silt loam Sand Clay loam Clay loam
Origin Ohio, USA Oregon,
USA
Germany Ohio, USA Montana,
USA
% organic
carbon 3.081 1.948 0.599
4.302 0.872
pH 5.6 6.2 5.7 7.9 8.1
Koc 5.53 10.11 5.04 4.53 12.11
The adsorption equilibrium was approximately 24-hour for all soils tested.
The percent of the test substance adsorbed to the soil ranged from 2.92 to
16.16%. The Koc ranged from 4.53 to 12.11.
The substance is highly mobile to mobile in soils according to the FAO
classification.
Conclusion Koc from 4.53 to 12.11. The substance is highly mobile to mobile in soils
Study type Batch equilibrium test
Flag Key study
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Test Substance Metabolite TFNA-OH with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Koc
Value 1.60 to 4.39
Reference
GJ Misich (2002) Adsorption and desorption of [14
C]-TFNA-OH in soil. Ricerca
LLC. Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio,
44077-1000. Report no 012063-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 106 (version not stated)
Dose Levels ca 5 µg/mL in 0.01 M calcium chloride solution
Analytical measurements HPLC
Study Summary
Screening test:
The adsorptive properties of the test substance were studied in 4 soils (3
European and 1 American soil): 2 loamy sands and 1 sandy loam and 1 sand with
organic matter content from 0.93 to 5.49% and pH from 5.7 to 7.2.
The percent of the test substance adsorbed to the soil using a 1:1 soil:test
substance solution (ca 5 µg/mL ) ratio ranged from 0.8 to 3.8%.
The concentration of the test substance in the aqueous phase after adsorption
was used to estimate the adsorption coefficients.
EFS-165 EFS-166 EFS-167 EFS-072
Soil type Loamy sand Sandy loam Sandy loam Loamy sand
Origin UK UK Germany Ohio, USA
% organic
carbon 0.727 3.19 0.599 0.541
pH 6.8 6.4 5.7 7.2
Koc 1.60 1.92 4.19 4.39
The substance is highly mobile in soils according to the FAO classification.
Conclusion Koc from 1.60 to 4.39. The substance is highly mobile in soils
Study type Batch equilibrium test
Flag Key study
Test Substance Metabolite TFNG with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Koc
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Value 0.20 to 4.05
Reference
KJ Sandacz Herczog & GJ Misich (2002) Adsorption and desorption of [14
C]-
TFNG in soil. Ricerca LLC. Environmental and metabolic fate. 7528 Auburn Road
Painesville Ohio, 44077-1000. Report no 012062-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 106 (version not stated)
Dose Levels ca 5 µg/mL in 0.01 M calcium chloride solution
Analytical measurements HPLC
Study Summary
Screening test:
The adsorptive properties of the test substance were studied in 4 soils (3
European and 1 American soil): 2 loamy sands and 1 sandy loam and 1 sand with
organic matter content from 0.93 to 5.49% and pH from 5.7 to 7.2.
The percent of the test substance adsorbed to the soil using a 1:1 soil:test
substance solution (ca 5 µg/mL ) ratio ranged from 0.10 to 2.15%.
The concentration of the test substance in the aqueous phase after adsorption
was used to estimate the adsorption coefficients.
EFS-165 EFS-166 EFS-167 EFS-072
Soil type Loamy sand Sandy loam Sandy loam Loamy sand
Origin UK UK Germany Ohio, USA
% organic
carbon 0.727 3.19 0.599 0.541
pH 6.8 6.4 5.7 7.2
Koc 0.20 1.05 1.29 4.05
The substance is highly mobile in soils according to the FAO classification.
Conclusion Koc from 0.2 to 4.05. The substance is highly mobile in soils
Study type Batch equilibrium test
Flag Key study
Test Substance Metabolite TFNG-AM with 14
C labelling at the C-3 carbon in the pyridyl ring
Endpoint Koc
Value 4.24 to 15.84
Reference W Song (2002) Adsorption and desorption of [14
C]-TFNG-AM in soil. Ricerca LLC.
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Environmental and metabolic fate. 7528 Auburn Road Painesville Ohio, 44077-
1000. Report no 012695-1
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study.
GLP Yes
Test Guideline/s OECD 106 (version not stated)
Dose Levels ca 5 µg/mL in 0.01 M calcium chloride solution
Analytical measurements HPLC
Study Summary
Screening test:
The adsorptive properties of the test substance were studied in 4 soils (3
European and 1 American soil): 2 loamy sands and 1 sandy loam and 1 sand with
organic matter content from 0.93 to 5.49% and pH from 5.7 to 7.2.
The percent of the test substance adsorbed to the soil using a 1:1 soil:test
substance solution (ca 5 µg/mL ) ratio ranged from 0.10 to 2.15%.
The concentration of the test substance in the aqueous phase after adsorption
was used to estimate the adsorption coefficients.
EFS-165 EFS-166 EFS-167 EFS-072
Soil type Loamy sand Sandy loam Sandy loam Loamy sand
Origin UK UK Germany Ohio, USA
% organic
carbon 0.727 3.192 0.599 0.570
pH 6.8 6.4 5.7 7.2
Koc 5.50 2.51 0.00 13.16
Adsorption equilibrium test
The adsorption equilibrium time was also investigated in four US soils and one
German soil.
EFS-116 EFS-150 EFS-167 EFS-170 EFS-200
Soil type Sandy
loam
Silt loam Sand Clay loam Clay loam
Origin Ohio, USA Oregon,
USA
Germany Ohio, USA Montana,
USA
% organic
carbon 3.081 1.948 0.599
4.302 0.872
pH 5.6 6.2 5.7 7.9 8.1
Koc 10.51 9.25 7.56 4.24 15.84
The adsorption equilibrium was approximately 24-hour for all soils tested.
The percent of the test substance adsorbed to the soil ranged from 4.54 to
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27.24%. The Koc ranged from 4.24 to 15.84.
The substance is highly mobile to mobile in soils according to the FAO
classification.
Conclusion Koc from 4.24 to 15.84. The substance is highly mobile to mobile in soils
Photodegradation in air
Study type Photodegradation in air (estimation)
Flag Key study
Test Substance IKI-220
Endpoint Half-life
Value 13.737 days (12-hour day)
Reference
A van der Gaauw (2001) Estimation of the degradation of IKI-220 by photo-
oxidation in air. Model calculation according to Atkinson. RCC Ltd Environmental
Chemistry & Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no
835176
Klimisch Score 1
Amendments/Deviations None
GLP yes
Test Guideline/s Calculation according to Atkinson method
Dose Levels Not relevant: calculation
Analytical measurements Not relevant: calculation
Study Summary
The rate constant for the atmospheric reaction between photochemically produced
hydroxyls radicals and the substance was estimated by AOPWIN.
An OH-concentration of 1.5 x 106 /cm
3 is used as an average concentration during
daylight, for the reaction with ozone, a concentration of 7 x 1011
molecules /cm3 is
assumed which is a concentration of ozone in unpolluted air in the lower
troposphere.
The half-life of the substance is estimated to be 13.737 days when a 12-hour day
is considered and 6.868 days when a 24-hour day is considered.
Conclusion Half-life = 13.737 days
General conclusion about environmental fate:
Flonicamid is persistent in aquatic environment according to HSNO criteria (DT50 > 16 days and not readily
biodegradable). But is not persistent in soil (DT50 < 30 days). The metabolites are also quickly degraded in
soils.
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Flonicamid and its metabolites are all mobile to highly mobile in soil according to their adsorption/desorption
coefficients.
Flonicamid bioconcentration properties are not fully characterised as no bioconcentration study is available
and the n-octanol-water partition coefficient is not a good predictor for surface active substances.
Metabolites to be considered for the risk assessment:
Soil: TFNA, TFNA-OH, TFNG-AM, TFNG, TFNA-AM
Water: TFNA, TFNA-OH
Ecotoxicity - Robust study summaries for the active ingredient and
metabolite(s)
Aquatic toxicity
Fish acute toxicity (Freshwater species)
Type of study Limit test
Flag Key study
Test Substance IKI-220 technical
Species Oncorhynchus mykiss (Rainbow trout)
Type of exposure Static for 96 hours
Endpoint LC50
Value > 100 mg/L
Reference
A. Peither (2001) Acute toxicity of IKI-220 technical to Rainbow trout (Oncorhynchus
mykiss) in a 96-hour static test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 807254.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium and at 48 and 96 hours, by HPLC
Study Summary
The acute toxicity of the test substance to rainbow trout was determined in a 96-hr
static test. A limit test was performed at 100 mg/L. The measured concentration in the
test medium was 98% of the nominal at the start and at the end of the test substance
so the results are expressed as nominal concentrations.
The pH values in the test medium and in the control ranged from 7.8 to 7.9, the
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oxygen concentration was always higher than 60% oxygen saturation (it was 8.7 mg/L
or above) and the temperature was constantly 15oC.
In the control and in the test concentration, no mortality or other signs of intoxication
were determined during the test period.
The LC50 is higher than 100 mg/L.
Conclusion The LC50 is higher than 100 mg/L. No mortality observed at 100 mg/L.
Type of study Limit test
Flag Key study
Test Substance IKI-220 technical
Species Lepomis macrochirus (Bluegill sunfish)
Type of exposure Static for 96 hours
Endpoint LC50
Value > 100 mg/L
Reference
A. Peither (2001) Acute toxicity of IKI-220 technical to bluegill sunfish (Lepomis
macrochirus) in a 96-hour static test. RCC Ltd, Environmental Chemistry &
Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no 807276.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium and at 48 and 96 hours, by HPLC
Study Summary
The acute toxicity of the test substance to bluegill sunfish was determined in a 96-hr
static test. A limit test was performed at 100 mg/L. The measured concentration in the
test medium was 99% of the nominal at the start and at the end of the test substance
so the results are expressed as nominal concentrations.
The pH values in the test medium and in the control ranged from 7.8 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 7.4 mg/L
or above) and the temperature was in the range of 22-23oC.
In the control and in the test concentration, no mortality or other signs of intoxication
were determined during the test period.
The LC50 is higher than 100 mg/L.
Conclusion The LC50 is higher than 100 mg/L. No mortality observed at 100 mg/L.
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Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA
Species Oncorhynchus mykiss (Rainbow trout)
Type of exposure Semi-static for 96 hours. Renewal after 48 hours
Endpoint LC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA to Rainbow trout (Oncorhynchus mykiss)
in a 96-hour semi-static test. RCC Ltd, Environmental Chemistry &
Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no 834208.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium on day 0 and at 48 h and from the old medium at 48 and
96 hours, by HPLC
Study Summary
The acute toxicity of the test substance to Rainbow trout was determined in a 96-
hr semi-static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 102-103% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.7 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 8.5
mg/L or above) and the temperature was in the range of 13-14oC.
In the control and in the test concentration, no mortality or other signs of
intoxication were determined during the test period.
The LC50 is higher than 100 mg/L.
Conclusion The LC50 is higher than 100 mg/L. No mortality observed at 100 mg/L.
Type of study Limit test
Flag Key study
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Test Substance Metabolite TFNA-AM
Species Oncorhynchus mykiss (Rainbow trout)
Type of exposure Semi-static for 96 hours. Renewal after 48 hours
Endpoint LC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA-AM to Rainbow trout (Oncorhynchus
mykiss) in a 96-hour semi-static test. RCC Ltd, Environmental Chemistry &
Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no 834952.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium on day 0 and at 48 h and from the old medium at 48 and
96 hours, by HPLC
Study Summary
The acute toxicity of the test substance to Rainbow trout was determined in a 96-
hr semi-static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 100-102% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.8 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 8.4
mg/L or above) and the temperature was in the range of 13-14oC.
In the control and in the test concentration, no mortality or other signs of
intoxication were determined during the test period.
The LC50 is higher than 100 mg/L.
Conclusion The LC50 is higher than 100 mg/L. No mortality observed at 100 mg/L.
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA-OH
Species Oncorhynchus mykiss (Rainbow trout)
Type of exposure Semi-static for 96 hours. Renewal after 48 hours
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Endpoint LC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA-OH to Rainbow trout (Oncorhynchus
mykiss) in a 96-hour semi-static test. RCC Ltd, Environmental Chemistry &
Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no 834996.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium on day 0 and at 48 h and from the old medium at 48 and
96 hours, by HPLC
Study Summary
The acute toxicity of the test substance to Rainbow trout was determined in a 96-
hr semi-static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 99-100% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.6 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 8.5
mg/L or above) and the temperature was in the range of 13-14oC.
In the control and in the test concentration, no mortality or other signs of
intoxication were determined during the test period.
The LC50 is higher than 100 mg/L.
Conclusion The LC50 is higher than 100 mg/L. No mortality observed at 100 mg/L.
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNG-AM
Species Oncorhynchus mykiss (Rainbow trout)
Type of exposure Semi-static for 96 hours. Renewal after 48 hours
Endpoint LC50
Value > 100 mg/L
Reference A. Peither (2002) Acute toxicity of TFNG-AM to Rainbow trout (Oncorhynchus
mykiss) in a 96-hour semi-static test. RCC Ltd, Environmental Chemistry &
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Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no 834974.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203 (1992)
No/Group 10
Dose Levels 100 mg/L
Analytical measurements Yes from fresh medium on day 0 and at 48 h and from the old medium at 48 and
96 hours, by HPLC
Study Summary
The acute toxicity of the test substance to Rainbow trout was determined in a 96-
hr semi-static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 105-107% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.7 to 8.0, the
oxygen concentration was always higher than 60% oxygen saturation (it was 8.4
mg/L or above) and the temperature was in the range of 13-14oC.
In the control and in the test concentration, no mortality or other signs of
intoxication were determined during the test period.
The LC50 is higher than 100 mg/L.
Conclusion The LC50 is higher than 100 mg/L. No mortality observed at 100 mg/L.
Fish acute toxicity (Marine species)
No study provided.
Invertebrates acute toxicity (Freshwater species)
Type of study Limit test
Flag Key study
Test Substance IKI-220 technical
Species Daphnia magna
Type of exposure Static, 48 hours
Endpoint EC50
Value > 100 mg/L
Reference
A. Peither (2001) Acute toxicity of IKI-220 technical to Daphnia magna in a 48-
hour immobilization test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 807311.
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Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202 (1984)
No/Group 2 replicates of 10
Dose Levels 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 48 h, by HPLC
Study Summary
The acute toxicity of the test substance to Daphnia magna was determined in a
48-hr static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 98-99% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.8 to 7.9, the
oxygen concentration was always higher than 8.1 mg/L and the temperature was
in the range of 20-21oC.
In the control and in the test concentration, no immobilisation or other signs of
intoxication were determined during the test period.
The EC50 is higher than 100 mg/L.
Conclusion The EC50 is higher than 100 mg/L. No immobilisation observed at 100 mg/L.
Type of study Full test
Flag Supporting study
Test Substance IKI-220 technical
Species Chironomus riparius (first instar larvae)
Type of exposure Static, 48 hours
Endpoint LC50
Value > 200 mg/L
Reference
U. Memmert (2002) Acute toxicity of IKI-220 technical to first instar larvae of
Chironomus riparius in a 48-hour immobilization test. RCC Ltd, Environmental
Chemistry & Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no
834074.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
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Test Guideline/s
No official test guideline available. The administration of the test item is based on
OECD 202 (1984) guideline about daphnia immobilisation test; the species and
age of organisms are based on the draft OECD 2019 (2001): “Sediment-water
chironomid toxicity test using spiked water”.
No/Group 4 replicates of 20 larvae
Dose Levels 2.0, 6.3, 20, 63 and 200 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 48 h, by HPLC. Only the samples of the
highest concentration were analysed as it is also the NOEC.
Study Summary
The acute toxicity of the test substance to first instar larvae of Chironomus riparius
was determined in a 48-hr static test. The test substance was dissolved in water
and the larvae were introduced into these test media. The survival rate of the
larvae and symptoms of intoxication were recorded. No sediment was added
since otherwise the small larvae, dwelt in sediment cannot be observed.
The nominal concentrations were 2.0, 6.3, 20, 63 and 200 mg/L. The measured
concentration in the test medium was 102-104% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.6 to 8.0, the
oxygen concentration was always higher than 7.8 mg/L and the temperature was
in the range of 22oC.
In the control and in the test concentrations, no mortality or other signs of
intoxication were determined during the test period.
The LC50 is higher than 200 mg/L.
Conclusion The LC50 is higher than 200 mg/L. No mortality observed at 200 mg/L.
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA
Species Daphnia magna
Type of exposure Static, 48 hours
Endpoint EC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA to Daphnia magna in a 48-hour
immobilization test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 834186.
Klimisch Score 1
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Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202 (1984)
No/Group 2 replicates of 10
Dose Levels 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 48 h, by HPLC
Study Summary
The acute toxicity of the test substance to Daphnia magna was determined in a
48-hr static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 99% of the nominal at the start and at the
end of the test substance so the results are expressed as nominal concentrations.
The pH values in the test medium and in the control ranged from 7.7 to 7.9, the
oxygen concentration was always higher than 8.2 mg/L and the temperature was
in the range of 21-22oC.
In the control and in the test concentration, no immobilisation or other signs of
intoxication were determined during the test period.
The EC50 is higher than 100 mg/L.
Conclusion The EC50 is higher than 100 mg/L. No immobilisation observed at 100 mg/L.
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA-AM
Species Daphnia magna
Type of exposure Static, 48 hours
Endpoint EC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA-AM to Daphnia magna in a 48-hour
immobilization test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 834895.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202 (1984)
No/Group 2 replicates of 10
Dose Levels 100 mg/L
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Analytical measurements Yes, from fresh medium on day 0 and at 48 h, by HPLC
Study Summary
The acute toxicity of the test substance to Daphnia magna was determined in a
48-hr static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 99% of the nominal at the start and at the
end of the test substance so the results are expressed as nominal concentrations.
The pH values in the test medium and in the control ranged from 7.8 to 7.9, the
oxygen concentration was always higher than 8.0 mg/L and the temperature was
in the range of 21-22oC.
In the control and in the test concentration, no immobilisation or other signs of
intoxication were determined during the test period.
The EC50 is higher than 100 mg/L.
Conclusion The EC50 is higher than 100 mg/L. No immobilisation observed at 100 mg/L.
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA-OH
Species Daphnia magna
Type of exposure Static, 48 hours
Endpoint EC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA-OH to Daphnia magna in a 48-hour
immobilization test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 834930.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202 (1984)
No/Group 2 replicates of 10
Dose Levels 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 48 h, by HPLC
Study Summary
The acute toxicity of the test substance to Daphnia magna was determined in a
48-hr static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 101% of the nominal at the start and at the
end of the test substance so the results are expressed as nominal concentrations.
The pH values in the test medium and in the control ranged from 7.6 to 7.9, the
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oxygen concentration was always higher than 8.1 mg/L and the temperature was
in the range of 21-22oC.
In the control and in the test concentration, no immobilisation or other signs of
intoxication were determined during the test period.
The EC50 is higher than 100 mg/L.
Conclusion The EC50 is higher than 100 mg/L. No immobilisation observed at 100 mg/L.
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNG-AM
Species Daphnia magna
Type of exposure Static, 48 hours
Endpoint EC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNG-AM to Daphnia magna in a 48-hour
immobilization test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 834917.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202 (1984)
No/Group 2 replicates of 10
Dose Levels 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 48 h, by HPLC
Study Summary
The acute toxicity of the test substance to Daphnia magna was determined in a
48-hr static test. A limit test was performed at 100 mg/L. The measured
concentration in the test medium was 96-99% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium and in the control ranged from 7.8, the oxygen
concentration was always higher than 8.1 mg/L and the temperature was 21oC.
In the control and in the test concentration, no immobilisation or other signs of
intoxication were determined during the test period.
The EC50 is higher than 100 mg/L.
Conclusion The EC50 is higher than 100 mg/L. No immobilisation observed at 100 mg/L.
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Invertebrates acute toxicity (Marine species)
No study provided.
Algae acute toxicity (Freshwater species)
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Pseudokirchneriella subcapitata
Type of exposure Static, 72 hours
Endpoint ErC50 and EbC50
Value > 100 mg/L
Reference
A. Peither (2001) Acute toxicity of IKI-220 technical to Pseudokirchneriella
subcapitata (formerly Selenastrum capricornutum) in a 72-hour algal growth
inhibition test. RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-
4452 Itingen Switzerland. Report no 807298.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201 (1984)
No/Group 3 replicates for test concentrations and 6 for the control of 104 cells/mL
Dose Levels 4.6, 10, 22, 46 and 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 72 h of the 2 highest concentrations, by
HPLC
Study Summary
The influence of the test substance on the growth of the green algal species
Pseudokirchneriella subcapitata was determined in a 72-hr static test. The nominal
concentrations were 4.6, 10, 22, 46 and 100 mg/L. The measured concentration in
the test medium was 94-97% of the nominal at the start and at the end of the test
substance so the results are expressed as nominal concentrations.
The pH values in the test medium and in the control ranged from 7.9 at the start of
the study to 9.0 at the end, and the temperature was 22oC.
The growth rate in the control was valid according to the guideline (at least a factor
of 16).
The test substance had a statistically significant inhibitory effect on the growth
(biomass and growth rate) at the concentration of 100 mg/L. The NOEC was 46
mg/L and both ErC50 and EbC50 are > 100 mg/L.
Conclusion The ErC50 and EbC50 are higher than 100 mg/L.
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Type of study Full test
Flag Key study
Test Substance Metabolite TFNA
Species Pseudokirchneriella subcapitata
Type of exposure Static, 72 hours
Endpoint ErC50 and EbC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA to Pseudokirchneriella subcapitata
(formerly Selenastrum capricornutum) in a 72-hour algal growth inhibition test.
RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 834164.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201 (1984)
No/Group 3 replicates for test concentrations and 6 for the control of 104 cells/mL
Dose Levels 1.0, 3.2, 10 and 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 72 h of the 2 highest concentrations, by
HPLC
Study Summary
The influence of the test substance on the growth of the green algal species
Pseudokirchneriella subcapitata was determined in a 72-hr static test. The
nominal concentrations were 1.0, 3.2, 10 and 100 mg/L. The measured
concentration in the test medium was 90-97% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values in the test medium were adjusted with diluted sodium hydroxide
solution at the start of the test. They ranged from 7.8 to 8.0 (after adjustment) at
the start of the study and from 8.8 to 9.0 at the end, and the temperature was
23oC.
The growth rate in the control was valid according to the guideline (at least a
factor of 16).
The test substance had a statistically significant inhibitory effect on the growth
(biomass and growth rate) at the concentration of 100 mg/L. The NOEC was 32
mg/L and both ErC50 and EbC50 are > 100 mg/L.
Conclusion The ErC50 and EbC50 are higher than 100 mg/L.
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Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA-AM
Species Pseudokirchneriella subcapitata
Type of exposure Static, 72 hours
Endpoint ErC50 and EbC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA-AM to Pseudokirchneriella subcapitata
(formerly Selenastrum capricornutum) in a 72-hour algal growth inhibition test.
RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 834838.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201 (1984)
No/Group 3 replicates for test concentrations and 6 for the control of 104 cells/mL
Dose Levels 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 72 h of the 2 highest concentrations, by
HPLC
Study Summary
The influence of the test substance on the growth of the green algal species
Pseudokirchneriella subcapitata was determined in a 72-hr static test. A limit test
was performed at 100 mg/L. The measured concentration in the test medium was
96-98% of the nominal at the start and at the end of the test substance so the
results are expressed as nominal concentrations.
The pH values ranged from 8.1 at the start of the study to 9.4-9.5 at the end, and
the temperature was 23oC.
The growth rate in the control was valid according to the guideline (at least a
factor of 16).
The test substance had no statistically significant inhibitory effect on the growth
(biomass and growth rate) at the concentration of 100 mg/L. The NOEC was 100
mg/L and both ErC50 and EbC50 are > 100 mg/L.
Conclusion The ErC50 and EbC50 are higher than 100 mg/L.
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Type of study Full test
Flag Key study
Test Substance Metabolite TFNA-OH
Species Pseudokirchneriella subcapitata
Type of exposure Static, 72 hours
Endpoint ErC50 and EbC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNA-OH to Pseudokirchneriella subcapitata
(formerly Selenastrum capricornutum) in a 72-hour algal growth inhibition test. RCC
Ltd, Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 834873.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201 (1984)
No/Group 3 replicates for test concentrations and 6 for the control of 104 cells/mL
Dose Levels 2.2, 4.6, 10, 22, 46 and 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 72 h of the 2 highest concentrations, by
HPLC
Study Summary
The influence of the test substance on the growth of the green algal species
Pseudokirchneriella subcapitata was determined in a 72-hr static test. The nominal
concentrations were 2.2, 4.6, 10, 22, 46 and 100 mg/L. The measured
concentration in the test medium was 91-106% of the nominal at the start and at the
end of the test substance so the results are expressed as nominal concentrations.
The pH values in the test medium were adjusted with diluted sodium hydroxide
solution at the start of the test. They ranged from 7.7 to 7.9 (after adjustment) at the
start of the study and from 8.6 to 9.1 at the end, except at 100 mg/L where it was
5.5, and the temperature was 23oC.
The growth rate in the control was valid according to the guideline (at least a factor
of 16).
The test substance had a statistically significant inhibitory effect on the growth
(biomass and growth rate) at the concentration of 10 mg/L and above. The NOEC
was 4.6 mg/L, ErC50 is > 100 mg/L and EbC50 is 29 mg/L.
Conclusion ErC50 > 100 mg/L and EbC50 = 29 mg/L.
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Type of study Full test
Flag Key study
Test Substance Metabolite TFNG-AM
Species Pseudokirchneriella subcapitata
Type of exposure Static, 72 hours
Endpoint ErC50 and EbC50
Value > 100 mg/L
Reference
A. Peither (2002) Acute toxicity of TFNG-AM to Pseudokirchneriella subcapitata
(formerly Selenastrum capricornutum) in a 72-hour algal growth inhibition test.
RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 834851.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201 (1984)
No/Group 3 replicates for test concentrations and 6 for the control of 104 cells/mL
Dose Levels 4.6, 10, 22, 46 and 100 mg/L
Analytical measurements Yes, from fresh medium on day 0 and at 72 h, by HPLC
Study Summary
The influence of the test substance on the growth of the green algal species
Pseudokirchneriella subcapitata was determined in a 72-hr static test. The
nominal concentrations were 4.6, 10, 22, 46 and 100 mg/L. The measured
concentration in the test medium was 99-108% of the nominal at the start and at
the end of the test substance so the results are expressed as nominal
concentrations.
The pH values ranged from 7.9 to 8.0 at the start of the study and from 8.4 to 8.6
at the end, and the temperature was 22oC.
The growth rate in the control was valid according to the guideline (at least a
factor of 16).
The test substance had a statistically significant inhibitory effect on the growth
(biomass and growth rate) at the concentration of 22 mg/L and above. The NOEC
was 10 mg/L, both ErC50 and EbC50 are > 100 mg/L.
Conclusion ErC50 and EbC50 > 100 mg/L.
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Algae acute toxicity (Marine species)
No study provided.
Aquatic plants acute toxicity (Freshwater species)
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Lemna gibba G3
Type of exposure Semi-static, 7 days, renewal on days 3 and 5
Endpoint EC50
Value > 119 mg/L
Reference
D Desjardins et al (2002) IKI-220 technical: a 7-day static-renewal toxicity test with
duckweed (Lemna gibba G3). Wildlife International Ltd, 8598 Commerce drive
Easton, Maryland 21601 USA. Report no 272A-108.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OPPTS 850.4400 (draft) and OECD 221 (draft)
No/Group 3 replicates of 5 plants, totalling 15 fronds
Dose Levels 7.5, 15, 30, 60 and 120 mg/L (nominal)
7.3, 15, 30, 59 and 119 mg/L (mean measured)
Analytical measurements Yes, from fresh medium and old medium on day 0, 3 and 5 and at the end of the
test, by HPLC
Study Summary
The influence of the test substance on the growth of the duckweed Lemna gibba
was determined in a 7-day semi-static test. The nominal concentrations were 7.5,
15, 30, 60 and 120 mg/L. The mean measured concentrations in the test medium
were 7.3, 15, 30, 59 and 119 mg/L.
The pH values ranged from 8.0 to 8.2 at the start of the study and from 9.3 to 9.4
at the end, and the temperature was 25.1-26.1oC. The light intensity ranged from
4480 to 5340 lux.
The toxicity was determined by evaluating the production of plants and fronds and
their general health over 7 days.
Inhibitions of frond growth and percent of growth rates were not statistically
significantly different from the control.
The NOEC was 119 mg/L, EC50 is > 119 mg/L.
Conclusion EC50 > 119 mg/L.
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Fish chronic toxicity (Freshwater species)
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Pimephales promelas (fathead minnow)
Type of exposure Flow-through, 33 days (5 days pre-hatch and 28 days post-hatch)
Endpoint NOEC for time of hatch, hatching success, growth and survival
Value 10 mg/L (based on growth)
Reference
SJ Palmer et al (2002) IKI-220 technical: an early life stage toxicity test with the
fathead minnow (Pimephales promelas). Wildlife International Ltd, 8598
Commerce drive Easton, Maryland 21601 USA. Report no 272A-104.
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OPPTS 850.1400 (draft) and OECD 210 (1992)
No/Group 4 replicates of 20 embryos
Dose Levels 1.3, 2.5, 5.0, 10 and 20 mg/L (nominal)
1.2, 2.6, 4.9, 9.5 and 20 mg/L (mean measured)
Analytical measurements Yes, from samples collected at the beginning of the test, on day 5 and weekly
thereafter and at the study termination, by HPLC
Study Summary
The influence of the test substance on the growth of the fathead minnow early-life
stages was determined in a 33-day flow-through test. The nominal concentrations
were 1.3, 2.5, 5.0, 10 and 20 mg/L. The mean measured concentrations in the test
medium were 1.2, 2.6, 4.9, 9.5 and 20 mg/L.
The pH values ranged from 8.1 to 8.4, and the temperature was in the range 25 ±
1oC. Dissolved oxygen concentration remained > 7.5 mg/L (92% saturation).
All embryos hatched between days 4 and 5, there was no apparent difference
among the experimental groups.
Hatching success in the control was 99%, and varied from 95 to 100% in the test
substance groups. There was no statistically significant difference.
Larval survival in the control was 91% at the end of the test; and varied from 92 to
96% in the test substance groups. There was no statistically significant difference.
Biological observations of sublethal effects during the 28-day post-hatch period
were the presence of curved spine and organisms that appeared smaller or weak
in comparison to the other fish in the replicate. However, these observations were
infrequent and did not occur in a concentration related pattern. Consequently they
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were not considered to be treatment-related.
Growth was evaluated at the end of the test by measuring the total length, wet
weight and dry weight of each surviving fish. Fish in the 20 mg/L group exhibited a
slight yet statistically significant reduction in total length and dry weight
measurements in comparison to the control.
The NOEC was 10 mg/L based on growth
Conclusion NOEC = 10 mg/L based on growth.
Fish chronic toxicity (Marine species)
No study provided.
Invertebrates chronic toxicity (Freshwater species)
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Daphnia magna
Type of exposure Semi-static, 21 days. Renewal on days 2, 5, 7, 9, 12, 14, 16, 19.
Endpoint NOEC for survival rate, reproduction rate and body length of the adults
Value 3.1 mg/L based on reproduction rate
Reference
A Peither (2002) Influence of IKI-220 technical on survival and reproduction of
Daphnia magna in a semi-static test over three weeks. RCC Ltd, Environmental
Chemistry & Pharmanalytics Division. CH-4452 Itingen Switzerland. Report no
834052
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 211 (1998)
No/Group 10
Dose Levels 3.1, 6.3, 12.5, 25, 50 and 100 mg/L
Analytical measurements
Yes, from samples collected at the beginning of the test, on day 12 and 16 of
fresh medium and on days 14 and 19 for old medium incubated in the same
conditions but without daphnia and food particles, by HPLC. Only the samples
from the 3.1 and 6.3 concentrations (NOEC and LOEC) were analysed.
Study Summary
The influence of the test substance on the reproduction and survival of Daphnia
magna was determined in a 21-day semi-static test. The nominal concentrations
were 3.1, 6.3, 12.5, 25, 50 and 100 mg/L. The measured concentrations in the test
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medium were in the range of 93 to 101% of the nominal, so the results are
expressed as nominal concentrations.
The pH values ranged from 7.6 to 8.2, and the temperature was in the range 20-
21oC. Dissolved oxygen concentrations remained > 8.0 mg/L.
The survival rate was above 90% in the control and in concentrations up to 25
mg/L. At 50 mg/L, 60% of mortality was observed and 100% in the 100 mg/L
concentration.
The first offspring was observed on day 9 in the control and in concentrations up
to 25 mg/L. In the 50 mg/L group the observation was delayed on day 12.
The mean reproduction rate of the control was 78.7 ± 9.7 alive offspring per adult.
No effect was observed at 3.1 mg/L but there was a statistically significant
difference at 6.3 mg/L and above.
The mean body length in the control was 4.0 ± 0.06 mm, no effect was observed
at 6.3 mg/L but there was a statistically significant difference at 12.5 mg/L and
above.
The NOEC was 3.1 mg/L based on reproduction rate.
Conclusion NOEC = 3.1 mg/L based on reproduction rate.
Invertebrates chronic toxicity (Marine species)
No study provided.
General conclusion about aquatic toxicity classification:
Flonicamid exhibits a very low level of acute and chronic toxicity on aquatic organisms. No classification is
triggered except a 9.1D for biocidal activity which doesn’t apply because flonicamid is also classified as
9.3C.
All relevant metabolites have also been tested on aquatic organisms. They show a very low toxicity as well.
Consequently no specific quantitative risk assessment will be performed for these metabolites.
Sediment toxicity (freshwater and/or marine)
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Chironomus riparius
Type of exposure Water-sediment (spiked water), 27 days, static.
Endpoint NOEC for development time/rate, emergence ratio and toxicity effects
Value 25 mg/L
Reference U Memmert (2002) Effects of IKI-220 technical on the development of sediment-
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dwelling larvae of Chironomus riparius in a water-sediment system. RCC Ltd,
Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 841621
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 219 (draft, 2001)
No/Group 20 larvae/group
Dose Levels 6.3, 12.5, 25, 50 and 100 mg/L (results are expressed as nominal concentrations
in the water phase)
Analytical measurements
Yes. Water, pore water and sediment samples were taken on day 0 (1 hour after
substance application), on day 7 and at termination (day 27) from control, and 25
and 100 mg/L systems.
Study Summary
The influence of the test substance on the development of sediment-dwelling
larvae of the midge Chironomus riparius was determined in a 27-day water-
sediment test. The nominal concentrations were 3.1, 6.3, 12.5, 25, 50 and 100
mg/L.
The mean measured concentrations of the substance one hour after application
was 93% of the nominal concentration in both analysed test concentrations. The
concentrations decreased only slightly in the water of the water-sediment systems
during the test period. Seven days after application, the concentrations in the
water column were 86% of the nominal and at the study termination they
corresponded to 77-79%.
In the pore water and sediment, the concentration of the active substance
continuously increased during the study period. At 25 mg/L, the concentration in
pore water was in maximum 14.2 mg/L and 6.6 mg/kg in the sediment at study
termination. At 100 mg/L, a maximum of 57.5 mg/L were found in the pore water
and 26.2 mg/kg in the sediment (based on dry sediment).
A statistically significant effect on the mean emergence ratio was observed at 50
and 100 mg/L. The mean development rates of the female midges were
statistically significantly reduced at 50 mg/L and higher, the mean development
rates of the males first at 100 mg/L.
The NOEC is 25 mg/L based on the emergence ratios, development rates and
toxicity symptoms.
Conclusion The NOEC is 25 mg/L based on the emergence ratios, development rates
and toxicity symptoms.
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General conclusion about sediment toxicity:
Flonicamid show a rather low toxicity on the reproduction of sediment dwelling arthropods.
Soil toxicity
Soil macro-invertebrates acute toxicity
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Eisenia fetida
Type of exposure Static, for 14 days
Endpoint LC50
Value > 1000 mg/kg dry soil
Reference
R Baetscher (2001) Acute toxicity of IKI-220 technical to the earthworm Eisenia
fetida in a 14-day test. RCC Ltd, Environmental Chemistry & Pharmanalytics
Division. CH-4452 Itingen Switzerland. Report no 807333
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 207 (1984)
No/Group 4 replicates of 10
Dose Levels 56, 100, 178, 316, 562, 1000 mg/ kg dry soil
Analytical measurements Not required
Study Summary
The acute toxicity of the substance to the earthworm Eisenia fetida was
determined in a 14-day test with artificial substrate containing 10% of peat.
After 7 and 14 days of exposure, no mortality was observed at any of the test
concentrations up to and including the highest test concentration of 1000 mg ai/kg
dry soil. Moreover, no abnormal behaviour or symptoms of toxicity were recorded.
The body weight was not affected up to the highest concentration.
Conclusion LC50> 1000 mg/kg dry soil
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA
Species Eisenia fetida
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Type of exposure Static, for 14 days
Endpoint LC50
Value > 100 mg/kg dry soil
Reference
R Baetscher (2002) Acute toxicity of TFNA to the earthworm Eisenia fetida in a
14-day test. RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-
4452 Itingen Switzerland. Report no 834221
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 207 (1984)
No/Group 4 replicates of 10
Dose Levels 100 mg/ kg dry soil
Analytical measurements Not required
Study Summary
The acute toxicity of the substance to the earthworm Eisenia fetida was
determined in a 14-day test with artificial substrate containing 10% of peat.
After 7 and 14 days of exposure, no mortality was observed at the test
concentration of 100 mg ai/kg dry soil. Moreover, no abnormal behaviour or
symptoms of toxicity were recorded. The body weight was not affected.
Conclusion LC50> 100 mg/kg dry soil
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA-AM
Species Eisenia fetida
Type of exposure Static, for 14 days
Endpoint LC50
Value > 100 mg/kg dry soil
Reference
R Baetscher (2002) Acute toxicity of TFNA-AM to the earthworm Eisenia fetida in
a 14-day test. RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-
4452 Itingen Switzerland. Report no 835018
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 207 (1984)
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No/Group 4 replicates of 10
Dose Levels 100 mg/ kg dry soil
Analytical measurements Not required
Study Summary
The acute toxicity of the substance to the earthworm Eisenia fetida was
determined in a 14-day test with artificial substrate containing 10% of peat.
After 7 and 14 days of exposure, no mortality was observed at the test
concentration of 100 mg ai/kg dry soil. Moreover, no abnormal behaviour or
symptoms of toxicity were recorded. The body weight was not affected.
Conclusion LC50> 100 mg/kg dry soil
Type of study Limit test
Flag Key study
Test Substance Metabolite TFNA-OH
Species Eisenia fetida
Type of exposure Static, for 14 days
Endpoint LC50
Value > 100 mg/kg dry soil
Reference
R Baetscher (2002) Acute toxicity of TFNA-OH to the earthworm Eisenia fetida in
a 14-day test. RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-
4452 Itingen Switzerland. Report no 835042
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 207 (1984)
No/Group 4 replicates of 10
Dose Levels 100 mg/ kg dry soil
Analytical measurements Not required
Study Summary
The acute toxicity of the substance to the earthworm Eisenia fetida was
determined in a 14-day test with artificial substrate containing 10% of peat.
After 7 and 14 days of exposure, no mortality was observed at the test
concentration of 100 mg ai/kg dry soil. Moreover, no abnormal behaviour or
symptoms of toxicity were recorded. The body weight was not affected.
Conclusion LC50> 100 mg/kg dry soil
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Type of study Limit test
Flag Key study
Test Substance Metabolite TFNG-AM
Species Eisenia fetida
Type of exposure Static, for 14 days
Endpoint LC50
Value > 100 mg/kg dry soil
Reference
R Baetscher (2002) Acute toxicity of TFNG-AM to the earthworm Eisenia fetida in
a 14-day test. RCC Ltd, Environmental Chemistry & Pharmanalytics Division. CH-
4452 Itingen Switzerland. Report no 835031
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 207 (1984)
No/Group 4 replicates of 10
Dose Levels 100 mg/ kg dry soil
Analytical measurements Not required
Study Summary
The acute toxicity of the substance to the earthworm Eisenia fetida was
determined in a 14-day test with artificial substrate containing 10% of peat.
After 7 and 14 days of exposure, no mortality was observed at the test
concentration of 100 mg ai/kg dry soil. Moreover, no abnormal behaviour or
symptoms of toxicity were recorded. The body weight was not affected.
Conclusion LC50> 100 mg/kg dry soil
Soil macro-invertebrates chronic toxicity
No study provided.
Since flonicamid is rapidly degraded in soil, continued exposure of earthworms to the active ingredient is not
likely to occur, even with 2 applications at 7-day interval. Moreover, the relevant metabolites have been
tested for acute toxicity and no effect was observed up to 100 mg/kg dry soil.
Non-target plants toxicity
No study provided with the active ingredient but tests were performed with Mainman.
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Nitrogen transformation test
Flag Supporting study (endpoint already covered by the formulation test which is of
better quality)
Test Substance Flonicamid technical
Species Soil micro-flora
Type of exposure Static, 28 days
Endpoint Nitrate content
Value No effect
Reference
M das Gracas S. Raposo (2003) Side effects of Flonicamid tecnico on soil
microorganisms: nitrogen transformation test. Bioagri Laboratorios Ltda
Piracicaba/SP Brazil. Report no RF-1213.218.027.02
Klimisch Score 2 report is not very detailed, results are poorly described
Amendments/Deviations None that had an impact on the results
GLP yes
Test Guideline/s OECD 2016
No/Group 3 replicates
Dose Levels 0.21 and 1.03 µg/g soil corresponding to 100 and 500 g ai /ha
Analytical measurements Not required
Study Summary
The objective of this study was to assess the effects of the substance on the
nitrogen transformation in soil, determined by the nitrate content. The substance
was applied on soils at concentrations of 0.21 and 1.03 µg/g soil corresponding to
100 and 500 g ai /ha. 2 different types of soil were used, they are classified as
Rhodic Hapludox and Typic Hapludox according to the USDA soil taxonomy
(1990). Samples were taken at 0, 7, 14 and 28 days to determine the nitrate
content.
There was no significant effect of the substance at any time, on any soil and at
any concentration.
Conclusion No effect on the N transformation in soil at concentrations up to 500 g ai/ha
Carbon transformation test
Flag Supporting study (endpoint already covered by the formulation test which is of
better quality)
Test Substance Flonicamid technical
Species Soil micro-flora
Type of exposure Static, 28 days
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Endpoint CO2 release
Value No effect
Reference
M das Gracas S. Raposo (2003) Side effects of Flonicamid tecnico on soil
microorganisms: carbon transformation test. Bioagri Laboratorios Ltda
Piracicaba/SP Brazil. Report no RF-1213.201.153.02
Klimisch Score 2 report is not very detailed, results are poorly described
Amendments/Deviations None that had an impact on the results
GLP Yes
Test Guideline/s OECD 217
No/Group 2 soils
Dose Levels 0.21 and 1.03 µg/g soil corresponding to 100 and 500 g ai /ha
Analytical measurements Not required
Study Summary
The objective of this study was to assess the effects of the substance on the
carbon transformation in soil, determined by the CO2 production rate by the
microbial respiration. The substance was applied on soils at concentrations of
0.21 and 1.03 µg/g soil corresponding to 100 and 500 g ai /ha. 2 different types of
soil were used, they are classified as Rhodic Hapludox and Typic Hapludox
according to the USDA soil taxonomy (1990). Samples were taken at 0, 7, 14 and
28 days to determine the 14
CO2 production after addition of 14
C-glucose.
There was no significant effect of the substance at any time, on any soil and at
any concentration.
Conclusion No effect on the C transformation in soil at concentrations up to 500 g ai/ha
General conclusion about soil toxicity classification:
Flonicamid does not trigger any 9.2 classification. The relevant metabolites have also been tested on
earthworms and do not show any toxicity.
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Terrestrial vertebrate toxicity
For effects on terrestrial vertebrates other than birds, refer to the mammalian toxicity section.
Oral acute toxicity
Type of study Limit test extended to a full test
Flag Supporting study
Test Substance IKI-220 technical
Species Colinus virginianus (Bobwhite quail)
Type of exposure Single dose, observation period of 14 days
Endpoint LD50
Value > 2000 mg/kg bw
Reference
R Burri (2002) IKI-220 technical: Acute oral toxicity test in the bobwhite quail. RCC
Ltd, Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 809043
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OPPTS 850.2100 (1996)
No/sex/Group 5 males and females for 2000 mg/kg bw, only 5 females for the lower doses.
Dose Levels 260, 432, 720, 1200 and 2000 mg/kg bw
Analytical measurements Not required
Study Summary
Initially, a limit test was performed with one dose group of 5 males and 5 females
treated with 2000 mg/kg bw. Because 40 % of mortality occurred in the females at
this concentration, additional groups of females were treated with 260, 432, 720 and
1200 mg/kg bw. No mortality was observed in these additional groups.
Clinical symptoms were exclusively observed in females exposed to 2000 mg/kg bw.
There was a significant influence of the treatment on the body weights of males and
females. It was caused by reduced food consumption. This effect was reversible for
the males and also for the females treated by doses up to 1200 mg/kg bw but the
body weight of the females treated with 2000 mg/kg bw was still reduced at the end of
the observation period (14 days).
A lower food consumption was observed during the first week after administration for
the animals treated by 2000 mg/kg bw, and during the first 3 days for the other doses.
Average food consumption was back to normal level during the 2nd
observation week.
Conclusion LD50 > 2000 mg/kg bw
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Type of study Limit test extended to a full test
Flag Key study
Test Substance IKI-220 technical
Species Mallard duck (Anas platyrhynchos)
Type of exposure Single dose, observation period of 14 days
Endpoint LD50
Value 2621 mg/kg bw (males) and 1591 mg/kg bw (females)
Reference
R. Burri (2002) IKI-220 technical: acute oral test in the Mallard duck. RCC Ltd
Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 809054
Klimisch Score 1
Amendments/Deviations None
GLP yes
Test Guideline/s OPPTS 850.2100 (1996)
No/sex/Group 5
Dose Levels 260, 432, 720, 1200 and 2000 mg/kg bw
Analytical measurements Not required
Study Summary
Initially, a limit test was performed with one dose group of 5 males treated with
2000 mg/kg bw. Because mortality occurred at this concentration, additional
groups of males and females were treated with 260, 432, 720 and 1200 mg/kg bw.
40 and 60 % of mortality were observed at 2000 mg/kg in males and females,
respectively; 0 % at 1200 mg/kg, 20 in males and females at 720, 20% in males at
432 and 20% in females at 260 mg/kg bw.
Clinical symptoms were exclusively ataxia, dyspnea and ventral recumbency.
Most of the animals that died developed symptoms rapidly and died within 1 to 3
hours after administration.
There was no influence of the treatment on the body weights of the surviving
males and females and no general test substance related effect on the food
consumption.
The NOED is 260 mg/kg bw (males) and 432 mg/kg bw (females)
Conclusion LD50 = 2621 mg/kg bw (males) and 1591 mg/kg bw (females)
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Dietary acute toxicity
Type of study Limit test
Flag Supporting study
Test Substance IKI-220 technical
Species Colinus virginianus (Bobwhite quail)
Type of exposure 5 days in diet followed by 3 days of observation
Endpoint LC50
Value > 5000 mg/kg diet corresponding to > 411.1 mg/kg bw/day
Reference
R Burri (2001) IKI-220 technical: avian dietary toxicity test in the bobwhite quail.
RCC Ltd Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 809065
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 205 (1984) and OPPTS 850.2200 (1996)
No/Group 10 animals of undetermined sex because of their size and age (14 days)
Dose Levels 5000 mg/kg diet
Analytical measurements Concentration of ai in diet was confirmed, stability and homogeneity were checked
Study Summary
A limit test was performed. IKI-220Technical was administered to one group of 10
Bobwhite quails by dietary ingestion at a nominal dose level of 5000 mg test
item/kg diet for 5 treatment days followed by 3 observation days where the
animals obtained regular diet.
No mortality was observed in the main study at a dose level of 5000 mg test
item/kg diet therefore the LC50 of IKI-220 Technical was > 5000 mg/kg diet.
No symptoms and no macroscopic findings were observed for the treated animals.
Therefore, the NOEC was > 5000 mg/kg diet.
During treatment, a significantly lower average body weight was observed for the
treated animals as compared to the control groups, reflecting a reduced food
consumption of the treated animals.
During treatment, individual and relative food consumption was lower for the
treated animals as compared to the control animals. This indicated a repellency
potential of the test item.
The LC50 based on test item intake was > 411.0 mg test item/kg body weight/day.
Conclusion LC50 > 5000 mg/kg diet corresponding to > 411.1 mg/kg bw/day
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Type of study Limit test
Flag Key study
Test Substance IKI-220 technical
Species Mallard duck (Anas platyrhynchos)
Type of exposure 5 days in diet followed by 3 days of observation
Endpoint LC50
Value > 5000 mg/kg diet corresponding to > 301.8 mg/kg bw/day
Reference
R Burri (2001) IKI-220 technical: Avian dietary toxicity test in the Mallard duck.
RCC Ltd Environmental Chemistry & Pharmanalytics Division. CH-4452 Itingen
Switzerland. Report no 809076
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 205 (1984) and OPPTS 850.2200 (1996)
No/Group 10 animals of undetermined sex because of their size and age (14 days)
Dose Levels 5000 mg/kg diet
Analytical measurements Concentration of ai in diet was confirmed, stability and homogeneity were checked
Study Summary
A limit test was performed. IKI-220 Technical was administered to one group of 10
ducks by dietary ingestion at a nominal dose level of 5000 mg test item/kg diet for
5 treatment days followed by 3 observation days where the animals obtained
regular diet.
No mortality was observed in the main study at a dose level of 5000 mg test
item/kg diet therefore the LC50 of IKI-220 Technical was > 5000 mg/kg diet.
No symptoms and no macroscopic findings were observed for the treated animals.
Therefore, the NOEC was > 5000 mg/kg diet.
During treatment, a significantly lower average body weight was observed for the
treated animals as compared to the control groups, reflecting a reduced food
consumption of the treated animals.
During treatment, individual and relative food consumption was lower for the
treated animals as compared to the control animals. This indicated a repellency
potential of the test item.
The LC50 based on test item intake was > 301.8 mg test item/kg body weight/day.
Conclusion LC50 > 5000 mg/kg diet corresponding to > 301.8 mg/kg bw/day
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Chronic toxicity (reproduction)
Type of study Full test
Flag Supporting study
Test Substance IKI-220 technical
Species Colinus virginianus (Bobwhite quail)
Type of exposure Dietary exposure for 21 weeks
Endpoint NOEC for reproduction and toxicity on adults
Value 1000 mg/kg diet corresponding to 90 mg ai /kg bw /day
Reference
LR. Mitchell et al (2002) A reproduction study with the Northern bobwhite (Colinus
virginianus). Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland
21601 USA. Report no 272-125
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study
GLP Yes
Test Guideline/s OECD 206 (1984)
No/sex/Group 16
Dose Levels 0, 160, 400 and 1000 ppm a.i.
Analytical measurements Concentration of ai in diet was confirmed, stability and homogeneity were checked
Study Summary
The objective of this study was to evaluate the effects upon the adult northern
bobwhite (Colinus virginianus) of dietary exposure to IKI-220 over a period of
approximately 21 weeks. Effects on adult health, body weight, and feed consumption
were evaluated. In addition, the effects of adult exposure to IKI-220 on the number of
eggs laid, fertility, embryo viability, hatchability, offspring survival, and egg shell
thickness were evaluated.
Each treatment and control group contained 16 pairs of birds with one male and one
female per pen. Three treatment groups were fed diets containing either 160, 400, or
1000 ppm a.i. of IKI-220 for approximately 21 weeks. The control group was fed diet
comparable to the treatment groups, but without the addition of the test substance. All
adult birds were observed daily throughout the test for signs of toxicity or abnormal
behaviour. Adult body weights were measured at test initiation, on Weeks 2, 4, 6, 8,
and at adult termination and feed consumption was measured weekly throughout the
test. At the beginning of Week 8, the photoperiod was increased to induce egg
production. Following the start of egg production, eggs were set weekly for
incubation. Weekly, eggs were selected by indiscriminate draw for egg shell thickness
measurement and all remaining eggs were candled prior to incubation to detect egg
shell cracks or abnormal eggs. Eggs were also candled twice during incubation to
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detect infertile eggs or embryo mortality. On Day 21 of incubation, the eggs were
placed in a hatcher and allowed to hatch. Once hatching was completed, hatchlings
were removed from the hatcher and the group body weight of the hatchlings by pen
was determined. At 14 days of age, the average body weight by parental pen of all
surviving offspring was determined.
The estimated test substance intakes for northern bobwhite during Weeks 1 through
21 of the test were 13, 33 and 90 mg IKI-220/kg body weight/day for the 160, 400 and
1000 ppm a.i. treatment groups, respectively. There were no treatment-related
mortalities, overt signs of toxicity or treatment-related effects upon adult body weight
or feed consumption at any of the concentrations tested. Additionally, there were no
treatment-related effects upon any of the reproductive parameters measured at the
160, 400 or 1000 ppm a.i. test concentrations. The no-observed-effect concentration
for northern bobwhite exposed to IKI-220 in the diet during the study was 1000 ppm
a.i. (90 mg a.i./kg body weight/day), the highest concentration tested.
Conclusion NOEC = 1000 mg/kg diet corresponding to 90 mg ai /kg bw/day
Type of study Full test
Flag Key study
Test Substance IKI-220 technical
Species Mallard duck (Anas platyrhynchos)
Type of exposure Dietary exposure for 20 weeks
Endpoint NOEC for reproduction and toxicity on adults
Value 400 mg/kg diet corresponding to 59 mg/kg bw/ day
Reference LR. Mitchell et al (2002) A reproduction study with the Mallard. Wildlife International,
Ltd. 8598 Commerce Drive Easton, Maryland 21601 USA. Report no 272-126
Klimisch Score 1
Amendments/Deviations None that impacted the results of the study
GLP Yes
Test Guideline/s OECD 206 (1984)
No/sex/Group 16
Dose Levels 0, 160, 400 and 1000 ppm a.i.
Analytical measurements Concentration of ai in diet was confirmed, stability and homogeneity were checked
Study Summary
The objective of this study was to evaluate the effects upon the adult mallard (Anas
platyrhynchos) of dietary exposure to IKI-220 over a period of approximately 20
weeks. Effects on adult health, body weight, and feed consumption were evaluated.
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In addition, the effects of adult exposure to IKI-220 on the number of eggs laid,
fertility, embryo viability, hatchability, offspring survival, and egg shell thickness were
evaluated.
Each treatment and control group contained 16 pairs of birds with one male and one
female per pen. Three treatment groups were fed diets containing either 160, 400, or
1000 ppm a.i. of IKI-220 for approximately 20 weeks. The control group was fed diet
comparable to the treatment groups, but without the addition of the test substance. All
adult birds were observed daily throughout the test for signs of toxicity or abnormal
behaviour. Adult body weights were measured at test initiation, on Weeks 2, 4, 6, 8,
and at adult termination and feed consumption was measured weekly throughout the
test. At the beginning of Week 9, the photoperiod was increased to induce egg
production. Following the start of egg production, eggs were set weekly for
incubation. Weekly, eggs were selected by indiscriminate draw for egg shell thickness
measurement and all remaining eggs were candled prior to incubation to detect egg
shell cracks or abnormal eggs. Eggs were also candled twice during incubation to
detect infertile eggs or embryo mortality. On Day 24 of incubation, the eggs were
placed in a hatcher and allowed to hatch. Once hatching was completed, hatchlings
were removed from the hatcher and the group body weight of the hatchlings by pen
was determined. At 14 days of age, the average body weight by parental pen of all
surviving offspring was determined.
The estimated test substance intakes for mallards during Weeks 1 through 20 of the
test were 25, 59 and 138 mg IKI-220/kg body weight/day for the 160, 400 and 1000
ppm a.i. treatment groups, respectively. There were no treatment-related mortalities,
overt signs of toxicity or treatment-related effects upon adult body weight or feed
consumption at any of the concentrations tested. Additionally, there were no
treatment-related effects upon any of the reproductive parameters measured at the
160 or 400 ppm a.i. test concentrations. However, there were treatment-related
effects upon both viability and hatchability at the 1000 ppm a.i. treatment group that
resulted in statistically significant (p < 0.05) reductions in hatchlings and 14-day old
survivors as percentages of the number of eggs set. Based on the effects noted in the
1000 ppm a.i. treatment group, the no-observed-effect concentration for mallards
exposed to IKI-220 in the diet during the study was 400 ppm a.i. (59 mg a.i./kg body
weight/day).
Conclusion NOEC = 400 mg/kg diet corresponding to 59 mg/kg bw/ day
General conclusion about terrestrial vertebrate classification:
Flonicamid is classified as 9.3C on the basis of its acute effects on birds.
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Ecotoxicity to terrestrial invertebrates
Bees - Laboratory tests (acute oral and contact)
Type of study Limit test (contact), full test (oral)
Flag Key study
Test Substance IKI-220 technical
Species Apis mellifera
Type of exposure 48 h (contact) and 96 h (oral)
Endpoint LD50
Value > 60.5 µg ai/bee (oral)
> 100 µg ai/bee (contact)
Reference
S Schmitzer (2001) Laboratory testing for toxicity (acute contact and oral) of IKI-
220 TGAI on Honey bees (Apis mellifera L.). IBACON GmbH Arheilger Weg 17
64380 Rossdorf Germany. Report no 10731036
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s EPPO 170 (1992)
No/Group 3 replicates of 10 (oral) and 5 replicates of 10 (contact)
Dose Levels Contact test: 100 µg ai/bee
Oral: 6.4, 11.6, 22.2, 47.7, 60.5 µg ai/bee (actual intake)
Analytical measurements Not required
Study Summary
In the contact toxicity test, 8.0% mortality was observed at 100 µg ai/bee after 48
hours. Behavioural impairments like nervousness (during the first 4 hours),
discoordinated movement and apathy were observed during the 48 hours of the
experiment.
In the oral test, honey instead of syrup was used as a carrier. Since pre-tests have
demonstrated that the substance had a strong repellent effect, honey was used
because it is more attractive to bees. The maximum nominal test level of 100 µg
ai/bee corresponded to an actual intake of 60.5 µg ai/bee. After a starving period
of 90 min, the desired nominal dosage could not be applied since honeybees
rejected to ingest the full volume of treated honey even when offered over 6
hours. The observation period was extended to 96 hours because delayed
mortality occurred in 3 or the 5 groups.
Oral dose of 60.5 µg ai /bee led to 36.7% mortality at test end; 20, 26.7, 26.7 and
6.7% of mortality were observed at 47.7, 22.2, 11.6 and 6.4 µg ai/bee,
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respectively.
Behavioural impairments like nervousness, discoordinated movement and apathy
was observed during the first 48 hours of observation. No further behavioural
abnormality was observed during the 72 and 96 hours check.
Conclusion Oral LD50 (48 + 96h) > 60.5 µg ai/bee
Contact LD50 (48h) > 100 µg ai/bee
Other non-target arthropods
No studies performed with the active ingredient itself, but tests are available with Mainman.
General conclusion about toxicity to terrestrial invertebrates classification:
Flonicamid does not trigger any 9.4 classification.
Appendix E: Staff’s risk identification
To assess the risks posed by the substance throughout its lifecycle, the most common potential sources of
risk to the environment and human health and safety were identified (Table 12). These are shown in Tables
13 and 14 with an assessment of risk that assumes an absence of controls and/or regulations.
Table 12 Potential sources of risks associated with hazardous substances
Lifecycle Activity Associated Source of Risk
Manufacture / Import
An incident during the manufacture or importation of the substance resulting in
a spill and the subsequent exposure of people or the environment to the
substance
Packing An incident during the packing of the substance resulting in a spill and the
subsequent exposure of people or the environment to the substance
Transport or storage
An incident during the transport, storage or handling or the substance, resulting
in a spill and the subsequent exposure of people or the environment to the
substance. For example, inadequate storage and security on farms resulting in
access by unauthorised persons (including children)
Use Application of the substance resulting in exposure of users or bystanders or the
environment
Disposal
Inadequate, incorrect or unlawful disposal of substance, excess tank mix or
empty packaging resulting in exposure of people or the environment to the
substance
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Appendix F: Qualitative risk assessment
Staff also carried out a qualitative assessment to assess risks to human health and the environment from the lifecycle of Mainman. A summary of the
results of the qualitative assessment are shown in Section 6 of this report and are detailed below. The process by which the risk assessment of
substances is undertaken is specified in the on risk assessment is provided on the EPA website. Methodology Guidance
Table 13 Qualitative assessment of human health risks
Lifecycle Description Likelihood Magnitude Level of risk
(matrix)
Comment Level of
risk
(EPA)
Manufacture
and
packaging
Eye irritation Highly
improbable
Moderate Negligible Manufacturing and packaging facilities in New Zealand will be
required to meet the HSNO requirements for equipment, emergency
management and WorkSafe New Zealand’s health and safety
requirements.
Negligible
Importation,
transport,
storage
Eye Irritation Very Unlikely Minimal Negligible Workers and bystanders will only be exposed to the substance
during this part of the lifecycle in isolated incidents if a spill occurs,
therefore only risks from acute exposure are considered here.
Compliance with HSNO controls (e.g. labels, SDS) and adherence
to the Land Transport Rule 45001, Civil Aviation Act 1990 and
Maritime Transport Act 1994 (as applicable) will make the likelihood
of exposure very unlikely Any exposure is only expected to have a
localised, reversible effect and is therefore considered minimal.
Negligible
Disposal Use Eye irritation Unlikely Minimal Negligible Eye irritation could occur during mixing, loading or application of
Mainman, or handling it for disposal. However, assuming the HSNO
controls and good practice for handling agrichemicals is followed
(e.g. use of eye protection when mixing or using chemicals) then it is
unlikely that someone will be affected by the substance. Any effects
are expected to be reversible and impact individuals handling the
substance therefore the impact of the effect is considered minimal.
All cases of disposal are required to be in accordance with the
requirements of the Hazardous Substances (Disposal) Regulations
2001. Compliance with these regulations will both reduce the
likelihood of adverse events occurring.
Negligible
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Table 14 Qualitative assessment of risks to the environment
Lifecycle Description Likelihood Magnitude Level of risk
(matrix)
Comment Level of
risk
Manufacture,
importation,
transport
and storage
Death or
adverse
effects to
terrestrial
vertebrates
Highly
improbable
Minimal Negligible The terrestrial environment will only be exposed to Mainman
during this part of the lifecycle in isolated incidents if a spill occurs.
Given adherence to the Controls, the Land Transport Rule 45001,
Resource Management Act 1991, Civil Aviation Act 1990 and the
Maritime Transport Act 1994 (as applicable), the staff consider a
spill to be highly improbable, and a spill is only likely to lead to
localised effects.
Negligible
Use
(application)
Death or
adverse
effects to
terrestrial
vertebrates
Very Unlikely Minimal Negligible Terrestrial invertebrates, such as bees, may be affected by
Mainman if it is applied to plants where bees are foraging.
Prohibiting application of Mainman where bees are foraging, will
reduce the likelihood of Mainman adversely affecting beneficial
invertebrates.
Negligible
Disposal Death or
adverse
effects to
terrestrial
vertebrates
Highly
improbable
Minimal Negligible Users will in most cases use all of the substance through normal
use as an insecticide. All cases of disposal are required to be in
accordance with the requirements of the Hazardous Substances
(Disposal) Regulations 2001. Compliance with these regulations
will both reduce the likelihood of adverse events occurring, and
diminish the quantity of substance released to the environment in
such an event. Accordingly, the overall level of risk to aquatic
organisms and terrestrial invertebrates will be assessed as
negligible.
Negligible
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Appendix G: Human health risk assessment
Quantitative worker (operator) risk assessment
Critical endpoint definition
The EFSA peer review previously established an AOEL for flonicamid of 0.025 mg/kg bw/day, based on what
were considered adverse effects (malformations) in the rabbit developmental toxicity study. However, as
noted above, these effects are not considered to be a direct adverse effect of flonicamid (the incidences of
malformations were within the historical control range, were not consistent nor were they dose related). This
conclusion is consistent with a recent review by the European Chemicals Agency’s Risk Assessment
Committee. Therefore the staff have derived a new AOEL for flonicamid.
Table 15: Deriving an AOEL for flonicamid
Key systemic
effect
NOAEL
mg/kg
bw/day
Uncertainty
factors
Absorption
factor18
AOEL
mg/kg
bw/day
Justification
Micropathological
changes in the
kidney (females):
90-day study in
dogs
Mild anemia: 52-
week study in
dogs
8 100 1 0.08
Lowest NOAEL from the sub-
chronic toxicity studies in mice,
rats and dogs; also consistent
with the NOAEL for maternal
toxicity in the rabbit
developmental toxicity study (7.5
mg/kg bw/day)
18
100
n% Absorptio
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Other inputs for human worker (operator) and re-entry exposure modelling19
Table 16 Derivation of dermal absorption value in humans
Active
ingredient
Physical
form
Concentration
of each active
(g/L or g/kg)
Maximum
application rate
(for each active, for
each method of
application)
g (ai)/ha
Dermal absorption (%)
AOEL
mg/kg
bw/day Concentrate Spray
Flonicamid
Water
dispersible
granule
500 g/kg
80 (boom)
70 (high volume
directed, aerial)
7 13 0.08
Comments on inputs for human worker (operator) exposure modelling input parameters:
The dermal penetration of IKI-220 formulated as IKI-220 50% WG in human skin was 7% at the lowest field
use dilution of 0.4 g IKI-220/L. At the highest field use dilution of 0.07 g IKI-220/L, the dermal absorption in
human skin was 13%. Therefore it is proposed to use 7% for the concentrate and 13% for the spray. This
was also the approach taken by EFSA.
19 The Staff has undertaken an assessment of risks to operator health using the United Kingdom Pesticide Safety Directorate’s
interpretation of the German BBA Model to estimate operator exposure. This model estimates the exposure of workers to a
pesticide during mixing, loading and during spray application, in mg/kg person/day (http://www.pesticides.gov.uk/index.htm).
The derived values consider both dermal and inhalation exposure routes. The Staff typically uses the geometric mean model.
The BBA model provides for a range of different spray applications (tractor-mounted/trailed sprayers and hand-held sprayers)
and formulation types (liquid, wettable powder and wettable granule). Additionally, the BBA model also allows flexibility to vary
protective clothing (hands, head and body).
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Output of human worker (operator) mixing, loading and application exposure modelling
Table 17 Risk Quotients for Mainman: Potato – 80 g ai/ha, 2 applications at 7 days, ground based
Exposure Scenario Estimated operator
exposure (mg/kg bw/day) Risk Quotient
Boom
No PPE20
during mixing, loading and application 0.0095 0.12
Gloves only during mixing and loading 0.0063 0.08
Gloves only during application 0.0083 0.10
Full PPE during mixing, loading and application (excluding
respirator)
0.0005 0.01
Full PPE during mixing, loading and application (including
respirator)
0.0003 0.004
Table 18 Risk Quotients for Mainman: Confidential use 1 – 70 g ai/ha, 2 applications at 14 days,
ground based
Exposure Scenario Estimated operator
exposure (mg/kg bw/day) Risk Quotient
Boom
No PPE21
during mixing, loading and application 0.0083 0.10
Gloves only during mixing and loading 0.0055 0.07
Gloves only during application 0.0073 0.09
Full PPE during mixing, loading and application (excluding
respirator)
0.0004 0.01
Full PPE during mixing, loading and application (including
respirator)
0.0003 0.003
Confidential use 2 – 70 g ai/ha, 2 applications at 14 days, aerial
The EPA’s exposure model does not include an evaluation of operator exposure during aerial application.
However, it is anticipated that exposure during mixing and loading prior to aerial application will be similar to
20 Full” PPE includes: gloves, hood/visor, coveralls, and heavy boots during application. The model only provides for use of
gloves at mixing loading.
21 Full” PPE includes: gloves, hood/visor, coveralls, and heavy boots during application. The model only provides for use of
gloves at mixing loading.
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that during mixing and loading before ground based application. Operator exposure during aerial application
is expected to be lower than that during ground based application. Therefore overall operator exposure for
confidential use 2 is expected to be lower than operator exposure for confidential use 1.
Table 19 Risk Quotients for Mainman: Confidential use 3– 70 g ai/ha, 2 applications at 7 days, ground
based
Exposure Scenario Estimated operator
exposure (mg/kg bw/day) Risk Quotient
Boom
No PPE22
during mixing, loading and application 0.0083 0.10
Gloves only during mixing and loading 0.0055 0.07
Gloves only during application 0.0073 0.09
Full PPE during mixing, loading and application (excluding
respirator)
0.0004 0.01
Full PPE during mixing, loading and application (including
respirator)
0.0003 0.003
Confidential use 4 – 70 g ai/ha, 2 applications at 7 days, aerial
The EPA’s exposure model does not include an evaluation of operator exposure during aerial application.
However, it is anticipated that exposure during mixing and loading prior to aerial application will be similar to
that during mixing and loading before ground based application. Operator exposure during aerial application
is expected to be lower than that during ground based application. Therefore overall operator exposure for
confidential use 4 is expected to be lower than operator exposure for confidential use 3.
Table 20 Risk Quotients for Mainman: Confidential use 5 – 70 g ai/ha, 3 applications at 21 days
Exposure Scenario Estimated operator
exposure (mg/kg bw/day) Risk Quotient
Airblast
No PPE during mixing, loading and application 0.0133 0.17
Gloves only during mixing and loading 0.0122 0.15
22 Full” PPE includes: gloves, hood/visor, coveralls, and heavy boots during application. The model only provides for use of
gloves at mixing loading.
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Gloves only during application 0.0126 0.16
Full PPE during mixing, loading and application (excluding
respirator)
0.0008 0.01
Full PPE during mixing, loading and application (including
respirator)
0.0006 0.01
Outcomes of the worker (operator) exposure assessment
The results of the quantitative risk assessment indicate that PPE is not required to protect against adverse
effects resulting from systemic exposure to the active ingredient23
Output of the re-entry exposure assessment:
Table 21 Risk Quotients for Mainman: Re-entry exposure modelling24
Re-entry Activity
Internal (absorbed) dose available for
systemic distribution
(mg/kg bw/8 hours)
AOEL
(mg/kg bw/day)
Risk Quotient at
24 hours without
gloves
Potatoes
Vegetables – reach/pick
0.013 0.08 0.17
Confidential use 1 0.004 0.08 0.05
Confidential use 2 0.004 0.08 0.05
Confidential use 3 0.005 0.08 0.06
Confidential use 4 0.005 0.08 0.06
Confidential use 5 0.02 0.08 0.21
23 The staff considers that, while the ‘no PPE’ exposure model leads to an acceptable level of risk, it is appropriate to retain
requirements for PPE since the use of PPE when handling agrichemicals is good practice. The Staff notes that the HSNO PPE
requirements are not prescriptive allowing users to select an appropriate level of PPE.
24 The staff assessed the re-entry worker exposures using the generic exposure model for “Maintenance and harvesting
activities: Dermal exposure” provided by the UK Health & Safety Executive chemical Regulation Directorate, on the following
web site:
http://www.pesticides.gov.uk/applicant_guide.asp?id=1246&link=%2Fuploadedfiles%2FWeb%5FAssets%2FPSD%2FRe%2Den
try%2520worker%2520guidance%5Ffinal%2520version%2Epdf.
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Outcomes of the re-entry exposure assessment
Estimated re-entry worker exposures 24 hours following application are below the AOEL, even without the
use of gloves. As the estimated exposures are very low it is anticipated that risks for re-entry workers will
also be low immediately following application. However, to promote good practice staff consider that a label
statement along the following lines should be considered:
‘Do not allow entry into treated areas until the spray has dried, unless wearing cotton overalls buttoned to
the neck and wrist (or equivalent clothing) and chemical resistant gloves. Clothing must be laundered
after each day’s use.’
Quantitative bystander risk assessment25
Critical endpoints definition
The AOEL derived for operator and re-entry worker assessment above is also used for the bystander
assessment calculations.
Output of human bystander mixing, loading and application exposure modelling26
Table 22 Risk Quotients for Mainman: Use on Potatoes
Exposure Scenario
Estimated exposure of 15 kg toddler exposed
through contact to surfaces 8 m from an
application area (µg/kg bw/day)
Risk Quotient
Boom
High boom, fine droplets 0.78 0.0097
25 The Staff considers that the main potential source of exposure to the general public for substances of this type (other than via
food residues which will be considered as part of the registration of this substance under the Agricultural Compounds and
Veterinary Medicines (ACVM) Act 1997) is via spray drift. In terms of bystander exposure, toddlers are regarded as the most
sensitive sub-population and are regarded as having the greatest exposures. For these reasons, the risk of bystander exposure
is assessed in this sub-population. EPA has agreed that the AOEL used for operator and re-entry worker exposure assessment
should be used for the bystander assessment, as the use of an oral chronic reference dose (CRfD) is usually likely to be over
precautionary.
26 Exposure is estimated using the equations from the UK Health & Safety Chemical Regulation Directorate which account for
dermal exposure, hand-to-mouth exposure and object-to-mouth exposure
(http://www.pesticides.gov.uk/uploadedfiles/Web_Assets/PSD/Bystander%20exposure%20guidance_final%20version.pdf
Accessed 27/01/2010a). In addition, incidental ingestion of soil is taken into account using a modified exposure equation from
the United States Environmental Protection Agency (USEPA, 2007, Standard Operating Procedures (SOPs) for Residential
Exposure Assessments, Contract No. 68-W6-0030, Work Assignment No. 3385.102). Spray drift is estimated using models
specific to the type of application equipment. For pesticides applied by ground boom or air blast sprayer, the AgDrift model is
used. Spray drift deposition from aerial application is estimated using the AGDISP model along with appropriate New Zealand
input parameters
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High boom, coarse droplets 0.12 0.0015
Low boom, fine droplets 0.26 0.0033
Low boom, coarse droplets 0.06 0.0008
Table 23 Risk Quotients for Mainman: Confidential uses 1 and 2 – 70 g ai/ha, 2 applications at 14
days, ground based and aerial
Exposure Scenario
Estimated exposure of 15 kg
toddler exposed through
contact to surfaces 8 m from
an application area
(µg/kg bw/day)
Risk Quotient
Confidential use 1 – Boom
High boom, fine droplets 0.58 0.0073
High boom, coarse droplets 0.09 0.0012
Low boom, fine droplets 0.20 0.0025
Low boom, coarse droplets 0.05 0.0006
Confidential use 2 – Aerial – agriculture
Swath width 20 m, Med-coarse droplet size 0.80 0.0100
Swath width 20 m, coarse- v. coarse droplets 0.59 0.0074
Swath width 20 m, extremely coarse droplets 0.40 0.0050
Swath width 24 m, v. fine-fine droplets 2.39 0.0299
Swath width 24 m, fine-med. droplets 1.19 0.0148
Swath width 24 m, med.-coarse droplets 0.80 0.0099
Table 24 Risk Quotients for Mainman: Confidential uses 3 and 4 – 70 g ai/ha, 2 applications at 7 days,
ground based and aerial
Exposure Scenario
Estimated exposure of 15 kg toddler
exposed through contact to surfaces 8 m
from an application area
(µg/kg bw/day)
Risk Quotient
Confidential use 3 – Boom
High boom, fine droplets 0.68 0.0085
High boom, coarse droplets 0.11 0.0013
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Low boom, fine droplets 0.23 0.0029
Low boom, coarse droplets 0.05 0.0007
Confidential use 4 – Aerial – agriculture
Swath width 20 m, Med-coarse droplet size 0.94 0.0117
Swath width 20 m, coarse- v. coarse droplets 0.69 0.0087
Swath width 20 m, extremely coarse droplets 0.47 0.0059
Swath width 24 m, v. fine-fine droplets 2.80 0.0350
Swath width 24 m, fine-med. droplets 1.39 0.0174
Swath width 24 m, med.-coarse droplets 0.93 0.0116
Table 25 Risk Quotients for Mainman: Confidential use 5 – 70 g ai/ha, 3 applications at 21 days
Exposure Scenario
Estimated exposure of 15 kg
toddler exposed through
contact to surfaces 8 m from
an application area
(µg/kg bw/day)
Risk Quotient
Airblast
Airblast sparse orchard 1.57 0.0196
Airblast dense orchard 0.52 0.0065
Outcomes of the bystander exposure assessment
The risks to bystanders are acceptable for the use scenarios evaluated.
Summary and conclusions of the human health risk assessment
Estimated exposures for operators, re-entry workers and bystanders are below the AOEL for all of the use
scenarios assessed. Operator exposures are below the AOEL even without the use of personal protective
equipment (PPE); however, staff consider that it is appropriate to retain requirements for PPE since the use
of PPE when handling agrichemicals is good practice. In addition, although the exposure assessment
indicates acceptable risks for re-entry workers even without the use of PPE, staff consider that it is
appropriate to include a label statement along the following lines:
‘Do not allow entry into treated areas until the spray has dried, unless wearing cotton overalls
buttoned to the neck and wrist (or equivalent clothing) and chemical resistant gloves. Clothing must
be laundered after each day’s use.’
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Appendix H: Environmental risk assessment
Summary of the data on the active ingredient and its metabolites
A summary of the environmental fate and ecotoxicological data is provided in the following tables
Table 26 Summary of environmental fate data on flonicamid and its metabolites – Values in bold are
used for the risk assessment.
Test Flonicamid TFNA TFNA-AM TFNA-OH TFNG-AM TFNG
Hydrolysis
DT50 = 204 d
at pH 9, stable
at pH 7 and
below
Stable - - - -
Aqueous photolysis DT50 = 267 d - - - - -
Aerobic degradation
(water/sediment)
DT50 =43.6
and 35.7 d
(whole system)
DT50 = 59 d
whole
system
- - - -
Bioaccumulation Log Kow = 0.3 - - - - -
Aerobic degradation in soil
(laboratory)
DT50 = 1.0, 0.7,
0.7, 1.8 days
DT50 = 0.5,
0.3, 0.5
days
DT50 = 1.2,
1.0, 3.82
days
DT50 = 1.0,
1.3, 3.4
days
DT50 = 1.0,
0.3, 0.2
days
-
Soil photolysis
DT50 = 22 days
in continuous
illumination
- - - - -
Adsorption/desorption
Koc = 7.9
(sand), 11.2,
13.4, 20.7
(sand)
Koc = 0.35
(sand),
0.00, 3.05,
2.67 (sand)
Koc = 5.53,
10.11, 5.04
(sand),
4.53, 12.11
Koc = 1.60
(sand),
1.92, 4.19,
4.39 (sand)
Koc =
10.51, 9.25,
7.56
(sand),
4.24, 15.84
Koc = 0.20
(sand),
1.05, 1.29,
4.05 (sand)
Photodegradation in air DT50 = 13.74 d - - - - -
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Table 27 Summary of ecotoxicological data on flonicamid and its metabolites – Values in bold are
used for the risk assessment.
Test Flonicamid Mainman TFNA TFNA-AM TFNA-OH TFNG-AM
Acute / fish
LC50 > 100
mg/L (trout and
bluegill sunfish)
LC50 > 100 mg/L
(trout)
LC50 > 100
mg/L (trout)
LC50 > 100
mg/L (trout)
LC50 > 100
mg/L (trout)
LC50 > 100
mg/L (trout)
Acute / aquatic
invertebrates
EC50 > 100
mg/L
(Daphnia)
EC50 > 200
mg/L
(Chironomus)
EC50 > 100 mg/L
(Daphnia)
EC50 > 100
mg/L
(Daphnia)
EC50 > 100
mg/L
(Daphnia)
EC50 > 100
mg/L
(Daphnia)
EC50 > 100
mg/L
(Daphnia)
Algae ErC50 > 100
mg/L ErC50 > 100 mg/L
ErC50 > 100
mg/L
ErC50 > 100
mg/L
ErC50 > 100
mg/L
ErC50 > 100
mg/L
Aquatic plant EC50 > 119
mg/L - - - - -
Chronic / fish
NOEC = 10
mg/L (fathead
minnow)
- - - - -
Reproduction
daphnia
NOEC = 3.1
mg/L - - - - -
Reproduction
Chironomus
NOEC = 25
mg/L
Acute / earthworm LC50 > 1000
mg/kg soil -
LC50 > 100
mg/kg soil
LC50 > 100
mg/kg soil
LC50 > 100
mg/kg soil
LC50 > 100
mg/kg soil
Terrestrial plants -
No effects at 150
g ai/ha (seedling
emergence and
vegetative
vigour)
- - - -
Soil micro-
organisms
No effects on C
and N
transformation
at 500 g ai/ha
< 25% effects on
C and N
transformation at
0.274 mg
- - - -
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Test Flonicamid Mainman TFNA TFNA-AM TFNA-OH TFNG-AM
(1.03 mg ai/kg
soil)
formulation /kg
soil
Acute / bird
LD50 = 1591
mg/kg bw
(duck, females)
LD50 > 2000
mg/kg bw
(quail)
- - - - -
Short-term / bird
LC50 > 5000
ppm (quail and
duck)
- - - - -
Reproduction bird
NOEC = 59
mg/kg bw/d
(1000 ppm)
(duck)
NOEC = 90
mg/kg bw/d
(1000 pm)
(quail)
- - - - -
Acute / bees
LD50 > 100
µg/bee
(contact)
LD50 > 60.5
µg/bee (oral)
LD50 > 100
µg/bee (contact) - - - -
Non target
arthropods
(laboratory)
-
A. rhopalosiphi:
55.5% mortality
at 210 g ai/ha
T. pyri: 100%
mortality at 80 g
ai/ha
C.
septempunctata:
30% mortality at
- - - -
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Test Flonicamid Mainman TFNA TFNA-AM TFNA-OH TFNG-AM
80 g ai/ha
P. cupreus: 3.3
mortality at 45 g
ai/ha
Non target
arthropods
(extended
laboratory)
-
A. rhopalosiphi:
4.4% mortality at
85 g ai/ha
T. pyri: 23.3%
mortality at 85 g
ai/ha
C.
septempunctata:
6.1% mortality at
85 g ai/ha
C carnea: 18.8
mortality at 85 g
ai/ha
E. balteatus:
2.6% mortality at
85 g ai/ha
- - - -
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Appendix I: Controls applying to Mainman
Notes: The controls for this substance apply for the indefinite duration of the approval of this
substance. Please refer to the Hazardous Substances Regulations and the modifications listed below
for the requirements of each control. The regulations can be found on the New Zealand Legislation
website http://www.legislation.co.nz.
Table 28: Controls for Mainman
Hazardous Substances (Classes 6, 8, and 9 Controls) Regulations 2001
Code Regulation Description Variation
T1 11 – 27
Limiting exposure to toxic
substances through the
setting of TELs
No TEL values are set for any component of the
substance at this time. The following ADE and
PDE values have been set for flonicamid
ADE = 0.073 mg/kg bw/day
PDEfood = 0.0511 mg/kg bw/day
PDEdrinking water = 0.0146 mg/kg bw/day
PDE other = 0.0073 mg/kg bw/day
T2 29 – 30
Controlling exposure in
places of work through the
setting of WESs
WorkSafe New Zealand WES values exist for
component G but these should not apply to
Mainman due to the low concentration of these
components and/or the form of the substance
T4 7
Requirements for
equipment used to handle
substances
T7 10
Restrictions on the
carriage of toxic or
corrosive substances on
passenger service vehicles
E1 32 – 45
Limiting exposure to
ecotoxic substances
through the setting of EELs
No EEL values are set for this substance at this
time and the default EELs are deleted
E2 46 – 48
Restrictions on use of
substances in application
areas
A Maximum application rate has been set under
section 77A for this substance
E3 49
Controls relating to
protection of terrestrial
invertebrates e.g.
beneficial insects
E6 7
Requirements for
equipment used to handle
substances
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Hazardous Substances (Identification) Regulations 2001
Code Regulation Description Variation
I1 6, 7, 32 – 35,
36(1) – (7)
Identification requirements,
duties of persons in charge,
accessibility,
comprehensibility, clarity and
durability
I9 18 Secondary identifiers for all
hazardous substances
I11 20 Secondary identifiers for
ecotoxic substances
I16 25 Secondary identifiers for
toxic substances
I19 29 – 31
Additional information
requirements, including
situations where substances
are in multiple packaging
I21 37 – 39,
47 – 50
General documentation
requirements
I28 46
Specific documentation
requirements for toxic
substances
I29 51 – 52 Signage requirements
Hazardous Substances (Packaging) Regulations 2001
Code Regulation Description Variation
P1 5 – 7(1), 8 General packaging requirements
P3 9
Criteria that allow substances to be
packaged to a standard not meeting
Packing Group I, II or III criteria
P13 19 Packaging requirements for toxic
substances
PS4 Schedule 4 Packaging requirements as specified
in Schedule 4
Hazardous Substances (Disposal) Regulations 2001
Code Regulation Description Variation
D4 8 Disposal requirements for toxic and
corrosive substances
D5 9 Disposal requirements for ecotoxic
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substances
D6 10 Disposal requirements for packages
D7 11, 12
Information requirements for
manufacturers, importers and
suppliers, and persons in charge
D8 13, 14
Documentation requirements for
manufacturers, importers and
suppliers, and persons in charge
Hazardous Substances (Emergency Management) Regulations 2001
Code Regulation Description Variation
EM1 6, 7, 9 – 11 Level 1 information requirements for
suppliers and persons in charge
EM6 8(e) Information requirements for toxic
substances
EM7 8(f) Information requirements for ecotoxic
substances
EM8 12 – 16,
18 – 20
Level 2 information requirements for
suppliers and persons in charge
EM13 42 Level 3 emergency management
requirements: signage
Hazardous Substances (Tank Wagon and Transportable Containers) Regulations 2004
Code Regulation Description
Tank Wagon 4 to 43
as applicable Controls relating to tank wagons and transportable containers
Additional controls
Code Regulation Description Variation
Water Section 77A The substance must not be applied
into, onto or over water.
The substance must not be applied into
or onto water27
App Rate Section 77A A maximum application rate is set for
this substance.
The maximum application rate of this
substance is 80 g of flonicamid/ ha.
27 where ‘water‘ means water in all its physical forms, whether flowing or not, and whether over or under ground, but does not
include water in any form while in a pipe, tank or cistern or water used in the dilution of the substance prior to application or
water used to rinse the container after use
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Code Regulation Description Variation
Max
Impure
Section 77A May not contain greater than a set
amount of a toxicologically
significant impurity
The flonicamid component of this
substance must not contain more than
3 g/kg of toluene.
Label Section 77A Additional label information has been
specified.
The following statements, or words to
this effect must appear on the product
label and safety data sheet
“Do not allow entry into treated areas
until the spray has dried, unless wearing
cotton overalls buttoned to the neck and
wrist (or equivalent clothing) and
chemical resistant gloves. Clothing
must be laundered after each day’s
use.”
“Do not apply this product to any plant
likely to be visited by bees at the time of
application; or immediately after
application until spray has dried; or in
areas where bees are foraging"
“The maximum application rate of this
substance is 80 g of flonicamid/ ha”
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Appendix J: Confidential information
This appendix contains the confidential information provided by the applicant and is not publically
available.
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Appendix K: Standard terms and abbreviations
ai active ingredient
ALD50 approximate median lethal dose,
50%
AOEL acceptable operator exposure level
ARfD acute reference dose
as active substance
BCF bioconcentration factor
bfa body fluid assay
BOD biological oxygen demand
BSAF biota-sediment accumulation factor
bw body weight
c centi- (x10-2
)
CA controlled atmosphere
CI confidence interval
CL confidence limits
CNS central nervous system
COD chemical oxygen demand
DFR dislodgeable foliar residue
DO dissolved oxygen
DOC dissolved organic carbon
DT50 period required for 50 percent
dissipation (define method of
estimation)
DT90 period required for 90 percent
dissipation (define method of
estimation)
dw dry weight
ED50 median effective dose
ERC environmentally relevant
concentration
F Field
F0 parental generation
F1 filial generation, first
F2 filial generation, second
fp freezing point
G glasshouse
GAP good agricultural practice
GC gas chromatography
GC-EC gas chromatography with electron
capture detector
GC-FID gas chromatography with flame
ionization detector
GC-MS gas chromatography-mass
spectrometry
GC-MSD gas chromatography with mass-
selective detection
GLC gas liquid chromatography
GLP good laboratory practice
GM geometric mean
H Henry’s Law constant (calculated
as a unitless value) (see also K)
ha hectare
Hb haemoglobin
HCG human chorionic gonadotropin
Hct haematocrit
HPLC high pressure liquid
chromatography or high
performance liquid
chromatography
HPLC-MS high pressure liquid
chromatography - mass
spectrometry
I indoor
I50 inhibitory dose, 50%
IC50 median immobilization
concentration or median inhibitory
concentration 6
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Application for approval to import Mainman for release (APP202145)
Feburary 2015
ID ionization detector
Im Intramuscular
inh Inhalation
ip Intraperitoneal
IPM integrated pest management
iv Intravenous
IVF in vitro fertilization
K Kelvin or Henry’s Law constant (in
atmospheres per cubic meter per
mole) (see also H)
Kads adsorption constant
Kdes apparent desorption coefficient
Koc organic carbon adsorption
coefficient
Kom organic matter adsorption
coefficient
kg Kilogram
LC liquid chromatography
LC-MS liquid chromatography- mass
spectrometry
LC50 lethal concentration, median
LCA life cycle analysis
LC-MS-MS liquid chromatography with tandem
mass spectrometry
LD50 lethal dose, median; dosis letalis
media
LDH lactate dehydrogenase
LOAEC lowest observable adverse effect
concentration
LOAEL lowest observable adverse effect
level
LOD limit of detection
LOEC lowest observable effect
concentration
LOEL lowest observable effect level
LOQ limit of quantification
(determination)
LPLC low pressure liquid
chromatography
LSC liquid scintillation counting or
counter
LSS liquid scintillation spectrometry
LT lethal threshold
M molar
μm micrometer (micron)
MDL method detection limit
MFO mixed function oxidase
μg microgram
MLT median lethal time
MLD median lethal dose
mol Mole(s)
MOS margin of safety
mp melting point
MS mass spectrometry
MSDS material safety data sheet
NAEL no adverse effect level
nd not detected
NEL no effect level
ng nanogram
nm nanometer
NOAEC no observed adverse effect
concentration
NOAEL no observed adverse effect level
NOEC no observed effect concentration
NOEL no observed effect level
NR not reported
OC organic carbon content
ODP ozone-depleting potential
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Application for approval to import Mainman for release (APP202145)
Feburary 2015
OM organic matter content
Pa pascal
PEC predicted environmental
concentration
PECS predicted environmental
concentration in soil
PECSW predicted environmental
concentration in surface water
PECGW predicted environmental
concentration in ground water
PHI pre-harvest interval
pKa negative logarithm (to the base 10)
of the dissociation constant)
PNEC predicted no effect concentration
POW partition coefficient between n-
octanol and water
ppb parts per billion (10-9
)
PPE personal protective equipment
ppm parts per million (10-6
)
ppp plant protection product
ppq parts per quadrillion (10-24
)
ppt parts per trillion (10-12
)
PTDI provisional tolerable daily intake
r correlation coefficient
r2 coefficient of determination
REI restricted entry interval
Rf retardation factor
RfD reference dose
RL50 median residual lifetime
RP reversed phase
RRT relative retention time
RSD relative standard deviation
sc subcutaneous
SD standard deviation
se standard error
SF safety factor
SIMS secondary ion mass spectroscopy
SOP standard operating procedures
sp species (only after a generic name)
SPE solid phase extraction
spp subspecies
SSD sulphur specific detector
STEL short term exposure limit
t½ half-life (define method of
estimation)
TCLo toxic concentration, low
TER toxicity exposure ratio
TIFF tag image file format
TOC total organic carbon
TWA time weighted average
UF uncertainty factor (safety factor)
ULV ultra low volume
UV ultraviolet
v/v volume ratio (volume per volume)
w/v weight per volume
ww wet weight
w/w weight per weight