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RAPID SCIENCE MEMO
www.epa.govt.nz
13 JUNE 2019
Summary
Substance AG-M4-700 OF1
Application code APP203824
Application sub-type Rapid reduced
Applicant Adama New Zealand Limited
Purpose of the application To obtain approval for AG-M4-700 OF1, a herbicide containing 700g/L
metamitron as a suspension concentrate
Date application formally received 28 May 2019
Page 2 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
1. Key Points
1.1. AG-M4-700 OF1 is a herbicide in the form of a suspension concentrate (SC) containing 700 g/L of the
active ingredient metamitron. It is intended for the control of weeds in beets (red, fodder, sugar) and
applied using both ground-based and aerial application methods at rates of up to 2.8 kg metamitron/ha
in a single application and up to 4.2 kg metamitron/ha per crop per season.
1.2. AG-M4-700 OF1 is intended to be imported in New Zealand.
1.3. At the time of this evaluation, Adama New Zealand Limited, was already putting a similar substance on
the market (“Goltix Herbicide”) using the approval number HSR000535, which contains 700 g/kg
metamitron in the form of a water dispersible granule, for the control of weeds in beets (red, fodder,
sugar) and mangolds.
2. Status Of Substance (SOS) or statutory determination history
2.1. No SOS or statutory determinations were issued for AG-M4-700 OF1.
3. Identification of substance and reference
3.1. The reference proposed by the applicant and identified by the EPA are the same (see Table 1).
Table 1: Identified references for the rapid assessment of AG-M4-700 OF1
Substance to be approved Reference
Name AG-M4-700 OF1 AGPRO Metamitron
Substance database ID 49711 44396
HSNO Approval number - HSR100753
Substance physical form Suspension concentrate (SC) Suspension concentrate (SC)
Active ingredient(s) and
concentration (g/L) 700 g/L metamitron 700 g/L metamitron
4. RAPID assessment criteria
Active ingredient
4.1. This substance meets the active ingredients criteria. The concentration of the active ingredient in AG-
M4-700 OF1 is the same as that of the reference substance.
Physical form
4.2. AG-M4-700 OF1 is in the same physical form as the reference substance, namely as a suspension
concentrate.
Page 3 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Use pattern
4.3. This substance meets the use pattern criteria. Both AG-M4-700 OF1 and the reference substance are
herbicides used in a similar manner (see Table 2).
Table 2: Use pattern of AG-M4-700 OF1 in comparison to its reference substance
Substance to be approved Reference
Target pest / condition
Susceptible weeds: Amaranthus spp.,
Buttercups (seedlings), Chickweed,
Docks (seedling) Fathen, Galinsoga,
Groundsel, Hawksbeard, Henbit,
Matricaria spp. (Stinking mayweed and
Chamomiles), Nettles, Pennycress,
Annual poa, Wild portulaca, Scotch
thistle (seedling), Shepherd’s purse,
Sow thistle, Speedwells, Spurrey, Twin
cress, White clover, Yarrow (seedling).
Moderately susceptible weeds: Black
nightshade, Field pansy, Fumitory,
Hedge mustard, Scarlet pimpernel,
Thorn apple, Wireweed.
Susceptible weeds: Amaranthus spp.,
Buttercups (seedlings), Chickweed,
Docks (seedling) Fathen, Galinsoga,
Groundsel, Hawksbeard, Henbit,
Matricaria spp. (Stinking mayweed and
Chamomiles), Nettles, Pennycress,
Annual poa, Wild portulaca, Scotch
thistle (seedling), Shepherd’s purse,
Sow thistle, Speedwells, Spurrey, Twin
cress, White clover, Yarrow (seedling).
Moderately susceptible weeds: Black
nightshade, Field pansy, Fumitory,
Hedge mustard, Scarlet pimpernel,
Thorn apple, Wireweed.
Target animal / crop Red beet, fodder beet, sugar beet,
mangolds
Red beet, fodder beet, sugar beet,
mangolds
Application rate (kg ai/ha)
Up to 2.8 kg metamitron/ha in a single
application and up to 4.2 kg
metamitron/ha per crop per season
4.2 kg metamitron/ha, three times per
year
Comment on any differences There is no difference between the use pattern of AG-M4-700 OF1 and the
reference substance
Are the differences
insignificant in terms of risk of
adverse effects?
As there is no difference between the use pattern of AG-M4-700 OF1 and the
reference substance, there is no difference in terms of adverse effects associated
with the use of the substances.
Major Hazardous Components
4.4. AG-M4-700 OF1 meets the major hazardous components criteria. The major hazardous components in
AG-M4-700 OF1 constitute a similar proportion as in the reference substance.
Adverse Effects
4.5. AG-M4-700 OF1 meets the adverse effects criteria, as the hazards of this substance are reduced
compared to the reference substance (see Table 3).
4.6. AG-M4-700 OF1 has a reduced hazard classification for acute toxicity via the inhalation route [6.1D
(inhalation) instead of 6.1B (inhalation)] and dermal route [6.1E (dermal) instead of 6.1D (dermal)].
Page 4 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
However, the 6.1D (inhalation) classification is primarily derived from a recent change in our
classification database for the active ingredient metamitron. Were the reference substance to be
assessed by mixture rules with the new classification, it would likely receive a 6.1D classification.
4.7. AG-M4-700 OF1 also has a reduced hazard classification for aquatic toxicity (9.1B instead of 9.1A) and
for soil toxicity (9.2B instead of 9.2A).
Table 3: Comparison of the respective classifications of AG-M4-700 OF1 and its reference substance
Classification comparison
Substance 6.1D (oral, inhalation), 6.1E (dermal), 9.1B, 9.2B, 9.3C
Reference 6.1B* (inhalation), 6.1D (oral, dermal), 9.1A, 9.2A, 9.3C
* The 6.1D (inhalation) classification is triggered by the active ingredient, metamitron. The classification for metamitron on the EPA
register is incorrect and will be reviewed to include the 6.1D (inhalation) classification.
5. Controls
EPA Notice controls
5.1. The Labelling, Safety Data Sheet (SDS), Packaging, Disposal and Hazardous Property Controls (HPC)
Part 1, Part 3, Part 4A and Part4B Notices apply to AG-M4-700 OF1.
5.2. The name and concentration of the following components need to be specified on the label and SDS
(see Table 4).
Table 4: Components required on the label and SDS of AG-M4-700 OF1
Labelling requirement SDS requirement
Metamitron 6.1D (oral, inhalation)
Metamitron 6.1D (oral, inhalation), 9.1A
1,2-Propanediol (WES)
Benzene, dimethyl-, mixed isomers (WES)
Benzene, ethyl- (WES)
Sodium hydroxide (WES)
5.3. No Tolerable Exposure Limit (TEL) value has been set previously for the active ingredient in AG-M4-700
OF1 because it is considered that exposure to this substance is not likely to result in an appreciable
toxic effect to people, provided controls on use are followed.
5.4. No Environmental Exposure Limit (EEL) value is set at this time, or has been set previously for the
active ingredient in AG-M4-700 OF1, as the level of risk of adverse effects to the environment has been
qualitatively assessed as being negligible, with controls in place.
5.5. AG-M4-700 OF1 has been assessed based on the use pattern described by the applicant. Additional
adverse environmental effects (which are not addressed by the prescribed controls) could occur if this
product was used outside of its intended use pattern, for example, if it was applied at higher application
Page 5 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
rates. Therefore, it is considered necessary to set a maximum application rate of 4 L/ha AG-M4-700
OF1 (equivalent to 2.8 kg/ha metamitron) per application, with a maximum of 6 L/ha AG-M4-700 OF1
(equivalent to 4.2 kg/ha metamitron) per season.
5.6. A maximum application rate was also set for the reference substance, which is 4.2 kg/ha metamitron,
three times per year.
Page 6 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Appendix: study summaries
Summary of mammalian toxicology data for AG-M4-700 OF1
Endpoint
(Test Guideline)
Klimisch
score Result
HSNO
Classification Reference
Acute oral toxicity
(OECD TG 423; EC
method B.1)
1 300 < LD50 <
2000 mg/kg 6.1D
(2007) - Acute Oral
Toxicity Study of Goltix OF in Rats.
Acute dermal toxicity
(OECD TG 402; EC
method B.3.)
1 LD50 >2000
mg/kg bw 6.1E
(2007) - Acute Dermal
Toxicity Study of Goltix OF in CD Rats.
Acute inhalation
toxicity
(OECD TG 403;
OPPTS 870.1300;
EC method B.2;
JMAFF 12 NohSan
No. 8147 (2-1-3))
1 LC50 >3.15 mg/L 6.1D 2008) - Acute Inhalation
Toxicity (Nose Only) Study in the Rat.
Skin
irritation/corrosion
(OECD TG 404; EC
method B.4)
1 Non-irritating
Mean score (24,
48, and 72 hrs)
– - Erythema:
0.0 Oedema: 0.0
No (2007) - Acute Dermal
Irritation/ Corrosion test (Patch test) of
Goltix OF in Rabbits.
Acute Eye
Irritation/Corrosion
(OECD TG 405; EC
method B.5)
1 Non-irritating
Mean Draize
Score (24, 48,
and 72 hrs) –
Cornea
-Opacity: 0.0
Conjunctiva
-Redness: 0.0
-Chemosis: 0.0
No (2007) - Acute Eye
Irritation/Corrosion Test of Goltix OF In
Rabbits.
Skin Sensitisation
(OECD TG 406; EC
method B.6.)
1 Non-sensitising No (2007) - Examination Of
Goltix OF in the Skin Sensitisation Test in
Guinea Pigs According to Magnusson
and Kligman (Maximisation Test).
Page 7 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Ecotoxicity study summaries for AG-M4-700 OF1
Aquatic toxicity
Table 1: Acute toxicity to Fish
Study type Acute Fish study
Flag Key Study
Test Substance Goltix OF
Exposure Static 96 hour
Test species Rainbow trout Oncorhynchus mykiss
Endpoint LC50
Value 283 mg/L
Reference 2007 Goltix OF Fish (Rainbow trout), Acute Toxicity Test, Static, 96
h
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 203
Dose Levels 25, 50, 100, 200 and 400 mg/L (nominal)
Analytical measurements Yes via HPLC
Study Summary
This study was carried out to examine the toxicity of Goltix OF to Rainbow trout.
The trout were nominally exposed to 25, 50, 100, 200 and 400 mg Goltix OF/L.
The study was carried out as per OECD 203 with no deviations from the protocol.
All of the test validity criteria were met. Measured concentrations of the active
ingredient metamitron were in the range of 89 -102% of the expected nominal
concentration throughout the test, hence it is appropriate to express the endpoints
in terms of the nominal concentrations. Metamitron is known to hydrolyse forming
the metabolite Desamino-Metamitron. Therefore recovery rates of desamino-
metamitron were analytically verified, however, it was only detected at values
below LOQ under test conditions. The LC50 was determined to be 283 mg/L
(equivalent to 169 mg ai/l). There were no mortalities up to 200 mg/L and the
NOEC was determined to be 50 mg/L based on the behaviour of the fish.
Conclusion LC50 = 283 mg/L
Table 2: Acute toxicity to aquatic invertebrates
Study type Acute aquatic invertebrate study
Flag Key Study
Test Substance Goltix OF
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Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Exposure Static 48 hour
Test species Daphnia magna
Endpoint EC50
Value 99.1 mg/L
Reference 2007 Goltix OF Acute Immobilization Test (Static, 48 h) to Daphnia
magna
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 202
Dose Levels 50, 75, 113, 169 and 253 mg/L (nominal)
Analytical measurements Yes via HPLC
Study Summary
This study was carried out to examine the toxicity of Goltix OF to Daphnia magna. The daphnia were nominally exposed to 50, 75, 113, 169 and 253 mg Goltix OF/L. The study was carried out as per OECD 202 with no deviations from the protocol. All of the test validity criteria were met. Measured concentrations of the active ingredient metamitron were in the range of 90 -106% of the expected nominal concentration throughout the test, hence it is appropriate to express the endpoints in terms of the nominal concentrations. Metamitron is known to hydrolyse forming the metabolite Desamino-Metamitron. Therefore recovery rates of desamino-metamitron were analytically verified, however, it was only detected at values below LOQ under test conditions.
The EC50 was determined to be 99.1 mg/L (equivalent to 57.2 mg ai/L). The NOEC
was determined to be 50 mg/L (corresponding to 29.9 mg ai/L based on the
behaviour of the fish.
Conclusion EC50 = 99.1 mg/L
Table 2: Acute toxicity to algae
Study type Acute algae study
Flag Key Study
Test Substance Goltix OF
Exposure Static 72 hour
Test species Pseudokirchneriella subcapitata
Endpoint ErC50
Value 5.32 mg/L
Reference 2007 Goltix OF Alga, Growth Inhibition Test with
Pseudokirchneriella subcapitata, 72 h
Klimisch Score 2 (see comments for more details)
Page 9 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 201
Dose Levels 0.10, 0.32, 1.0, 3.2, 10, 32, 100 mg/ L (nominal)
Analytical measurements Yes via HPLC
Study Summary
This study was carried out to examine the toxicity of Goltix OF to
Pseudokirchneriella subcapitata. The algae were nominally exposed to 0.10, 0.32,
1.0, 3.2, 10, 32 mg Goltix OF/L. The study was carried out as per OECD 201 with
no deviations from the protocol. All of the test validity criteria were met (see
comments section for details on the recovery rates). The EC50 for growth rate was
determined to be 5.32 mg/L (equivalent to 3.18 mg ai/L). The EC10 (for growth rate)
was determined to be 1.39 mg/L (corresponding to 0.83 mg ai/L.
Comments
Measured concentrations of the active ingredient metamitron decreased
significantly during the test period duration. While the initial concentrations were in
the range of 92-115% of the expected nominal concentration at the end of the test
period the recovery rate varied from 41% to 85%. This corresponds to an increase
in the concentration of the metabolite desamino-metamitron (present at the end of
test duration at between 7-43% of the nominal concentration of the active
ingredient). Based on the low recovery of the active ingredient at the test
completion it may have been more appropriate to express the results in terms of
the measured concentrations. Despite this, however, the EPA staff feel that the
study is sufficient for classification since the conclusion is the same irrespective of
which value is used, but the value may not be appropriate to use for risk
assessment.
Conclusion EC50 = 5.32 mg/L (equivalent to 3.18 mg a.i/L)
Table 4: Acute toxicity to aquatic plants
Study type Acute aquatic plant study
Flag Key Study
Test Substance AG-M4-700 OF1
Exposure Semi-static 7-day
Test species Lemna gibba L.
Endpoint ErC50
Value 1.49 mg/L – EPA calculated value for the formulation based on the measured
active ingredient EC50 value of 1.04 mg ai/L
Reference
2017 Effects of AG-M4-700 OF1 on Lemna gibba in a growth inhibition
test under semi-static test conditions
Klimisch Score 2
Page 10 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Amendments/Deviations None that would affect the results
GLP Yes
Test Guideline/s OECD 221
Dose Levels 0.72, 1.22, 2.08, 3.53, 6.00 mg/L test item (nominal)
Analytical measurements Yes via HPLC
Study Summary
This study was carried out to examine the toxicity of AG-M4-700 OF1 to Lemna
gibba. The algae were nominally exposed to 0.72, 1.22, 2.08, 3.53, 6.00 mg/L of
the test substance which corresponds to 0.42, 0.71, 1.21, 2.06, 3.50 mg/L ai.
The study was carried out as per OECD 221 with no significant deviations from the
protocol. All of the test validity criteria were met.
The most sensitive ErC50 was 3.27 mg/L test item nominal (1.91 mg/L ai nominal;
1.04 mg/L ai mean measured) based on the assessment of dry weight.
Comments
Measured concentrations of the active ingredient metamitron decreased
significantly during the test period duration. At test start and test solution renewal
processes nominal concentrations were analytically confirmed in fresh test
solutions. Recoveries between 86 and 93% were found. In spent test solutions
taken at test solution renewals and test end, recoveries ranging from 22 to 54%
were determined.
Results are expressed in terms of the nominal concentrations and the measured
concentration of active ingredient. Based on the low recovery of the active
ingredient. The EPA have estimated the ErC50 based on the measured active
ingredient ErC50 value of 1.04 mg ai/L (1.04 x 1000/700 = 1.49 mg AG-M4-700
OF1).
Conclusion ErC50 = 1.49 mg/L – EPA calculated value for the formulation based on the
measured active ingredient ErC50 value of 1.04 mg ai/L
Conclusion about aquatic toxicity
The formulated product triggers a 9.1B HSNO classification based on its toxicity to algae and aquatic plants
and the fact that the active ingredient is not rapidly biodegradable. It is also harmful to crustaceans.
Soil toxicity
Table 5: Earthworm chronic toxicity test
Study type Earthworm chronic toxicity test
Flag Key Study
Test Substance AG-M4-700 OF1
Endpoint NOEL
Value 180 mg/kg soil dry weight
Reference 2016 AG-M4-700 OF1 – Determination of chronic toxicity to the
earthworm Eisenia andrei in an artificial soil substrate
Page 11 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 222
Dose Levels 31, 56, 100, 180, 324, 583, 1050 and 1890 mg product/kg soil dry weight
Analytical measurements None
Study Summary
The aim of the study was to determine under laboratory test conditions whether AG-
M4-700 OF1 had harmful effects on the survival, growth or reproductive capacity of
adults of the earthworm Eisenia andrei.
AG-M4-700 OF1 was evaluated at eight concentrations, equivalent to 31, 56, 100,
180, 324, 583, 1050 and 1890 mg test item/kg soil dry weight. These were
compared to a control treatment of purified water. Treatments were incorporated
into an artificial soil substrate containing 5% w/w peat, which was placed within
polystyrene boxes (17.1 cm x 11.3 cm in area, by 6 cm deep) with ventilated lids.
There were 8 replicate boxes for the control and 4 for each test-item treatment
concentration. For all treatments, the soil moisture content was maintained at 50%
(± 10%) of the maximum water-holding capacity throughout the bioassay. Ten adult
E. andrei (approx. 5.5 months old, each 300-600 mg fresh weight and with a visible
clitellum) were weighed and then placed on the surface of the soil in each arena
immediately after treatment application. One day after treatment (DAT), hydrated
oat flakes were placed on the soil surface and this food source was replenished
weekly for the first four weeks of the bioassay. At 28 DAT, the numbers of the
original worms still surviving and their fresh weights were recorded. Any apparent
change in the behaviour or physical condition of the confined worms was noted.
These adult worms were then removed and the test soil and any egg cocoons or
juvenile worms were returned to the test chambers. A final supply of oat flakes was
provided on the soil surface. After a further 28 days (ie. 56 DAT), the number of
juvenile worms that had developed in each replicate arena was recorded.
After 28 days, there was no significant mortality in any of the AG-M4-700 OF1
treatments (Fisher’s Exact Test, α = 0.05). All of the worms appeared healthy and
no abnormal behaviour was recorded. In terms of changes in mean body weight,
the only treatment to differ significantly from the control was 1890 mg test item/kg
soil dry weight (one-way ANOVA and Dunnett’s t-test, α = 0.05; or Mann-Whitney U-
test, α = 0.05). Thus, the lowest-observed-effect concentration (LOEC) for direct
effects on adult worms was 1890 mg/kg soil dry weight and the no-observed-effect
concentration (NOEC) was 1050 mg/kg soil dry weight.
In terms of the mean numbers of juveniles, statistically significant effects were
observed at treatment concentrations of 1890, 1050, 583 and 324 mg test item/kg
soil dry weight (t-test for independent samples, α = 0.05). Thus, with respect to
reproduction, the LOEC was 324 mg test item/kg soil dry weight and the NOEC was
180 mg test item/kg soil dry weight.
Conclusion NOEL = 180 mg/kg soil dry weight
Page 12 of 21
Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Table 6: Predatory mite Hypoaspis aculeifer chronic toxicity test
Study type Predatory mite chronic toxicity test
Flag Key Study
Test Substance AG-M4-700 OF1
Endpoint NOEC
Value 1000 mg/kg soil dry weight
Reference
2016 AG-M4-700 0F1 (Metamitron 700 SC) - A laboratory test to
determine the effects of fresh residues on the predatory mite Hypoaspis aculeifer
(Acari, Laelapidae)
Klimisch Score 1
Amendments/Deviations None that affect the conclusions
GLP Yes
Test Guideline/s OECD 226
Dose Levels 1000 mg/kg soil dry weight
Analytical measurements None
Study Summary
The aim was to determine under laboratory test conditions whether AG-M4-700
OF1 has harmful effects on the predatory soil mite Hypoaspis aculeifer (Canestrini)
(Acari, Laelapidae).
Following a preliminary range finding test, AG-M4-700 OF1 was evaluated in a
definitive bioassay at a single concentration, equivalent to 1000 mg test item/kg soil
dry weight. This was compared to an untreated control and a toxic reference
treatment of Perfekthion (nominally 400 g/L dimethoate), which was applied at a
concentration of 14 mg ai/kg soil dry weight. Treatments were incorporated into an
artificial soil substrate (containing 5% w/w peat), aliquots of which were then
transferred into small, ventilated, glass jars (n = 8 for control; n = 8 per test-item
treatment concentration; n = 5 for toxic reference). Ten female soil mites
(approximately 7-14 days after becoming adult) were then introduced into each
arena. Cheese mites (Tyrophagus putrescentiae (Schrank)) and springtails
(Folsomia Candida (Willem)) were provided as food for the predatory soil mites and
were replenished ad libitum. At 14 days after treatment (DAT), both the numbers of
surviving adult predatory soil mites and of their offspring were recorded.
The results were used to determine the no-observed-effect concentration (NOEC)
for the test item, with respect to the assessments of both mortality and reproduction.
In addition, values for key effect concentrations (EC50, EC20 and EC10) for
reproduction were estimated, as was the median lethal concentration (LC50) for the
mites originally introduced.
Based on the numbers of offspring produced, the 14-day EC50 and the EC20 and
EC10 values for AG-M4-700 OF1 were all estimated as > 1000 mg test item/kg soil
dry weight. The LC50 was also estimated to be > 1000 mg test item/kg soil dry
weight. The overall NOEC with respect to both mite survival and reproduction was
1000 mg test item/kg soil dry weight.
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Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Conclusion NOEC = 1000 mg/kg soil dry weight
Table 7: Predatory mite toxicity test
Study type Predatory mite toxicity test
Flag Key Study
Test Substance AG-M4-700 0F1
Test species Predatory mite Typhlodromus pyri
Endpoint NOER
Values NOER = 9200 mL product/ha
Reference
2016 AG-M4-700 0F1 (Metamitron 700 SC) - A rate-response
laboratory bioassay ofthe effects of fresh residues on the predatory mite
Typhlodromus pyri (Acari: Phytoseiidae)
Klimisch Score 1
Amendments/Deviations None noted
GLP Yes
Test Guideline/s Blumel et al. (2000)
Dose Levels 1200, 2300, 4600, 9200 and 13800 mL/ha.
Analytical measurements None
Study Summary
The aim of this study was to determine the effects of fresh-dried residues of AG-
M4-700 OF1 on the predatory mite Typhlodromus pyri Scheuten (Acari:
Phytoseiidae), under worst-case laboratory test conditions. The test substance
was applied to glass plates at the doses outlined above. Twenty protonymphal T.
pyri were placed in each replicate unit, with three replicates (ie. 60 mites) prepared
per treatment. The mites were fed regularly with untreated pollen for food. Their
survival was assessed after a 7-day period, by which time the mites in the control
treatment were adult. A check was then made for sub-lethal treatment effects on
reproduction. For this, the mites were left in-situ and the numbers of eggs
produced per female were recorded over a further 7-day period. The mean number
of eggs produced per female from 7-14 days after treatment (DAT) was calculated
for each treatment. These reproduction assessments were made for mites from the
control and from the highest three treatment rates that had resulted in < 50%
corrected mortality.
Based on statistical comparison with the control, the NOER with respect to mite
survival was 13800 mL product/ha and the NOER for reproduction was 9200 mL
product/ha. A 10.2% effect on reproduction was noted at the highest dose 13800
mL/ha. The 7-day LR50 was > 13800 mL product/ha.
Conclusion NOER = 9200 mL product/ha (based on reproduction)
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Rapid Science Memo for approval to import or manufacture AG-M4-700 OF1 for release (APP203824)
Table 8: Collembola toxicity test
Study type Collembola toxicity test
Flag Key Study
Test Substance AG-M4-700 0F1
Test species Collembola
Endpoint LC50
Values 0.442 mg test item/kg soil
Reference
2017 AG-M4-700 OF1 (Metamitron 700 SC) – A laboratory test to
determine the effects of fresh residues on the springtail Folsomia candida
(Collembola, Isotomidae)
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 232
Dose Levels 0.8, 0.4, 0.2, 0.1, 0.05, 0.025, 0.0125 and 0.00625 mg/kg soil dry weight
Analytical measurements None
Study Summary
The aim of this study was to determine under laboratory test conditions whether
AG-M4-700 OF1 has harmful effects on the springtail Folsomia candida
(Collembola, Isotomidae).
Following a preliminary range finding test, AG-M4-700 OF1 was evaluated in a
definitive bioassay at eight concentrations, equivalent to 0.8, 0.4, 0.2, 0.1, 0.05,
0.025, 0.0125 and 0.00625 mg test item/kg soil dry weight. This was compared to
an untreated control and a toxic reference treatment of Betosip 114 (nominally 114
g/L phenmedipham), applied at a concentration of 200 mg product/kg soil dry
weight. Treatments were incorporated into an artificial soil substrate (containing
5% w/w peat), aliquots of which were then transferred into small glass jars (n = 8
for control; n = 4 per test-item treatment concentration; n = 5 for toxic reference).
Ten juvenile F. candida (10 days old) were introduced into each jar and dry
granulated yeast was provided as food. This food was replenished at 14 days. The
jars were ventilated every 1-4 days. At 28 days, the numbers of the springtails
originally introduced that still survived and the numbers of their offspring were
recorded.
There was a statistically significant effect on adult springtail survival in the 0.8 and
0.4 mg test item/kg soil dry weight treatments, and also in the 0.025 and 0.0125
mg test item/kg soil dry weight treatments. Since there were no significant effects
observed on mortality at concentrations between these, the effects at the lower
concentrations were taken to be an anomaly and the NOEC for mortality was
assumed to be 0.2 mg/kg soil dry weight. The 28-day LC50 was estimated to be
0.442 mg test item/kg soil dry weight.
There was a statistically significant effect on reproduction at 0.025, 0.2, 0.4 and 0.8
mg/kg soil. While the effects at 0.025 mg/kg soil are statistically significant effects
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were not seen at higher doses and the % change in numbers of progeny relative to
the control (14%) is within the allowable variation for the control according the
OECD guidance (20%). The NOEC for reproduction can therefore be considered
to be 0.1 mg test item/kg soil dry weight.
Conclusion LC50 = 0.442 mg test item/kg soil
Table 9: Collembola field test
Study type Collembola field test
Flag Key Study
Test Substance AG-M4-700 0F1
Test species Collembola
Endpoint NOER
Values NOER = 2 product/ha
Reference
2017 A field trial to determine the effects AG-M4-700 OF1 (Metamitron
700 SC) on natural populations of Collembola
Klimisch Score 1
Amendments/Deviations None noted
GLP Yes
Test Guideline/s
Candolfi et al. (2001). Guidance document on regulatory testing and risk
assessment procedures for plant protection products with non-target arthropods.
Barrett et al. (1994). Guidance document on regulatory testing procedures for
pesticides with non-target arthropods.
Dose Levels 2 L/ha
Analytical measurements None
Study Summary
The aim of this field study was to determine the effects of three applications of AG-
M4-700 OF1 at 2 L/ha, with a nominal 7-day spray interval, on the numbers of
Collembola (springtails) present in the soil.
AG-M4-700 OF1 was evaluated at a single application rate, equivalent to 2 L
product/ha. This was applied on three consecutive occasions (times T1, T2 and
T3), with 7- and 8-day intervals in-between the respective applications. This
treatment regime was compared to a water control (applied at time T3 only) and a
toxic reference treatment of chlorpyrifos (an EC formulation nominally containing
480 g ai/L, applied as a spray at a rate equivalent to 1.5 L product/ha at time T3
only).
Twenty-four plots, each approximately 5 m x 5 m in area and separated from each
other by a 2-m-wide buffer strip were marked out in a line alongside a fallow arable
field in southern England. The soil type was characterised as being a sandy clay
loam. Each treatment was applied to eight replicate plots (arranged in a
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randomised block design), with applications being made using a boom sprayer at a
volume rate equivalent to 200 L spray solution/ha.
The Collembola present in each plot were sampled using both a soil-corer and a
motorised soil-surface suction-sampler (both being density-dependent sampling
techniques). The samples were taken within the central sampling area of each
treatment plot. They were collected in the week prior to the initial (T1) application
of the test-item treatment, in order to determine the level of any homogeneity
between the plots, and then again at 1, 4 and 8 weeks after the final (T3) treatment
application.
In the laboratory, the Collembola in the samples were extracted over a four- to six-
day period using banks of Tullgren funnels (one funnel per replicate plot and
sampling technique). The extracted springtails were stored in methyl alcohol prior
to sorting and counting of the main taxonomic groups.
The two most abundant taxa that were identified were the super-family
Entomobryoidea and the order Symphypleona. Statistical comparisons for the pre-
treatment samples did not indicate that numbers for either of these groups differed
significantly between treatments (α = 0.05). After application, there were no
statistically significant effects detected in the AG-M4-700 OF1 (3 x 2 L product/ha.)
Comment
It is noteworthy that the application rate is less than that proposed in New Zealand
(up to 4 litres of product per hectare). Also the applicant has proposed up to 6
applications per season.
Conclusion NOER = 2 product/ha
Table 10: Terrestrial plant toxicity, seedling emergence
Study type Terrestrial plants toxicity, seedling emergence
Flag Key study
Test Substance Goltix OF
Test species Oats, onion, sugar beet, rape, carrot and soybean
Endpoint Effective Dose (ED)50
Values > 5 L/ha
Reference 2007 Goltix OF Terrestrial Plants Toxicity, Seedling Emergence
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 208
Dose Levels 5.0, 2.5, 1.25, 0.625 and 0.313 L/ha
Analytical measurements Yes- HPLC
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Study Summary
The phytotoxicity of the test item Goltix OF to six terrestrial plant species was
determined each over a period of at least 21 days. According to the results of a
range-finder, the test was conducted with the application rates listed above. Test
plants were two monocotyledons (oats, Gramineae; onion, Liliaceae) and four
dicotyledons (sugar beet, Chenopodiaceae; rape, Brassicaceae; carrot,
Umbelliferae; soybean, Leguminosae). The test item was applied once on the soil
surface after the seeds were sown.
The test containers were bottom watered with nutrient solution as needed. The
toxic effects of the test item were determined on day 7, 14 and 21 (onion and
carrot additionally on day 24) by visual observations and on day 21 and 24,
respectively by shoot height and shoot fresh weight determination.
The ED50 values for all plants were > 5 L/ha. NOEL values were 5 L/ha for all
plants with the exception of carrots which had a NOEL of 2.5 L/ha.
Conclusion ED50 > 5 L/ha
Table 11: Terrestrial plants toxicity, vegetative vigour
Study type Terrestrial plants toxicity, vegetative vigour
Flag Key study
Test Substance Goltix OF
Test species Oats, onion, sugar beet, rape, carrot and soybean
Endpoint Effective Dose (ED)50
Values 2.27 L/ha
Reference 2007 Goltix OF Terrestrial Plants Toxicity, Vegetative Vigour
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 227
Dose Levels 0.313, 0.625, 1.25, 2.5 and 5.0 L/ha
Analytical measurements Yes- HPLC
Study Summary
The phytotoxicity of the test item Goltix OF to six terrestrial plant species was
determined each over a period of at least 21 days.
The test was conducted with the application rates 5.0, 2.5, 1.25, 0.625 and 0.313
L/ha. For the plant species carrot 0.156 L/ha was tested additionally. Test plants
were two monocotyledons (oats, Gramineae; onion, Liliaceae) and four
dicotyledons (sugar beet, Chenopodiaceae; rape, Brassicaceae; carrot,
Umbelliferae; soybean, Leguminosae). The test item was applied at test start onto
the foliage of the plants that reached the 2-4 leaf stage. The plants were watered
from the bottom and fertilised during the test with nutrient solution as needed. The
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toxic effects of the test item were assessed on day 7, 14 and 21 by visual
observations (phytotoxic effects and number of dead plants) and on day 21 by
shoot height and shoot fresh weight determination.
The lowest ED50 value was 2.27 L/ha for carrots. The lowest NOEL was 0.625 L/ha
for carrots.
Conclusion ED50 = 2.27 L/ha
Table 12: Nitrogen transformation test
Study type Nitrogen transformation
Flag Key study
Test Substance GOLTIX OF
Endpoint NOER
Value > 20 L/ha
Reference 2007 Effects of GOLTIX OF on the activity of soil microflora (Nitrogen
transformation test)
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 216
Dose Levels 2 L/ha and 20 L/ha
Analytical measurements None
Study Summary
This test examined the effects of the test substance on the soil microflora involved
in nitrogen transformation. The study was carried out in full accordance with the
OECD study guideline and fulfilled all the validity criteria. The test substance
caused no adverse effects (deviation from control <25%, OECD 216) on the soil
nitrogen transformation (measured as NO3-N production) at the end of the 28-day
incubation period up to an application rate of 20 L test item/ha.
Conclusion NOER = 20 L/ha
Table 13: Carbon transformation test
Study type Carbon transformation
Flag Key study
Test Substance GOLTIX OF
Endpoint NOER
Value > 20 L/ha
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Reference 2007 Effects of GOLTIX OF on the activity of soil microflora (Carbon
transformation test)
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 217
Dose Levels 2 L/h and 20 L/ha
Analytical measurements None
Study Summary
This test examined the effects of the test substance on the soil microflora involved
in carbon transformation. The study was carried out in full accordance with the
OECD study guideline and fulfilled all the validity criteria. The test substance
caused no adverse effects (deviation from control <25%, OECD 216) on the soil
carbon transformation at the end of the 28-day incubation period up to an
application rate of 20 L test item/ha.
Conclusion about soil toxicity
The formulated product triggers a 9.2B classification (ecotoxic in the soil environment). This is based on the
ED50 (equivalent to an ER50) for vegetative vigour of 2.27 L/ha. If the density of the product is 1.197 this
would be equivalent to 2.72 kg/ha. By using the EPA default values for converting field application rates to
units of milligrams of active ingredient per kilogram of dry soil this equates to an EC50 of 3.63 mg/kg soil.
Terrestrial invertebrate toxicity
Table 6: Honey bee acute toxicity test
Study type Honey bee acute oral and contact test 48 hour
Flag Key Study
Test Substance GOLTIX OF
Test species Honeybee Apis mellifera L.
Endpoint LD50
Values Oral LD50 > 181.55 ug/bee equivalent to >108 ug ai/bee
Contact LD50 >168.04 ug/bee equivalent to >100 ug ai/bee
Reference 2007 Acute toxicity of GOLTIX OF to the honeybee Apis mellifera L. under
laboratory conditions
Klimisch Score 1
Amendments/Deviations None
GLP Yes
Test Guideline/s OECD 213 and OECD 214
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Dose Levels Contact test: 168.04 ug product/bee
Oral test (nominal): 201.64 ug product/bee
Analytical measurements None
Study Summary
This test examined the oral and contact of the test substance on honey bees (see
dose levels above). The study was carried out in full accordance with the OECD
study guideline and fulfilled all the validity criteria. The test substance caused no
adverse effects at any of the doses tested.
The Oral LD50 was therefore determined to be > 181.55 ug/bee (equivalent to >108
ug ai/bee) while the contact LD50 was determined to be >168.04 ug/bee equivalent
to >100 ug ai/bee.
Conclusion Oral LD50 > 181.55 ug/bee equivalent to >108 ug ai/bee
Contact LD50 >168.04 ug/bee equivalent to >100 ug ai/bee
Table 8: Extended laboratory bioassay: Rove Beetles
Study type Extended laboratory bioassay
Flag Key Study
Test Substance AG-M4-700 OF1
Exposure 66 day exposure to fresh residues
Test species Rove Beetle Aleochara bilinerata
Endpoint NOER
Value 6000 mL/ha
Reference
2016 AG-M4-700 OF1- An extended laboratory bioassay of the effects of
fresh residues on the rove beetle Aleochara bilinerata (Coleoptera; Staphylinidae)
Klimisch Score 1
Amendments/Deviations None reported
GLP Yes
Test Guideline/s Grimm et al. (2000)
Dose Levels 500, 1000, 2000, 4000 and 6000 ml/ha
Analytical measurements None
Study Summary
This test examined the effects of fresh residues of the test item on adults of the
rove beetle Aleochara bilinerata and their larval offspring. The test item was
applied at rates of 500, 1000, 2000, 4000 and 6000 mL/ha onto replicate arenas
containing a natural sandy soil. Immediately after application 20 adult beetles of
equal sex ratio were introduced into each arena. The beetles were assessed at 1,
7 and 28 days after treatment. The success of the larval offspring of the original
beetles was assessed by the provision of ca. 500 onion fly pupae in each replicate
box on three weekly occasions i.e. at 7, 14 and 21 DAT. The original adult beetles
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were removed from the arenas at 28 days and the soil was left to dry for a week.
The fly pupae were sieved from the soil and the number of new adult beetles (in
the F1 progeny) that developed from the fly pupae was recorded over a further 31
days up until 66 DAT.
The results indicated that the NOER was 6000 mL/ha and that the ER50 >6000
mL/ha.
Conclusion NOER = 6000 mL/ha
Conclusion about terrestrial invertebrate toxicity
The formulated product does not trigger 9.4 classification