MarkRogers.pdf

7/21/2019 MarkRogers.pdf

http://slidepdf.com/reader/full/markrogerspdf 1/75

Copyright © 2014 STERIS Corporation. All Rights Reserved. CONFIDENTIAL and PROPRIETARY to STERIS Corporation.

Marc Rogers, Ph.D.

Technical Service Specialist

Life Sciences

Formulated Chemistries

St. Louis, MO USA

AEBioS

Madrid

11 March 2014

Biological Indicators (BIs) &Chemical Indicators (CIs)



Copyright © 2014 STERIS Corporations. All Rights Reserved. CONFIDENTIAL and PROPRIETARY to STERIS Corporation.

Agenda

• Sterilization Methods• Biological Indicators (56*)

– Discs, Strips, Suspensions

– SCBI, PCD, Media, Custom BIs• Chemical Indicators (48*)

– CI Classification, examples

– Air Removal Tests

• ISO and EP Guidelines

* Number of items not including VHP®




Modes of Sterilization

• High Temperature – Steam (Moist Heat)

– Dry Heat

– SOP, SIP, Steam-Air

– Superheated water • Gas / Vapor

– Ethylene Oxide

– VHP, VHP Plasma

– Chlorine Dioxide, NO2

– Formaldehyde – Peracetic Acid

– Ozone

• Ionizing Radiation – Gamma Radiation

– E-Beam

– X-ray

• UV light (non-ionizing)• Mechanical (High

Pressure, Filtration)

• Liquid Chemical(oxidizers, aldehydes)




Terminal Sterilization Methods

Medical Device Manufacture

EO

50%

Gamma

42%

E-beam

4%

Heat and

Other

4%




Pharma/ Biopharm Drug

Sterilization

Aseptic

75%

Steam/Steam-

air mixture

Superheated

Water 23%

Other

2%

Terminal Sterilization

Aseptic Processing




Ethylene Oxide Sterilization

• Critical process parameters: – TIME (3-5 hours typical, depends on DEO value)

– TEMPERATURE (120 – 135 °F typical)

– GAS CONCENTRATION (400-800 mg/L)

– HUMIDITY (most 35-80% RH)• Where used:

– Terminal sterilize heat / moisture sensitive medical devices (e.g.catheters, implants, etc.)

– Usually done off site at large contract sterilization facilities (i.e.

ISOMEDIX, Griffith Microscience, Ethox, etc.)• Frequency

– Accounts for approx. 65% of medical devices sterilized in U.S.

– Validated using thermocouples, biological indicators, etc.


http://slidepdf.com/reader/full/markrogerspdf 7/75Copyright © 2014 STERIS Corporations. All Rights Reserved. CONFIDENTIAL and PROPRIETARY to STERIS Corporation.

Ethylene Oxide Sterilization

• Inactivation of Spores by EO:

– Reaction of EO with nucleic acids is the primary

cause of sporicidal activity

– Alkylation of DNA causes cells to lose ability toreproduce

http://en.wikipedia.org/wiki/File:Ethylene-oxide-from-xtal-3D-balls.png




VHP®

• Vaporized Hydrogen Peroxide – STERIS VHP®

– Room Decontamination

– Chambers

http://en.wikipedia.org/wiki/File:Hydrogen-peroxide-3D-balls.png




Vaporized Hydrogen Peroxide

• Critical process parameters: – TIME

– CONCENTRATION

– HUMIDITY

– SATURATION – TEMPERATURE

• Where used: – Transfer isolator or sterile environment within a barrier

isolator (VHP 1000ED)

• Newer application - terminal sterilization of heatsensitive medical devices (similar to EO) utilizingdeep prevacuum phase (VHP MD and V-Pro)




Dry Heat Sterilization

• Critical Process Parameters

– Temperature

– Time

• Commonly used for: – Glass, metal items

– Depyrogenation

• Generally any dry heat process > 220 °C

• Rather than utilize BIs, compendia recommend

demonstrating 3-log reduction of endotoxins

Initial Validation with TC and BI




Inactivation of Spores by Dry Heat (B. atrophaeus)

Destruction of microorganisms primarily by oxidation

Damage to DNA to induce mutations

Dry Heat Sterilization




Moist Heat / Steam Sterilization

• Equipment – Steam In Place (SIP) equipment

– Autoclaves

• Pre-Vacuum

• Gravity




Steam Sterilization

• Critical Process Parameters: – Time

– Temperature

– Saturated Steam

• Common uses: – Non heat/moisture-sensitive product components and

supplies

– Terminal sterilization of non-heat sensitive drugs

– Sterilization of equipment / components used in aseptic

manufacture – Research or test materials, supplies, glassware, culture

media, cages, bedding, etc.

– Decontamination of biohazardous or biological wastes

NOTE: FDA method of choice for the

sterilization of equipment and transfer lines

is saturated clean steam under pressure.




Anatomy of a Sterilization Cycle




Critical parameters of an

Autoclave Cycle

• Load Configuration

• Temperature Mapping

• Sterile Barrier System (Packaging)

– Penetration will depend on Density and Penetrability of

materials

• Time, Pressure & Temperature parameters

– Typical Parameters

• 121 °C for 15 minutes

• 134 °C for 3.5 minutes




Agenda



– SCBI, PCD, Media, Custom BIs

• Chemical Indicators (48*)








Sterilization – What is Sterility?

• Sterility Assurance Level

– In microbiology, it is impossible to prove that all

organisms have been destroyed/inactivated

– SAL: Probability of a single viable microorganismoccurring on an item after sterilization (e.g., 10-6)

– PNSU: Probability of a Non-Sterile Unit

From PDA Technical Report #1 (Revised 2007), Validation of Moist Heat

Sterilization Processes: Cycle Design, Development, Qualification, and

Ongoing Control

EN556-1:2001 Requirements for medical devices to be designated

“STERILE” – defines sterilization as an SAL = 10-6

17




Sterility Assurance

• European approach vs. U.S. approach• EU based on parametric release (declares

product is sterile based on records that process

parameters were delivered within specifiedtolerances (ISO 11139)

• U.S. - based on BI monitoring of

each load (CIs support BIs)

• CIs are an intrinsic part of each

– process indicators, in-pack indicators,

– Bowie Dick tests




What is Sterility Assurance? A way of evaluating or monitoring a sterilization process/ products

to ensure that all pathogens (disease causing microbes) havebeen inactivated/ destroyed and to provide protection from

recontamination; used as a supplement to physical monitoring.

Chemical Indicators

H i g h e r m o r e

q u a n t i f i a b

l e l e v e l o f

s t e r i l i t y a

s s u r a n c e

3 Levels of Sterility Assurance

Chemical Integrators

Biological Indicators

19





• What are they?

– Living microorganisms intended to present a worst-

case for a sterilization process (i.e. steam)

• Working principle – If a high number (population) of the most resistant

organisms to a particular sterilant can be inactivated

by sterilization , there is a high PROBABILITY that all

other organisms at that location in the sterilizer willalso be inactivated

20





• Common BI Uses: – Qualification testing of sterilization equipment.

– Validation of a sterilization process or cycle, or for

periodic revalidation, in conjunction with type “T”

thermocouples, Kaye Validators®, data loggers,recorders, etc.

– Routine monitoring of a sterilization cycle.

21




Biological Indicator Definitions

Spores The dormant stage of some microorganisms – most resistant to

environmental stress

Media The perfect food to grow spores (i.e., convert them into bacteria

that can reproduce)Incubation Spores usually grow fastest within fairly narrow temperature

ranges. The process of heating the media and spore to grow is

incubation

Population The number of spores on a BI – usually measured as 106 (i.e.,

1,000,000)

Controls Positive control (not sterilized) & negative control (sterile, so

should have no growth) used in validation

22




What spores are used?

• STEAM (& VHP®) – Geobacillus stearothermophilus

(GST, Gram positive)

– Formerly Bacillus stearothermophilus

– Thermophile (heat loving) – Growth at 55° to 60°C (131° to

140°F)

– No growth at 37°C (98.6°F)

– Not pathogenic to humans

• EO & Dry Heat – Bacillus atrophaeus (formerly

B. subtilis var. niger )

G. stearothermophilus

23Plomp M et al. 2005. Langmuir 21(23):10710-6.

Singh MK et al. 2009. NewJ Phys 11:115027




0.1

1

10

100

1,000

10,000

0.01

0 1 2 3 4 5 6 7 8 9 10 11 12

Number of Survivors Log Number of Survivors

4

3

2

1

0

-1

-2

D-Value

D-Value is the time to reduce

the microbial population by

90% or 1 log

Exposure Time (Minutes) 24

BI Survivor Curve




BI Formats Pros/Cons

SCBI Strips Discs Suspensions

Advantages •Complete

system

•No transfer

required

•No micro labrequired

•Broad

applicability

•Cost

•Size (6.3 mm

diameter)

•Cost

•Flexibility in

applications

•Allows for

direct

inoculation

Disadvantages •Not

submersible

•Size

•Requires

aseptic

transfer

•Not

individually

packaged

•Requires

aseptic

transfer

•D-value may

need to be

determined for

different

substrates

25




Suspensions

• 10 mL vial of bacterial spores in water – Comes with syringe for use in inoculation of product

– One vial is good for approximately 100 product inoculations

• Spores – GST & BA

• Population 103

to 109

/0.1 mL – 3,4,5,8,9 logs are Special Order

• Resistance – Available for:

– Steam Sterilization – 1.5 to 2.5

– Dry Heat Sterilization – 1.0 to 3.0 – EO Sterilization – 2.6 to 4.5

• Expiration – 18 mo. from DOM

• Storage at 2-8ºC (No RH req.)

26




Suspensions

• Applications

– Steam, Dry Heat and EO

– To directly inoculate a medical device to determine

the worst-case location where a spore strip or disccannot be used

– To directly inoculate a product in the places least

accessible to the sterilization agent

– To manufacture biological indicators using a substratesimilar or the same as the product being sterilized

– To monitor sterilization of liquids

27




Strips

• 1.5" X .25" strip of filter paper packaged in aglassine envelope

• Spores - GST & BA

– Spordex – single species

– Spordi – dual species

• Population - 1.0 X 104 to 5 X 106

• Resistance – D value 1.0 to 4.5 min

• Expiration -18 mo. from DOM

• Storage at 2-24ºC and 30-80% RH

– No Direct Sunlight or oxidative chemistry exposure

28




Strips

• Applications

– Most widely used format

– Validation and routine monitoring of steam, EO, and

Dry Heat sterilization cycles – Used in PCDs

– EO and Dry Heat standards

• USP, ANSI/AAMI/ISO/EP

– Steam standards• USP and ISO

29




Discs

• .25” round unpackaged filter paperdisc inoculated with spores


• Population - 1.0 X 106

to 5 X 106

• Resistance – D value 1.0 to 4.5

minutes

• Expiration – 18 mo. from DOM

• Storage at 2-24ºC and 30-80% RH

– No direct sunlight or oxidative

chemistry

30




Discs

• Applications

– Dry heat, steam and EO sterilization

– Commonly used in development of PCDs

– Because of their size they may be used in areaswhere strips will not physically fit

– Tool in determining worst case location within a

product

– USP

31




BI Media (Spordex®)

• Sterile modified soybean

casein broth

• Each lot validated for use with

Spordi and Spordex strips

• Meet current USPrequirements for growth

promotion

• All D-values for the BIs are

based on testing using

Spordex Media

– To duplicate our D-value, our

media should be used




SCBIs

• BI system designed to eliminatethe challenges and time needed

for aseptic handling


• Population – 1 X 105 to 5 x 106

• Resistance – D value ≥1.5 to

5.8 minutes

• Expiration – 12 mo from DOM• Storage – 20-25ºC and 30-80%

RH

33




SCBI Configuration

– Plastic vial and cap

– Paper disc inoculated with bacterial spores

– Crushable glass ampoule containing culture

media and pH indicators (bromthymol blue,

bromocresol purple)

– Process chemical indicator for steam or EOsterilization

– Dual species(S3060, S3061) and single

species (S3111, S3112)

– S3111 and S3112 are tested in accordance

with International Standard ISO 11138 andEU Standard EN866; Facilitates

harmonization of BIs between global

customer sites

Colors are an approximation

Positive

(Failure)Caramelized




SCBI Processing

1) Place the SCBI in the special

tool (Activator Set S3075)

3) Push or pull the Verify SCBI completely

through the gap on the side of the activator

to crush the media ampoule.

2) Press down

the blue cap to

close vial to

protect SCBI

from outsidecontaminates

4) Place the

Verify SCBI in

an incubatorat the

appropriate

temperature.

35




Incubating the SCBI

Incubation Temperature: 37°C (35°C - 39°C) EO

57°C (55°C - 60°C) Steam

Media does not evaporate if incubated for 7 days- 24 hour reduced incubation time for steam (S3060,S3061)

- 48 hour reduced incubation time for EO (S3060, S3061)

- Customers can validate a reduced incubation time (less than 7 days) for S3111, S3112

Available in 120 VAC

and 220 VAC




PCD

• Spores - GST & BA

• Population – 1 X 105 to 5 x 106

• Resistance - ≥ 1.4 to 5.8

minutes

• Expiration – 12 mo. from DOM• Storage at 20-25ºC and 30-

80% RH

• Accessories – Incubator

– Activator tool – S3075

S3065 Steam

EO PCD




Custom BI Products

– Pinecone Biologics can offer

a variety of custom products

– Pricing and timing are given

once the form in submitted

– Form available on the

intranet

• Life Sciences

• Sales and Marketing

• FC Pricing




Agenda












What are chemical indicators

(CIs)?

• A chemical indicator is any substance that gives

a visible sign, usually by a color change, of the

presence or absence of a threshold

concentration of a chemical species or physicalparameter

• In terms of steam sterilization, chemical

indicators may indicate:

– Time

– Temperature

– Steam/Water

40




What are chemical indicators

(CIs)?

• Chemical indicators do not:

– Indicate sterility: CIs provide a visual indication that

the indicator has been exposed to the physical

parameters (time, temperature, steam) required to

achieve sterility

– React based on lethality: Although microorganisms

may be killed at sub lethal temperatures (<121°C), a

CI will not indicate a pass unless ALL parameters

have been met

41




Classifications of CIs

• The ANSI/AAMI/ISO 11140-1:2005 documentdefines six classes of chemical indicators

• The indicators are subdivided within these 6classes based upon the information they relay tothe user

• The classification structure is used solely todenote the characteristics and intended use ofeach type of indicator as defined by themanufacturer. The classification has nohierarchical significance.”

42





Class ANSI/AAMI/ISO 11140-1:

2005 Definition

Practical Appl ication

Class 1:

Process Indicators

(PCC003, PCC004)

Verify EO

(PCC009, PCC010)

Verify Steam

(260010) steam tape

(410100) EO tape

“Process indicators are

intended for use with

individual units, (e.g.,

packs, containers) toindicate that the unit has

been directly exposed to

the sterilization process

and to distinguish

between processed and

unprocessed units. Theyshall be designed to react

to one or more of the

critical process

variables.”

Indicator tapes, indicator

labels, and load cards

are

examples of externallyvisible Chemical

Indicators that

are Process Indicators

used for exposure control

– VHP and Dry Heat

chemical indicatorscommonly fall into this

category

43






2005 Definition

Practical Application

Class 2:

Indicators

for use

in SpecificTests

“Class 2 indicators are

intended for use in specific

test procedures as defined

in relevantsterilizer/sterilization

standards.”

Bowie Dick type tests are specific

tests used for equipment control to

evaluate the sterilizer

performance.

44

e.g. EQC003 Steraffirm™ Bowie Dick Test Pack; EQC009, EQC010 Verify®

Bowie Dick Test Packs; NB113, NB125 DART ® Daily Air Removal Test; S3095

Verify ® Bowie Dick Test Card






2005 Definition


Class 3:

Single

Variable

Indicators

“A single variable indicator

shall be designed to react to

one of the critical variables

and is intended to indicateexposure to a sterilization

process at a stated value

(SV) of the chosen

variable.”

An example of a Single Variable

Indicator is a

temperature tube that contains a

chemical pellet that melts at aspecific temperature. Single

variable indicators may be used for

pack control monitoring but would

not provide as much information

as a Class 4 or Class 5 Chemical

Indicator. Single VariableIndicators may also be used for

exposure control monitoring.

45





Class ANSI/AAMI/ISO

11140-1: 2005

Definition


Class 4:

Multi-

variable

Indicators

“A multi-variable

indicator shall be

designed to react

to two or more ofthe critical

variables and is

intended to indicate

exposure to a

sterilization cycle at

SVs of the chosenvariable.”

Multi-variable Chemical Indicators are used

for pack control. These internal Chemical

Indicators are usually paper strips printed

with a Chemical Indicator.

46

e.g. Steraffirm™ Sterilizer Control Tube 121°C 15 min (PCC024), and 12 min

(PCC025); at 124°C for 3 min (PCC026)




Classifications of CIsClass ANSI/AAMI/ISO

11140-1: 2005Definition


Class 5:

Integrating

Indicators

PCC039

VERIFY®

Integrating

Indicator

for Steam

“Integrating

indicators shall be

designed to react

to all criticalvariables. The SVs

are generated to

be equivalent to, or

exceed the

performance

requirements givenin the ISO 11138

series for BIs.”

Integrating Indicators are the most accurate

of the internal Chemical Indicators.

Integrating Indicators are used for pack

control monitoring. They can also be used asan additional monitoring tool to release loads

that do not contain implants. For this

additional monitoring the Class 5 Integrating

Indicator must be used in the appropriate

challenge test pack or Process Challenge

Device (PCD). These indicators must nowhave SVs at 121°C/250°F, 135°C/276°F, and

at least one more temperature in between.

Also, the SV at 121°C MUST be greater than

16.5 minutes to ensure performance is

comparable to BIs in saturated steam.

47




Classifications of CIsClass ANSI/AAMI/ISO

11140-1: 2005Definition


Class 6:

Emulating

Indicators

Steraffirm™Sterilizer

Control

Tubes

PCC027,

PCC028

Steraffirm™Steam Class

6 Emulating

Indicators

PCC012

PCC013

PCC014

“Emulating

indicators are cycle

verification

indicators which

shall be designed toreact to all critical

variables for

specified

sterilization cycles.

The SVs are

generated from the

critical variables of

the specified

sterilization

process.”

Emulating Indicators can be used as

internal Chemical Indicators for pack

control. These indicators are indentified for

specific sterilization cycles which means an

end user will need to inventory a differentClass 6 Emulating Indicator for each

sterilization cycle time and temperature

(i.e., 3 min, 5 min, 10 min, 18 min, 40

min,etc.) performed in the facility. The

response of a Class 6 Emulating Indicator

does not necessarily correlate to a BI so the

indicator cannot be used as an additional

monitoring tool to release loads that do not

contain implants. (See Class 5 definition)

48




Chemical Indicators

• May consist of sensitivechemical ink/dye applied to

paper or in liquid

• Immediate visual indication of

exposure to a specific sterilant(e.g. steam, EO, dry heat)

• Assist detection of sterilizer

malfunctions and personnel

errors

• Many different types

available




Steraffirm Class 6 Emulating

Indicators

• Cycle Specific – Steam sterilization

• Measures temperature, saturated steam and time

– 121C/250F for 15 minutes

– Use inside individual packs, trays or pouches

– Other indicators available for different cycles

50




Verify® Integrating Class 5

Indicator (PCC039)

– Steam sterilization

• Measures temperature, saturated steam and time

– Stated Values are 121ºC (18 min), 132ºC (3 min), and

134ºC (2.5 min)

– Placed into each pack to be steam sterilized

– No ink color interpretation

• Clear and unambiguous

– Correlated to bacterial spore death

51




Difference between Class 5 & 6

• Class 6 emulating indicators

– Cycle specific

– React to all critical variables within a specific cycle

• Class 5 integrating indicators

– Not cycle specific

– React to all critical variables, designed to meet or

exceed the BI performance requirements

52

S ff S C




• Glass tube with a heat sensitive liquidwhich will change color from red to green

• Used routine monitoring of gravity,prevacuum or liquid cycles

• Product Range:• Steam (ISO Class 4) - dry goods or liquids

– 134C 3 mins

– 121C 15 mins

– 121C 12 mins

• Dry Heat (ISO Class 6)

– 160C 120 mins

– 160C 60 mins

(Note: can vary times and temperaturesbased on Tech Data –e.g. low temp, raisetime)

Steraffirm™ Sterilization Control

Tubes




VHP® BIs and CIs

• Spordex® S24 SCBI (NA340), 24-h readout

• Verify ® S24 SCBI Activator (LCB004)

• Steraffirm® Process Indicator (PCC049, PCC050)

• Steraffirm

®

Process Indicator PCC051

PCC049




VHP® BIs and CIs




VHP® BIs and CIs




VHP® BIs and CIs

NA117, cultureMedia for use with

NA333 BIs, 24-h

RIT

NA333, E6 GST 12980 on SS in

Tyvek®

Steraffirm® PCC060 for

Intermediate to high

H2O2 vapor concentrations




Agenda












Testing for Air Removal

• Bowie and Dick Type tests for air removal

– Utilize a steam sensitive chemical indicator and

porous barrier or challenge material (e.g., paper,

cloth)

– Test pack serves as a “heat sink” in otherwise empty

chamber

– Any air not removed during prevac cycles will be

concentrated in test pack

– Residual air prevents steam contact with chemical

indicator, preventing color change

59

H th B i Di k T t D t t




How the Bowie Dick Test Detects

Air (Illustrated)

CHAMBER TRAP

STERILIZER DRAIN

BD TEST

STEAM

AIR

AIR

AIR

AIR AIR

60




Air Removal Indicators

• Steraffirm™ Bowie DickTest Pack

– Tests air removal of pre-

vacuum sterilizer cycles

with cycles of 121-124 ºC – Most popular

– Easy to use - single use

– Diagnostic capability –

pattern of failure can helpisolate problem

– Lead free ink




Available Bowie Dick Test Packs

A range of preassembled, single use Bowie Dick Test Packs:

134°C-137°C (273°F-279°F) for 3.5 minutes

132°C (270°F) for 3 to 4 minutes

121°C- 124°C (250°F-255°F) for 8 – 8.3 minutes*

*For use by any LS customer using a prevacuum steam sterilization cycle

Steraffirm™ Bowie Dick Test




AIR REMOVAL NON-CONDENSABLES WET STEAM

SUPERHEAT

Steraffirm™ Bowie Dick Test

Packs

DART® TEST




DART® TEST

Dart Temperature Range:

For prevacuum cycles at 132-134°C for 3.5-4 minutes only

Preassembled, Single Use Test




Agenda









Regulatory Guidance




Regulatory Guidance

Standards Hierarchy




EN Standards




EN ISO 11138 Series

• EN ISO 11138-1* was first published 1994,revised 2006

• It specifies requirements for the labeling and

performance of BIs

• Subsequent parts give performance

requirements of BIs for specific sterilization

methods, e.g. ethylene oxide (Part 2), moist heat

(Part 3), dry heat (Part 4), formaldehyde (Part 5)• EN ISO 11138-6 hydrogen peroxide vapor

processes in development

*Sterilization of health care products – Biological Indicators – Part 1: General requirements




EN ISO 11140 Series

• EN ISO 11140-1* first published 1995, revised 2005

• Specifies requirements for labeling and performance of

CIs

• Subsequent parts give performance requirements for

Bowie Dick type indicators

• Class 2 indicator systems for use in the Bowie and Dick-

type steam penetration test (Part 3)

• Alternatives to B-D, European (Part 4)

• Alternatives to B-D, USA (Part 5)• Part 6 in development – Class 2 indicators and process

challenge devices for use in performance testing of

steam sterilizers*Sterilization of health care products – Chemical indicators – Part 1: General requirements

Requirements for BI Spore




Requirements for BI Spore

Population (per unit)

Requirements for BI D-Value




Sterilization Guidance




EN ISO 14161

• First published in 2000• Guidance on selection, use, and interpretation of

results of BIs when used to develop, validate

and monitor sterilization processes

•First published 2003

•Gives guidance for users on the selection,use, and interpretation of results of CIs

EN ISO 15882




Process Validation Guidance




Agenda











QUESTIONS?

[email protected]

Documents

MarkRogers.pdf