ASTM E3002-15(2023)
(Practice)Standard Practice for Assessing the Comparative Efficacy of Products Used for the Decontamination of Chemical Warfare Agents (CWAs) on Skin
Standard Practice for Assessing the Comparative Efficacy of Products Used for the Decontamination of Chemical Warfare Agents (CWAs) on Skin
SIGNIFICANCE AND USE
4.1 This practice specifies an in-vivo measurement of CWA decontamination on the skin.
4.2 CWA skin decontaminants will have different modes of action including absorption, adsorption, removal, chemical neutralization or some combination of the above. There is, therefore, no single representative in-vitro method for validation of decontamination efficacy of products for skin decontamination. For example, measuring the presence of a radiolabelled chemical warfare agent after chemical neutralization, may give a false positive results. It has been shown that if the agent has been chemically neutralized, the radiolabel may still be present in a non-toxic molecule. In addition, some chemical neutralization methods may break down the original agent, but the breakdown product is highly toxic. In the case of VX, hydrolysis produces a highly toxic product, EA2192 (S-(2-diisopropylaminoethyl) methylphosphonothioic acid (8).
4.3 This standard practice is of significance in that efficacy is thoroughly evaluated to the extent possible to represent use on human skin. In-vivo studies have demonstrated that simple chemical monitoring for disappearance of the chemical agent may not be sufficient to measure decontamination and neutralization effectiveness. A standard practice is needed for determining actual decontamination and neutralization by measuring the decrease in mortality or lesion size caused by the agent.
SCOPE
1.1 This practice establishes an in-vivo method for assessing the comparative efficacy of products used for the decontamination of chemical warfare agents (CWAs) on the skin.
1.2 This practice provides a quantitative efficacy comparison of different skin decontamination products.
1.3 To minimize the number of animals used, this in-vivo practice should be performed only after rigorous in-vitro studies of the candidate decontaminant, which can show the implied claims including chemical neutralization, decontamination studies on surfaces and appropriate testing such as cytotoxicity.
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.5 This standard does not purport to address all of the safety concerns, if any, associated with the use of decontamination products or CWAs. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
General Information
Relations
Standards Content (Sample)
This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E3002 − 15 (Reapproved 2023)
Standard Practice for
Assessing the Comparative Efficacy of Products Used for
the Decontamination of Chemical Warfare Agents (CWAs)
on Skin
This standard is issued under the fixed designation E3002; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 2.1.1.1 Discussion—The most common chemical warfare
agents are: (a) nerve agents—tabun (GA), sarin (GB), soman
1.1 This practice establishes an in-vivo method for assessing
(GD), cyclosarin (GF), VX; and (b) blister agents (or
the comparative efficacy of products used for the decontami-
vesicants)—mustard and lewisite. (1 and 2)
nation of chemical warfare agents (CWAs) on the skin.
2.1.2 decontamination, n—the process of physical removal
1.2 This practice provides a quantitative efficacy compari-
or chemical neutralization, or both, of CWAs to decrease or
son of different skin decontamination products.
prevent health effects due to a dermal contamination.
1.3 To minimize the number of animals used, this in-vivo
2.1.3 in-vitro study, n—study or protocol performed outside
practice should be performed only after rigorous in-vitro
of a living organism, either with or without the use of a
studies of the candidate decontaminant, which can show the
biological material.
implied claims including chemical neutralization, decontami-
2.1.4 in-vivo study, n—study using a whole living organism.
nation studies on surfaces and appropriate testing such as
cytotoxicity. 2.1.5 Organophosphate Agent (OP), n—the general name
for esters of phosphoric acid that are toxic through inhibition of
1.4 The values stated in SI units are to be regarded as
the enzyme acetylcholinesterase.
standard. No other units of measurement are included in this
2.1.6 Protective Ratio (PR), n—the LD of the decontami-
standard.
nated animals divided by the LD of the positive control
1.5 This standard does not purport to address all of the
(exposed to CWAs and not decontaminated) animals (3-5).
safety concerns, if any, associated with the use of decontami-
2.1.7 vesicant agent—a chemical agent that causes burns
nation products or CWAs. It is the responsibility of the user of
and destruction of tissue.
this standard to establish appropriate safety, health, and
environmental practices and determine the applicability of
2.2 Acronyms:
regulatory limitations prior to use.
2.2.1 GA—common name: Tabun; IUPAC name: (Ethyl
1.6 This international standard was developed in accor-
dimethylphosphoramidocyanidate): Organophosphate nerve
dance with internationally recognized principles on standard-
agent.
ization established in the Decision on Principles for the
2.2.1.1 Discussion—This nerve agent is the easiest to manu-
Development of International Standards, Guides and Recom-
facture. Consequently, it is more likely that developing coun-
mendations issued by the World Trade Organization Technical
tries start their CW arsenal with this nerve agent whereas
Barriers to Trade (TBT) Committee.
industrialized countries consider Tabun to be out-of-date and of
limited use.
2. Terminology
2.2.2 GB—common name: Sarin; IUPAC name: ((RS)-
2.1 Definitions of Terms Specific to This Standard:
Propan-2-yl methylphosphonofluoride) Organophosphate
2.1.1 Chemical Warfare Agents (CWA), n—toxic chemicals
nerve agent.
that have been used as chemical weapons, or have been
2.2.2.1 Discussion—GB is a volatile substance mainly taken
developed for use as chemical weapons.
up through inhalation.
2.2.3 GD—common name: Soman; IUPAC name: (O-
1 Pinacolyl methylphosphonofluoridate Organophosphate nerve
This practice is under the jurisdiction of ASTM Committee E54 on Homeland
Security Applications and is the direct responsibility of Subcommittee E54.01 on
agent.
CBRNE Detection and CBRN Protection.
Current edition approved Sept. 1, 2023. Published October 2023. Originally
approved in 2015. Last previous edition approved in 2015 as E3002 – 15. DOI: The boldface numbers in parentheses refer to a list of references at the end of
10.1520/E3002-15R23. this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E3002 − 15 (2023)
2.2.3.1 Discussion—A moderately volatile substance which 4. Significance and Use
can be taken up by inhalation or skin contact.
4.1 This practice specifies an in-vivo measurement of CWA
2.2.4 GF—common name: Cyclohexyl sarin; IUPAC name: decontamination on the skin.
(Cyclohexyl methylphosphonofluoridate) Organophosphate
4.2 CWA skin decontaminants will have different modes of
nerve agent.
action including absorption, adsorption, removal, chemical
2.2.4.1 Discussion—A substance with low volatility which
neutralization or some combination of the above. There is,
is taken up through skin contact and inhalation of the substance
therefore, no single representative in-vitro method for valida-
either as a gas or aerosol.
tion of decontamination efficacy of products for skin decon-
tamination. For example, measuring the presence of a radiola-
2.2.5 HD—common name: Mustard or Distilled Sulfur
belled chemical warfare agent after chemical neutralization,
Mustard; IUPAC name: (bis(2-chloroethyl) sulfide); Vesicant.
may give a false positive results. It has been shown that if the
2.2.5.1 Discussion—In its pure state, mustard agent is col-
agent has been chemically neutralized, the radiolabel may still
orless and almost odorless.
be present in a non-toxic molecule. In addition, some chemical
2.2.6 L—common name: Lewisite: IUPAC name: (2-
neutralization methods may break down the original agent, but
chloroethenylarsonous dichloride). Vesicant.
the breakdown product is highly toxic. In the case of VX,
hydrolysis produces a highly toxic product, EA2192 (S-(2-
2.2.7 LD —a standard measure of toxicity. The individual
dose required to kill 50 % of the animals in a test population. diisopropylaminoethyl) methylphosphonothioic acid (8).
4.3 This standard practice is of significance in that efficacy
2.2.8 VX—common name: VX, IUPAC name: (O-ethyl
is thoroughly evaluated to the extent possible to represent use
S-[2-(diisopropylamino)ethyl] methylphosphonothioate).
on human skin. In-vivo studies have demonstrated that simple
2.2.8.1 Discussion—Organophosphate nerve agent, a persis-
chemical monitoring for disappearance of the chemical agent
tent substance which can remain on material, equipment and
may not be sufficient to measure decontamination and neutral-
terrain for long periods. Update is mainly through the skin but
ization effectiveness. A standard practice is needed for deter-
also through inhalation of the substance as a gas or aerosol.
mining actual decontamination and neutralization by measur-
ing the decrease in mortality or lesion size caused by the agent.
3. Summary of Practice
3.1 Due to the extreme hazards of the chemical warfare
5. Reagents
agents, the efficacy of decontamination products cannot be
5.1 All the CWAs for these experiments shall be synthesized
evaluated in a human clinical study. This practice has been
3 in the laboratory where the experiments will be performed or
used to support FDA clearance (for example, RSDL® (6 and
obtained from a legitimate external source which shall be
7) 510k – K023969) for decontamination devices for use on
included in the report.
human skin (3-5).
5.2 All test materials shall be at the same temperature as that
3.2 Determination of the efficacy of decontamination prod-
of the room where the test is conducted at the time of
ucts for use on the skin against these toxic compounds requires
application to the animals.
in-vivo data, which are more physiologically relevant than
5.3 Appropriate solvents shall be purchased from legitimate
in-vitro studies. This practice provides a methodology for
vendors as required and disclosed in the report.
obtaining comparative in-vivo data.
5.4 The standard decontaminant used for comparison should
3.3 The practice is used in order to calculate the Protective
be the product currently accepted for use by the majority of
Ratio of the decontaminant. The Protective Ratio is the LD of
Defense Forces and First Responders in the world, which at
animals treated with the chemical agent and decontaminated,
this point in time is the RSDL® skin decontamination system.
divided by the LD of control animals (animals treated with
the contaminant and not decontaminated) measured 24 h after
6. Procedure
exposure to the CWA.
6.1 A reputable laboratory animal supplier must be used for
3.4 This practice is based on decontamination efficacy of
any specified species or strain. Only a single constant source of
decontamination products against two nerve agents and one
supply for each species should be used by the testing laboratory
blister agent, for a total of three CWAs. The nerve agents are
to maintain genetically homogenous test subjects.
either G-agents or V-agents based on their chemical structures.
The two nerve agents included in the practice are GD (from
6.2 Animal Care—All animal care should conform to the
G-agents) and VX (from V-agents). The blister agent included
appropriate standards (Association for Assessment and Ac-
in this practice is HD.
creditation of Laboratory Animal Care International AAALAC;
http://www.aaalac.org/) and all animal test protocols must have
appropriate approvals.
The sole source of supply of RSDL known to the committee at this time is
6.3 As required by the Association for Assessment and
Emergent BioSolutions, 400 Professional Drive, Suite 400, Gaithersburg, MD,
Accreditation of Laboratory Animal Care (AAALAC), animal
20879. If you are aware of alternative suppliers, please provide this information to
care and recording of the information including, but not limited
ASTM International Headquarters. Your comments will receive careful consider-
ation at a meeting of the responsible technical committee, which you may attend. to, the following will be expected:
E3002 − 15 (2023)
6.3.1 Condition of the Animals—Only animals that show no 6.5 A patient monitor shall be used to continually monitor
signs of ill health shall be used for these studies. animals for physiological parameters such as heart rate, blood
pressure, arterial oxygen saturation, respiration rate, and tem-
6.3.2 Pre-test Conditioning—All animals to be used in these
perature and these parameters shall be recorded.
studies shall be quarantined for a designated period of time
dependent on species and strain and source of the animal to
7. Test Methods
ensure good health prior to use. The period of quarantine
7.1 This practice utilizes the clipped hair guinea pig model
should be appropriate for the species and strain of animal. An
(3-5). With proper documentation, other animal models with
adequate diet for the species and fresh water shall be allowed
skin similar to human skin, such as hairless guinea pig, rabbit,
with free access. Each animal shall be identified, and food
minipig, or swine, can be used.
consumption should also be noted. Laboratory animal housing
and test environmental conditions shall be maintained accord-
7.2 Animals should be anaesthetized during the study.
ing to the acceptable animal care accreditation. Any significant
Anesthesia should be appropriate to the animal model selected.
deviations shall be noted and reported.
7.3 Exposure to the nerve agents is done by applying a
6.3.3 Weight of Test Animal—Pretest weights of the test
single dose to the application site 5 min after anesthesia. If
animal shall be taken five days before and just prior to
needed to improve dosing accuracy, diluted agent may be used.
application of the test material. Only animals that either
NOTE 1—Use caution when handling nerve agents.
maintain their weight or show an increase equivalent to others
7.4 Exposure to the blister agent is done by applying three
of the same age, sex, and starting weight shall be used for the
doses to three application sites 5 min after anesthesia. If
studies. In the selection of the test subjects an attempt should
required, to improve dosing accuracy, diluted agent may be
be made to produce as nearly identical groups as possible.
used.
6.3.4 Number of Test Animals—All of the studies designed
to measure efficacy of decontaminants need two (2) sets of
NOTE 2—Use caution when handling blister agents.
animals. The first set of animals is used to identify the
7.5 Two minutes after agent application, the dosing site
percutaneous 24 h LD for nerve agents or the appropriate
shall be untreated (control) or decontaminated (per manufac-
dosage range to create measurable lesion sizes. The second set
turer instructions for use) with the test and standard or
of animals is used to test the comparative efficacy of the
comparator decontamination product.
decontamination products. For determining LD values of
7.6 Application method for the decontamination product is
nerve agents without decontamination, 3 animals per dose, 5
to be carefully thought out prior to application to ensure that
doses per agent will be tested. Thus for the two nerve agents
manufacturer’s instructions can be scaled down to the appro-
(GD and VX), 3 animals per dose × 5 doses per agent × 2 nerve
priate treatment area.
agents = 30 animals are required. For determining LD values
with decontamination, three stages, 1 to 2 animals per dose, 4 7.7 Determination of LD of nerve agents a process which
to 5 doses will be tested using sequential stage wise methods (9 is essentially identification of the dose response curve, should
and 10). For the blister agent, 2 sets of animals × 3 animals per be measured by using doses varying from non-lethal to lethal
set × 1 blister agent = 6 animals required. All test groups by establishing 24 h dose response curves in sets of animals
should be evenly divided by sex. receiving various doses of agent spanning the non-lethal to
lethal dose range using sequential stage wise methods (9 and
6.3.5 Animal Preparation—Each animal should be exam-
10). Randomly assign the animals to each treatment group and
ined 24 h prior to testing. No animals showing signs of ill
assess mortality at 24 h. The LD estimate value is complete
health shall be sued for the testing. The experimenter should 50
when the difference between the upper and lower 95 %
carefully remove the hair from the backs and sides of
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