ASTM E1482-92(1998)
(Test Method)Standard Test Method for Neutralization of Virucidal Agents in Virucidal Efficacy Evaluations
Standard Test Method for Neutralization of Virucidal Agents in Virucidal Efficacy Evaluations
SCOPE
1.1 This laboratory test method is intended for use in conjunction with evaluations of the virucidal efficacy of disinfectant solutions or pressurized disinfectant spray products intended for use on inanimate nonporous environmental surfaces or for other special applications. The test method may be employed with all viruses and host systems.
1.2 This test method should be performed only by persons trained in microbiology and virology.
1.3 This test method utilizes gel filtration technology. The effectiveness of the test method is dependent on the ratio of gel bed volume to sample size and uniformity in the preparation of columns and centrifugation conditions. The effectiveness of this test method is maximized by investigator practice and experience with gel filtration techniques.
1.4 This test method will reduce, but not necessarily eliminate, disinfectant toxicity while preserving the titer of input virus.
1.5 The values stated in SI units are to be regarded as the standard.
1.6 This standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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Designation: E 1482 – 92 (Reapproved 1998)
Standard Test Method for
Neutralization of Virucidal Agents in Virucidal Efficacy
Evaluations
This standard is issued under the fixed designation E 1482; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope centrifuged to separate the virus from the disinfectant by gel
filtration. The filtrate (the column flow-through that contains
1.1 This laboratory test method is intended for use in
the virus) is assayed in the appropriate host system. The
conjunction with evaluations of the virucidal efficacy of
untreated virus control suspension is similarly gel-filtered, and
disinfectant solutions or pressurized disinfectant spray prod-
the virus titer of the filtrate is determined by assay of
ucts intended for use on inanimate nonporous environmental
infectivity. The residual cytotoxicity of the disinfectant is
surfaces or for other special applications. The test method may
determined by gel filtration of the disinfectant control under the
be employed with all viruses and host systems.
same conditions. Results for the virus inactivation and disin-
1.2 This test method should be performed only by persons
fectant cytotoxicity of gel filtrates are recorded in the same
trained in microbiology and virology.
manner as described in Test Methods E 1052 and E 1053. The
1.3 This test method utilizes gel filtration technology. The
gel filtration procedures described in this test method are a
effectiveness of the test method is dependent on the ratio of gel
modification of the method of Blackwell and Chen.
bed volume to sample size and uniformity in the preparation of
columns and centrifugation conditions. The effectiveness of
4. Significance and Use
this test method is maximized by investigator practice and
4.1 This test method is to be used for the removal of
experience with gel filtration techniques.
virucidal agents from agent-virus mixtures, or from agent-
1.4 This test method will reduce, but not necessarily elimi-
neutralizer-virus mixtures, after the contact period and before
nate, disinfectant toxicity while preserving the titer of input
the inoculation of these mixtures into host systems for assay of
virus.
infectivity.
1.5 The values stated in SI units are to be regarded as the
4.2 The purpose of the test method is to reduce the concen-
standard.
tration of agents and neutralizers in order to permit the
1.6 This standard does not purport to address all of the
evaluation of viral infectivity at dilutions that would otherwise
safety problems, if any, associated with its use. It is the
be toxic to the host.
responsibility of the user of this standard to establish appro-
4.3 The test method is applicable to the testing of liquid and
priate safety and health practices and determine the applica-
pressurized disinfectant products.
bility of regulatory limitations prior to use.
4.4 This test method is compatible with organic soil loads,
2. Referenced Documents hard water, disinfectants containing organic solvents, and
chemical neutralizers.
2.1 ASTM Standards:
E 1052 Test Method for Efficacy of Virucidal Agents In-
5. Reagents and Materials
tended for Special Applications
5.1 Reagents:
E 1053 Test Method for Efficacy of Virucidal Agents In-
2 5.1.1 Purity of Reagents—Reagent grade chemicals shall be
tended for Inanimate Environmental Surfaces
used in all tests. Unless otherwise indicated, it is intended that
3. Summary of Test Method all reagents shall conform to the specifications of the Commit-
tee on Analytical Reagents of the American Chemical Society,
3.1 After the exposure of a virus to a disinfectant, the
where such specifications are available. Other grades may be
virus-disinfectant suspension is applied to a column of Sepha-
dex LH60-120. The column is placed in a centrifuge and
Blackwell, H. H., and Chen, J. H. S., “Effects of Various Germicidal Chemicals
on H.EP.2 Cell Culture and Herpes simplex Virus,” Journal of the AOAC, Vol 53,
This test method is under the jurisdiction of ASTM Committee E-35 on 1970, pp. 1229–1236.
Pesticides and is the direct responsibility of Subcommittee E35.15 on Antimicrobial “Reagent Chemicals, American Chemical Society Specifications,” Am. Chemi-
Agents. cal Soc., Washington, DC. For suggestions on the testing of reagents not listed by
Current edition approved June 15, 1992. Published August 1992. the American Chemical Society, see “Analar Standards for Laboratory U.K.
Annual Book of ASTM Standards, Vol 11.05. Chemicals,” BDH Ltd., Poole, Dorset, and the “United States Pharmacopeia.”
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E 1482
used, provided it is first ascertained that the reagent is of approxima
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