ASTM E1482-92(2004)
(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 test method covers the 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 concerns, 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 2004)
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 E 1053 Test Method for Efficacy of Virucidal Agents In-
tended for Inanimate Environmental Surfaces
1.1 This test method covers the use, in conjunction with
evaluations of the virucidal efficacy, of disinfectant solutions or
3. Summary of Test Method
pressurized disinfectant spray products intended for use on
3.1 After the exposure of a virus to a disinfectant, the
inanimate nonporous environmental surfaces or for other
virus-disinfectant suspension is applied to a column of Sepha-
special applications. The test method may be employed with all
dex LH60-120. The column is placed in a centrifuge and
viruses and host systems.
centrifuged to separate the virus from the disinfectant by gel
1.2 This test method should be performed only by persons
filtration. The filtrate (the column flow-through that contains
trained in microbiology and virology.
the virus) is assayed in the appropriate host system. The
1.3 This test method utilizes gel filtration technology. The
untreated virus control suspension is similarly gel filtered, and
effectiveness of the test method is dependent on the ratio of gel
the virus titer of the filtrate is determined by assay of
bed volume to sample size and uniformity in the preparation of
infectivity. The residual cytotoxicity of the disinfectant is
columns and centrifugation conditions. The effectiveness of
determined by gel filtration of the disinfectant control under the
this test method is maximized by investigator practice and
same conditions. Results for the virus inactivation and disin-
experience with gel filtration techniques.
fectant cytotoxicity of gel filtrates are recorded in the same
1.4 This test method will reduce, but not necessarily elimi-
manner as described in Test Methods E 1052 and E 1053. The
nate, disinfectant toxicity while preserving the titer of input
gel filtration procedures described in this test method are a
virus.
modification of the method of Blackwell and Chen.
1.5 The values stated in SI units are to be regarded as the
standard.
4. Significance and Use
1.6 This standard does not purport to address all of the
4.1 This test method is to be used for the removal of
safety concerns, if any, associated with its use. It is the
virucidal agents from agent-virus mixtures, or from agent-
responsibility of the user of this standard to establish appro-
neutralizer-virus mixtures, after the contact period and before
priate safety and health practices and determine the applica-
the inoculation of these mixtures into host systems for assay of
bility of regulatory limitations prior to use.
infectivity.
4.2 The purpose of the test method is to reduce the concen-
2. Referenced Documents
2 tration of agents and neutralizers in order to permit the
2.1 ASTM Standards:
evaluation of viral infectivity at dilutions that would otherwise
E 1052 Test Method for Efficacy of Antimicrobial Agents
be toxic to the host.
Against Viruses in Suspension
4.3 The test method is applicable to the testing of liquid and
pressurized disinfectant products.
4.4 This test method is compatible with organic soil loads,
This test method is under the jurisdiction of ASTM Committee E35 on
hard water, disinfectants containing organic solvents, and
Pesticides and Alternative Control Agents and is the direct responsibility of
chemical neutralizers.
Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved April 1, 2004. Published April 2004. Originally
approved in 1992. Last previous edition approved in 1998 as E 1482 - 92 (1998).
2 3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or Sephadex is a registered trademark of Amersham Biosciences.
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Blackwell, H. H., and Chen, J. H. S., “Effects of Various Germicidal Chemicals
Standards volume information, refer to the standard’s Document Summary page on on H.EP.2 Cell Culture and Herpes simplex Virus,” Journal of the AOAC, Vol 53,
the ASTM website. 1970, pp. 1229–1236.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E 1482 – 92 (2004)
5. Reagents and Materials (0.6 mL of sample to be added); a 10-cc syringe is used for 6-cc
columns (1.2 mL of sample to be added). Remove the cap from
5.1 Reagents:
the syringe tip, remove the plunger from the syringe, and place
5.1.1 Purity of Reagents—Reagent grade chemicals shall be
the syringe in an 18 by 150-mm test tube or in a suitable tube
used in all tests. Unless otherwise indicated, it is intended that
holder above a sink or liquid receptacle.
all reagents shall conform to the specifications of the Commit-
6.3 Place a small wad of glass wool in the syringe to cover
tee on Analytical Reagents of the American Chemical Society,
the internal tip opening. The wad should have a diameter
where such specifications are available. Other grades may be
approximately the same size as the internal syringe diameter,
used, provided it is first ascertained that the reagent is of
and it should
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