ASTM E2805-18

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: E2805 − 18
Standard Practice for
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Measurement of the Biological Activity of Ricin
This standard is issued under the fixed designation E2805; 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.
INTRODUCTION
Ricin is a member of the protein toxins that cause their physiological effect by inactivation of
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ribosomes.Ricinisamemberoftheclass2ribosomeinactivatingproteins (1). Othermembersofthis
class of toxins include the proteins abrin and Shiga toxin.
Ricinus communis (Ricin) consists of two chains, the A-chain is responsible for the N-glycosidase
enzymatic activity and the B-chain is needed for cell binding and intra-cellular processing. Ricin is a
heterogeneous protein with molecular weights ranging from approximately 62 to 64 kilodaltons (kDa)
(2). Both chains are glycosylated and of similar size (approximately 32 kDa). There are several genes
encoding putative ricin and ricin-like proteins in the genome of R. communis (3) resulting in
differences in the amino acid sequence of the subunits. The differences in amino acid sequence and
glycosylation both contribute to the heterogeneity of ricin. Various research reports exist describing
different means of detecting ricin activity.
1. Scope 1.5 Appendix X1 and Appendix X2 also provide guidance
for the measurement of the biological activity of ricin using
1.1 This guide is intended for the manufacturers and users
cell-based assays and the use of synthetic enzyme substrates.
of ricin reference material. Ricin reference materials are
well-characterized materials that can be used to test detection 1.6 Ricin is a category 2 select agent and acquisition of the
devicesandcalibratelaboratorymeasurements.Itisanticipated ricin standard must adhere to the Center for Disease Control
that ricin reference materials will be characterized by bio- and Prevention (CDC) regulations. Ricin is listed on the select
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chemical methods in addition to the measurement of biological agent list (42 CFR Part 72). The possession, transfer, and use
activity. of ricin are restricted under the Public Health Security Pre-
paredness Act (CRS Report RL31263 Public Health Security
1.2 This practice details the measurement of ricin biological
and Bioterrorism Preparedness and Response Act (P.L. 107-
activity using a cell-free translation (CFT) assay (4).
188): Provision and Changes to Preexisting law). Access to
1.3 The CFT assay has been developed for use in any
stores of ricin is limited (USA Patriot Act, P.L. 107-56). Ricin
biotechnology laboratory where determination or confirmation
is also a prohibited substance under the Biological Weapons
of ricin biological activity is required.
Convention and the Chemical Weapons Convention (CRS
1.4 The CFT assay has been validated by the U.S. Army Report RL31559 Proliferation Control Regimes: Background
and Status).
Medical Research Institute of Infectious Diseases (USAM-
RIID) VP-016 Validation of Cell-Free Translation Assay for
1.7 The values stated in SI units are to be regarded as
the Detection of Ricin Toxin Biological Activities in compli-
standard. No other units of measurement are included in this
ance (5) with Good Laboratory Practices (GLP) Regulations of
standard.
the Food and Drug Administration (21 CFR Part 58). Strict
1.8 This standard does not purport to address all of the
adherence to the protocol is necessary for validity of the test
safety concerns, if any, associated with its use. It is the
results.
responsibility of the user of this standard to establish appro-
priate safety, health, and environmental practices and deter-
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This practice is under the jurisdiction of ASTM Committee E54 on Homeland
mine the applicability of regulatory limitations prior to use.
Security Applications and is the direct responsibility of Subcommittee E54.01 on
Ricin is an extremely dangerous toxin. See Section 9 for
CBRNE Sensors and Detectors.
specific hazards information.
Current edition approved Dec. 1, 2018. Published December 2018. Originally
approved in 2011. Last previous edition approved in 2011 as E2805 – 11. DOI:
10.1520/E2805-18.
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The boldface numbers in parenthesis refer to the list of references at the end of
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this standard. Available at https://emergency.cdc.gov/agent/agentlist.asp.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
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