ASTM E1566-00

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation:E1566–00
Standard Guide for
Handling Hazardous Biological Materials in Liquid Nitrogen
This standard is issued under the fixed designation E1566; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
Hazardous biological materials maintained at liquid nitrogen temperatures require special handling
and storage procedures. The penetration of vials by liquid nitrogen during storage can result in
contamination of the nitrogen itself or contamination of personnel resulting from improperly sealed
3,4
vials exploding. Ensuring safe storage conditions and taking precautions during the retrieval of
cultures from liquid nitrogen can help reduce the potential hazards. To avoid the penetration of vials
by liquid nitrogen and resulting hazards, they should be filled to a point that minimizes the air space
in the vial, and they shall be sealed completely. Vials may be examined for leaks by immersing them
in an aqueous methylene blue (0.05%) solution at 4°C.
1. Scope E 1342 Practice for Preservation by Freezing, Freeze-
Drying, and Low Temperature Maintenance of Bacteria,
1.1 This guide covers recommended procedures for main-
Fungi,Protista,Viruses,GeneticElements,andAnimaland
taining and handling hazardous biological materials at liquid
Plant Tissues
nitrogen temperatures.
1.2 This guide covers the safety precautions recommended
3. Terminology
when handling material stored in liquid nitrogen.
3.1 Definitions:
1.3 Thisguidedoesnotcoverthemaintenanceandhandling
3.1.1 cryogenic temperatures—temperaturesbeloworequal
of hazardous biological materials maintained at cryogenic
to−100°C.
temperatures in systems other than liquid nitrogen.
3.1.2 hazardous biological materials—livingbiologicalma-
1.4 The values stated in SI units are to be regarded as the
terials, and products derived therefrom, that pose a potential
standard.
threat to human health.
1.5 This standard does not purport to address all of the
3.1.3 liquid nitrogen storage—storage directly in liquid
safety problems, if any, associated with its use. It is the
nitrogen or in the vapor phase above liquid nitrogen.
responsibility of the user of this standard to establish appro-
priate safety and health practices and determine the applica-
4. Significance and Use
bility of regulatory limitations prior to use.
4.1 This guide is intended for use by individuals maintain-
2. Referenced Documents ing and handling hazardous biological material in liquid
5 nitrogen freezers.
2.1 ASTM Standards:
4.2 This guide does not cover all aspects of every situation
that may be encountered in maintaining hazardous biological
ThisguideisunderthejurisdictionofASTMCommitteeE48onBiotechnology material in liquid nitrogen; each situation must therefore be
and is the direct responsibility of Subcommittee E48.02 on Characterization and
assessed individually using these guidelines.
Identification of Biological Systems.
4.3 This guide is not intended for use with systems other
Current edition approved Sept. 10, 2000. Published December 2000.
than liquid nitrogen storage.
Originally published as E1566-93. Last previous edition E1566-93.
Shafer, T. W., Everett, J., Silver, G. H., and Came, P. E., “Biohazard:
4.4 This guide does not cover practices for preservation by
Virus-Contaminated Liquid Nitrogen,” Science, Vol 191, 1976, pp. 24-26.
freezing which are covered in Practice E1342.
Simione, F. P., Jr., Daggett, P.-M., McGrath, M. S., andAlexander, M.T., “The
Use of PlasticAmpoules for Freeze Preservation of Microorganisms,” Cryobiology,
5. Procedures
Vol 14, 1977, pp. 500-502.
Grieff, D., Melton, H., and Rowe, T. W., “On the Sealing of Gas-Filled Glass
5.1 All Liquid Nitrogen Freezers:
Ampoules,” Cryobiology, Vol 12, 1975, pp. 1-14.
5.1.1 Liquid nitrogen freezers vary from all-liquid storage,
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
to liquid and vapor storage, to all-vapor storage. Freezers with
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on all-liquid storage present the greatest hazard, and freeze
...