1、Designation: E 1566 00 (Reapproved 2006)Standard Guide forHandling Hazardous Biological Materials in Liquid Nitrogen1This standard is issued under the fixed designation E 1566; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the ye
2、ar of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.INTRODUCTIONHazardous biological materials maintained at liquid nitrogen temperatures require special handlingand storage p
3、rocedures. The penetration of vials by liquid nitrogen during storage can result incontamination of the nitrogen itself2or contamination of personnel resulting from improperly sealedvials exploding.3,4Ensuring safe storage conditions and taking precautions during the retrieval ofcultures from liquid
4、 nitrogen can help reduce the potential hazards. To avoid the penetration of vialsby liquid nitrogen and resulting hazards, they should be filled to a point that minimizes the air spacein the vial, and they shall be sealed completely. Vials may be examined for leaks by immersing themin an aqueous me
5、thylene blue (0.05 %) solution at 4C.1. Scope1.1 This guide covers recommended procedures for main-taining and handling hazardous biological materials at liquidnitrogen temperatures.1.2 This guide covers the safety precautions recommendedwhen handling material stored in liquid nitrogen.1.3 This guid
6、e does not cover the maintenance and handlingof hazardous biological materials maintained at cryogenictemperatures in systems other than liquid nitrogen.1.4 The values stated in SI units are to be regarded as thestandard.1.5 This standard does not purport to address all of thesafety concerns, if any
7、, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:5E 1342 Practice for Preservation by Freezing, Freez
8、e-Drying, and Low Temperature Maintenance of Bacteria,Fungi, Protista,Viruses, Genetic Elements, andAnimal andPlant Tissues3. Terminology3.1 Definitions:3.1.1 cryogenic temperaturestemperatures below or equalto 100C.3.1.2 hazardous biological materialsliving biological ma-terials, and products deriv
9、ed therefrom, that pose a potentialthreat to human health.3.1.3 liquid nitrogen storagestorage directly in liquidnitrogen or in the vapor phase above liquid nitrogen.4. Significance and Use4.1 This guide is intended for use by individuals maintain-ing and handling hazardous biological material in li
10、quidnitrogen freezers.4.2 This guide does not cover all aspects of every situationthat may be encountered in maintaining hazardous biologicalmaterial in liquid nitrogen; each situation must therefore beassessed individually using these guidelines.4.3 This guide is not intended for use with systems o
11、therthan liquid nitrogen storage.1This guide is under the jurisdiction ofASTM Committee E48 on Biotechnologyand is the direct responsibility of Subcommittee E48.02 on Characterization andIdentification of Biological Systems.Current edition approved Nov. 1, 2006. Published December 2006.Originally ap
12、proved in 1993. Last previous edition approved in 2000 asE 1566 00.2Shafer, T. W., Everett, J., Silver, G. H., and Came, P. E., “Biohazard:Virus-Contaminated Liquid Nitrogen,” Science, Vol 191, 1976, pp. 24-26.3Simione, F. P., Jr., Daggett, P.-M., McGrath, M. S., and Alexander, M. T., “TheUse of Pla
13、stic Ampoules for Freeze Preservation of Microorganisms,” Cryobiology,Vol 14, 1977, pp. 500-502.4Grieff, D., Melton, H., and Rowe, T. W., “On the Sealing of Gas-Filled GlassAmpoules,” Cryobiology, Vol 12, 1975, pp. 1-14.5For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact
14、ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.4.4 This guide does not co
15、ver practices for preservation byfreezing which are covered in Practice E 1342.5. Procedures5.1 All Liquid Nitrogen Freezers:5.1.1 Liquid nitrogen freezers vary from all-liquid storage,to liquid and vapor storage, to all-vapor storage. Freezers withall-liquid storage present the greatest hazard, and
16、 freezers withall-vapor storage present the least hazard. The type of systemin use should be assessed before attempting to apply theseguidelines.5.1.2 When working in freezers with all liquid storage, theoperator shall work in or near the liquid nitrogen wheninserting and retrieving material.5.1.3 F
17、reezers with both liquid and vapor storage reduce theproximity of the operator to the liquid nitrogen, but the risk ofliquid splashing remains.5.1.4 In all vapor freezers, the operator is protected from theliquid by a barrier.5.2 Liquid Storage Freezers:5.2.1 Hazardous biological material should nev
18、er be storeddirectly in liquid nitrogen. However, if a situation occurs inwhich hazardous biological materials are maintained in liquidnitrogen, the following guidelines should be followed:5.2.2 Wear a laboratory coat over a layer of protectiveclothing, such as a heavy jacket, that provides some pro
19、tectionfrom the splashing of liquid nitrogen.5.2.3 Wear insulated gloves over rubber surgical gloves.5.2.4 Wear a full face shield extended to cover the neck.5.2.5 Immerse the material slowly into liquid nitrogen inorder to avoid splashing.5.2.6 Remove the material from liquid nitrogen slowly.5.2.7
20、Immediately place the retrieved material into dry icefor transport, or into a metal canister containing a liquiddisinfectant for immediate thawing.5.3 Liquid/Vapor Storage Freezers:5.3.1 Protective clothing should include a laboratory coatand surgical gloves. A face shield should be worn whenretriev
21、ing from the liquid, and eye protection should be usedwhen retrieving from the vapor.5.3.2 Immerse the material slowly into liquid nitrogen inorder to avoid splashing.5.3.3 Remove the material from the liquid slowly, and allowit to warm to the temperature of the vapor phase prior toremoving it from
22、the freezer.5.3.4 Material stored in the vapor phase may be removedfrom the freezer immediately for transport in dry ice orthawing.5.3.5 Precautions shall be taken to ensure that the liquidnitrogen does not come into contact with material stored in thevapor phase.5.4 Vapor Storage Units:5.4.1 Protec
23、tive clothing should include a laboratory coat,surgical gloves, and eye protection.5.4.2 Material may be removed from the freezer immedi-ately for transport in dry ice or for thawing.5.5 Decontamination:5.5.1 Whenever a liquid nitrogen freezer, used to storehazardous biological material, is emptied
24、and warmed it shouldbe decontaminated. Decontamination procedures should bebased on the nature of the hazardous material maintained in theunit.5.5.2 When vial breakage occurs, involving hazardous bio-logical material, an assessment of the nature of the hazard shallbe made. If decontamination is requ
25、ired, the remaining mate-rial should be surface decontaminated rapidly in order to avoidwarming and moved to a different freezer. Before decontami-nating the empty liquid nitrogen freezer, it should be warmedto room temperature.6. Keywords6.1 cryogenic; hazardous biologicals; liquid nitrogenASTM Int
26、ernational takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the riskof infringement of such rights, are entirel
27、y their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five years andif not revised, either reapproved or withdrawn. Your comments are invited either for revision of this standard or for additional standardsand sh
28、ould be addressed to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of theresponsible technical committee, which you may attend. If you feel that your comments have not received a fair hearing you shouldmake your views known to the ASTM Committee on St
29、andards, at the address shown below.This standard is copyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959,United States. Individual reprints (single or multiple copies) of this standard may be obtained by contacting ASTM at the aboveaddress or at 610-832-9585 (phone), 610-832-9555 (fax), or serviceastm.org (e-mail); or through the ASTM website(www.astm.org).E 1566 00 (2006)2
