1、Designation: E1566 00 (Reapproved 2014)Standard Guide forHandling Hazardous Biological Materials in Liquid Nitrogen1This standard is issued under the fixed designation E1566; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year
2、 of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.INTRODUCTIONHazardous biological materials maintained at liquid nitrogen temperatures require special handlingand storage proc
3、edures. 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 ni
4、trogen 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 methy
5、lene 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 guide d
6、oes 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 asstandard. No other units of measurement are included in thisstandard.1.5 This standard does not
7、 purport to address all of thesafety concerns, if any, 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
8、:5E1342 Practice for Preservation by Freezing, Freeze-Drying,and Low Temperature Maintenance of Bacteria, Fungi,Protista, Viruses, Genetic Elements, andAnimal and PlantTissues (Withdrawn 2011)63. Terminology3.1 Definitions:3.1.1 cryogenic temperaturestemperatures below or equalto 100C.3.1.2 hazardou
9、s biological materialsliving biologicalmaterials, and products derived 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 indiv
10、iduals maintain-ing and handling hazardous biological material in liquidnitrogen freezers.4.2 This guide does not cover all aspects of every situationthat may be encountered in maintaining hazardous biological1This guide is under the jurisdiction of ASTM Committee E55 on Manufactureof Pharmaceutical
11、 Products and is the direct responsibility of Subcommittee E55.04on General Biopharmaceutical Standards.Current edition approved Dec. 1, 2014. Published January 2015. Originallyapproved in 1993. Last previous edition approved in 2006 as E1566 00 (2006).DOI: 10.1520/E1566-00R14.2Shafer, T.W., Everett
12、, J., Silver, G.H., and Came, P.E., “Biohazard: Virus-Contaminated Liquid Nitrogen,” Science, Vol 191, 1976, pp. 2426.3Simione, F.P., Jr., Daggett, P.-M., McGrath, M.S., and Alexander, M.T., “TheUse of Plastic Ampoules for Freeze Preservation of Microorganisms,” Cryobiology,Vol 14, 1977, pp. 500502.
13、4Grieff, D., Melton, H., and Rowe, T. W., “On the Sealing of Gas-Filled GlassAmpoules,” Cryobiology, Vol 12, 1975, pp. 114.5For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, re
14、fer to the standards Document Summary page onthe ASTM website.6The last approved version of this historical standard is referenced onwww.astm.org.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1material in liquid nitrogen; each situat
15、ion must therefore beassessed individually using these guidelines.4.3 This guide is not intended for use with systems otherthan liquid nitrogen storage.4.4 This guide does not cover practices for preservation byfreezing which are covered in Practice E1342.5. Procedures5.1 All Liquid Nitrogen Freezer
16、s: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 freezers withall-vapor storage present the least hazard. The type of systemin use should be assessed before attempting to
17、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 Freezers with both liquid and vapor storage reduce theproximity of the operator to the liquid nitrogen, but the risk ofliqui
18、d 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 never be storeddirectly in liquid nitrogen. However, if a situation occurs inwhich hazardous biological materials are maintain
19、ed 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 protectionfrom the splashing of liquid nitrogen.5.2.3 Wear insulated gloves over rubber surgical gloves.5.2.4 Wear a full face
20、 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 Immediately place the retrieved material into dry icefor transport, or into a metal canister containing a liquiddisinfectan
21、t 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 whenretrieving from the liquid, and eye protection should be usedwhen retrieving from the vapor.5.3.2 Immerse the material slowly into
22、 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 the freezer.5.3.4 Material stored in the vapor phase may be removedfrom the freezer immediately for transport in dry ice or
23、thawing.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 Protective clothing should include a laboratory coat,surgical gloves, and eye protection.5.4.2 Material may be removed from the f
24、reezer 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 and warmed it shouldbe decontaminated. Decontamination procedures should bebased on the nature of the hazardous material ma
25、intained 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 required, the remaining mate-rial should be surface decontaminated rapidly in order to avoidwarming and moved to a different fr
26、eezer. Before decontami-nating the empty liquid nitrogen freezer, it should be warmedto room temperature.6. Keywords6.1 cryogenic; hazardous biologicals; liquid nitrogenASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin
27、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 entirely their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must
28、 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 should be addressed to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of ther
29、esponsible 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 Standards, at the address shown below.This standard is copyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700,
30、 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). Permission rights to photocopy the standard may also be secured from the Copyright Clearance Center, 222Rosewood Drive, Danvers, MA 01923, Tel: (978) 646-2600; http:/ 00 (2014)2
