1、Designation: E1564 00 (Reapproved 2014)Standard Guide forDesign and Maintenance of Low-Temperature StorageFacilities for Maintaining Cryopreserved BiologicalMaterials1This standard is issued under the fixed designation E1564; the number immediately following the designation indicates the year oforig
2、inal adoption or, in the case of revision, the year 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.INTRODUCTIONStorage facilities for maintaining cryopreserved biological mat
3、erials are generally comprised of twotypes of low-temperature systems: (1) freezers that are cooled by mechanical refrigeration and (2)freezers that are cooled passively with liquid nitrogen. Either system can be used for the storage offrozen biological materials as long as it meets the criteria spe
4、cific to the material being stored forensuring stability. Each system has its own unique handling requirements.1. Scope1.1 This guide covers recommended procedures for devel-oping and maintaining low-temperature storage facilities forfreezers with mechanical refrigeration.1.2 This guide covers recom
5、mended procedures for devel-oping and maintaining low-temperature storage facilities forfreezers cooled with liquid nitrogen.1.3 This guide does not cover practices for preservation byfreezing which are covered in Practice E1342.1.4 The values stated in SI units are to be regarded asstandard. No oth
6、er units of measurement are included in thisstandard.1.5 This standard does not purport to address all of thesafety problems, 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 o
7、f regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2E1342 Practice for Preservation by Freezing, Freeze-Drying,and Low Temperature Maintenance of Bacteria, Fungi,Protista, Viruses, Genetic Elements, andAnimal and PlantTissues (Withdrawn 2011)3E1565 Guide for Inventory Co
8、ntrol and Handling of Bio-logical Material Maintained at Low TemperaturesE1566 Guide for Handling Hazardous Biological Materialsin Liquid Nitrogen3. Terminology3.1 Definitions:3.1.1 cryogenic temperaturestemperatures below or equalto 100C.3.1.2 mechanical refrigerationa refrigeration system inwhich
9、cooling is provided by mechanical means such as acompressor.3.1.3 passive refrigerationa refrigeration system in whichcooling is provided by a refrigerant such as liquid nitrogen.4. Significance and Use4.1 The proper design of low-temperature storage facilitiesensures that sensitive biological mater
10、ials are maintained underconditions providing maximum storage stability.4.2 Properly designed and operated low-temperature storagefacilities ensure that the handling of sensitive biologicalmaterials at low temperatures does not compromise stability(see Guide E1565).4.3 Properly designed low-temperat
11、ure storage facilitiesensure that adequate safeguards are provided to prevent un-toward events from compromising the stability of sensitivebiological materials.1This guide is under the jurisdiction of ASTM Committee E55 on Manufactureof Pharmaceutical Products and is the direct responsibility of Sub
12、committee E55.04on General Biopharmaceutical Standards.Current edition approved Dec. 1, 2014. Published January 2015. Originallyapproved in 1993. Last previous edition approved in 2006 as E1564 00 (2006).DOI: 10.1520/E1564-00R14.2For referenced ASTM standards, visit the ASTM website, www.astm.org, o
13、rcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3The last approved version of this historical standard is referenced onwww.astm.org.Copyright ASTM International, 100 Barr Harbor Dr
14、ive, PO Box C700, West Conshohocken, PA 19428-2959. United States15. Procedure5.1 Low-Temperature Mechanical Freezers:5.1.1 Ensure that adequate dedicated electrical power isavailable to support the number of mechanical freezers re-quired in the facility.5.1.2 Provide for emergency electrical power
15、to maintainthe freezers when commercial power disruptions occur.5.1.3 Place the freezers in a temperature- and humidity-controlled area with adequate ventilation.5.1.4 Obtain a source of service support for the refrigerationsystems that is reliable and can respond to mechanical prob-lems rapidly. An
16、 alternative source should also be available.5.1.5 When selecting mechanical freezers, be sure that thelowest attainable temperature of the unit is cold enough toensure the stability of the biological material to be stored inthem.5.1.6 The temperature of mechanical freezers shall be moni-tored daily
17、 to ensure that consistently low temperatures aremaintained. Most units have temperature-sensing devices builtin that allow both control and monitoring of the freezertemperature. These internal units can malfunction and foolalarm systems. A temperature detecting probe should thereforebe inserted int
18、o each mechanical freezer, and the temperatureshould be read manually, or recorded automatically, on aremote sensing device.5.1.7 The temperature monitoring device shall be calibratedannually against a standard traceable to the National Institutefor Standards and Technology (NIST).45.2 Liquid Nitrog
19、en Freezers:5.2.1 Liquid nitrogen freezers are passive cooling systemsthat use liquid nitrogen as the refrigerant. A reliable source ofliquid nitrogen shall be available to maintain liquid nitrogenfreezers.5.2.2 Liquid nitrogen freezers provide the greatest assur-ance of long-term stability because
20、of the low temperatures thatcan be attained (150 to 196C).5.2.3 Choose a liquid nitrogen freezer that is compatiblewith inventory needs. Small units with large openings for easyaccess are less expensive to purchase and maintain, but theyrequire more careful monitoring and attention. Larger unitsmay
21、be more efficient. Most of these units have a workingopening that allows adequate access but does not compromisethe working temperature of the unit.5.2.4 Install liquid level sensing probes and controllingdevices on the liquid nitrogen freezer, if possible, to monitorliquid levels and to allow autom
22、atic and manual filling. Manualdipstick measurements should also be performed to ensure thatthe monitoring equipment is working properly.5.2.5 If possible, do not store vials directly in the liquidphase since improperly sealed vials can leak, leading topotential contamination and exploding vials upo
23、n retrieval.Maintain the vials in the vapor phase above the liquid. Whenvials are stored directly in the liquid, they should be filled to apoint that minimizes the air space in the vial, and they shall besealed completely. Vials may be examined for leaks byimmersing them in an aqueous methylene blue
24、 (0.05 %)solution at 4C.5.2.6 Storing material in the vapor phase of the liquidnitrogen freezer requires validating the unit to ensure thatsufficient liquid nitrogen is kept in the bottom of the unit tomaintain a working temperature at the top of the unitof 130C or below. To validate the freezer, me
25、asure thetemperature at the top with the freezer open, and raise the levelof the liquid nitrogen until the temperature remains at orbelow 130C for at least 1 h.5The requirements for storingmaterial in the vapor phase shall be determined prior topurchasing a liquid nitrogen freezer since not all free
26、zerdesigns are amenable to all vapor storage.5.2.7 Avoid using small liquid nitrogen units for permanentstorage since they require constant surveillance and increasethe probability of problems occurring.5.2.8 To ensure a constantly available source of liquidnitrogen, a bulk storage tank should be de
27、dicated to liquidnitrogen storage only.This can be a portable unit located withinthe facility, or an external stand tank with insulated piping fortransporting the liquid nitrogen to the freezers. Solenoids andpressure safety valves should be located in the piping to allowthe control of nitrogen flow
28、 and ventilation of excess pressure.Care shall be taken to avoid moisture accumulation on thevalves that could freeze and cause a malfunction.5.3 Safety:5.3.1 Precautions shall be taken to provide personnel pro-tection from the extreme cold encountered at cryogenic tem-peratures. Extremities shall b
29、e protected at all times usinggloves. Thin nylon gloves provide some protection whenhandling ampules; however, insulated gloves designed forworking at cryogenic temperatures should be used whenpossible.5.3.2 When vials are stored directly in the liquid, theyshould be filled to a point that minimizes
30、 the air space in thevial, and they shall be sealed completely. Vials may beexamined for leaks by immersing them in an aqueous methyl-ene blue (0.05 %) solution at 4C.5.3.2.1 Because of the potential for exploding vials whenretrieving from liquid nitrogen, precautions shall be taken toprotect the op
31、erator from glass shards.6,7Heavy gloves, labo-ratory coat, and face shield should be mandatory wheneverretrieving vials from liquid nitrogen. In addition, glass vialsshould be placed inside a metal canister before removing thevial to warmer temperatures.5.3.3 Hazardous biological materials should n
32、ot be main-tained directly in liquid nitrogen, because nitrogen will dis-place oxygen in confined areas with poor ventilation, espe-cially during fill operations. Care shall be taken to ensure4Available from National Institute of Standards and Technology (NIST), 100Bureau Dr., Stop 1070, Gaithersbur
33、g, MD 20899-1070, http:/www.nist.gov.5Simione, F.P., and Karpinsky, J.Z., “Points to Consider Before Validating aLiquid Nitrogen Freezer”, In: Validation Practices for Biotechnology Products,ASTM STP 1260, Shillenn, J.K., ed., ASTM, 1996, pp. 2430.6Simione, F.P. Jr., Daggett, P.M., McGrath, M.S., an
34、dAlexander, M.T., “The Useof Plastic Ampules for Freeze Preservation of Microorganisms”, Cryobiology,Vol14, 1977, pp. 500502.7Grieff, D., Melton, H., and Rowe, T.W., “On the Sealing of Gas-Filled GlassAmpoules”, Cryobiology, Vol 12, 1975, pp. 114.E1564 00 (2014)2proper ventilation of all areas housi
35、ng liquid nitrogen freezers,and an oxygen monitor with alarm should be installed in thearea if possible.5.3.4 Hazardous materials should not be maintained directlyin liquid nitrogen because of the potential for leaking andexploding vials (see Guide E1566).5.3.5 When freezers are no longer used, they
36、 should bedecontaminated after warming before removal or reuse.6. Keywords6.1 biological materials; cryopreserved; facilitiesASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard. Users of this standard are ex
37、pressly 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 be reviewed every five years andif not revi
38、sed, 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 theresponsible technical committee, which you ma
39、y 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, West Conshohocken, PA 19428-2959,United Sta
40、tes. 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)3
