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本文(EN 14031-2003 en Workplace atmospheres - Determination of airborne endotoxins《工作场所空气 空气中内毒素的测定》.pdf)为本站会员(arrownail386)主动上传,麦多课文库仅提供信息存储空间,仅对用户上传内容的表现方式做保护处理,对上载内容本身不做任何修改或编辑。 若此文所含内容侵犯了您的版权或隐私,请立即通知麦多课文库(发送邮件至master@mydoc123.com或直接QQ联系客服),我们立即给予删除!

EN 14031-2003 en Workplace atmospheres - Determination of airborne endotoxins《工作场所空气 空气中内毒素的测定》.pdf

1、BRITISH STANDARD BS EN 14031:2003 Workplace atmospheres Determination of airborne endotoxins The European Standard EN 14031:2003 has the status of a British Standard ICS 13.040.30 BS EN 14031:2003 This British Standard was published under the authority of the Standards Policy and Strategy Committee

2、on 17 April 2003 BSI 17 April 2003 ISBN 0 580 41594 5 National foreword This British Standard is the official English language version of EN 14031:2003. The UK participation in its preparation was entrusted by Technical Committee EH/2, Air quality, to Subcommittee EH/2/2, Workplace atmospheres which

3、 has the responsibility to: A list of organizations represented on this subcommittee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Catalogue under the sec

4、tion entitled “International Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online. This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application.

5、Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor re

6、lated international and European developments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the EN title page, pages 2 to 14, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when the do

7、cument was last issued. Amendments issued since publication Amd. No. Date CommentsEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM EN14031 January2003 ICS13.040.30 Englishversion WorkplaceatmospheresDeterminationofairborneendotoxins AtmosphresdeslieuxdetravailDterminationdes endotoxinesensuspensiondan

8、slair ArbeitsplatzatmosphreBestimmungvonluftgetragenen Endotoxinen ThisEuropeanStandardwasapprovedbyCENon21November2002. CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatel

9、istsandbibliographicalreferencesconcernings uchnational standardsmaybeobtainedonapplicationtotheManagementCentreortoanyCENmember. ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguage

10、andnotifiedtotheManagementCentrehasthesamestatusasthe official versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,CzechRepublic,Denmark,Finland,France,Germany,Greece, Hungary,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal,Slovakia,Spain,Sweden,SwitzerlandandUn ite

11、d Kingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISATION EUROPISCHESKOMITEEFRNORMUNG ManagementCentre:ruedeStassart,36B1050Brussels 2003CEN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.EN14031:2003EEN14031:2003(E) 2 Contents page Fore

12、word3 Introduction .4 1 Scope 4 2 Normativereferences 4 3 Termsanddefinitions . 4 4 Abbreviations.5 5 Overalldemandsinendotoxinwork. .6 5.1 General6 5.2 Demandsonoperator. .6 5.3 Demandsonequipment 6 5.4 Demandsonglassware .6 5.5 Demandsonextractionliquids,wateranddetergents. .6 5.6 Demandsonfilters

13、 6 5.7 Transportationofsamples. 6 5.8 Storageofsamplesatthelaboratory 7 5.9 Testforcontamination .7 5.10 Samplingdocumentation. 7 6 Extraction .7 6.1 General7 6.2 Equipment .7 6.3 Howtoextract .8 6.4 Storage8 6.5 Extractiondocumentation 8 7 Analyticalmethods 8 7.1 General8 7.2 Equipment .8 7.3 Proce

14、dure9 7.4 Validation.9 7.5 Analyticalprotocol . 9 8 Expressionofresults . .10 9 Precision.10 10 Testreport 10 AnnexA (informative) Propertiesofendotoxinsandsourcesofexposure11 A.1 Identityandphysicalandchemicalproperties. .11 A.2 Sourcesofoccupationalexposure .11 AnnexB (informative) PrincipleofLALa

15、ssays .12 B.1 General .12 B.2 Detectionofinhibitionorenhancementofmeasuredendotoxinsusingspikedsamples.12 B.3 Detectionofinhibitionorenhancementofmeasuredendotoxinsusingparallellineassay.12 Bibliography 14EN14031:2003(E) 3 Foreword Thisdocument(EN14031:2003)hasbeenpreparedbyTechnicalCommitteeCEN/TC1

16、37“Assessmentof workplaceexposure“,thesecretariatofwhichisheldbyDIN. ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublicationofanidenticaltextor byendorsement,atthelatestbyJuly2003,andconflictingnationalstandardsshallbewithdrawnatthelatestby July2003. AnnexesAandBareinformati

17、ve. AccordingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizationsofthefollowing countriesareboundtoimplementthisEuropeanStandard:Austria,Belgium,CzechRepublic,Denmark,Finland, France,Germany,Greece,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal,Spain, Sweden,Swit

18、zerlandandtheUnitedKingdom.EN14031:2003(E) 4 Introduction Endotoxinsareintegralcomponentsoftheoutermembraneofgramnegativebacteriawhicharecomposedof proteins,lipids,andlipopolysaccharides.Thetermendotoxinreferstothetoxinaspresentonthebacterialcell wall.Lipopolysaccharidesofgramnegativebacteriareferto

19、aclassofpurelipidcarbohydratemolecules(freeof proteinandothercellwallcomponents)thatareheldresponsibleformostofthebiologicalpropertiescharacteristic ofbacterialendotoxins.InannexAabriefoverviewisgivenwithrespecttophysicalandchemicalpropertiesof endotoxinsandsourcesofexposureintheoccupationalenvironm

20、ent. Endotoxinsarebelievedtoplayanimportantroleinthedevelopmentoforganicdustrelateddiseasesinexposed workers. Itisimportanttoassessoccupationalexposuretoairborneendotoxinsinarepresentativewaytoevaluatethe exposure.Thereare,however,atpresentnogenerallyacceptedproceduresforthemeasurementofenvironmenta

21、l endotoxins,thusdifferentpracticescontinuetoexist.Rigorousstandardizationwithrespecttosamplingmedia, extractionmedia,analyticalmethodsandstorageconditionsforendotoxinsareneededtoobtainresultsthatare comparablebetweenstudies.Byadheringtotherecommendationsoutlinedinthisstandardforchoiceofsampling, st

22、orageofsamples,extractionandanalyticalproceduresuncertaintiesinexposureassessmentcanbereduced andcontrolled,allowingcomparableandrepresentativemeasurementstobemade. 1Scope ThisEuropeanStandardprovidesguidelinesfortheassessmentofworkplaceexposuretoairbornebacterial endotoxins.Thestandardprovidesmetho

23、dsforsampling,transportation,andstorageofsamplesand determinationofendotoxins. 2 Normativereferences ThisEuropeanStandardincorporatesbydatedorundatedreference,provisionsfromotherpublications.These normativereferencesarecitedattheappropriateplaceinthetext,andthepublicationsarelistedhereafter.Fordated

24、 references,subsequentamendmentstoorrevisionsofanyofthesepublicationsapplytothisEuropeanStandardonly whenincorporatedinitbyamendmentorrevision.Forundatedreferences,thelatesteditionofthepublication referredtoapplies(includingamendments). EN481:1993, WorkplaceatmospheresSizefractiondefinitionsformeasu

25、rementofairborneparticles. EN1232, WorkplaceatmospheresPumpsforpersonalsamplingofchemicalagentsRequirementsandtest methods. EN12919, WorkplaceatmospheresPumpsforthesamplingofchemicalagentswithavolumeflowrateofover 5l/minRequirementsandtestmethods. 3 Termsanddefinitions ForthepurposesofthisEuropeanSt

26、andard,thefollowingtermsanddefinitionsapply. 3.1 bioaerosol airborneparticleswithbiologicalorigin NOTE SeeEN13098:2000,3.3,notes. 3.2 controlstandardendotoxin standardthatistraceabletotheRSEEN14031:2003(E) 5 3.3 endotoxin seeEN13098:2000. 3.4 endotoxinfreeliquids waterandsolutionswith0,2EU/ml NOTE E

27、ndotoxinfreeandpyrogenfreeareusedinterchangeableintheliterature. 3.5 endotoxinunit seeEN13098:2000 NOTE Endotoxinconcentrationsinsamplesarereferencedtoadefinedreferencestandardendotoxin(RSE)and subsequentlyexpressedinunitsbasedontheactivityofthereferencematerialinaLALassay. 3.6 exposure(byinhalation

28、) situationinwhichachemicalorbiologicalagentispresentinairwhichisinhaledbyaperson EN1540:1998 3.7 inhalableparticlesizefraction inhalablefractionwhichconstitutesthemassfractionoftotalairborneparticleswhichisinhaledthroughthenose andmouth EN481:1993 3.8 LALassay functionalassaytomeasureendotoxinconce

29、ntrations NOTE Themethodishighlysensitiveandbasedontheactivationofaclottingenzymepresentinthelysateofhemolymphof the Limuluspolyphemus (Horseshoecrab). 3.9 lipopolysaccharides LPSpresentingramnegativebacteriaarewatersolubleandstablemoleculescomposedoflipidand polysaccharide NOTE ThelipidpartofLPSist

30、ermedlipidAandisessentialforthetoxicpropertiesofLPS.Theterms endotoxins and lipopolysaccharides areoftenusedinterchangeablyinthescientificliterature. 3.10 referencestandardendotoxin purifiedLPSfrom Escherichiacoli thatservesasaninternationalreferencestandard 4Abbreviations CSE controlstandardendotox

31、in EUendotoxin units LAL limulusamoebocytelysate LPSlipopolysaccharidesEN14031:2003(E) 6 RSE referencestandardendotoxin 5 Overalldemandsinendotoxinwork 5.1General Environmentalmonitoringofendotoxinsisperformedbysamplinginhalableaerosols.Aerosolsaresampledona filterusingapumptodrawairthroughthefilter

32、.Forassessingoccupationalexposurepersonalsamplingshallbe usedforexamplewhencomparingwithexposureguidelines.Areasamplingcanbeusedtoidentifysourcesof exposure. 5.2 Demandsonoperator Theoperatorthatperformsthesamplingshallbetrainedinasepticworkconditions,haveknowledgeofsampling equipmentandknowhowtocar

33、ryoutthesampling.Theoperatorshallavoidcontaminationofthesampleduringall phasesofsampling. 5.3 Demandsonequipment Thepumpsshallfulfiltherequirementsofthestandardsforpumpsforpersonalandstaticsampling(EN1232and EN12919).PersonalsamplersshallcollecttheinhalableaerosolfractionasdefinedbyEN481. Thefilterh

34、olderorcassetteshouldberenderedendotoxinfree.Priortosampling,reusablesamplersshouldbe cleanedthoroughlysothatcontaminationofthesamplesareavoided.Cleaningagentsshallberemovedby subsequentwashinginendotoxinfreewater.Disposablesamplersdonotneedtobesterilisedwhenonecan assumethemanufacturingprocesstobef

35、airlyendotoxinfree.Possiblecontaminationshallbecontrolledbyusing blankfilters(see5.9). 5.4 Demandsonglassware Allothermaterialsusedduringsampling,transportationandstorageshallberenderedendotoxinfree.Glassware canberenderedendotoxinfreebyheatingatleastat180Cfor4h. 5.5 Demandsonextractionliquids,water

36、anddetergents Allliquidsusedshallbeendotoxinfree. 5.6 Demandsonfilters Binderfreeglassfibrefiltersshouldbeusedforendotoxinsamplingwhilethesefiltersusuallygivethehighest recoveries.Othertypesoffilterscanbeused.Ifanyotherfilterisuseditshallbevalidatedtowardfiltersofglass fibre. NOTE Severaltypesoffilt

37、ers(celluloseesters,polyvinylchloride,polytetrafluoroethyleneandpolycarbonate)arecommonly usedbutsomematerialadsorbendotoxinsandtherebygivefalselowvalues. 5.7 Transportationofsamples Aftersamplingfiltersshallbepreparedfortransportbysealinginacontainerorsamplercassette.Thesampleshall betransportedind

38、ryconditions,preferablywithadehumidifier.Samplescanbetransportedatroomoroutside temperaturebutconditionsfortransportshallbedocumentedandincludetransportationtime,temperatureetc.If transportationtimeisover24hthesamplesshallbetransportedinaconditionthatpreventsthefiltersfromgetting moistduringtranspor

39、tation,i.e.withadehumidifierorfrozen.EN14031:2003(E) 7 5.8 Storageofsamplesatthelaboratory Samplesshallneverberepeatedlyfrozenandthawed,asthismayeffectthedetectableendotoxincontentofthe sample.Endotoxinsareverystablewhenfrozen.Ifthesampleisnotextractedwithinafewdaysafterarrivaltothe laboratorythesam

40、pleshallbestoredatapproximately20Corbelow. 5.9 Testforcontamination Totestforcontaminationaseriesofatleast2blankfiltersshallbeincludedineachsamplingsession.Exceptfor theactualsampling,blankfiltersshallbetreatedequaltotheotherfiltersthatareusedforsampling. 5.10 Samplingdocumentation Alistofsamplingop

41、erationsshallbedocumentedtoobtaincomparableandreliableconcentrationvaluesof endotoxins.Thesamplingdocumentshallincludeatleastthefollowingparameters: nameoftheorganizationandoperatorperformingthesampling; dateofsampling; auniqueidentifiercodeforthesample; nameandaddressoftheplantwheresamplingwascarri

42、edout,orauniqueidentifiertopreserveconfidentiality; typeofsampling(personalorstatic)andincaseofpersonalsampling:nameandfunctionofthepersonwhowas involvedinthesampling,orauniqueidentifiertopreserveconfidentiality;orincaseofstaticsampling:locationof sampling; typeofsamplerandtypeoffilterused; startand

43、endtimeofsampling; flowratebeforeandaftersamplingperiod; sampledvolume; detailsaboutsampletransportationandstorage(temperature,duration,humidity); otherrelevantobservationsfromtheoperator. 6Extraction 6.1General Thefollowingparagraphsdescribeastandardmethodfortheextractionofendotoxinsfromfilters.Ifo

44、ther equipmentandmethodsareusedtheyshallbevalidatedagainstthisdescribedmethod. 6.2Equipment Astandardshakerapparatusabletovigorouslyshakethesampleinextractionfluidshouldbeused.Inadditiona standardtablecentrifugeisneededthatisabletocentrifugeat1000 G.Endotoxinfreeglassorpolypropylene tubesshallbeusedthroughoutt

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