1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS EN ISO 9439:2001 BS 60
2、68-5.12:2001 The European Standard EN ISO 9439:2000 has the status of a British Standard ICS 13.060.70 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Water quality Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium Carbon dioxide evolution
3、testThis British Standard, having been prepared under the direction of the Health and Environment Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 March 2001 BSI 03-2001 ISBN 0 580 34729 X BS EN ISO 9439:2001 Amendments issued since publicati
4、on Amd. No. Date Comments National foreword This British Standard is the official English language version of EN ISO 9439:2000. It is identical with ISO 9439:1999. It supersedes BS 6068-5.12:1993 which is withdrawn. The UK participation in its preparation was entrusted by Technical Committee EH/3, W
5、ater quality, to Subcommittee EH/3/5, Biological methods, which has the responsibility to: 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 relat
6、ed international and European developments and promulgate them in the UK. A list of organizations represented on this committee can be obtained on request to its secretary. This is one of a series of standards on water quality, others of which have been, or will be, published as Sections of BS 6068.
7、 The various Sections of BS 6068 are comprised within Parts 1 to 7, which, together with Part 0, are listed below. Part 0 Introduction Part 1 Glossary Part 2 Physical, chemical and biochemical methods Part 3 Radiological methods Part 4 Microbiological methods Part 5 Biological methods Part 6 Samplin
8、g Part 7 Precision and accuracy NOTE The tests described in this British Standard should only be carried out by suitably qualified persons with an appropriate level of biological expertise. Standard biological procedures should be followed throughout. Cross-references Attention is drawn to the fact
9、that CEN and CENELEC Standards normally include an annex which lists normative references to international publications with their corresponding European publications. The British Standards which implement these international or European publications may be found in the BSI Standards Catalogue under
10、 the section entitled “International Standards Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct app
11、lication. Compliance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, the EN ISO title page, the EN ISO foreword page, the ISO title page, pages ii to iv, pages 1 to 17 and a back cover.
12、The BSI copyright notice displayed in this document indicates when the document was last issued.EUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM ENISO9439 April2000 ICS13.060.70 Englishversion WaterqualityEvaluationofultimateaerobicbiodegradabilityof organiccompoundsinaqueousmediumCarbondioxide evolut
13、iontest(ISO9439:1999) QualitdeleauEvaluationdelabiodgradabilitarobie ultimeenmilieuaqueuxdescompossorganiquesEssai dedgagementdedioxydedecarbone(ISO9439:1999) WasserbeschaffenheitBestimmungdervollstndigen aerobenbiologischenAbbaubarkeitorganischerStoffeim wrigenMediumVerfahrenmitKohlenstoffdioxid Me
14、ssung(ISO9439:1999) ThisEuropeanStandardwasapprovedbyCENon11March2000. CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatelistsandbibliographicalreferencesconcernings uchnat
15、ional standardsmaybeobtainedonapplicationtotheCentralSecretariatortoanyCENmember. ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheCentralSecretariathasthesamestat
16、usast heofficial versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,CzechRepublic,Denmark,Finland,France,Germany,Greece, Iceland,Ireland,Italy,Luxembourg,Netherlands,Norway,Portugal,Spain,Sweden,SwitzerlandandUnitedKingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISAT
17、ION EUROPISCHESKOMITEEFRNORMUNG CentralSecretariat:ruedeStassart,36B1050Brussels 2000CEN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.ENISO9439:2000EForeword ThetextoftheInternationalStandardfromTechnicalCommitteeISO/TC147 “Waterquality“ oftheInternati
18、onalOrganizationforStandardization(ISO)hasbeentakenoverasan EuropeanStandardbyTechnicalCommitteeCEN/TC230 “Wateranalysis“,thesecretariat ofwhichisheldbyDIN. ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublication ofanidenticaltextorbyendorsement,atthelatestbyOctober2000,andc
19、onflictingnational standardsshallbewithdrawnatthelatestbyOctober2000. AccordingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizations of thefollowingcountriesareboundtoimplementthisEuropeanStandard:Austria,Belgium, CzechRepublic,Denmark,Finland,France,Germany,Greece,Iceland,Ireland,It
20、aly, Luxembourg,Netherlands,Norway,Portugal,Spain,Sweden,SwitzerlandandtheUnited Kingdom. Endorsementnotice ThetextoftheInternationalStandardISO9439:1999hasbeenapprovedbyCENasa EuropeanStandardwithoutanymodification. ENISO9439:2000 A Referencenumbe r ISO9439:1999 (E) INTERNATIONAL STANDARD ISO 9439
21、Secondedition 19990301 WaterqualityEvaluationofultimate aerobicbiodegradabilityoforganic compoundsinaqueousmediumCarbon dioxideevolutiontest Qualitdeleauvaluationdelabiodgradabilitarobieultimeen milieuaqueuxdescompossorganiquesEssaidedgagementde dioxydedecarbone ENISO9439:2000 ii Contents 1Scope 1 2
22、Definitions 1 3Principle3 4Testenvironment.3 5Reagents.3 6Apparatus .4 7Procedure .5 8Calculation7 9Validityofresults.8 10Testreport 9 AnnexA (informative) Principleofatestsystemformeasuringcarbondioxide(example) .10 AnnexB (informative) Examplesofthedeterminationofreleasedcarbondioxide.11 AnnexC (i
23、nformative) Exampleofabiodegradationcurve 13 AnnexD (informative) CombineddeterminationofcarbondioxideandDOC 14 Bibliography17 ENISO9439:2000 OSI )E(9991:9349OSI iii Foreword ISO(theInternationalOrganizationforStandardization)isaworldwidefederationofnationalstandardsbodies(ISO memberbodies).Theworko
24、fpreparingInternationalStandardsisnormallycarriedoutthroughISOtechnical committees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteehasbeenestablishedhas therighttoberepresentedonthatcommittee.Internationalorganizations,governmentalandnongovernmental,in liaisonwithISO,alsotakepartinthewo
25、rk.ISOcollaboratescloselywiththeInternationalElectrotechnical Commission(IEC)onallmattersofelectrotechnicalstandardization. InternationalStandardsaredraftedinaccordancewiththerulesgivenintheISO/IECDirectives,Part3. DraftInternationalStandardsadoptedbythetechnicalcommitteesarecirculatedtothememberbod
26、iesforvoting. PublicationasanInternationalStandardrequiresapprovalbyatleast75%ofthememberbodiescastingavote. InternationalStandardISO 9439hasbeenpreparedbyTechnicalCommitteeISO/TC 147, Waterquality, SubcommitteeSC5, Biologicalmethods. Thissecondeditioncancelsandreplacesthefirstedition(ISO9439:1990),
27、whichhasbeentechnicallyrevised. AnnexesAtoDofthisInternationalStandardareforinformationonly. ENISO9439:2000 )E(9991:9349OSI OSI iv Introduction TheconditionsdescribedinthisInternationalStandarddonotalwayscorrespondtotheoptimalconditionsfor allowingthemaximumdegreeofbiodegradationtooccur.Withthistest
28、system,themicrobiallyderivedcarbon dioxide(CO 2 )ismeasuredinthetrapsthroughwhichgasexhaustedfromthetestvesselsispassed.Someofthe CO 2 remainsinthemediuminthevesselsasdissolvedinorganiccarbon(DIC),theconcentrationofwhichmay increaseasbiodegradationproceeds.Astheorganiccarbonapproachescompleteremoval
29、,theconcentrationof DICgraduallyfallsandtendstoreachzerobytheendofincubation.Itisthusnecessarytoacidifythemediumat theendofthetesttomeasurethebiogenicallyformedCO 2 completely.ThemeasurementofCO 2 intheexternal trapsmaydifferfromthetrueproductionofCO 2 andthekineticratemayalsobelowerthanaratebasedon
30、DOC removalmeasurement.TheconsequencemaybethatthebiodegradationcurvesbasedonthetrappedCO 2 may notfullyrepresentthetruemicrobialkineticrate.Foralternativebiodegradationmethods,seeISO15462andin particularISO14593,whichisbasedonCO 2 productionaswellbutdoesnothavethisdefect. ENISO9439:2000 DRADNATSLANO
31、ITANRETNI OSI )E(9991:9349OSI 1 WaterqualityEvaluationofultimateaerobicbiodegradabilityof organiccompoundsinaqueousmediumCarbondioxide evolutiontest WARNINGActivatedsludgeandsewagemaycontainpotentiallypathogenicorganisms.Appropriate precautionsshouldbetakenwhenhandlingthem.Toxictestcompoundsandthose
32、whosepropertiesare unknownshouldbehandledwithcare. 1Scope ThisInternationalStandardspecifiesamethod,bydeterminationofcarbondioxide(CO 2 ),fortheevaluationinan aqueousmediumoftheultimatebiodegradabilityoforganiccompoundsatagivenconcentrationbyaerobic microorganisms. Themethodappliestoorganiccompounds
33、whichare: a) watersolubleundertheconditionsofthetest,inwhichcaseremovalofDOCmaybedeterminedasadditional information(seeannexD); b) poorlywatersolubleundertheconditionsofthetest,inwhichcasespecialmeasuresmaybenecessaryto achievegooddispersionofthecompound(see,forexample,ISO10634); c) nonvolatileorwhi
34、chhaveanegligiblevapourpressureundertheconditionsofthetest; NOTE ForvolatilesubstancesuseforexampleISO9408orISO14593. d) notinhibitorytothetestmicroorganismsattheconcentrationchosenforthetest. NOTE Thepresenceofinhibitoryeffectscanbedeterminedasspecifiedin8.3,orbyusinganyothermethodfor determiningth
35、einhibitoryeffectofacompoundonbacteria(see,forexample,ISO8192). 2 Termsanddefinitions ForthepurposesofthisInternationalStandard,thefollowingtermsanddefinitionsapply. 2.1 ultimateaerobicbiodegradation breakdownofachemicalcompoundororganicmatterbymicroorganismsinthepresenceofoxygentocarbon dioxide,wat
36、erandmineralsaltsofanyotherelementspresent(mineralization)andtheproductionofnewbiomass 2.2 primarybiodegradation structuralchange(transformation)ofachemicalcompoundbymicroorganismsresultinginthelossofaspecific property 2.3 activatedsludge biomassproducedintheaerobictreatmentofwastewaterbythegrowthof
37、bacteriaandothermicroorganismsinthe presenceofdissolvedoxygen ENISO9439:2000 )E(9991:9349OSI OSI 2 2.4 concentrationofsuspendedsolids amountofsolidsobtainedbyfiltrationorcentrifugationofaknownvolumeofactivatedsludge anddryingatabout105 o Ctoconstantmass 2.5 dissolvedorganiccarbon DOC thatpartoftheor
38、ganiccarboninawatersamplewhichcannotberemovedbyspecifiedphaseseparation NOTE Forexample,bycentrifugationat40000m s 2 for15minorbymembranefiltrationusingmembraneswithpores ofdiameter0,2mto0,45m. 2.6 totalinorganiccarbon TIC allthatinorganiccarboninthewaterderivingfromcarbondioxideandcarbonate 2.7 dis
39、solvedinorganiccarbon DIC thatpartoftheinorganiccarboninwaterwhichcannotberemovedbyspecifiedphaseseparation NOTE Forexample,bycentrifugationat40000m s 2 for15minorbymembranefiltrationusingmembraneswithpores ofdiameter0,2mto0,45m. 2.8 theoreticalamountofformedcarbondioxide ThCO 2 theoreticalmaximumam
40、ountofcarbondioxideformedafteroxidizingachemicalcompoundcompletely NOTE Itiscalculatedfromthemolecularformulaandexpressedinthiscaseasmilligramscarbondioxidepermilligram(or gram)testcompound. 2.9 lagphase timefromthestartofatestuntiladaptationand/orselectionofthedegradingmicroorganismsareachievedandt
41、he biodegradationdegreeofachemicalcompoundororganicmatterhasincreasedtoabout10%ofthemaximum levelofbiodegradation NOTE Itisnormallyrecordedindays. 2.10 maximumlevelofbiodegradation maximumbiodegradationdegreeofachemicalcompoundororganicmatterinatest,abovewhichnofurther biodegradationtakesplaceduring
42、thetest NOTE Itisnormallyrecordedinpercent. 2.11 biodegradationphase timefromtheendofthelagphaseofatestuntilabout90%ofthemaximumlevelofbiodegradationhasbeen reached NOTE Itisnormallyrecordedindays. 2.12 plateauphase timefromtheendofthebiodegradationphaseuntiltheendofthetest NOTE Itisnormallyrecorded
43、indays. ENISO9439:2000 OSI )E(9991:9349OSI 3 2.13 preexposure preincubationofaninoculuminthepresenceofthetestchemicalcompoundororganicmatter,withtheaimof enhancingtheabilityofthisinoculumtobiodegradethetestmaterialbyadaptationand/orselectionofthe microorganisms 2.14 preconditioning preincubationofan
44、inoculumundertheconditionsofthesubsequenttestintheabsenceofthetestchemical compoundororganicmatter,withtheaimofimprovingtheperformanceofthetestbyacclimatizationofthe microorganismstothetestconditions 3Principle Thebiodegradabilityoforganiccompoundsbyaerobicmicroorganismsisdeterminedusingastaticaqueoustest system.Thetestmixturecontainsaninorganicmedium,theorganiccompoundasthenominalsolesourceof carbonandenergyataconcentrationof10mg/lto40mg/lorganiccarbonandamixedinoculumobta
