Test No. 302C: Inherent Biodegradability: Modified MITI Test (II) [electronic resource]

2009·0.6 MB·English

by  Organisation for Economic Co-operation and Development

302C
Adopted:;.
OECDGUIDELINEFORTESTIN^GOFCHEMICALCSorrected8Se1p2teMmabeyr1'2908019
"InherentBiodegradabaity:
ModifiedMITITest(II)"
1.TNTROnITCTORY INFORMATION
• Prerequisites
illthetestsolution.
- Theempiricalformulaofthetestmaterialisrequiredsothatthetheoieticalojqfgcndemand
(TOD)maybecalculated.
• Oiiidance information
- Iunseffourlmaitniionntoernprdeiteirneglatthieverepsruoltpsorotbitoanisneodf,thpearmtaijcuolrarcloymipnontehonstescoafstehsewtheesrtmeattheeriraelsu-lmtillibees
closetothe"passlevel".
- Informationmthetoxicityofthechemicalrwybeusefultotheinterpretationoflowresults
andintheselectionofappropriatetestconcentrations.
• Qualifyingstatements
Themethodisonlyapplicabletothoseorganictestmaterials.which,attheconceiitration
usedinthetest,
*- havenegligiblevapourpressure,
--adroennoottrienahcihbiatonrdyrteoacbtacwtietrhia,thaenCdOj®dSorhant. .
fhoarsdbeeteeTnrhmitisenstitenesgtdhtiahnestbihreeiehOnerEfenoCluDbnidoLsdauebigotrraaabdtlaeobrtiyylitIthnyeteoOrfEcooCrngD^aanErixicpsceohrnetmTGiercsaotluPspruDonegdgreraramadmeareotb(ii1oc9^7cA8oc-n1cd9ul8d^0oa)nt.si.oInt
• Recommendations
useofuIlfttrhaestoeusntdmdaitsepreiraslioisn,nmotaysohlaubvleetaotbteheetmepsltocyoencdetnotraacthiioenv,esapegcoioaldmdeiasspuerrseiso,nsoufcthhaestfeislte
ntaterial.
UsersoftihnispsTrefsjtbuGluairrpfaetriangersahpohusl3d,c4o,n7s.ualntdth8e.Pmtace,
302C : ' ; : -
^ "InhereiitBiodegradabillty:
ModifiedMITITest(liy^
• Standarddocumeats '
ThisT^tGuidelinewasbasedonanorderprescribingtestingrelatedtohewchemical
substancesoftheQienucalSubstanceControlI^w(LawNo.117,1973),Order-oftheJapanes®'
PrimeMinister,theMinisterofHealtharidWelfare,andtheMinisterofInternationalTradeand
Industry,No.1(Jufy13,1974). ‘ .
2,ivrETHOD
A.IANPTERUOCDAUTCTmIAOmN_.IPJUMRIPTO_SES.J3SFCOTPEES.RELEVANCR;
The.purposeofthistestisdiemeasuremehtoftheBiochemicalO^genDemand(BOD)
achnedmitchealansaulbysstiasncoefsrwehsiicdhuahlacvheembieceanlsfoiunnodrbdyerttboeleMvoadliuaftieedtihMeIiHnMheer&enotdbi(oI)detgorainddaibcialtietyloowf
degradability!. .
• Defini11onsandunits
Pmeni^edegradation- ~—Sx100(%)
or ^ ^
Percemagedegradaddn= So 100(%)
BBTOODD::: BBOiHciionjOoemoylQpDcorglngeeechitntuuciermacclvalyoIelsonoxsoaxyujdingyddpegietnesdiendodme(ne^tanh(seae-onxurdpretee(irdecixampoleen,nrtitmamhgele)n,tBdanOle^,Dmacnoufdrobvrfaeesqtauhliertceeudslt:t\cusorhemenpmot/euhd'enid,tuemmsetatcsoouw.rmhepidocb.uhnn.dtdhiiees
Sa : bRieosdiedgruaMdaabmioluitnytt(eesxtperimental,mg),ofdietestcon^xjundafterconplctionofthe
Sb : wRaestiedrutaolwahmiocuhnton(leyxptehreimteesnttaclo,nifriogu)nodfhthaes,tbeesetnCaodn<f^oundintheblanktestwith
302C
"InherentBiodegr,adability: ^
ModifiedMITITest(II)"
• KRfRTencesubstances
hascootwdieviviuetmyrI,nboefssnoptzehmoceeaitifcneiacosmceurasleyuf'm^«b,eh-eentnuchseeienudsvseuesfbotsoritfgatacnhtcmiiestnsrgpouaclrapnnsoenusobwets.stuyabentsctebasenicsredeercseoifmreamrbeelnnecd.eeAdsnu.iblsiItnnaeno,cresd^soerdrittKuo5mfcbaheceeVtcBaketfietihloe;r
ttotexcIefetdhe4p0erpceerncteangteadfetgerrad7atdaiyosn,ofanadnil6i5nepecarlccuelnattaefdtefrro1m4tdh^eso,xytgheentecsotnsisumrpetgiaordneddoeass
invalid.
• Princinleofthetestmethod
Thistestmethodisbasedonthefollowingcmdltions;’
.-testchemicalsassoleorganiccarbonsources
-noadaptationofmicro-iffganismstotestchemicals
utmssheueetcdeht.re.asCstBAhlpimeelomerdiaaiecusogatudrol,ramsetdahmtateeobenibbdthietoctoyclefhositestshmeeecidda-cdalsaircylseussohitoixneltoymvecgedouedxlonayontdrgetgeedhanmeniacinbncoatdsnhciseaisusrtonmbepfseottBnaiisnOcougonDrnvmeceaeedsnnastcdesrolusansrtitiwipinoipnngtlu,heoamucnpesoiplnncaytcrraoeahl-ntyotcurrmhsgaeea(tmnaiBiinoOcsnsDma-sloo:mffa.ernatDeaueBlsyirrOsdi)iDunsa-ig,ls
chemicals,etc.
• Ona1itVcriteria
Reproducibility
ThereproducibiUtyofthismethodisgenerallygood,especiallyforchemicalssolublefar
waterover100ppm.
Semidvity
-Oiygenconsuii^tion: detectionlimit=1mg(oi^genconsmqitianbymicro-organisms)
-rbamiedanalysis ; dqiendsonthesensitivityoftheanalyticalmethodsused
- .
302C
"InherentBiodegradabiUty:
ModifiedMITITest(II)"
• Specificity
Applicabletoeveiy,kindofchemical,foryiiicli
ForvolatilechemicalsamodifiedBOD-meter**shouldbeused.
PqssWUifyofstandardisaiwn
ThismethodhasbeenauthorisedbytheJapaneseGovenunent-asa“M^odfor.TMting
theBiodegmdabihtyofChemical'SubstancestyMicroorganisms'-.
l^ssibUityofautoinadott
ByusingaBOD-meter*^oxygenconsurptionbymicro-organisms(intheclosed
^stem)isrecordedautomatically.
B.DESGRIPtlONOFTHETESTPROCEDURE
• Preparations
Apparatus:BQD-meterequippedwith6bottlesf3QQmleach!
bottle1. : deionisedwater***,300mi+testchemical,9mg-
bottles2,3,4: bcahs^ailcaclul,tu9remmgediu,m,300mi+activatedsludge,30mg(diybasis)-+test
;bottles : b3a0samlgculturemedium,300.ml+activatedsludge,;9mg(drybasis)+aniline,'
bottle6 :-basalciituremedium,300mg+activatedsludge,30mg(drybasis)
\•**• CTipl=Aw.lcmioonadaci^f1b'e.a)dliBoiO,iDHMeteristtunposedofc^illaiytubingand3-noOTialBOD^eter.(Seetf,iedesaiptioiigivsi
•**.ThewaterusedmustnevercontednmoreSian10percentoftheoigatiiccaibonintroducedby4ietestmaterial.
302C
^^InherentBiodegradahility; ^
ModifiedMITITest(II)”
.Preireatmentoftestchemical
Incasetiietestcompoundisnotsolubleinwateratthedesiredtestconcentration,the
testcompoundpulverisedasfinelyaspossibleisemployed.
Incasethetestconfoundisvolatile,testchemicalsshouldbewelbcooledtoprevent
evaporation..
Theidentificationoftestsan^ile,ifnecessary,shouldbemade.
Basalculturemedium
3mleachofsolutionA,solutionB,solutionCandsolutionD,andwateraremadeup
to10(K)ml.(Deionisedwaterisusedthroughout.)
SolutionA: 4c2h41l..o67r5igdgeoofafrddeiibdpaiosstisacoslssvheoiddmiihunymwdarptohegoresanpnhpadhtoteshpedhvoaodtelec,uam8he.y5disrgamotafedpeaontudapsst1oi.u71m0ga0c0oifdmlpa.hmo(msTpohhneaitpeuH,m
ofthesolutionis7.2)
SolutionB: 2v2o.l5umgeoifsmmaagdneeuspiutmos1u0l0p0hamtlehq)tahydratearedissolvedinwaterandthe
SolutionC:.1207.050gmlo.fcalciumchloridearedissolvedinwaterandthevolumeismadeupto
SolutionD: 0m.a2d5eguopftfoer1ri0c00chnliolr.idehexahydr•atearedissolvedinwaterandthevolumeis
Activatedsludge
throughSoluutdtgheescaomupnltirnyg,scihtieesf:lySliund^geosseamaprleiansgwihsemraedea,vairnipertiyncoifpcleh,emaitcnaoltsluesbssttahnacnes10mpelyacbees
consideredtobeconsumedanddiscarded.
takenuFporfreoxmantphlee,fosltlaonwdianrgdpalcatcievsataenddslmuidxgeed.oftheJapaneseChemicalBiotestingCenteris
- Citysewageplant:3plantslocatedinthenorfliem,centralandsouthernpMtofJapan.
- Industrysewageplant:oneplantusedforthewastewatertreatmentofchemicalindustries.
302C
^ "InherentBiodegradabiUty:
ModifiedMITITest(II)"
-River:3riverslocatedinthenorthern,centralandsoiihempartofJapan.
-Lake:Onelake-locatedinthemiddleofJapan.
-Sea:2inlandseasofJapan.
ayear,FirneqMuaerncchy,nJfusnlei,TdSpeepstaemmpbleirnga:nSdluDdegceemsabne^rj.lingshouldbemade,inprinciple,fourtimes
SludgesampUngmethods
-Citysewage:1litreofreturnsludgeatasewagedisposalplant.
-Rivers,lakesandmarshesorsea:1litreofsurfacewaterand1litreofsurfacesoilonthe
beachwhichisincoitactwiththeatmosphere.
iwanintdahtshsieoPnsdr^ueieppueacmrronanhattytaidaiornnnote:xriTi,shdfeeilsotreltruhepddelgiterohrssripaxohntcu^urgrIiheecNsiosac.caoi2lldll,foeiwctl'ttreeearddnpstfaforpeerosrrtm.eatndThdieh.nestFoalfmoialpattlcriiauntnlegtguifssroieatrdeetsjiaugansnrkteemamdanittdtxoeeaprdeHribasty7re.des0.tmi±orvr1ie.nd0g
a0mo.upitp1xrtp3ouexr1rie2Cmcu5iaels*tnteat2uelr^rsye^ya:nt01Aie/hdb3eotaouigfactitsn3h.ee0wTwamhlgiiinesou*ltpeerisvsocoaaeldfdudtumeerredectoieosfatsrthieheipe^ersaetqtmhieaeediranaceainrtncaagetnipteoovnirestroiyrfoendtmahooyefv.etsTdho.heleuAstciinuo^lneetmquoaurbtaitelnangivtni,oseldacunamadrberoiteohvdefe
adjustmCeonnttsroalre:Fmoardceo;ntroloftheculturingstep,thefollowingitemsarecheckedandnecessary
-Appearanceofsupernatant:thesupernatantofactivesludgeshouldbeclear.
-Precipitabilityofactivesludge:theactivesludge,beinginlargeflocks,musthavehigh
precipitability.
* Oi.nJ1pleirtreceoaftwsajt'nefrefaitnidcstteiewasogleuti1ongoisfagdljuucsotseed,to1pgHof7.p0q±UDInDeswiatnhdsIodgioufmihnyodntoopxoitdea.ssiumphosphatearedissolved•
302C
"InherentBiodegradability; ^
ModifiedMITITest(II)"
- vStoaltuemoefoffOT0m.a1tipoenrocefnatptsiyvnethseltuidcges:e.wWahgeeretogrboewt^heofdfloorcktsheisfrneoqtueonbcsyervoefd,adediitthieornthoef
syntheticsewageisincreased.
- pH:ThepHofthesupernatantis7.0±1.0.
- Temperature:Thetemperaturefercultivationofactivesludgeis25°±2°C.
- Amountofaeration:Inreplacingthesvpemataatwithsyntheticsewage,thesolutioninthe
culturingtankmustbesufficientlyaeratedtomaintainthedissolvedoxygenconcentration
oftilesolutionabove5ppm.
-M1i0c0ro-fl4o0r0aXomfagancitfiivcaatteidons)lu,dagen:umWbheernoftphreotaoctziovaeofsdliufdfgeereinstnsipcercoisecsoptiocgaeltlhyerowbistehrvceldou(dayt
flocksmustbeseen.
-Mixingoffreshandoldactivatedsludge:Inordertomaintainfreshandoldactivated
sludgesatthesameactivity,thefiltrateofthesupernatantofanactivatedslu^einactual
useinthetestis'mixedwithanequalvolumeofthefiltrateofthesupernatantofafreshly
collectedactivatedsludgeandthenixtureiscultured.
- Checking,ofactivityofactivatedsludge:Activityofactivatedsludgeshouldbechecked
periodically(atleastonceeverythreemonths)withstandardssubstances, applyingthe
testmethodprovidedbelow.Especiallyviheufreshandoldactivatedsludgesanplesare
mixed,carefulcheckingmustbedoneinrelationtotheoldactivatedsludge.
ExanqfleofPreparationofActivated^udgeSandiesandPeriodofUse
Dec. Jan. Feb.
Culture Periodofuse
Mar. Apr.May
Mixing,culture Periodofuse
JuneJulyAug.
Mixing,culture Periodofuse
SepLOct.Nov.
Mixing,culture Periodofuse
(Thesamepatternofpreparationandusefollows.)
302C
8 "InherentBiodegradability:
ModifiedMITITest(II)"
Additionoftestcompoundandpreparationfortest
Thefollowingtestsvesselsareprovidedandadjustedtothetesttemperature;
(1)Acotmepsotuvnesds;elthceonptTaIionfintghitshseoblautsiaolnciusltaudrjeumsteeddituom,7btoefvodrieicthhiesiandodceudla3t0iopnpofma(ctWifvWe)solufdtgees,t
ifnecessary.
(2)Atestvesselforthecontrolblanktest,containingonlythebasalculturemedium
(3)Atestvesselcontainingwatertowhichisadded30ppm(WAOofthetestcompound.
(4)Aortaesntyveostsheelrcoofnttahiencicnngtbraoslaslucbuslttaunrceesm.ediumtowhichisadded100ppm(W/V)ofaniline
Inoculationofactivesludge
suspendIendocmualtutmeris(1a0d0dpedpmtoVtAhOetreesqtuiversesdeilnst1heaJnadpa2neasbeovIend,usstoritahlatStthaendcaorndcdeentsrcartiiboendoinf
Annex2isachieved.
Forvessel4,therequiredconcentrationofsmpendedmatteris30ppuL
• Testconditions
" Concentrationoftestcheinicals : 30ppm
- Concentrationofactivatedsludge: 100ppm(WAO
- Testtemperature : 25°C±2°C
- Period : 14to28days
- Performindarkness.Thetenperatureandthechangeincolourofthecententsofthe
culturingvesselshouldbecheckedeveryday.
- StirvigorouslywiOimechanicalstirrer.
• Performanceoftest
TheBODcurveisobtainedcoitinuouslyandautonaticallyfor14to28days(see
Figure1).
^
302C
"InherentBiodegradability; ^
ModifiedMITITest(II)"
Figure1:Degradationcurveofareadily,degradableconfound
a = adaptationperiod
1mrt =—== pmrteeiaqrmxuieioirwdmeiuodnfmdlleodvoewegglarroaidftahdtmeiigocrnagrdraaottweilohn
Afterthe14to28daysoftesting,pH,residualchemicalsandinteimediatesinthetesting
vesselsareanalysed.
Thetestchemicalsinthetestingvesselwithoutactivatedsludgearealsoanalysedin
ordertodeterminewhetherthereisanychangeinthelestchemicalduringthetestingperiod,
oranylossoftheoriginaltestchemicalbyevaporationorbyadsorptionbythewallsofthetest
vessels,etc.
• Analytical means
Ifthetestcompoundissolubleinwater,theresidualamountoftotalorganiccarbonis
alsodetermined.
(a) fWrhoemrethaetteosttalveosrsgealniccacrebnotnrtimfaulgy^seart3is00us0edg:fo1r0fmivleomfintuhteest.estTehdesroelsuitdiuoanliasmsoaunn^tleodf
thetotalorganiccarboninthesupernatantisdeterminedonatotalorganiccarbon
analyser.
302C
"InherentBiodegradability:
ModifiedMITITest(U)"
(b) Wscoohnlecvreennettroatsthuieiortna,abnltaehleyfsroeerrssidtauhraeeluatsmeesodt:unctthoemofptoottuhalendtceosntatnecdno,trroagffttoaeurntedspitrsionpdgeevrteesrpsmreielnteirsdeaeotxnmteranancttaensdaul-cywhistihnagsa
instrument(gaschromatcgraphy,absorptionspectrometry,massspectrottetry,atomic
absorptionspectrophotometry,etc.)
Inthecaseofvolatilechemicals,thetemperatureconftolbathoftheBOD-metershould
becooledto10°Candthistemperatureheldforatleast30min,inordertopreventevapcration.
Theanalyticalprocedures(a)and(b)shouldfiienbestarted.
3.I>ATAAND REPORTING
• Treatmentofre.suits
a) Methodforcalculatingthepercentagedegradationfromtheoxygenconsumption;
Percentagedegt’odaUon — —x100(%)
BOD: Boino.ltohgeiBcaOlDojcyugr\e'nedemand(experimental,mg)ofthetestconfoundmeasured
B : Oinxoycguelnucmoinssmardpdteidomne(aesxuprereidmeonntatlh,emBg)ODofbcausravleculturemediumtoMvhichthe
TOD: Tishecoorneqt)ilceatleioyxyogxiednidseedmand(theoretical,mg)required\s4ienthetestcompoimd
b) Methodforcalculatingthepercentagedegradationfromtheresultofdirectanalysis:
^ ^
Percentagedegrada^on= aP x100
Sa : Rtehesibdiuoadlegarmaoduabnitli(teyxpteersitmental,mg)ofthetestconfoundafterconpletionof
Sb : bAlvaenrkagteesrtsesvirdiutahlwaamtoeurntto(^e^xipcerhimoennltyalt,hemgte)stofctohne4)toesutncdohnaqsiobuenedninadtdhee.dt.wo