Existing Chemical
Secondary Notification Assessment STD/1021S
Complex Soap TH17
October 2009
NationalIndustrialChemicalsNotificationandAssessmentScheme
GPO Box 58, Sydney NSW 2001, Australia
ISBN978-0-9803124-9-2
©CommonwealthofAustralia2009
Thisworkiscopyright.Youmaydownload,display,printandreproducethismaterialinunalteredformonly(retainingthisnotice)foryourpersonal,non-commercialuseorusewithinyourorganisation.ApartfromanyuseaspermittedundertheCopyrightAct1968,allotherrightsarereserved.RequestsandinquiriesconcerningreproductionandrightsshouldbeaddressedtoCommonwealthCopyrightAdministration,AttorneyGeneral’sDepartment,NationalCircuit,BartonACT2600orpostedat
Contents
PREFACEVI
OVERVIEWAND RECOMMENDATIONSVIII
SECONDARY NOTIFICATIONXII
ACRONYMSABBREVIATIONSXIII
1.INTRODUCTION1
1.1Background1
1.2Declaration2
1.3Objectives2
1.4Internationalperspective2
1.5Peer review3
1.6Applicants3
2.CHEMICAL IDENTITY, PHYSICAL AND CHEMICAL PROPERTIES4
2.1Chemical identity4
2.2Physical and chemical properties4
3.MANUFACTUREANDUSE8
3.1Manufacture and importation8
3.2Usesof Complex SoapTH178
4.OCCUPATIONAL EXPOSURE9
4.1Routesof exposure9
4.2Sources ofoccupationalexposure9
4.2.1Importation anddistribution9
4.2.2Endusers9
4.3Measuresandestimates of occupationalexposure9
4.4Conclusions10
5.PUBLICEXPOSURE11
6.ENVIRONMENTAL EXPOSURE12
6.1Sourcesof environmental exposure12
6.1.1Release fromuse of ready-to-use formulated products
containing Complex SoapTH1712
6.1.2Release from wastewater, sewerage systems and landfill12
6.1.3Release due tounintentionalincidents13
6.2Fate and transformation processes13
6.2.1Fate13
6.2.2Persistenceandbioaccumulation13
6.3Conclusions13
7.HUMAN HEALTH HAZARDASSESSMENT14
7.1Toxicokineticsand metabolism(ND)14
7.2Effects on laboratory animalsand other test systems15
7.2.1Acute toxicity15
7.2.2Irritation andsensitisation15
7.2.3Repeat dose toxicity16
7.2.4Genotoxicity16
7.2.5Other end points16
7.3Effectsobserved in humans16
7.4Regulatoryclassificationsbasedonhazard16
7.4.1Physicochemical hazards16
7.4.2.Health hazards17
7.5Conclusions17
8.ENVIRONMENTAL HAZARD ASSESSMENT18
8.1Terrestrial organisms18
8.2Aquatic organisms18
8.2.1 / Amphibians / 188.2.2 / Fish / 18
8.2.3 / Aquatic invertebrates / 18
8.2.4 / Algae / 18
8.2.5 / Microorganisms / 19
8.3Predicted-no-effectconcentrations(PNEC)19
8.4Conclusions19
9.HUMAN HEALTH RISK CHARACTERISATION20
9.1Critical health effects20
9.2Occupationalhealth risk estimates20
9.3Public health riskestimates20
9.4Conclusions20
10.ENVIRONMENTAL RISKCHARACTERISATION21
10.1Critical environmental effects21
10.2Environmental risk estimates21
10.3Conclusions21
APPENDIX122
A.1.TOXICOLOGICAL INVESTIGATIONS23
A.1.1. / Acute toxicity–oral / 23A.1.2. / Acute toxicity-dermal / 23
A.1.3. / Acute toxicity- inhalation / 24
A.1.4. / Irritation–skin / 24
A.1.5. / Irritation- eye / 24
A.1.6. / Skin sensitisation / 25
A.1.7. / Repeat dose toxicity / 26
A.1.8. / Genotoxicity -bacteria / 27
A.1.9. / Genotoxicity – invitro / 28
A.2.ENVIRONMENT 29
A.2.1. / Environmental fate / 29
A.2.2. / Ecotoxicological investigations / 29
GLOSSARY / 33
REFERENCES / 38
Preface
ThisassessmentwascarriedoutundertheNationalIndustrialChemicalsNotificationandAssessmentScheme(NICNAS).ThisSchemewasestablishedbytheIndustrialChemicals(NotificationandAssessment)Act1989(Cwlth)(theAct),whichcameintooperationon17July1990.
TheprincipalaimofNICNASistoaidintheprotectionofpeopleatwork,thepublicandtheenvironmentfromtheharmfuleffectsofindustrialchemicals.
NICNASassessmentsarecarriedoutinconjunctionwiththeAustralianGovernmentDepartmentoftheEnvironment,Water,HeritageandtheArts,whichcarriesouttheenvironmentalassessmentforNICNAS.
NICNAShastwomajorassessmentprograms:theassessmentofhumanhealthandsafetyandenvironmentaleffectsofnewindustrialchemicalspriortoimportationormanufacture;andtheotherfocussingontheassessmentofchemicalsalreadyinuseinAustralia,inresponsetospecificconcernsabouttheirhealth/orenvironmentaleffects.
ChemicalsthathavebeenassessedasneworexistingchemicalsmayrequireareassessmentoftheriskofthechemicalunderthesecondarynotificationprovisionsoftheAct.
ThisassessmentreporthasbeenpreparedbytheDirectorofNICNAS,inaccordancewiththesecondarynotificationprovisionsoftheAct.UndertheActmanufacturers/importersofthechemicalarerequiredtonotifytheDirectorof newinformation andapplyfor assessment.Newinformationcanincludeanincreaseinquantityimported,thecommencementofAustralianmanufacture,increasedenvironmentalexposure,and/oradditionalinformationbecomingavailableonhazards.
Applicantsforassessmentaregivenadraftcopyofthereportand28daystoadvisetheDirectorofanyerrors.Followingthecorrectionofanyerrors,theDirectorprovidesapplicantsandotherinterestedpartieswithacopyofthedraftassessmentreportforconsideration.Thisisaperiodofpubliccommentlastingfor28daysduringwhichrequestsforvariationofthereportmaybemade.WherevariationsarerequestedtheDirector’sdecisionconcerningeachrequestismadeavailabletoeachrespondentandtootherinterestedparties(forafurtherperiodof28days).NoticesinrelationtopubliccommentanddecisionsmadeappearintheCommonwealthChemicalGazette.
InaccordancewiththeAct,publicationofthisreportrevokesthedeclarationofthischemicalforsecondaryassessment,thereforemanufacturersandimporterswishingtointroducethischemicalinthefutureneednotapplyforassessment.However,manufacturersandimportersneedtobeawareoftheirdutytoprovideanynewinformationtoNICNAS,asrequiredunderSection64oftheAct.
ForthepurposesofSection78(1)ofthe Act,copiesofassessmentreportsfornewandexistingchemicalassessmentsarefreelyavailablefromtheweb( arepublishedintheCommonwealthChemicalGazette(
CopiesofthisandotherpriorityexistingchemicalreportsareavailableontheNICNASwebsite.HardcopiesareavailablefreeofchargefromNICNASfromthefollowingaddress:
NICNAS
GPOBox58,Sydney,NSW2001,AUSTRALIA
Tel:+61(2)85778800
Fax:+61(2)85778888
Freecall:1800638528
OtherinformationaboutNICNAS(alsoavailableonrequestandontheNICNASwebsite)includes:
–NICNASServiceCharter;
–InformationsheetsonNICNASCompanyRegistration;
–InformationsheetsonthePriorityExistingChemicalsandNewChemicalassessmentprograms;
–SafetyinformationsheetsonchemicalsthathavebeenassessedasPriorityExistingChemicals;
–DetailsfortheNICNASHandbookforNotifiers;and
–DetailsfortheCommonwealthChemicalGazette.
MoreinformationonNICNAScanbefoundattheNICNASwebsite:
OtherinformationonthemanagementofworkplacechemicalscanbefoundatthewebsiteofSafeWorkAustralia:
Overview and Recommendations
Overview
ComplexSoapTH17wasassessedasSTD/1021undertheNICNASNewChemicalsprogramin2002inthestandardnotificationcategory.Asaresultofnewinformation,referredtoasnewdata(ND)below,relatingtophysico-chemicalandtoxicologicalpropertiesofComplexSoapTH17becomingavailable,ComplexSoapTH17hasnowbeenreassessedunderthesecondarynotificationprovisionsoftheIndustrialChemicals(NotificationandAssessment)Act1989(theAct)relevanttoexistingchemicals,asmorethanfiveyearshaveelapsedsincetheoriginalassessment.
ComplexSoapTH17belongstothegroupofUnknownorVariableComposition,ComplexReactionProductsorBiologicalMaterials(UVCB).Itisawhitesolidcomplexbariumsaltsynthesizedin,andneverseparatedfromanoilbase.Itisstableundernormalconditionsandathightemperature.TheoverallwatersolubilityoftheComplexSoapTH17UVCBislow.Initialassessmentofwatersolubilitybyvisualexaminationofundissolvedparticlesindicatedsolubilityoflessthan1g/L.DuetothevariablecompositionoftheUVCB,differentcomponentshavedifferentpropertiesrelatedtothewatersolubilityandpartitioncoefficient.Based onmeasurementsofdissolvedcarboncontent(DOC) overallwatersolubilitywasdeterminedtobe219mgDOC/Lat20oC.Then-octanol/waterpartitioncoefficient(logKow)wasestimatedtovarybetween0.9and18forthedifferentcomponentsoftheUVCBcomplex.Inaddition,newdatasubmittedduringthesecondarynotificationassessmentdemonstratethatwaterextractabilityofbariumionsfromtheUVCBcomplexislow.
PartitioncoefficientwasestimatedthroughQuantitativeStructureActivityRelationships(QSAR)andindicatedthattwomaincomponentsofComplexSoapTH17willbeimmobileinsoilwhileoneofthemajorthreecomponentsisexpectedtoexhibithighermobilityinsoil.EstimatesofdissociationconstantsuggestthatComplexSoapTH17iscompletelydissociatedintheenvironmentallyrelevantpHrange.
Manufactureandimportation
ComplexSoapTH17isnotmanufacturedinAustralia.Itisintroducedasacomponent(<35%)ofavarietyofready-to-usegreaseproductsandnotinitsnativestate.
Importationvolume of ComplexSoap TH17 hasnotexceeded 5tonnes per annumin the past4years.
Uses
ImportationanduseofComplexSoapTH17isthesameasthatnotifiedfortheoriginalassessmentasanewchemical.ThegreaseproductscontainingtheComplexSoapTH17areusedforlongtermandlifelonglubricationofrollingbearingsinsealedcomponentsintheautomotiveindustry.SincetheintroductionofComplexSoapTH17in2002,sixteendifferentproductsarebeingintroduced.Importedproductscontain30%ofComplexSoapTH17asnotifiedintheoriginalassessmentexceptonewhichcontains<35%ofthechemical.
MorethanhalfoftheimportedComplexSoapTH17isusedinnewautomotive,machineryandequipmentmanufacturingsitesandtherestisusedinrepairsandmaintenance.
Healtheffects
ToxicokineticsdatawerenotavailablefortheassessmentofComplexSoapTH17asanewchemical.Duringthesecondarynotificationassessmentnewinformationwasprovided.Inparticular,invitroCaco-2cellspermeabilityassaywasprovidedassupportingdataforassessmentofthebioavailabilityofthischemicalviatheoralroute.Thestudyreportcontainedreportingerrorsrenderingthestudyunreliable.Inaddition,therecoveryofthetestmaterialatdifferentstagesofthestudywashighlyvariable,probablyduetothelowwatersolubilityofComplexSoapTH17inthisaqueousbasedcellsystem,whichcontributedfurthertothelowreliabilityofthestudy.
Basedonthephysico-chemicalpropertiesofComplexSoapTH17whichindicatelowwaterextractabilityofthebarium ioncomponentoftheUVCB,thebioavailabilityofthiscomponentviatheoralrouteisalsoconsideredtobelow.ThisissupportedbythelowacuteoralanddermaltoxicityofComplexSoapTH17experimentallydeterminedtobegreaterthan2000mg/kgbw.Noacuteinhalationdataareavailable.
ComplexSoapTH17wasonlyslightlyirritatingtotheeyesinrabbitsandhadnoirritatingorsensitisingeffectswhenapplieddermallyinrabbitsandguineapigs.
RepeatedoraltreatmentofWisterratswithComplexSoapTH17for28dayscausednosignificanttoxiceffectsatthemid-doselevelof150mg/kgbw/dwhichwasdeterminedtobetheno-observed-adverse-effectlevel(NOAEL).Atthelowest-observed-adverse-effectlevel(LOAEL)of750mg/kgbw/d(thehighdose),signsoftoxicityincluded:decreasesinfoodconsumptionandbodyweightgainsinhigh-dosemalerats,anddecreasesofthemeanrelativespleenandadrenalweightsinhigh-dosefemalerats.
ComplexSoapTH17wasnegativeinbacterialmutationtestsandwasnotclastogenicinachromosomalaberrationstudy inChinesehamster V79cells.The latterstudywashampered bypoorsolubility,sotheconcentrationstestedwerelowerthanusualforthistypeoftest.Noothertoxicologicalstudieswereavailable.
ComplexSoapTH17wasclassifiedasahazardoussubstancefollowingtheassessmentofthischemicalasanewchemicalin2002(NICNAS,2002).TheassessmentindicatedalikelihoodthatbariumionsfromtheComplexSoapTH17mayhavesimilarhazardouseffectsassolublebariumsalts,listedontheHazardousSubstancesInformationSystem,withtheriskphrasesR20/22(Harmfulbyinhalationandifswallowed).
Basedonthedataavailableatthetimeoftheoriginalassessmentandthenewdataprovidedforthesecondarynotification,itisestablishedthatComplexSoapTH17haslowacutetoxicityandthelikelihoodofsystemicexposuretobariumionsfromComplexSoapTH17throughtheoralrouteislow.
Basedontheavailabledata,ComplexSoapTH17isnotclassifiedashazardousunderthe
ApprovedCriteriaforClassifyingHazardousSubstances(NOHSC,2004).
Occupationalexposureandhealthrisk
TherisktooccupationalhealthandsafetyislowbasedonthelowtoxicityprofileofComplexSoapTH17andthelimitedexposureofworkersundertheconditionsoftheoccupationalsettingsdescribedbytheapplicant.
DermalexposuretoproductscontainingComplexSoapTH17isthepredominantrouteofoccupationalexposure.Thelikelihoodforexposureishighestforworkersatmaintenancesitesandsitesmanufacturingnewmachineryfortheautomotiveindustry.Exposureofimportation
anddistributionworkersisnegligibleandisonlyexpectedintheunlikelyeventofbreachedpackaging.
GiventhatComplexSoapTH17isnotaskinirritantoraskinsensitiser,therisktoworkersresultingfromcontactwiththischemicalislow.However,repeatedorprolongedskincontactwithlubricantandgreaseproductsshouldbeavoidedsincehumanexperiencehasshownthatprolongedskincontactwithlubricantorgreaseproductsmaycauseskinirritationand/ordermatitis(oilacneorfolliculitis).
Tofurtherminimise therisk ofexposure,personalprotective equipmentsuchas gloves,eyewearandprotectiveclothingarereportedtobewornwhenthepossibilityofexposuretodripsandspillsexistsduringtheprocessesofpreparation,cleaningandmaintenance.
Publicexposureandhealthrisk
ComplexSoapTH17isintendedforindustrialuseonlyandwillnotbeavailabletotheAustralianpublic.GiventhelikelylowexposureofthepublicandthelowtoxicityprofileofComplexSoapTH17,therisktopublichealthisconsideredtobenegligible.
Environmentaleffects
DatawereavailableonlyforalimitedsetofenvironmentaltoxicityendpointsforComplexSoapTH17.Thischemicalisnottoxictofish,daphniaandmicro-organismsuptothelimitofitssolubility.Lowtoxicitywasobservedinatestwithalgaebelowthesolubilitylimit.
Dataonbioaccumulationarenotavailable,butduetothelimitedexposureoftheaquaticcompartmenttoComplexSoapTH17,bioaccumulationisnotexpected.
Environmentalexposureandrisks
EnvironmentalexposuretoComplexSoapTH17isexpectedtobelowasthemajorityofthegreasecontainingComplexSoapTH17willbecollectedanddisposedofbyincineration.Thewastesresultingfromspillageandresiduallubricantinimportcontainerlinersanddiscardedmachinerywillalsobedisposedofinlandfill.
Althoughitisnotconsideredtobereadilybiodegradable,ComplexSoapTH17isexpectedtobiodegradetoacertainextentinlandfill.Thehighoctanol-waterpartitioncoefficientcalculatedforComponentsIIandIIIofthechemicalUVCBandtheexpectedlowwatersolubilities,indicatethattheywillpartitiontosoilandsedimentandbeimmobileintheenvironment.Thethirdcomponentofthe UVCB,ComponentI,is moderately soluble,hasalowpartitioncoefficientandispotentiallymobileinsoil.However,asaconsequenceofitsanionicnatureitisexpectedtoassociatewithmetalionsonthesurfaceofsoilandbeimmobile.
Basedontheexpectedlowenvironmentalexposure toComplex SoapTH17theriskofadverseeffectsfortheenvironmentisconsideredtobelow.
Recommendations
ThissectionprovidestherecommendationsarisingfromthesecondarynotificationassessmentofComplexSoapTH17.RecommendationsaredirectedprincipallytoimportersofComplexSoapTH17.Implicitintheserecommendationsisthatbestpracticeisimplementedtominimiseoccupationalandpublicexposureandenvironmentalimpact.
Recommendationstoindustry:
Recommendationsrelatedtooccupationalcontrols:
•Thefollowingsafeworkpracticesshouldbeimplementedtominimiseoccupationalexposureduringhandlingofproductscontainingthenotifiedchemical:
–avoidrepeatedorprolongeddermalexposure.
–avoidgenerationofanyoilmistoraerosolofthenotifiedchemical.
•Thefollowingpersonalprotectiveequipmentshouldbeusedbyworkerstominimiseoccupationalexposuretoproductscontainingthenotifiedchemical:
–gloves
–safetyeyewear,and
–protective clothing.
•In the casethat anyoilmist oraerosol of Complex Soap TH17 is generatedatworkplaces,theExposureStandardsforoilmist(5mg/m3,TWA)mustbeapplied(NOHSC,1995).
Secondary Notification
UnderSection65oftheAct,thesecondarynotificationofComplexSoapTH17mayberequiredwhereanapplicantorotherintroducer(importer)ofComplexSoapTH17,becomesawareofanycircumstancesthatmaywarrantareassessmentofitshazardsandrisks.Specificcircumstancesinclude:
a.The use of Complex Soap TH17 has changed, or is likely to changesignificantly.
b.Manufacture of Complex Soap TH17 has begun, or is likely to begin inAustralia.
c.Additional information has become available on the adverse health and/orenvironmentaleffectsofComplexSoapTH17.
TheDirectormustbenotifiedwithin28daysoftheintroducerbecomingawareofanyoftheabovecircumstances.
Acronyms & Abbreviations
ATMApicaltransportmedium
BTMbasolateraltransportmedium
bwbodyweight
Ccelsius
CaCO3calciumcarbonate
CASChemicalAbstractsService
dday
DMSODimethylsulfoxide
DOCdissolvedorganiccarbon
EbC50medianeffectiveconcentration(biomass)ECEuropeanCommission
ECBEuropeanChemicalsBureau
EC50medianeffectiveconcentration
EgC50medianeffectiveconcentration(growth)
EUEuropeanUnion
ggram
GIgastro-intestinal
GLPGoodLaboratoryPractice
hhour
HBSSHanks'BalancedSaltSolution
HPLCHighperformanceliquidchromatography
ICP-MSInductivelycoupledplasmamassspectroscopykgkilogram
Kocorganiccarbonabsorptioncoefficient
Kowoctanol/waterpartitioncoefficient
Llitre
LC50medianlethalconcentration
LD50medianlethaldose
LOAELlowest-observed-adverse-effectlevel
LOEClowest-observed-effectconcentrationm3cubicmetre
mgmilligram
mg/cm3milligramspercubiccentimetremg/kgbwmilligramsperkilogrambodyweightmg/kgbw/d mg/kgbodyweight/day
minminute
mLmillilitre
μgmicrogram
μMmicromolar
MSDSMaterialSafetyDataSheet
NDnewdata
NICNASNationalIndustrialChemicalsNotificationandAssessmentSchemeNOAELno-observed-adverse-effectlevel
NOECno-observed-effectconcentration
NOHSCNationalOccupationalHealthandSafetyCommissionOECDOrganisationforEconomicCooperationandDevelopmentOECDTG OECDTestGuideline
PaPascals
PECPredictedenvironmentalconcentration
PNECPredictednoeffectconcentration
QSARquantitativestructure-activityrelationship
ThODNH4theoretical oxygen demand calculated on the basis of formation ofammonium
ThODNO3theoreticaloxygendemandcalculatedonthebasisofformationofnitrateUSFDA UnitedStatesFoodandDrugAdministration
UVCBSubstancesofunknownorvariablecomposition,complexreactionproductsorbiologicalmaterial
lessthan
1.Introduction
1.1Background
ComplexSoapTH17wasassessedasanewchemicalunderSection23oftheIndustrialChemicals(NotificationandAssessment)Act1989(theAct)inthestandardnotificationcategoryandisnowlistedontheAustralianInventoryofChemicalSubstances(AICS).
TheNewChemicalsAssessmentReport(STD/1021)waspublishedinAugust2002.NoexperimentaldatawereprovidedformostofthechemicalandphysicalpropertiesofComplexSoapTH17fortheassessment.Variationoftheschedulefordatarequirementswasclaimedforvapourpressure,watersolubility,hydrolysisasfunctionofpH,partitioncoefficient,adsorption/desorption,dissociationconstant,particlesize,flashpoint,explosivepropertiesandreactivity.Theseendpointswereeitherestimatedbased onQSARor limitedexperimentaldata.In addition notoxicological studieswereprovidedforacuteinhalationtoxicityandchromosomeaberrationinvivo.
Fromthedataprovidedatthattime,ahazardassessmentwasconductedanditwasdeterminedthatthenotifiedchemical(ComplexSoapTH17)wouldnotbeclassifiedashazardousundertheApprovedCriteriaforClassifyingHazardousSubstances(NOHSC,1999).However,ComplexSoapTH17wasdeterminedtocorrespondtothegroupofsolublebariumsalts,whichislistedontheHazardousSubstancesInformationSystemwiththeriskphrasesR20/22(Harmfulbyinhalationandifswallowed),andwasthereforeclassifiedasahazardoussubstanceandassignedthesameriskphrases.
InformationonthewatersolubilityofComplexSoapTH17includedaverycrudeprotocolofmixingandvisualassessment.Watersolubilitybythismethodwasdeterminedtobelessthan1g/L.Anothertest,asimplifiedflaskmethodandanalysisofdissolvedorganiccarboncouldnotprovideinformationaboutthesolubilityorextractabilityofthebariumcomponentofComplexSoapTH17.
Readersare referred to the New Chemicals Full Public Report onComplex SoapTH17availableontheNICNASwebsite:
Recommendationsweremaderelatingtosafeworkpractices,useofpersonalprotectiveequipmentandMaterialSafetyDataSheet(MSDS). TheserecommendationswerebasedontheintendeduseofComplexSoapTH17asacomponentoflubricatingproductsintheautomotiveindustry.
ComplexSoapTH17wasoriginallyassessedforuseasacomponentofaready-to-usegreaseforlongtermandlifelonglubricationofrollingbearingsinsealedcomponents.Theusepatternremainsthesame.
IntheperiodbetweenFebruary2008andOctober2008,additionalinformationwithregardtothebariumextractabilityandtoxicokineticpropertiesinvitrowassuppliedforComplexSoapTH17.Thenewdatawarrantreassessment,andasfiveyearshaveelapsedsincetheoriginalassessment, thischemicalisnowbeingassessedasan
existingchemicalunderSection68AoftheAct,coveringsecondarynotificationsofexistingchemicals.
Datasubmittedfortheoriginalassessmentonuse,exposure,animaland humantoxicityaresummarisedinthisreportintherelevantsections.DetailsofthestudiesprovidedforassessmentasanewchemicalarereproducedinAppendixI.NewdatasubmittedforthisassessmentarediscussedindetailandidentifiedbytheabbreviationND.
1.2Declaration
DeclarationasasecondarynotificationwasinitiatedwhenNICNASreceivedadditionalinformationregardingthewaterextractabilityofbariumfromComplexSoapTH17andaninvitrostudyofitstoxicokineticproperties.Theseadditionaldatawerenotavailableduringitsassessmentasanewchemical.Theinformationincludes:
•anEnglishtranslationandadditionalinformationontechnicalaspectsofatestregardingthewaterextractabilityofbarium(thisinformationisexemptfrompublication),and
•Caco-2cellsPermeabilityAssay.
ItwasconsideredthatasecondarynotificationforComplexSoapTH17wasrequiredbecausetheadditionalinformationprovidedhaverelevancetothehazardousnatureofthechemical.
AnoticewaspublishedintheChemicalGazetteof6January2009(AustralianGovernment,2009)requiringallpersonswhointroduceComplexSoapTH17intoAustraliaeitherbymanufactureorimport,toapplyforsecondarynotification.
1.3Objectives
Theobjectivesofthisassessmentweretoreviewthenewinformationmadeavailablesincethepublicationofthe2002NewChemicalAssessmentReport,andwhereappropriate,revisetheoriginalassessmentto:
•characterisethehazardsofComplexSoapTH17tohumanhealth;
•characterisepotentialoccupational,andpublicexposuretoComplexSoapTH17;
•characterisetherisksofadverseeffectsresultingfromexposuretoworkers,andthegeneralpublic;and
•makeappropriaterecommendationstocontrolexposuresand/orreducepotentialhealthrisksforworkersandthegeneralpublic.
NonewenvironmentalstudiesorusepatternswerereportedforComplexSoapTH17.Henceanenvironmentalassessmentwasnotconducted.TheenvironmentalassessmentconductedforComplexSoapTH17asanewchemicalhasbeenreproducedhere.
1.4Internationalperspective
ComplexSoapTH17wasassessedasanewchemicalbytheGermanCompetentAuthorityintheEU.TheEuropeanChemicalBureau(ECB)ClassificationandLabellingsectionconfirmedthatnolabellingisrequiredforComplexSoapTH17intheEUandthatnoEUharmonisedclassificationisproposed.(ND).
1.5Peerreview
Duringallstagesofpreparation,thisreporthasbeensubjecttointernalpeerreviewinNICNAS.
1.6Applicants
FollowingtheSecondaryNotificationdeclarationofComplexSoapTH17,onecompanyappliedforassessmentofthischemical.
InaccordancewiththeIndustrialChemicals(NotificationandAssessment)Act1989,NICNASprovidedthe applicantwith a draft copyof thereportfor commentduring thecorrections/variationsphaseoftheassessment.Theapplicantdetailsareasfollows:
KLUBERLUBRICATION(KL)AUSTRALIAPTYLTD
1stFloor,3BrandDriveThomastown,VIC3074
2.Chemical Identity, Physical and Chemical Properties
2.1 / ChemicalidentityName: / ComplexSoapTH17
ChemicalName: / Confidential
Marketingname(s): / ComplexSoapTH17KomplexSeifeTH17
IsoflexNBU(formulatedgrease)IsoflexTopasNB(formulatedgrease)StaburagsNBU(formulatedgrease)AltempQNB(formulatedgrease).
CASNumber: / Confidential
MolecularFormula / Confidential
MolecularWeight: / Confidential
StructuralFormula: / Confidential
Degreeofpurity: / High
Additives/Adjuvants: / Confidential
2.2Physicalandchemicalproperties
PhysicalandchemicaldatapreviouslyassessedbyNICNASfortheoriginalassessmentofComplexSoapTH17asanewchemical(STD/1021)isreproducedinthisreport.NewinformationrelatingtowaterextractabilityofbariumfromComplexSoapTH17was providedinconfidencetoNICNAS.The conclusionfromtheanalysis ispresentedasnewdata(ND)inthissection.
ComplexSoapTH17isathreecomponentUVCBbariumsaltsynthesizedin,andneverseparatedfrom,anoilbase.
ThephysicalpropertiesofComplexSoaparegiveninTable2.1.
Table2.1PhysicalpropertiesofComplexSoapTH17
PropertyValueMethod/Comment
Physical stateWhite solid
Specificgravity1219kg/m3at 20ºCEC Directive 92/69/EEC
A.3RelativeDensity and OECDTG109, Density of liquids andSolids.
Melting point270ºC (decomposition)EC Directive 92/69/EEC
A.1 Melting Freezing Temperature and OECD TG102, Melting point/ Melting Range.
Vapour pressure8 × 10-8Pa at 25oCEstimated basedon the
calculatedboiling point and using theModified WatsonCorrelation.
Water solubilityExtractabilityofbariumfromthe
Complex Soap in water isvery low,10mg/L (ND).
Water solubility determined by visual assessment was less than 1g/L.
Water solubility determined by measuringDissolvedOrganic Carbon (DOC) wasdetermined tobe219 mgDOC/L at20oC
BasedonEECdirective 92/69,AnnexVA6and OECD TG105Water
Solubility (DOC measurement).
Partitioncoefficient (LogKow)
logPow =0.9(ComponentI) logPow =9.2(ComponentII) logPow =18 (Component III)
OECDTG117; HPLC;
EEC Publication No. 383, A8
Hydrolysis asa Function of pH:
NotdeterminedGiven thelowwater solubility of the chemical it was not possible todetermine its propensity tohydrolyse.
Adsorption/ Desorption
logKoc=1.9 (Component I) logKoc=6.3 (Component II) logKoc= 11.2(ComponentIII)
Estimated basedon octanol/waterpartition coefficients
Dissociation constant (pKa)
pKa=4.9(Aliphatic carboxylic acid)
pKa=22.7 (Amide)
Estimated applying a free energyrelationship basedonTaftand Hammett correlations.
Particle Size / Notdetermined / The chemical issynthesisedin situ in base
oils and will not be
isolated fromthegrease.
Flash PointNotdeterminedThechemical is estimated to havevery lowvapourpressure.
FlammabilityLimits / Nothighly flammable / In contact with theignition source, the
chemical glowed and
coloured black.
However, thetest
substance couldnot
sustain a burning
reaction.
Autoignition Temperature
315oC92/69/EECA.16
Relative Self-Ignition Temperature for Solids.
SurfaceTension / 70mN/mat 20oC / OECD TG115 Surface Tension ofAqueous Solutions, andEC Directive 92/69/EEC
A.5 SurfaceTension.
Vapourpressure
Estimatedvapourpressureofthelowestboilingpointcomponentofthechemicalmixturewas8×10-8Pa.Aboilingpointof331ºCwascalculatedforthefreeacidcomponentutilisingMeissner’smethod.Thevapourpressureofthiscomponentwascalculatedtobe1×10-4Pa.
Watersolubility(ND)
Theexperimentaldataprovidedfortheinitialassessmentdidnotincludeinformationonthesolubilityoftheentirecomplexsubstancebutonlyontheorganiccarboncomponent.
New datameasuring bariumcations extractedfromwater suspension of Complex SoapTH17sonicatedfor15minutesandstirredfor48hatroomtemperaturedemonstratedthatthesolubilityofthebariumcomponentoftheComplexSoapTH17isverylow,
<10mg/L.
Octanol/waterpartitioncoefficient
SolubilityofComplexSoapTH17inbothwaterandoctanolwassolowthatexperimentaldeterminationofthepartitioncoefficientwasnotpossible.Therefore,thepartitioncoefficientwasdeterminedbytheoreticalfragmentationofthemoleculeaccordingtotheLeo-Hanschmethod.
Dissociationconstant
ThedissociationconstantwasestimatedbasedonapplyingafreeenergyrelationshipbasedonTaftandHammettcorrelations.Basedonthese,ComplexSoapTH17iscompletelydissociatedintheenvironmentallyrelevantpHrange.
Adsorption/desorption
EstimatedthroughQuantitativeStructureActivityRelationships(QSAR)usingtherelationshipslog10Koc=0.544logPow+1.377(basedonpartitioncoefficient).Theadsorption/desorptioncoefficientindicatesthatcomponentsIIandIIIofthechemicalwillbeimmobileinsoil.However,ComponentIisexpectedtoexhibithighermobilityinsoil.
Explosiveproperties
Thechemicaldoesnotcontainanychemicallyunstableorhighlyenergeticgroupsthatmightleadtoanexplosion.
Reactivity
Thechemicaldoesnotcontainanychemicalgroupsthatindicateitmightactasoxidisingagent.
PhysicalpropertiesofthegreaseproductscontainingtheComplexSoap TH17
Thedensityoftheformulatedgreaseproductscontaining<30%ofComplexSoapTH17isintherangeof800to990kg/m3.Thebaseoilviscosityat40°Cmayrangefrom20to130mm2/s,andthedroppointsarebetween170°Cand>240°C.
3.Manufacture andUse
Nonewinformationonusepatternswasprovidedduringthesecondarynotification.Theuseandmaximumimportationvolumeofthechemicalisasoriginallynotifiedforassessmentasanewchemicalandisreproducedherefromthenewchemicalassessmentreport.
3.1Manufactureandimportation
ComplexSoapTH17isintroducedinAustraliaasacomponentofaready-to-usegreasepackagedinplasticlined25kgpailsand180kgsteeldrums.
FifteendifferentproducttypesareimportedwithaComplexSoapTH17contentof
<30%. Thisisthe sameconcentrationreported forimportation of Complex SoapTH17intheassessmentasanewchemical.Onlyonenewproductwasintroducedinsignificantvolumeduring2008withaComplexSoapTH17contentof<35%(ND).
MaximumimportationvolumeofComplexSoapTH17inalltheseproductshasnotexceededfivetonnesperannuminthepastfouryears.
3.2UsesofComplexSoapTH17
ComplexSoapTH17isacomponentofaready-to-usegreaseusedforlongtermandlifelonglubricationofrollingbearingsinsealedcomponents.
MorethanhalfofthetotalquantityofimportedComplexSoapTH17isusedinnewautomotive,machineryandequipmentmanufacturingsites.ThegreasescontainingtheComplexSoapTH17areaddedviaautomaticmeteringdevices,dipfeeddevicesorcentralisedlubricationdevicesinaclosedsystemduringtheassemblyofautomotivecomponentsormachineandequipmentparts.
MaintenancefittersandothermechanicsatmaintenanceworkshopsuselessthanhalfoftheimportedComplexSoapTH17.Theyapplythegreasemanuallybybrush,spatula,greasegunorgreasecartridgetoexistingmachinery.
4.Occupational Exposure
Theoccupationalexposureassessmentisreproducedfromthenewchemicalassessmentreport(NICNAS,2002).
4.1Routesof exposure
Dermalcontactisexpectedtobethemainrouteofoccupationalexposure.InhalationexposureisexpectedtobenegligiblebecausetheproductcontainingComplexSoapTH17ishighlyviscousandthereforehasreducedpotentialtogenerateaerosols.Inaddition,ComplexSoapTH17hasaverylowvapourpressure,sovapouraccumulationintheworkplaceairisunlikely.Eyecontactispossiblebutalsounlikelyduetothehighlyviscousnatureofgreaseproducts.
4.2Sourcesofoccupationalexposure
4.2.1Importation and distribution
Theformulatedgreaseproductscontaining35%ofComplexSoapTH17areimportedfromoverseasanddistributedtoendusersincludingcarandenginemanufacturers,miningsites,engineeringsitesandmaintenanceworkshops.Transport,storageanddistributionofthelubricantsshouldinvolvelittleexposuretothechemical,exceptinthecaseofanaccidentalspill.
4.2.2End users
At the automotive, machineryand equipmentmanufacturingsites,the grease isappliedautomaticallybyautomaticdripfeedingsystems and centralautomated greasedeliverysystems.Occupational exposure during theautomaticoperation is expected to benegligible.Possibleoccupationalexposuremayoccurwhenopeningthecontainers,addingthegreaseintostoragecontainers,andduringequipmentcleaningandmaintenance.Theseoperationsgenerallywilllastforashortperiodoftime,anddermalcontaminationwouldbethemainrouteofoccupationalexposure.
Maintenancefittersandothermechanicsatmaintenanceworkshopsapplythegreasemanuallybybrush,spatulaandgreasegun.Dermalexposuremayoccurduringthesemanualoperations.However,theexposureisexpectedtobeinfrequent(monthlyoryearly)andofshortduration(<1hour),astheseproductsaredesignedtobelongtermlubricants.
Workersare reportedtowearimpermeablegloves,protectiveeyewear,protectiveclothing,andsafetybootswhenusingthegreaserepeatedlyorforprolongedperiods.
4.3Measuresandestimatesofoccupationalexposure
Intheassessmentofoccupationalexposure,itispreferabletousemeasureddataforeachexposurescenario,however,nomeasureddataareavailableforComplexSoapTH17fromtheAustralianindustry.
The applicant has provided estimates of the categories and number of workerspotentiallyexposedtoComplexSoapTH17atconcentrationof<35%.
CategoryofWorkerNumberExposure
Duration
ExposureFrequency
Importer2-3h/day10-15days/year
8-10
Distributor
Manufacturingworker<35
2-3h/day<50days/year
Maintenanceworker<230<1h/time
4.4Conclusions
ComplexSoapTH17isimportedinpailsanddrumsasacomponent(<35%)ofready-to-usegrease.Theexposureforimportationanddistributionworkersisnegligibleexceptintheeventthatthepackagingisbreached.
MorethanhalfoftheimportedComplexSoapTH17isusedatmanufacturingsites.Minimalexposureoccursbecausethesystemsforapplyingthegreasearegenerallyenclosedandautomated.However,thepossibilityofexposuretodripsandspillsexistsduringtheprocessesofpreparation,cleaningandmaintenance.Dermalexposurewouldbethepredominantrouteofoccupationalexposuretoworkersduringtheseactivities.
Fittersorothermechanicsatthe maintenancesitesapplythegrease manuallytoexistingmachinerybybrush,spatula,greasegunorgreasecartridge.Dermalexposuremayoccur.However,theexposureisconsideredtobeofshortdurationandintermittent.
5.Public Exposure
Lubricantscontainingthechemicalarenotavailabletothepublicfordomesticuse,andpublicexposurethroughtheintendedindustrialuseisnegligible.SignificantpublicexposuretoComplexSoapTH17isunlikely,exceptintheeventofanaccidentalspillduringtransport.Intheeventofanaccidentalspillage,spillswouldbecollectedandplacedinsuitablecontainersfordisposal.AllwastematerialswouldbedisposedofinaccordancewithStateregulationsforthedisposalofoilandgreasebylicensedoilwasteandregisteredwastedrumcompanies,wheretheproductwillbeeitherrecycledorincinerated.
6.Environmental Exposure
NonewdatawereprovidedforassessmentoftheenvironmentalexposureforthesecondarynotificationassessmentofComplexSoapTH17.ThissectioncontainsrelevantinformationfromtheassessmentofComplexSoapTH17asanewchemicalwithoutchangestothecontentandformattedtothestyleofthesecondarynotificationreport.
Theenvironmentispotentiallyexposedduringallstagesofachemical’slifecycle.Thefourstages,whichshouldbeconsideredseparately,areasfollows:
•Production(notrelevantforthisassessmentasnoproductionoccursinAustralia);
•Processing/formulation(notrelevantforthisassessmentasnoprocessing/formulationoccursinAustralia);
•Useofready-for-useproductsatmultiplesitesthroughAustralia;and
•Disposal.
Regardlessoftheenvironmentalmediabeingassessedforexposure,withintheexposureassessmenttherearethreemainsteps,theultimateonebeingtoderivepredictedenvironmentalconcentrations(PECs)inrelevantenvironmentalmedia.Thesestepsare:
1.Releaseestimation;
2.Considerationofenvironmentalfateandpartitioningbehaviour(distribution);and
3.PECderivation.
6.1Sourcesofenvironmentalexposure
ComplexSoapTH17isusedasacomponentofready-to-usegreaseusedatindustrialsitesthroughoutAustralia.
6.1.1Releasefromuseofready-to-useformulatedproductscontainingComplex SoapTH17
Based on thepattern ofuse of products,eventuallytheentire importvolume ofComplexSoapTH17isexpectedtoneeddisposal.
Thechemicalinwastesresultingfromspillageandresiduallubricantinimportcontainerlinersanddiscardedmachinerywouldbedisposedofinlandfill.Similarly,themajorityoftheusedgreasecontainingthechemicalwouldbecollectedanddisposedofbyincineration.
6.1.2Releasefromwastewater,seweragesystemsandlandfill
Aswaste,themajorityoftheimportvolumeiscollectedanddisposedofbyincineration,resultingintheformationofwatervapourandoxidesofcarbonandnitrogenandmetalsaltsinthesludge.Asmallamountisdiscardedinlandfillthrough
thedisposalofplasticdrumlinersandmachinerytowhichthegreasecontainingComplexSoapTH17hasbeenapplied.
6.1.3Releaseduetounintentionalincidents
ComplexSoapTH17isimportedintoMelbourneinplasticlined25kgpailsand180kgsteeldrumsbysea.Fromtheapplicant’ssiteinVictoriathegreaseistransportedtovariousindustrialsitesthroughoutAustralia.
ReleaseofComplexSoapTH17duringimportation,transportationandstorageislikelyonlyinthecaseofanaccident.However,insuchcasespillsarelikelytobecollectedandplacedinsuitablecontainersfordisposal.AllwastematerialsaredisposedofinaccordancewithStateregulationsforthedisposalofoilandgreasebylicensedoilwasteandregisteredwastedrumcompanieswheretheproductiseitherrecycledorincinerated.
6.2Fateandtransformationprocesses
6.2.1Fate
ComplexSoapTH17isnotconsideredtobereadilybiodegradableundertheconditionsofaReadyBiodegradabilityManometricRespirometryTest(OECDTG301F).However,itisexpectedtobiodegradetoacertainextentinlandfill.Thehighoctanol-waterpartitioncoefficientcalculatedforComponentsIIandIIIoftheComplexSoapTH17UVCBmixtureandtheexpectedlowwatersolubilitiesindicatethattheywillpartitiontosoilandsedimentandbeimmobileintheenvironment.Thethirdcomponentofthemixture,ComponentI,ismoderatelysoluble,hasalowpartitioncoefficientandispotentiallymobileinsoil.However,asaconsequenceofitsanionicnatureitisexpectedtoassociatewithmetalionsonthesurfaceofsoilandbeimmobile.
6.2.2Persistenceandbioaccumulation
DataonthebioaccumulationpotentialofComplexSoapTH17werenotprovidedfortheassessment.DuetolowaquaticexposureComplexSoapTH17isunlikelytobioaccumulate(Connell1990).
6.3Conclusions
ComplexSoapTH17isusedasacomponentofready-to-usegrease,andmostofitiscollectedandincinerated,resultingintheformationofwatervapourandoxidesofcarbonandnitrogen.Asmallamountisdiscardedinlandfillprimarilythroughthedisposalofplasticdrumliners.Giventhatitisnotreadilybiodegradable(<20%over28days)anditscomponentshaverelativelyhighpartitioncoefficientsandlowwatersolubility,ComplexSoapTH17wouldassociatewithsoilandsedimentandslowlydegradeovertimeindicatinglowexposuretoorganismsintheenvironment.
7.HumanHealthHazardAssessment
Thissectioncontainsashortsummaryofthedatarelevanttothehumanhealthhazardassessmentofthechemical.Robustsummariesofthedataavailable fortheassessmentofComplexSoapTH17asanewchemicalarereproducedfromtheNewChemicalreportinAppendix1ofthisreportwithoutanymodification.Adetaileddescriptionofthenewdatasubmittedforthesecondarynotificationisincludedhere.
7.1Toxicokineticsandmetabolism(ND)
Dataonmetabolismand/orkineticsofComplexSoapTH17werenotavailableforitsassessmentasanewchemical.
DuringthesecondarynotificationassessmentnewinformationwasprovidedconsistingofaCaco-2cellsinvitrointestinalpermeabilityassayassupportingdataforassessmentofthebioavailabilityofthechemicalviatheoralroute.ThisinvitroassayisusedforassessmentofintestinalpermeabilityofdrugsubstancesasapartoftheclassificationofdrugsubstanceswithintheUSFDABiopharmaceuticsClassificationSystem(BCS).UndertheBCSsystemawaiverforinvivotestingofthebioavailabilityofanewdrugformulationcanberequestedforhighlysolubleandhighlypermeablesubstancesinformulationwhichhaverapidorsimilardissolutionrateasapreviouslytestedformulation.
Caco-2cellspermeabilityassayutilizesatwocompartmentsystemseparatedbypolarizedhumancoloncarcinomacelllinemonolayerwhichexhibitsabsorptionpropertiesofhumanintestine.Theintegrityofthecellmonolayerswasconfirmedbeforeandaftertheassay.Apical(200μL)andbasolateral(600μL)transportmedium(ATMandBTM)consistedofHanks'BalancedSaltSolution(HBSS)adjustedtopH
6.5andpH7.4,respectively.PropranololandRanitidinewereusedasreferencedrugswithhighandlowpermeability,respectively.
Thetestsubstance,ComplexSoapTH17,orthereferencedrugs(50μMfinaleach)wereaddedtotheATMfordeterminationofAtoBtransportortotheBTMfordeterminationoftheBtoAtransport.Cellswereincubatedfor120minutesat370Cinorbitalshakersinanincubatorwith5%CO2supplementedatmosphere.
ThechemicalsintheATMandBTMweremeasuredat0and120minatthetestfacilitybyLC-MS1(referencechemicals)orsenttothesponsor(KlüberLubricationMünchenKG)foranalysisofbariumconcentrationbyICP-MS2(testchemical).NoGLPcompliancereportforthestudywasavailable.
Theaveragerecovery ofComplexSoap TH17frompooledsamplesatthebeginningoftheassaywasabout60%.Lowrecoveryismostlikelyduetolowsolubilityofthetestchemicalandpossibleadsorptiontothecellsandtheplasticsurfacesofthecultureplate.Recoverywasgreaterthan100%aftertheshakingincubationfor120minat370C.Nosignsofprecipitationofthetestchemicalwasnotedatanytimeoranyconcentration.
1Liquid chromatography-mass spectrometry
2Inductively coupled plasmamass spectroscopy
Meanapparentpermeability(Papp)andthepercentofchemicaltransportedthroughthecellmonolayerarepresentedinthetablebelow.Theauthorsofthestudyonlycalculated%transportthroughthecellmonolayer forthetestchemicalfromanexperimentalruninwhichrecoveryatt0was~60%.TheresultsindicatethatapparentpermeabilityAtoBislowerthanthatBtoA,whichisinconsistentwiththe%ofsubstancetransportedinthesametimeinterval(AtoBmorethanBtoA).Therefore,valueswererecalculatedusingrawdatainthereportbyapplyingacorrectionforvolumedifferenceinthedonorandreceptorcompartmentswhichhadapparentlynotbeendonebeforebytheauthorsandresultedintheinconsistency.
ChemicalMeanPapp
MeanPapp%
%
rtb
areportedvalues
brecalculatedvalues
Overall,intestinalpermeabilityofComplexSoapTH17asestimatedinthisinvitrosystemappearstobelowcomparedtothereferencedrugs.ItisnotedthatgiventhelowwatersolubilityofComplexSoapTH17,theHBSSbufferedCaco-2cellsystemmaynotbeoptimalfordeterminingitsintestinalpermeability.
Consideringthatbioavailabilityofasubstanceisdeterminedbyitssolubilityinadditiontotheintestinalpermeability,thebioavailabilityofComplexSoapTH17throughGItractislikelytobelow.
7.2Effectsonlaboratoryanimalsandothertestsystems
7.2.1Acutetoxicity
AcutetoxicityofComplexSoapTH17wasexaminedintworeliablestudieswithrats,afteroralanddermalexposure.LD50forbothroutesofexposurewasfoundtobegreaterthan2000mg/kgbw
Noacuteinhalationstudydataweresubmitted.
7.2.2Irritationandsensitisation
In aneyeirritationstudywithComplexSoapTH17,two rabbitshadDraizescoresof 1andonehadascoreof2forconjunctivalrednessatonehouraftertreatment.Allotherscoreswerezero.Thus,ComplexSoapTH17isconsideredtobeaslighteyeirritant.
ComplexSoapTH17wasnon-irritanttorabbitskinandwasnegativetoguinea-pigsinaskinsensitisationadjuvanttest.
7.2.3Repeatdosetoxicity
Inareliable28-dayoralrepeatstudywithrats,decreasesinfoodconsumptionandbodyweightgainsinhigh-dose(750mg/kgbw/d)malerats,anddecreasesofthemeanrelativespleenandadrenalweightsinhigh-dosefemaleratswereconsideredtobetreatment-related.Nosignificanthistopathologicalchangeswereobservedatanydose.Theno-observed-adverse-effectlevel(NOAEL)wasestablishedasthemid-doseof150mg/kgbw/d.
7.2.4Genotoxicity
Complex Soap TH17 was negative in an Ames test and foundnotto be clastogenic in achromosomalaberrationstudyinChinesehamsterV79cells.However,thelatterstudywashamperedbypoorsolubility,sotheconcentrationstestedwerelowerthanusualforthistypeoftest.Noinvivogenotoxicitydataweresubmitted.
7.2.5Otherendpoints
NostudieswereprovidedtoallowassessmentofthereproductivetoxicityandcarcinogenicpotentialofComplexSoapTH17.
7.3Effectsobservedinhumans
InformationwithregardtoeffectsinhumanswasnotavailablefortheassessmentofComplexSoapTH17asanewchemical.
Theinformationsubmitted from the applicantduringthe secondarynotificationdidnotindicate any adverseeffects tohumans fromtheuse ofComplex Soap TH17inoccupationalsetting.
7.4Regulatoryclassificationsbasedonhazard
ThissectiondiscussestheclassificationofthehealtheffectsofComplexSoapTH17accordingtotheApprovedCriteriaforClassifyingHazardousSubstances(NOHSC,2004)whichprovide mandatorycriteria for determiningwhether aworkplacechemicalishazardous.
Theclassificationforhealtheffectsisbasedonexperimentalstudies(animalandinvitrotests).TheassessmentconductedforSTD/1021determinedComplexSoapTH17tobehazardousandclassifieditastoxicifswallowedandbyinhalation(R20/22)basedonpresumedsimilaritytobariumsaltswhicharelistedastoxicifswallowedandbyinhalation(R20/22)intheHazardousSubstancesInformationSystem (
Thehazardclassificationbasedontheoveralldata,includingthenewdata(ND)
providedfortheSecondaryNotificationispresentedbelow.
7.4.1Physicochemicalhazards
NophysicochemicalhazardsareindicatedforComplexSoapTH17ortheproductsthatcontainitwhichwillbeimportedinAustralia.Thischemicaldoesnotcontainany
unstableorhighlyenergeticgroupsthatmightleadtoanexplosion.Alsoitdoesnotcontainanychemicalgroupsthatindicateitmightactasanoxidisingagent.
7.4.2. Healthhazards
Acutetoxicity
ComplexSoapTH17showedloworalanddermaltoxicityinacuteratstudiesperformedaccordingtoOECDTG423AcuteOralToxicity–AcuteToxicClassMethodandOECDTG402AcuteDermalToxicity–LimitTest.Noacuteinhalationstudydataweresubmitted.Inaddition,ComplexSoapTH17haslowwatersolubilityandshowslowextractabilityofbariumionsinaqueoussolutionindicatinglowbioavailabilityforthechemicalandbariumionsinthegastrointestinaltract(ND).
Classification:Basedontheavailabledata,ComplexSoapTH17isnotclassifiedashazardousforacutetoxicityundertheNOHSCApprovedCriteriaforClassifyingHazardousSubstances(NOHSC,2004).
7.5Conclusions
Basedontheavailabledata,ComplexSoapTH17isnotconsideredtobehazardousundertheNOHSCApprovedCriteriaforClassifyingHazardousSubstances(NOHSC,2004).
8.Environmental HazardAssessment
Thissectioncontainsashortsummaryofthedata relevanttotheenvironmentalhazardassessmentof thechemical.Robustsummariesofthe dataprovidedfortheassessmentofComplexSoapTH17asanewchemicalarereproducedinAppendix1ofthisreportwithoutanymodification.
Nonewdatawereprovidedforassessmentoftheenvironmentalhazardsforthesecondarynotificationassessment.
8.1Terrestrialorganisms
Nodatawereprovidedrelatingtoterrestrialvertebrates,invertebratesorplants.
8.2Aquaticorganisms
8.2.1Amphibians
Nodataareavailable
8.2.2Fish
AcutetoxicitytestwithZebraFishBrachydaniorerio(OECDTG203)wasconductedusingthefiltrateofasupersaturatedstocksuspensionofComplexSoapTH17.Nomortalitieswereobserved.A96-hourEC50wasnotdeterminedbutthe96-hourNOECisexpectedtobegreaterthanthelimitofitsolubility.
ThedataindicatethatComplexSoapTH17isnottoxictofishuptothelimitofitssolubility.
8.2.3Aquaticinvertebrates
AcuteImmobilisationTest(OECDTG202)withDaphniaspeciesDaphniamagnawasconductedusingthefiltrateofasupersaturatedstocksuspensionofComplexSoapTH17.Noeffectswereobservedat24or48hours.A48-hourEC50couldnotbedeterminedbutthe48-hourNOECisexpectedtobegreaterthanthelimitofitssolubility.
ThedataindicatethatComplexSoapTH17isnottoxictodaphniauptothelimitofitssolubility.
8.2.4Algae
Algalgrowthinhibitiontest(OECDTG201)wasconductedwithScenedesmussuspicatususingthefiltrateofasupersaturatedstocksuspensionofComplexSoapTH17.After72hours,therewasnosignificantinhibitionofalgalgrowthandbiomassatthenominalconcentrationsof6.25,12.5and25mg/L.Atanominalconcentrationof50and100mg/LbothalgalgrowthandbiomasswerereducedwithEgC50at72hof
>100mg/LandEbC50at72hof85.6mg/L,respectively.
ThedataindicatethatComplexSoapTH17hadsomebutnotsignificanttoxicitytoalgaebelowthelimitofitssolubility.
8.2.5Microorganisms
ToxicityofComplexSoapTH17tomicrobialactivitywasexaminedinActivatedSludge,RespirationInhibitionTest(OECDTG209)usingsludgeobtainedfromsewagetreatmentplantinGermany.Afterthreehours,activatedsludgeshoweddifferencesinrespirationofbetween–8.5to–18.6%comparedtocontrol.The3-hourEC50couldnotbequantifiedbutisexpectedtobegreaterthan1000mg/L.
ThedataindicatethatComplexSoapTH17isnottoxictoactivatedsludgemicroorganismsupto1000mg/Linsuspension.
8.3Predicted-no-effectconcentrations(PNEC)
ComplexSoapTH17isoflowtoxicityuptothelimitofitssolubilityandnoPNECwascalculated.
8.4Conclusions
ComplexSoapTH17isnottoxictofish,daphniaandmicro-organismsuptothelimitofitssolubility.However,itshowssometoxicitytoalgaebelowthislimit.Bioaccumulationisnotexpectedduetothelimitedexposuretotheaquaticcompartment.
9.Human Health Risk Characterisation
9.1Criticalhealtheffects
ComplexSoapTH17showsnosignificantsystemictoxiceffectsinanimalsafteracuteorrepeatedtreatment.ThisisconsistentwithloworalanddermalbioavailabilityofpossiblytoxicbariumionsdissociatingfromthiscomplexUVCB.
ComplexSoapTH17showsnosignificantlocaltoxiceffectsaftertopicaltreatmentoftheskinandeyesofanimals.
9.2Occupationalhealthriskestimates
DermalexposurewouldbethepredominantrouteofoccupationalexposuretoComplexSoapTH17.Althoughthechemicalisneitheraskinirritantnoraskinsensitiser,repeatedorprolongedskincontactwithlubricantandgreaseproductsshouldbeavoidedsincehumanexperiencehasshownthatprolongedskincontactwithlubricantorgreaseproductsmaycauseskinirritationand/ordermatitis(oilacneorfolliculitis).
Thehealthriskforimportationanddistributionworkersisexpectedtobenegligibleexceptintheeventthatthepackagingisbreached.
At sitesmanufacturing newmachinery, exposure is likelytobe minimal as the systemsareenclosedandautomated.However,gloves,eyewearandprotectiveclothingarereportedtobewornduringtheprocessesofpreparation,cleaningandmaintenance.Withtheapplicationofpersonalprotectiveequipment,theriskofadversehealtheffectsatthemanufacturingsitesislow.
Fittersorothermechanicsatmaintenancesitesapplythegreasemanuallytoexistingmachinery.Adverseskineffectsmayensueifdermalcontactisrepeatedorprolonged.
Overall,thereisalowconcernforoccupationalhealthandsafetyundertheconditionsoftheoccupationalsettings.
9.3Publichealthriskestimates
ComplexSoapTH17isnotavailableforusebytheAustralianpublicasitisonlyacomponentingreaseintendedforindustrialuse.GiventhelikelylowexposureofthepublicandthelowtoxicityprofileofComplexSoapTH17,therisktopublichealthisconsideredtobenegligible.
9.4Conclusions
ThecurrentconditionsofuseofComplexSoapTH17inAustraliaposesnosignificantrisksforpublicorworkerhealth.
10.Environmental Risk Characterisation
10.1Criticalenvironmentaleffects
ComplexSoapTH17isnottoxictofish,daphniaandmicro-organismsuptothelimitofitssolubility.However,itshowssometoxicitytoalgaebelowthislimit.Bioaccumulationisnotexpectedduetothelimitedexposuretotheaquaticcompartment.
10.2Environmentalriskestimates
ComplexSoapTH17isusedasacomponentofready-to-usegrease,andmostofitiseventuallycollectedandincinerated,whichresultsintheformationofwatervapourandoxidesofcarbonandnitrogen.Asmallamountisdiscardedinlandfillprimarilythroughthedisposalofplasticdrumliners.Here,givenitisnotreadilybiodegradable(<20%over28days),hasarelativelyhighpartitioncoefficientsandlowwatersolubility,thechemicalwouldassociatewithsoilandsedimentandslowlydegradeovertime.
TheaboveconsiderationsindicateminimalrisktotheenvironmentwhenComplexSoapTH17isusedinthemannerandlevelsindicatedbythenotifier.
10.3Conclusions
Onthebasisoftheavailableinformation,theoverallrisktotheenvironmentfromtheuseofComplexSoapTH17islow.
Appendix1
ThetoxicologicalassessmentofComplexSoapTH17asanewchemicalisreproducedherewithoutmodification.
A.1.TOXICOLOGICAL INVESTIGATIONS
A.1.1.Acutetoxicity–oral
TestSubstanceKomplexseifeTH17
MethodOECDTG423Acute OralToxicity– AcuteToxic Class Method.
Species/StrainRat/Wistar
Vehicle1% carboxymethylcellulose
Remarks - MethodGLPQA
Results
GroupNumberandSex ofAnimals
Dose mg/kg bw
Mortality
13 males20000
23females20000
LD50> 2 000mg/kgbw
Signs ofToxicityNone.
EffectsinOrgansNone.
Remarks - ResultsNone.
ConclusionThenotified chemical is of low toxicity via theoralroute.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999a).
A.1.2.Acutetoxicity-dermal
TestSubstanceKomplexseifeTH17
MethodOECDTG402Acute Dermal Toxicity – Limit Test.
ECDirective 92/69/EEC B.3Acute Toxicity (Dermal) –Limit Test.
Species/StrainRat/Wistar
Vehicle1% carboxymethylcellulose
Typeof dressingSemi-occlusive.
Remarks - MethodGLPQA.
Results
GroupNumberandSex ofAnimals
Dose mg/kg bw
Mortality
15/sex20000
LD50> 2 000mg/kgbw
Signs ofToxicity- LocalNone.Signs ofToxicity- SystemicNone.
EffectsinOrgansNone.
Remarks - ResultsNo skin irritation observed.
ConclusionThe notifiedchemical is of low toxicity via thedermal route.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999b).
A.1.3.Acutetoxicity-inhalation
No toxicity data were submitted.
A.1.4.Irritation–skin
TestSubstanceKomplexseifeTH17
MethodOECDTG404Acute DermalIrritation/Corrosion.
EC Directive 92/69/EEC B.4 AcuteToxicity (Skin Irritation).
Species/StrainRabbit/New Zealand White
Numberof Animals3
Vehicle1% carboxymethylcellulose
Observation Period72 hours
Typeof DressingSemi-occlusive.
Remarks - MethodGLPQA.
ResultsTheDraize scores for erythema/eschar andoedemawere zero for all animals throughout the observationperiod of72 hours after treatment.
Remarks - ResultsNo clinical signs of systemic toxicity were found.
ConclusionThenotifiedchemical is non-irritatingtoskin.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999c).
A.1.5.Irritation-eye
TestSubstanceKomplexseifeTH17
MethodOECDTG405Acute Eye Irritation/Corrosion.
EC Directive 92/69/EEC B.5 AcuteToxicity (EyeIrritation).
Species/StrainRabbit/New Zealand White
Numberof Animals3
Observation Period72 hours
Remarks - MethodGLPQA.
Results
LesionAnimalNo.
Mean Score*
Maximum Value
Maximum Durationof Any Effect
Maximum Value at Endof Observation Period
1 / 2 / 3Conjunctiva: redness / 1 / 2 / 3 / 2 / 1hour / 0
Conjunctiva: chemosis / 0 / 0 / 0 / 0 / - / 0
Conjunctiva: discharge / 0 / 0 / 0 / -
Cornealopacity / 0 / 0 / 0 / 0 / - / 0
Iridialinflammation / 0 / 0 / 0 / 0 / - / 0
*Calculated on thebasisof the scores at24,48, and 72 hours for EACH animal.
Remarks - ResultsConjunctivadischargewas notobserved.At1hour, theDraize scoresof
conjunctivalredness for the three animals were1,2 and1, respectively.
ConclusionThenotified chemical is slightly irritating to the eye.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999d).
A.1.6.Skinsensitisation
TestSubstanceKomplexseifeTH17
MethodOECD TG406 SkinSensitisation- Adjuvanttest.
EC Directive 96/54/EC B.6Skin Sensitization - Adjuvant test.
Species/StrainGuineapig/DH
PRELIMINARYSTUDYMaximumNon-irritating Concentration: intradermal:notstated.
topical:100%
MAINSTUDY
Numberof AnimalsTest Group: 10ControlGroup:5 induction phase Induction Concentration:
intradermalinjection25% (highest applicable
concentration).
topicalapplication100%
SignsofIrritationNone.
CHALLENGEPHASE
1stchallengetopicalapplication:100%
Remarks - MethodGLPQA.
Results
AnimalChallengeConcentrationNumberofAnimalsShowingSkinReactionsafter:
1stchallenge
24h48 h72h
TestGroup100%0/100/100/10
Control Group100%0/50/50/5
Remarks - ResultsHistoricdataofpositivecontrols wereprovided in thereport.
ConclusionThere wasno evidenceof reactions indicative of skin sensitisationto the notifiedchemicalunder the conditionsof the test.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999e).
A.1.7.Repeatdosetoxicity
TestSubstanceKomplexseifeTH17
MethodOECDTG407 Repeated Dose 28-dayOralToxicityStudyinRodents.
EC Directive 96/54/EC B.7Repeated Dose(28 Days) Toxicity (Oral).
Species/StrainRat/Wistar
Routeof AdministrationOral–gavage.
Exposure InformationTotal exposuredays:28days; Doseregimen:7daysperweek;
Vehicle1% carboxymethylcellulose
Remarks - MethodGLPQA.
Results
GroupNumberandSex ofAnimals
Dose mg/kgbw/day
Mortality
I(control)5/sex00
II(low dose)5/sex300
IIImiddose)5/sex1500
IV (highdose)5/sex7500
Mortality and Time to Death
None.
Clinical Observations
Adose-dependentdecrease in foodconsumption and bodyweightgains was observedin treated male rats. The reduced foodconsumption andbodyweightgainreachedstatisticalsignificance in thehigh-dosemales.
Onemale in the group IIIhaddiarrhoea foroneday.
Laboratory Findings– Clinical Chemistry,Haematology,Urinalysis
There wereno treatment-relatedchanges inclinicalchemistry,haematologyand urinalysis tests.
Pathology
Noabnormalfindings were observed.
Effects in Organs
The meanrelative spleenand adrenalvaluesinfemales of thegroup IVwere significantly lower than correspondingcontrolgroup.Therewere noother treatment-relateddifferences in relativeand absolute organ weight for both sexes and anyof thegroups.
Histopathology
Histopathologicalexamination showedthatnodifferencesinincidence orseveritybetween controland treatment groupswereconsidered tobe of toxicological significance.
Remarks– Results
Thefollowingchangeswere considered tobetreatment-related in thestudy:
•Decreases in food consumptionand bodyweightgain inhigh-dose male rats.
•Decreasesof the mean relative spleen andadrenalweights in high-dosefemale rats.
Conclusion
The NoObserved AdverseEffectLevel(NOAEL)wasestablishedas150 mg/kgbw/day inthis study,basedon the decreases ofbodyweightgainand foodconsumption inhigh-dose malerats and the meanrelative spleen andadrenal values in high-dose female rats.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999f).
A.1.8.Genotoxicity-bacteria
TestSubstanceKomplexseifeTH17
MethodOECD TG471 Bacterial Reverse Mutation Test.
Plate incorporationprocedurePre incubation procedure
Species/StrainS. typhimurium: TA1535, TA1537,TA98, TA100,TA102Metabolic Activation System S9-mix
Concentration Range in MainTest
a)With metabolic activation:0-5000 µg/plate.
b)Withoutmetabolic activation:0–5000 µg/plate.
VehicleDMSO
Remarks - MethodGLPQA.
Experiment Iwas an incorporationtest, andexperiment IIwas a pre- incubation test.
Results
Metabolic
Test Substance Concentration(µg/plate)Resulting in:
ActivationCytotoxicityin PreliminaryTest
Cytotoxicityin MainTest
PrecipitationGenotoxicEffect
Absent>5000≥1 000
Test 1Not seen≥1 000Notseen
Test 2Not seen≥1 000Notseen
Present>5000≥1 000
Test 1Not seen≥1 000Notseen
Test 2Not seen≥1 000Notseen
Remarks - ResultsNo significantincreases inrevertantcolonynumbersof any testedstrains
wereobserved following treatment withthenotifiedchemical either in
thepresenceor absenceofmetabolic activation in bothincorporationtest and pre-incubation test.
Thepositive controls induced adistinct increaseof inducedrevertant colonies.
ConclusionThenotified chemical was not mutagenic tobacteriaunderthe conditions of the test.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999g).
A.1.9.Genotoxicity–invitro
TestSubstanceKomplexseifeTH17
MethodOECD TG473 InvitroMammalianChromosomal AberrationTest.
Cell Type/CellLineChinesehamsterV79 cells Metabolic Activation SystemS-9 mix
VehicleDMSO
Remarks - MethodGLPQA.
Metabolic Activation
Absent
Test Substance Concentration(μg/mL)Exposure Period
Harvest Time
Test10, 5*, 50*and100*4h20h
Test20, 5*, 50*and100*20h20h
Present
Test10, 5*, 50*and100*4h20h
*Cultures selected for metaphase analysis.
Results
Metabolic
Test Substance Concentration(µg/mL)Resulting in:
ActivationCytotoxicityin PreliminaryTest
Cytotoxicityin MainTest
PrecipitationGenotoxicEffect
Absent
Test 1Not seen≥5Notseen
Test 2Not seen≥5Notseen
Present
Test 1Not seen≥5Notseen
Remarks - ResultsThenotifiedchemical didnotincrease thefrequencyof aberration in
ChineseV79 cells inthepresence and absence ofmetabolic activation.
Thepositive controls induced asignificantincreaseof cells with structuralchromosome aberrations above test laboratory’s historic control level.
The studywashamperedbythe lowsolubility of the test substance in DMSOand theculture medium.
ConclusionThe notified chemical was not clastogenictoChinesehamsterV79 treatedin vitrounder the conditionsof the test.
TestFacilityBioservice Scientific LaboratoriesGmbH(1999h).
A.2.ENVIRONMENT
A.2.1.Environmentalfate
A.2.1.1.Ready biodegradability
TestSubstanceComplexSoap TH17
MethodOECD TG301 F Ready Biodegradability:ManometricRespirometryTest.
Exposure Period28days
Remarks - MethodActivated sludgewas mixed withthe test substance togive final
concentrationof 20 mg/L and with the standard material, aniline at a
concentrationof 25.3mg/L.The studywascarried outin darknessat 22°C.
Results
Test substanceAniline
DayMean%degradationDay%degradation
1418.51472.0
2816.82896.0
Remarks - ResultsTheaniline standardattained96%biodegradationafter28 days,
indicatingthe test conditions were valid.After28days, themean
biodegradationof the test substancebased onThODNH4was 16.8% and basedonThODNO3was16.2%. Resultsfromthe toxicity control indicate that thenotifiedchemical does nothavean inhibitory effecton activated sludge micro-organisms.
ConclusionThe notified chemical is notconsidered tobereadilybiodegradableunder the conditionsofOECD TG 301F.
TestFacilityInstitutFürBiologischeAnalytikundConsultingIBACON GmbH (1999i).
A.2.1.2.Bioaccumulation
Data on the bioaccumulation potential of thenotified chemical werenotprovided for this notification. Due to low aquatic exposurethenotified chemicalit is unlikely to bioaccumulate(Connell1990).
A.2.2.Ecotoxicologicalinvestigations
A.2.2.1.Acute toxicitytofish
TestSubstanceComplexSoap TH17
MethodOECDTG203Fish,Acute ToxicityTest
SpeciesZebraFish (Brachydaniorerio)
Exposure Period96h
WaterHardness250mg CaCO3/L
Analytical MonitoringTestsolutionswere notmeasured.
Results
Concentration mg/LNumber ofFishMortality
Nominal2 h24h48h72h96h
0700000
100700000
LC50Notdetermined
NOEC (or LOEC)Notdetermined
Remarks– ResultsThedefinitivestudieswere conducted onthe filtrate of a super saturated
stocksuspension of thenotified chemicalata nominalconcentrationof
100mg/L.The results of thedefinitive study showedthatnomortalities were observed at this test substance concentration.A96-hour EC50forthe notified chemical to Brachydanioreriowasnotdetermined but the96h NOECis expected tobe greater than the limitof it solubility.
ConclusionThe ecotoxicity data indicates thenotified chemical is not toxic to fish up to the limit of its solubility.
TestFacilityInstitutFürBiologischeAnalytikundConsultingIBACON GmbH (1999j).
A.2.2.2.Acute/chronictoxicitytoaquatic invertebrates[Delete as appropriate]
TestSubstanceComplexSoap TH17
MethodOECD TG202 Daphnia sp. Acute ImmobilisationTest
SpeciesDaphniamagna
Exposure Period48 hours
WaterHardness250mg CaCO3/L
Analytical MonitoringTestsolutionswere notmeasured.
Results
Concentration mg/LNumber ofD.magnaNumber ImmobilisedNominal 24h 48h
02000
1002000
LC50Notdetermined
NOEC (or LOEC)Notdetermined
Remarks - ResultsThedefinitivestudieswere conducted onthe filtrate of a super saturated
stocksuspension of thenotified chemicalata nominalconcentrationof
100mg/L.The immobilisationtestswith daphniawereperformed in duplicate using 10daphnids per flaskwith observationsperformed at24
and48 hours.A48-hour EC50for thenotified chemical to Daphnia magnawas notdeterminedbut the 48h NOEC is expected to be greater thanthe limitof it solubility.
ConclusionThe ecotoxicity data indicates thenotified chemical is not toxic to daphnia uptothe limit of its solubility.
TestFacilityInstitutFürBiologischeAnalytikundConsultingIBACON GmbH (1999k).
A.2.2.3.Algalgrowthinhibitiontest
TestSubstanceComplexSoap TH17
MethodOECD TG201 Alga,Growth InhibitionTest.
SpeciesScenedesmussuspicatus
Exposure Period72 hours
Concentration Range Nominal6.25,12.5,25,50and 100mg/L WaterHardness24mgCaCO3/L
Analytical MonitoringTestsolutionswere notmeasured
Results
BiomassGrowth
EbC50 (mg/Lat 72 h)NOEC(mg/L)ErC50(mg/Lat 72 h)NOEC(mg/L)
85.65010025
Remarks - ResultsAlgaewere exposed tothe filtrate ofa supersaturated stock suspension of
the notified chemical atanominal concentrationof 100mg/Lunder
constant illumination and shaking. After72h, there was no significant inhibitionof algal growth andbiomass at the nominal concentrationsof 6.25,12.5and25mg/L. Atanominalconcentrationof 50 and 100 mg/L both algalgrowth andbiomass were significantlyreduced.
ConclusionThe ecotoxicity data indicates thenotifiedchemical shows some toxicity toalgaebelowthe limit of itssolubility.
TestFacilityInstitutFürBiologischeAnalytikundConsultingIBACON GmbH (1999l).
A.2.2.4.Inhibition ofmicrobialactivity
TestSubstanceComplexSoap TH17
MethodOECD TG209 Activated Sludge, Respiration Inhibition Test.
InoculumActivated sludge
Exposure Period3hours
Concentration Range Nominal10,32,100, 320, 1000mg/L
ResultsThe activated sludge studywas conductedusingsludge obtainedfrom sewage treatmentplantinGroβ-Zimmern,Germany. Thedefinitive studywas conductedon nominalconcentrations of10,32,100, 320 and
1000 mg/L. Amounts of test material(5, 16, 50, 160and 500mg) were added towater(284mL) and sewerage(16 mL) andsamples were stirred continuously. The referencematerialusedin the study was 3,5- dichlorophenol.Whencomparedto the control, activatedsludge after3 h experienceddifferences in respirationof between –8.5 to–18.6%. The 3-hour EC50for the notified substance to activated sludge couldnotbe quantified.However, the3-hour EC50for the notifiedsubstance to