Document kg11p8z5kXxZr7V5K2VwdQ7y

PHYSICAUCHEMICAL PROPERTIES TEST SUBSTANCE Identity:[2-(N-Ethylperfluoroocetsaunlfonamido) ethylacrylate;may also be referredtoas B1228, D-1, ETFOSEA, or FX-13.(2-Propenoicacid,2[ethy[lheptadecafluorooctyl)slufonyl]amnio]ethylester,CAS # 423-825) Remarks:Materiiaslan ambersolid. Remarks:Reportstatetshatsamplepuritiys99% ormorebasedon information suppliedby Sponsor. The pudtyfidentiotfythe testsubstance cannot be substantiated.As presentedinthe reportt,he structuraflormulaindicatesthat "pudt)('cannobte assigned as "99% ormore".The lotnumber was 101. METHOD Water Solubility: OECD 105, Shake FlaskMethod Hydrolysis as a Function of pH: OECD 111 Dissociation Constants inWater: OECD 112, Spectrophotometdc Method PartitionCoefficient(1-octanoltwaterb)y HPLC: OECD 117 Vapor Pressure: OECD 104, Gas SaturationMethod GLP (Y/N): Yes Year completed: 1994 Remarks: Detailfsromthesestudiecsan be foundinone reporetntitle"dD:eterminatiofn Physico-chemicalPropertiesofSample D-1". RESULTS Water Solubility0:.89 mg/L at250C Hydrolysis as a Function of pH: At 25"C the reactionrateconstantsofthe test. substance at pH 4,7,and 9 were 0.017,0.020,and 0.046 day-'withhalflife times of42, 35,and 15 days, respectively. Dissociation Constants inWater: No dissociatiocnonstantsofthetest substance were determinedbecause the spectraofthe testsubstance at differenptH values didnotsignificantdliyffefrrom one another. PartitionCoefficient:K. > 6 Vapor Pressure: 6.0x 10-3 Pa at 250C Remarks: The studyresultisndicattehdatthetestsubstancweas composed of differenctomponents which,insome cases,yieldedresultsdifferenftrom each other. Water solubility:Itissuggested thatthe water solubilitoyf each component ofthe testsubstance isdifferenftrom the others. Hydrolysis as a functionof pH: The chemicalstructureofthe test substance suggeststhatthe reactionrateofeach component issimilarto each other. Dissociation constants Inwater: The water solubilitoyfthetest substance istoo low tofindionizedand unionizedforms of thetest substance. The laboratorydidnot investigatietsconstantsfurtherby eitherthe titratimoenthod orthe conductometdc method inthe OECD Guidelinebecause both methods are considerednot suitablefor substances withsuch a low solubilityH.owever, the chemical structureof the testsubstance suggeststhatitsdissociatiopnotentiailsvery low. ParitioncoefficientT:he testsubstance was detectedas ten peaks or more by HPLC. Vapor Pressure: The chemicalstructureofthetestsubstance suggests thatthe vapor pressureofeach component isdifferenftrom the others. DATA QUALITY Water Solubility ReliabilityK:limischranking2. The reportstatesa purityofthe substance of"99% or more".Since the purity/identoiftythetestsubstance cannot be substantiatedt,he statedaverage solubilitoyfthe testsubstance isnotdefensible.As presented,the structural formulaindicatesthat"pudt)('cannobte assigned as "99 % or more".Since the method of detectionisnot compound specifica,nd was not validatedi,tcannot be determined ifthe peaks detectedinthesample chromatograms do indeed adse from the testsubstance. Hydrolysis ReliabilityK:limischranking2. Withoutthe purity/identintfyormatioonn thetestsubstance and compound specificanalyticatlechniques,the experimentalldyetermined rateconstant cannot be substantiated. Dissociation Constants inWater Reliability:Klirriisrcahnking1. Potentialfora compound ofthe givenstructureto ionizeatany pH islow. Octanol/Water PartitionCoefricient ReliabiliKtlyi:miscrhankin3g. Since thepudty/identiotfythetestsubstancecannotbe substantiatedt,hestated partitiocnoefficienitsnotdefensible.Itwas notestablishedinthisstudywhat the retentiotnime(s)ofthetestsubstance(si)s.The UV detectorisnotcompound specifiacnd ithas notbeen definitivelsytablishetdhatthe>10 peaks assigned as belongingtothetestcompound areactuallryelatedto thetestmaterial. Furthermore,ithas notbeen establishetdhatthismethod fordeterminingthe octanol/wateprartitiocnoefficieanptpliestofluorochemicaclompounds similarin structurteothe testmaterial. Vapor Pressure ReliabilityK:limischranking3. The pudty/identiotfythetestsubstancecannotbe substantiatedT.he chromatogram ofthe testsubstancehas a distinctdliyfferenptrofiltehatthatof the standard.Ithas notbeen establishetdhatunder theconditionsofthe experimentthe nitrogenpassed overthesubstancehas become saturatedwith gas phase testsubstance.The referencecompound (benzoicacid)has a measured vaporpressure- 16 timesthatdeterminedforthetestsubstance. It would be appropriatetochoose a referencecompound witha vapor pressure closertothatdeterminedforthetestcompound The lowermolecularweight constituentwsillhave a highervapor pressurethanthe n=8 component. REFERENCES Study conductedattherequestof Sumitomo 3M LTD by MitsubishCihemical SafetyInstitutLetd.,Yokohama, Japan OTHER Submifter: 3M Company, EnvironmentalLaboratoryP,.O. Box 33331, St.Paul, Minnesota,55133 Last changed: 5/22/00 M.S.I.Report No. 4B2234B227 Determinationof Physico-chemicalPropertiesof Sample D-1 Submittedto: SUMITOMO 3M LTD. Prepared by: MitsubishiChemical SafetyInstitutLetd. February 14, 1996 (Theoriginarleportwas submittedon August 8, 1994) Determinationof Physico-chemicalPropertiesof Sample D-1 (Englishversion) Study No.: Study Title: Sponsor: 4B223-4B227 Determinationof Physico-chemicaPlropertieosfSample D-1 SUMITOMO 3M LTD. This testwas conductedin Yokohama Laboratoryof MitsubishiChemicalSafety InstitutLetd.,1000 Kamoshida-cho,Aoba-ku, Yokohama 227, Japan. ThisreportistheEnglishversionof theoriginalw,hich was writtenin Japanese. The undersignedherebydeclarethatthisversionfaithfulrleyflecttsheoriginarleport tothebestofour knowledge. Translatedby Date: ShiroIWAMI, M.Sc. EnvironmentalScienceDivisionof theYokohama Laboratory Approved by -----------------D-at-e-:J-i-TTOoRIYA, c. Head of theYokohama Laboratory, ------- TestingFacility Study No.: 4B223-4B227 Study Title: Determinatioonf Physico-chemicaPlropertieosfSample D-1 Sponsor: SUMITOMO 3M LTD. Testing Fa@cflity: Yokohama LaboratoryM,itsubishCihemicalSafetyInstituLtted.t (M.S.I.), 1000 Kamoshida-cho,Aoba-ku,Yokohama 227,Japan(telephone0:45-963-3541) t ne conwany'sname was changedon October1,1994. (ne formername:Mitsubishi-kaIsnesitituotfeToxicologicaanldEnvironmentaSlciences) FacilityManagement: JunTORIYA, B.Sc. Head oftheYokohama Laboratory Sealeddate:August 8, 1994 Study Management: TadayoshiSHIGEOKA, Ph.D. Sealeddate:August 8, 1994 Chief ResearchScientist, EnvironmentalScienceDivisionoftheYokohnma Laboratory Study Director: Takemi NAKANOME, B.Sc. -Sealeddate:August 8, 1994 SeniorResearchScientist, EnvironmentalScienceDivisionof theYokohama Laboratory Experimental Scientist: Akiko TAKEDA, M.Sc. Sealeddate:August 8, 1994 EnvironmentalScienceDivisionof theYokohama Laboratory Contents page Abstract.................................................5............ 1 TestSubstan.c.e.........................................6............ 1.1 Identificat.i.o.n.....................................6............ 1.2 Source............................................6............ 2 WaterSolubili[tSytudyNo. 4B223].............................7............ 2.1 Materialasnd Methods...................................7............ 2.2 Resultsand Discussio.n.................................9............ Tablesand Figures........................................27........... 3 Hydrolysiassa functioonfpH [StudyNo. 4B224]....................1.0........... 3.1 Materialasnd Methods..................................1.0........... 3.2 Resultasnd Discussio.n.................................1.4........... Tablesand Figures........................................35........... 4 DissociatiCoonnstantisnWater [StudyNo. 4B225]....................16........... 4.1 Materialasnd Methods..................................1.6........... 4.2 Resultasnd Discussio.n.................................1.8........... Figure................................................5.0.......... 5 PartitioCnoefficie(n1t-octanol/wabtyerH)PLC Method(StudyNo. 4B226]....19. 5.1 Materialasnd Methods..................................1.9........... 5.2 Resultsand Discussio.n..................................21........... Figures...............................................5.1........... 6 Vaporpressur[eStudNyc@4.B227]..............................2.2........... 6.1 Materialasnd Methods..................................2.2.......... 6.2 Resultsand Discussio.n.................................2.6.......... Tablesand Figures..................................*...........@.5.3........... (59pagesinall) Abstract Study No.: Study Title: Sponsor: 4B223-4B227 Determinationof Physico-chemicalPropertieosf Sample D-1 SUMITOMO 3M LTD. 1. Test Substance Name: Sample D-1 Chemical name: 2-[N-ethyl-N-perfluoroa(lCk-y1l-8)sulfonylamino] ethylacrylate Structuraflormula:CnF2n+IS02N(C2H5)CH2cH20C(.O)CH=CH2 n=1-8 (n=8: approx.78 %; n=1-7: approx.21 %) 2 Water Solubilit[yStudyNo. 4B223] 2.1 Methods: No. 105 @ "Water Solubility"(Flask Method) 2.2 Results: Water solubUityof the testsubstancewas 0.89 mg/L at 25 C. 3 Hydrolysis as a function of pH [Study No. 4B2241 3.1 Methods: No. 111 @ "Hydrolysis as a function of pH", (testingat pH 4, 7, and 9 at250C) 3.2 Results: At 25 0C reactionrateconstantsof the testsubstanceatpH 4, 7, and 9 were 0.017, 0.020, and 0.046 day-I with half lifetime of 42, 35, and 15 days, respectively. 4 Dissociation Constants in Water [Study No. 4B225] 4.1 Methods: No. 112t "DissociationConstants in Water" (Spectrophotometric Method) 3.2 Results: No dissociationconstantsof the testsubstance were determined because the spectraof the testsubstance at differentpH did not significantldyifferfrom one another.(But the chemical structureof the testsubstance suggests that its dissociationpotentialis very low.) 5 PartitionCoefiriden(t1-octanol/watebry) BPLC Method [StudyNo. 4B2261 5.1 Methods: No.117@ "Partition Coefficient (n-octanol/water),High Performance Liquid Chromatography (HPLC) Method" 5.2 Results: The testsubstance was detectedas ten peaks or more by HPLC. The log Pow value of the main component was more than 6. 6 Vapor Premure [Study No. 4B227] 6.1 Methods: No. 104@ "Vapor Pressure Curve" (Gas SaturationMethod) 6.2 Results: Vapor pressureof the testsubstancewas 6.0 X 10-3 Pa at 25" C. t Number in OrganizationforEconomic Cooperationand Development (OECD) Guidelines for Testingof Chemicals Test Substance 1.1 Identifi*cation 1)Namet: Chemical namet: Sample D-1 2-[N-ethyl-N-perfTuoroalk=yl1(-C8)sulfonylamino] ethylacrylate 2)StructurfaolrmulatC:,,F2,,I+S02N(C2H5)CH2CH20C(=O)CH=CH2 n=1-8 (n=8: approx.78 %; n=1-7: approx.21 %) 3)Physico-chemicaplroperties: Solubilityt: water: acetone-' DMSO: insoluble 50 % ormore insoluble Meltingpointt: 27-42 *C Boilingpointt: approx.150*C (1nim Hg) t providedby thesponsor 1.2 Source I)Batcht: Lot No. 101 2)Supplier: Misao SHIBA, SUMITOMO 3)Suppliedquantityta:pprox.100 g 4)Purityt: 99 % or more 5)Appearance: amber likewax iM LTD.- t providedby thesponsor 2 Water Solubilit[yStudyNo. 4B223] A solutionis a homogeneous mixtureof differenstubstancesin a solvent.The particlesizesof thedispersedsubstancesare of the same magnitude as moleculesand ions.Therefore,the smallestvolumes which can be obtainedfrom a solutionare always ofuniform composition. Solubilitiynwater isa significanptarameterbecause 0 the spatiaalnd temporalmovement (mobilityo)f a substanceis largely determinedby itssolubilitiynwater; 0 water solublesubstancesgain ready accessto humans and otherliving organisms; 0 theknowledge of thesolubilitiynwaterisa prerequisitfeortestinbgiological degradationand bio-accumulatioinnwawr and forothertests. 2.1 Materialsand Methods This study was conducted in accordancewith the standardprocedure "Water Solubility("FlaskMethod) in the OECDT Guidelinesfor Testingof Chemicals No. 105 (1981).This method issummarized asfollows: The testsubstance(Solidsmust be pulverized.i)s dissolvedin water at a temperaturesomewhat above'thetestt'emperature.When saturatioinsachieved the mixtureiscooledand kept at the testtemperature,stirrinags long as necessaryto reach equilibriumS.ubsequently,mass concentratioonf the testsubstancein the aqueous solutionw,hich must not containany undissolvedparticlesi,sdeterminedby a suitableanalyticamlethod. t OrganizatiofnorEconomic C@operatioannd Development 2.1.1 Reagents n-hexane: acetone: Wako Pure Chemical IndustriesL,td., guaranteedreagent Wako Pure Chemical IndustriesL,td., guaranteedreagent 2.1.2 Apparatus incubator: TaitecCorporation, model M-100 centrifugasieparator: HitachiLtd., model SCT 15B gas chromatograph(GC):Shimadzu Corporation,model GC-14A integrator: ShimadziL.Corporation,-.modCe-lR3A, 2.1.3 Testprocedure Two hundremdilligraomfsthetesstubstanwcaesdissolviend10mL ofacetone in an Erlenineyerglassflask.The solventwas evaporatedwithstirrintgo deposita thinlayerof thetestsubstanceonto innersurfaceof theflask.The residualsolvent was thoroughlyremoved by stream of nitrogengas . Four hundred milliliteorfs distillewdater was added to the flask,which was then shaken at 400.2*C over a 3-day period.A 50-mL aliquotof thewaterphase was sampled after1,2, and 3 days.The aliquotwas subsequentlyshakenat250.2'C for 1 day or more toreach equilibriumof dissolutioonf thetestsubstance. The operationby theabove procedurewas repeatedonce more. 2.1.4 Analytical methods The concentrationof the testsubstancewas measured by gas chromatography (GC). Priorto GC analysise,achequilibrateadliquotwas treatedasfollows: The aliquotwas centrifugeadt 4000 rpm (3000 x g) at 25*C. Sodium chloride,1.2 g, was dissolvedin thesupernatantt,o which 4 mL of n-hexanewas then added They were shaken for 1 minutesa-nd then centrifugeadt4000 rpm (3000 X g) at 25* *C. The n-hexanephase (supernatantw)as analyzedby GC under the followingconditions: GC conditions column: Shimadzu Corporation,wide bore column CBP20-W25-100, 0.53 mm i.d.2,5 m inlength temperature: column: 120*C; injector2:00*C; detector2:40*C carrier: nitrogengas (flowrate:20 mL/min) detector: electroncapturedetector(ECD) injectiovnolume: 3 pL As informedby the sponsor,thecomponents ofthe testsubstancedifferinlength of the perfluoroalkyclhain (see'1.1)I.n fact,the testsubstancewas detectedas fourmajor peakswithretentiotnimeof 3.8,4.5,5.1,and 6.0minutesby GC under the conditionsdescribedabove.Therefore,the concentratioonf the testsubstance was based on totalareaof thefourpeaks. 2.1.5 Calibration curve Standardsolutionsof the testsubstancewere preparedto make concentrationsof 0, 0.2, 1.0,and 5.0 mg/L inn-hexane.These standardsolutionwsere analyzedby GC under the conditionsd.escribe@d-i2n,1.4,.Thetgtalpeak area of the fourpeaks was calculatedand plottedagainstthe concentratioonf the testsubstance.The calibratiocnurveyieldeda straighltinepassingthroughtheoriginand itscorrelation coefficienwtas calculatetdo be 0.999 by the leastsquare method describedin JapaneseIndustriaSltandards(JIS)Z 9041-1968. [Figure2.1and Figure2.2) At measurement of the testsubstanceconcentrationt,he standardsolution with 1.0 mg/L was analyzed.Concentrationineach testsample was calculatefdrom ratiooftotalpeak areaforthesample tothatfor thestandardsolution. 2.1.6 Recovery An acetonesolutionof the testsubstancewas preparedto make a concentration of 483 mg/L. Two milliliteorfsthesolutionwas dissolvedin 1000 mL of distilled water(finalconcentratioonf thetestsubstance0:.97 mg/L). Sodium chloride,1.2g, was dissolvedin4 mL of thewatersolutiont,owhich 4 mL of n-hexanewas then added. They were shaken for 1 minutesand then centrifugeadt4000 rpm (3000X g) at25 *C. The concentratioonf thetestsubstance in the n-hexane phase (supernatantw)as measured by GC under the conditions describedin 2.1.4toevaluaterecovery. [Table2.2 and Figure2.3] The recoveryof the testsubstancewas 94 %. The concentrationosf the test substancereportedbelow were correctedby thisfactor. 2.2 Resultsand Discussion Average watersolubilitoyf thetestsubstancewas 0.89 mg/L at25 *C. [Table2.1 and Figure2.4.1toFigure2.4.3] The component withGC retentiotnime of 4.5 minuteswas rathersolubleinwater compared with the others.Dissolutioonf thiscomponent at40 *C reachedequilibrium within1 day. Itissuggestedthatwatersolubilitoyf each component of the testsubstancediffers from one another. 3 Hydrolysisas a Functionof pH [StudyNo. 4B2241 The testingof substanceforhydrolysisisrelevanttotheirpersistenceH.ydrolysis isone of themost common reactionscontrouingabioticdegradationand istherefore one of themain degradationpathsof substancesintheenvironment. A procedureto determinehydrolysisisimportantalsoinindicatinwghether other testingshouldbe carriedout on a parentcompound or itshydrolysisproduct.Itisthe degradationproductsthatarecrucial. Hydrolysisbehaviorneeds to be examined at pH valuesnormallyfound -inthe environment(pH 4-9)and under more acidiccondition(spH 1-2)for physioloigcal purpose. Surface-controllerdeactionscan sometimes predominate over bulk solution hydrolysise,specialliynthesoilenvironment.This may resultindifferendtegradation ratesthan would be predictedfrom thismethods based upon ratesin homogeneous solutions. 3.1 Materialsand Methods This studywas conductedinaccordancewith the standardprocedure "Hydrolysis asa functionof pH" intheOECD GuidelinesforTestingofChemicalgNo.,l11 (1981). In theenvironimentc,hemicalsusuallyoccur in dilutesolutionw,hich means that water ispresentinlargeexcess,and thereforet,hekineticosfhydrolysisare generally pseudo-firsotrderatfixedpH and temperature. The hydrolysirseactionmay be influencebdy acidicor basicspeciesH30+ (H+) and OH-, inwhich caseitisreferredtoasspecifiaccidorspecifibcase catalysis. The concentratioonf thetestsubstanceisdeterminedas a functionof time.The logarithmsof the concentrationasre plottedagainsttime and theslopeof theresulting straightline(assumingfirst-ordeor pseudo-firsotrder behavior)gives the rate constant. 3.1.1 Reagentsand water acetone: n-hexane: sodium chloride: sodium hydroxide: Wako Pure Chemical IndustriesL,td., guaranteedreagent KishidaChemical Co.,Ltd. guaranteedreagent JunseiChemical Co., Ltd., guaranteedreagent JunseiChemical Co., Ltd., guaranteedreagent monopotassium phosphate: Wako Pure Chemical IndustriesL,td., guaranteedreagent monopotassium citrate:NacalaiTesque Inc., extraPure reagent boricacid: potassiumchloride: Wako Pure Chemical IndustriesL,td., guaranteedreagent JunseiChemical Co., Ltd. guaranteedreagent water: deionizedwaterpurifiedby MiUi-(@9 3.1.2 Apparatus ,incubator: pH meter: gas chromatograph(GC): integrator: TaitecCorporation, model M-100 Toa ElectronicLstd., model HM-50S Shimadzu Corporation, model GC-14A [equippedwithan electroncapturedetector(ECD)] Shimadzu Corporation, model C-R3A 3.1.3 Testprocedure 3.1.3.1 Preparationof buffersolutions Buffersolutionwsere preparedby thefollowingtwo methodsdescribedin the annex of theGuidelineNo. 111: Buffermixturesof Clarkand Lubs CitratbeufferofKolthoff-andVleeschhouwer Each buffersolutionofpH values4, 7, and 9 was preparedtobe 1000 n3L using thefollowingreagents: I I)pH4: 0.1 m monopotassium citrate and 0.1 N sodium hydroxide 2)pH7: 0.1 m monopotassium phosphate and 0. IN sodium hydroxide 3)pH9: 0.1 m potassium chloride, 0. 1 m boric acid and 0. 1 N sodium hydroxide The buffersolutionsobtainedwere passedthroughMilliporeefilteorf pore size 0.25 lim.The pH valuesof thefiltratweesre determinedtobe 4.05,7.02,and 9.05 fornominalvaluesof 4, 7,and 9, respectively. 3.1.3.2 Preparation of stocksolutionof the testsubstance The testsubstance,80 mg, was dissolvedand dilutedwith acetonetopreparea stocksolutionwith40 mg/L. 3.1.3.3Preliminatreys(tat50*C) Fourmilliliotfetrhsestocskoluti(opnrepariend3.1.3.w2a)saddedto400mL each of thethreebuffersolutiontsopreparetestsolutionosf pH values4, 7, and 9. The solutiontshuspreparedwere testedunder thefollowingconditions: 1) Conditions pH: concentration: temperature: testinpgeriod: testvolume: test'vessel: 4, 7, and 9 0.40mg/L 50.00.10c 5 dayswithcontinuousshaking 400 mL (containin1g% acetone) Erlenmeyerglassflask II)Analyticalmethods The concentrationof the testsubstancewas measured by GC. Priorto GC analysist,hetestsolutionwsere treatebdy thefollowingprocedure: Sodium chloride,128 g, was dissolveidn each of thethree400-mL solutions, from which thetestsubstancewas thenextractewdith 100 mL of n-hexane.The extractiownas repeatedtwice more. The threeextractswere combined and concentratetdo50 mL, which was thentransferreidntoa 200-mL glassvolumetric flaskand filletdo200 mL with n-hexane.The hexanesolutiownas analyzedby GC under thefollowingconditions: GC conditions column: Shim@dzu Corporationw,ide borecolumn CBP20-W25-100, 0.53mm i.d.2,5 m inlength temperature: column:120*C;injecto2r0:0'C; detector2:40'C carrier: nitrogegnas(flowrate:20 mL/min) detector: ECD injectiovnolume: 3 pL As describedin2.1.4,theconcentratioonf thetestsubstancewas basedon total area of the fourpeaksdetectedby GC (retentiotnime:3.6,4.3,4.9,and 5.8 minutes). III)Recovery Four milliliteorfsthestocksolutio(nprepareidn3.1.3.2)was addedto400 mL of waterin an Erienmeyerglassflask(fmalconcentratio0n.:4 mg/L). Sodium chloride,128 g, was dissolvedin thissolutionf,rom which thetestsubstancewas thenextractedwith 100 mL of n-hexane.The extractiownas reppatedtwicemore. The threeextractswere combined and concentratedto 50 mL, which was then transferreidntoa 200-mL glassvolumetricflaskand filletdo200 mL withn-hexane. The concentratioonf the testsubstancein the finalsolutionwas measured by GC under theconditiondsescribedin3.1.3.3.Il. Recovery was determinedto be 101 %. T"neconcentrationosf thetestsubstance reportedforthispreliminarytestwere correctedby thisfactor. [Table3.6 and Figure3.1] 3.1.3.4 Further investigddon (at25 OC) Four milliliteorfsthestocksolution(preparedin 3.1.3.2)was added to 400 mL each of thethreebuffersolutiontsopreparetestsolutionosf pH values4, 7, and 9. Six 5-mL aliquotsfrom each of thetestsolutionwsere transferreidntoglasstubes. These solutionwsere examined asa functionof time under thefollowingconditions: I) Conditions pH: concentration: temperature: testingperiod: testvolume: testvessel: 4, 7,and 9 0.40 mg/L 25.00.2 0C 33 days with no shaking 5 mL (containinIg % acetone) glasstube II)Analytical methods The concentrationof the test substance was measured by GC. analysis,thetestsolutionswere treatedby the followingprocedure: Prior to GC Sodium chloride,1.6 g, was dissolvedin the5-mL aliquottreatedin the tube. Two milliliteorfsn-hexanewas added to thissolutionT.hey were shaken for 1 minutes and thencentrifugeadt 3000 rpm. The n-hexanephase (supernatantw)as analyzedby GC asdescribedin3.1.3.3. As describedin 2.1.4,theconcentratioonf the testsubstancewas based on total area of the fourpeaks detectedby GC (retentiotnime:3.6,4.3, 4.9,and 5.8 minutes). III)Recovery Four milliliteorfsthestocksolution(preparedin3.1.3.2)was added to400 mL of water in an Erlenmeyerglassflask(thefinalconcentratio0n.:4 mg/L). Sodium chloride1,.6g, was dissolvienda 5-mL sampleofthissolutiotno,whichtwo milliliteorfs n-hexanewas then added. They were shaken for 1 minutes and then centrifugedat 3000 rpm. The n-hexanephase (supernatantw)as analyzedby GC under theconditionsdescribedin3.1.3.3. Recovery was determinedto be 95 %. The concentrationosf the testsubstance reportedforthisfurtherinvestigatiwoenre correctedby thisfactor. [Table3.7 and Figure3.3] 3.1.3.5 CaUbration curve TJie calibratiocnurve prepared in 2.1.5 (Determinationof Water Solubility) was used. [Figure2.1 and Figure2.2) At measurement of the testsubstanceconcentrationt,hestandardsolutionof the testsubstancewith 1.0 mg/L was analyzed.Concentrationin each testsample was calculatedfrom ratioof totalpeak area for the sample to thatfor the standard solution. 3.1.3.6 Calculafions Residualpercentof thetestsubstancewas calculateadsfollows: rt= 100 (Ct/cto) where rt: residualpercentof thetestsubstanceafter"t"days ct: concentratioonf thetestsubstanceafter"t"days (mg/L) cto: initiacloncentratioonf thetestsubstance(0.4mg/L) Assuming thatthelogarithmsof theresidualconcentrationasre firstorderas a functionof time,theirreactionrateconstantswere calculateads follows: k t-Iln(100/r) where k: reactionrateconstantof thetestsubstance(day-1) Using theaverageof theserateconstants,halflifetime of the testsubstancewas calculateads follows: tl/2 = k-I In 2 where tl/2: halflifetime of the testsubstanre(days) 3.2 ResultasndDiscussion 1)Preliminatreyst After5 daysat50'C theresidupaelrcentosfthetesstubstanactepH 4,7,and9 were 76, 76, and 52, respectively. (Table3.5 and Figure3.2] On thebasisof theresultsi,tissuggestedthatthetestsubstanceistransformedat leastby 24 Relatingto thepreliminarytestresultt,hereisthe followingdescriptioinn the OECD Guideline: Iflessthan 10 per centofthe reactionisobservedafter5 days (tII2> I year), thechemicalisconsideredhydrolyticalsltyableand no additionatlestingisrequired. Therefore,we decidedtoperformthefurtherinvestigatidoenscribedbelow. 2)Furtherinvestigation At 25'C thereactionrateconstantsof thetestsubstanceatpH 4, 7, and 9 were 0.017,0.020,and 0.046 day-I with halflifetime of 42,.35, and 15 days, respectively. [Table3.1toTable3.4and Figure3.4] The chemicalstructuroef the testsubstance(shown'in 2.2)suggeststhatthe reactionrateof each component issimilarto each other. 4 DissociatioCnonstantsin Water [StudyNo. 4B225] The dissociatioonf a chemicalin water isof importancein assessingitsimpact upon theenvironment.Itgovernstheform of thesubstancewhich inturndetermines itsbehaviorand transportI.tmay affecttheadsorptionof thechemicalon soilsand sedimentsand adsorptionintobiologicaclefls.- 4.1 Materiallasnd Methods This study was conductedinaccordancewith the standardprocedure"Dissociation Constantsin Water" (SpectrophotometrMiecthod) in the OECD Guidelinesfor TestingofChemicalsNo. 112 (1981).Generalsummary of thismethod isas.follows: A wavelengthisfound where the ionizedand unionizedforms of thecompound have appreciablydifferenetxtinctiocnoefficientTsh.e LJV-visiblaebsorptionspectrum isobtainedfrom solutionosf constantconcentratiounnder a pH conditionwhere the substanceisessentialluynionizedand fullyionizedand atseveralintermediatpeH's. This may be done, eitherby adding incrementsof concentrateadcid (base)to a relativellyargevolume of a solutionof the compound in a multicomponentbuffer, initialaltyhigh (low)pH, or by addingequalvolumes of a stocksolutioonf the compound in e.g.water,methanol,to constantvolumes of variousbuffersolutions coveringthe desiredpH range.From thepH and absorbancevaluesatthe chosen wavelength,a sufficienntumber of valuesfor thepKa iscalculateudsingdatafrom at least5 pH's where thecompound isat least10 per centand lessdm 90 per cent ionized. 4.1.1 Reagents and water methanol: JunseiChemical Co., Ltd., guaranteedreagent 0.1 N hydrochloricacid: KishidaChemicalCo., Ltd., reagentfortitration (dilutetdo0.01 N withwaterpriortouse) 0.1 N sodium hydroxide: KishidaChemicalCo., Ltd., reagentfortitration (dilutetdo0.01N withwaterpriortouse) water: Deionizedwater was distilled. 4.1.2 Apparatus UV-visiblespectrophotometer:'ShimadzuCorporationM,odel UV-260 4.1.3 Testprocedure 4.1.3.1Preparatioonfstocksolutioonfthetestsubstance A stocksolutionfthetesstubstanwcieth80 mg/L inmethanowlasprepared using20.0 mg ofthesubstance. 4.1.3.2 Preparationof testsolutions For measurement of UV-visiblespectraof thetestsubstancet,hreesolutionosf thetestsubstancweere preparedasfollows: 1)Acid solutionof thetestsubstance Two milliliteorfsthestocksolutio(npreparedin4.1.3.1.)and 2.4 ml'of 0.01 N hydrochloriaccidwere transferriendtoa 200-mL glassvolumetrifclaskw,hich was thenfilletdo200 mL withwater.(finacloncentratiofnthetestsubstance0:.8 mg/L) 2)Alkalinseolutioonfthetesstubstance Two millilitoefrtshestocksolutioand2.4mL of0.01N sodiumhydroxide were transferreidntoa 200-mL glassvolumetricflaskw,hich was thenfilletdo200 mL withwater.(finacloncentratioofnthetestsubstance0:.8 mg/L) 3) Neutralsolutioonf thetestsubstance Two milliliteorfs the stocksolutionwas transferreidntoa 200-niLglass volumetrifclaskw,hichwas thenfilletdo200 mL withwater.(finacloncentration of thetestsubstance0:.8 mg/L) 4.1.3.3 Measurement of UV-visiblespectrum The UV-visiblespectraof thetestsubstanceinthesolutionosf threedifferenptH valueswere measuredwiththeapparatushown in4.1.2. 4.2 Resultsand Discussion No 4issociatiocnonstantsof thetestsubstancewere determinedbecause the spectra of thetestsubstanceatdifferenptH did notsignificantdliyfferfrom one another. [Figure4.1 The water solubilitoyf the testsubstanceis too low to find an ionizedand unionizedforms of thetestsubstance.We did not investigatietsconstantsfurtherby either"TitratioMnethod" or "ConductometricMethod" in the OECD Guideline because theboth methods are considerednot suitablefor thesubstancehaving such a low solubilit(y0.89mg/L, reporteidn2.2). But the chemicalstructuroef the testsubstance(shown in 1.1.2) suggeststhatits dissociatiopnotentiailsvery low. 5 PartitiCoonefficie(n1t-octanol/water) by BPLC Method [StudyNo. 4B226] The partitiocnoefficie(nPt) is defmed as the ratioof the equilibrium concentrationosf a dissolvedsubstancein a two-phasesystem.consistinogf two largelyimmisciblesolventsI.n caseof 1-octafiaonld water, POW= co /cw where Pow: 1-octanol/watpearrtitiocnoefficient Co: concentratioinn 1-octanoplhase Cw: concentratioinnwaterphase The partitiocnoefficiebnetingthequotienotf two concentrationisusuallygiven intheform of itslogarithmtobaseten(logPow). Pow isa key parameterinstudiesof theenvironmentaflateof chemicalsubstances. A highly-significraenltationshibpetween the-Pow of substancesand their bioaccumulatioinnfishhas been shown. Ithas alsobeen shown thatPow isa useful parameterin thepredictioonf adsorptioonn soiland sedimentsand forestablishing quantitatisvteructure-activrietlyationshifposra wide rangeof biologicaelffects. 5.1 Materialsand Methods This studywas conductedin accordancewith the standardprocedure"Partition Coefficien(tn-octanol/wateHri)g,h PerformanceLiquidChromatography(HPLC) Method" in theOECD GuidelinefsorTestingof ChemicalsNo.117(1989).Principle of thismethod isas follows: HPLC isperformedon analyticaclolumns packed witha commerciallyavailable solidphasecontaininlgonghydrocarbonchains(e.g.C8, C18) chemicallybound onto silica. Chemicalsinjecteodntosuch a column move alongitby partitionibnegtween the mobile solventphase and the hydrocarbonstationarpyhase.The chemicalsare retainedin proportionto theirhydrocarbon-wateprartitiocnoefficienwti,th water solublechemicalselutedfirstand oil-solublcehemicalslast.T'his enablesthe relationshibpetween the retentiontime on a reverse-phasceolumn and the 1-octanol/watpearrtitiocnoefficienttobe established. 5.1.1 Reference compounds A referencecompounds whose Pow valuesare well known, we selectedfive substanr-esm:ethyl benzoate,bromobenzene, diphenyl,dibenzyl,and DDT. In additiont,hioureawas used fordeterminatioonf thedead time.A mixtureof thesix substanceswas preparedin acetonitrile. 5.1.2 Correlationbetween retentiontime and Pow The mixtureofthereferencceompounds(shownin5.1.1)was analyzebdy high performanceliquidchromatography(.HPLC)under the conditiondsescribedbelow: HPLC con&dons column: mobile phase: flow rate: wavelength: temperature: GL SciencesInc.,InertsiOlDS-2 4.6mm i.d.,250 mm inlength (Cl8 chemicallybound ontosilica) acetonitrile/wa=te7r5/25 (v/v) 1.0 mL/min. 210 run 25 OC The HPLC retentiotnime of thereferencecompounds were correctedas follows: Rt '=Rt' -Rto where Rt: correctedretentiotnime ofthecompound (minutes) Rt': Rto: retentiotnimeof thecompound (minutes) retentiotnimeof thiourea(minutes) Correlatioenquationbetween thecorrectedretentiotnime of the fivecompounds and theircorrespondingPow was computed by the leastsquaremethod, resultinign thefollowingequation: log Pow = 5.274 log Rt - 0.0219 (r 0.973) [Figure5.1 and Figure5.21 5.1.3 Retention time of the testsubstance A mixtureof the testsubstanceand thioureawas preparedin acetonitrilTeh.e mixturewas analyzedby HPLC under theconditionsdescribedin 5.1.2.Based on thechromatogram,retentiotnime(Rt',minutes)were determinedforcomponents of the testsubstance. 5.2 Resultsand Discussion The,t.estsubstancewas detectedas tenpeaks or more by HPLC. The correctedretentiotnime (Rt)of the main component was determinedto be 29.245 minutes,whilethatof.DDT (Log Pow isreportedto be 6.20.)was 13.263 minutes. Based on thecorrelatioenquationshown in 5.1.2,thelog Pow valueof themain component was more than 6. [Figure5.2) 6 Vapor pressure[StudyNo. 4B227] The envirorunentarlelevanceof vapor pressureisaccountedforby the following reasons: The vapor pressuregivesan indicatioonf the probabilitoyf the phase transitionlsi,quid/gaasnd solid/gas. The vapor pressure,togetherwith the solubilitiyn water,is the major auxiliarvyariableforcalculatintghevolatiliotfy a substancefrom an aqueous solution. Vapor pressureis thus a significanftactorfor predictinagtmospheric concentrations. The vapor pressureof a substancecan furthermorebe usefulas a basisfor decidingwhetheror nota photochemicallyinduceddegradationstudy(inthe homogeneous gas phase or inan absorbedphase) isnecessary. 6.1 Materialsand Methods This study was conductedin accordancewith the standardprocedure "Vapor PressureCurve" (Gas SaturatioMnethod) in the OECD Guidelinesfor Testingof ChemicalsNo. 104 (1981).This method issummarized asfollows: A streamof inertcarriergas (nitrogengas)ispassedover thesubstanceinsuch a way thatitbecomes saturatedwith vapor of the substanceand the vapor is then collectedina trapadsorbent.Measurement of theamount ofmaterialtransportebdy a known amount of carriergas is used to calculattehe vapor pressureat a given temperature. 6.1.1 Reagents acetonitrile: Wako Pure ChemicalIndustriesL,td., reagentforHPLC adsorbent: GL SciencesInc., Tenax GC'O, 60-80 mesh 6.1.2 Apparatus saturatcoorlumn: adsorbentcolilmn: glassbead: flow meter: gas chromatograph: data processor: SibataScientifiTcechnology Ltd., 12 mm i.d.,150 mm inlength SibataScientifiTcechnology Ltd., 12 mm i.d.,150 mm inlength IuchiSeieidoCo.,Ltd., 1 mm indiameter Shinagawa Corporation, model NWK-IC Shimadzu Corporation, model GC-14A Shimadzu Corporation, model C-R3A 6.1.3 Vapor pressurememuring apparatus For measurement of vapor pressureof thetestsubstance,an apparatusshown in Figure 6.5 was assembledand setin a room air-conditioneadt25 1*C. [Figure6.5] The glassbeads coatedwith the testsubstancewere packed intothe saturator column up to 7 cm in length.Nitrogengas,kept at a constantflow rateby the pressurecontrollearnd theflow gauge,was introducedto thiscolumn. The nitrogengas saturatedwith vapor of the testsubstancewas deliveredto the Tenax GCO-packed column to trapthe testsubstance.Totalamount of the nitrogen gas which passedthroughthistrapcolumn was measured by theflow meter. Preparation of saturatorcolumn 1) Ten milligramsof the testsubstancewas dissolvedin 10 mL of acetonitrilien a 100-mL round bottom glassflask. 2) Eight milliliteorfsglassbeads was added intothe flask.The solventwas removed by rotaryevaporation.The residualsolventwas thoroughlyeliminated with a vacuum pump atroom temperature.(The testsubstancewas thuscoated onto theglassbeads.) 3) The glassbeads coatedwith the testsubstancewas packed intothe saturator column up to7 cm inlength.Both open sidesof-thecolumn were heldby quartz glasswool, which was then incorporateidn the vapor pressuremeasuring apparatus. nitrogen gas 1) Nitrogen gas was deliveredintothevapor pressuremeasuring apparatusfor 70 hours.Totalvolume of the carriergas was measured with the flow meter and determinedtobe 1.017 m3. (averag.eflow..rat2e4:2 mumin) 2) The totalvolume of thegaswas correctedasfollows: V = Vr [(273/T)(760-/P7)601"2 where V: Vr: T: P: totaglasvolumecorrecte(dm3) amountofthegasmeasuredby'theflowmeter(1.017m3) temperatureof thenitrogengas(298 *K) pressuredifferencebetween at the flow meter and at the absorber-packedcolumn (0.4 mm Hg) The totalvolume of the carrierwas thus correctedby factorsof the temperature and pressureditference,resultingin 0.973 m3. Adsorbent column 1) Eight milliliterosf the adsorbent (Tenax GC!O) was packed into the absorbent column. Both open sides of the column were retainedwith quartz glass wool. The column thusprepared was connected to the saturatorcolumn. 2) The testsubstance trapped by the Tenax G(Z!O was desorbed by the procedure describedin 6.1.4. 6.1.4 Analyticalmethods The testsubstancetrappedby the adsorbentwas desorbed as follows: 1) The adsorbentpacked in -thecolumn was transferredinto a glass filterT.he insideof the vacant column was rinsedwith 20 mL of acetonitrilew,hich was then transferredin the glassfilterT.he rinseof the Tenax.GC was repeated twice more with 20 mL and 10 mL each of acetonitrile. 2) The threefiltratewsere combined and transferredintoa 50-mL volumetric glass flask,which was then filledto 50 mL with acetonitrilAen. aliquotof the solutionwas diluted50 fold in volume. 3) The concentrationof the testsubstancein the finalsolutionwas measured by GC under the followingconditions: GC conditions column: Shimadzu Corporation,wide bore column CBP20-W25-100, 0.53mm i.d.2,5 m inlength temperature: column 120 'C, injecto2r00 *C, detector240 0C carrier: nitrogengas(flowrate:20 mL/min) detector: ECD injectiovnolume: 3 liL Calibrationcurve Standardsolutionosfthetestsubstancweere preparedtomake concentratioonfs 0, 0.25,0.5, and 1.0 mg/L in acetonitrilTeh.ey were analyzedby GC under the above conditionsA. calibratiocnurvepreparedby the method describedin 2.1.5 generateda straighltinewhich crossestheoriginand itscorrelatiocnoefficienwtas calculatetdo be 0.999. [Figure6.1 and Figure6.2] At measurement of the testsubstanceconcentrationt,he standardsolution with 1.0 mg/L was analyzed.Concentrationineach testsample was calculatefdrom ratioof totalpeak areaforthesample tothatforthestandardsolution. Desorption efficiency(Recovery) Desorptionefficiencyof the testsubstancefrom Tenax GC'O was determined accordingtothe followingprocedure: 1) Twenty fivemilliliteorfsthesolutioonf thetestsubstancewith I mg/L in acetonitri(lmeass of the testsubstance2:5 pg) was added to 4 mL of the adsorbentin a 50-mL round bottom glassflask.The solventwas removed by rotaryevaporationto adsorbthetestsubstanceonto theTenax G(@e. 2) The testsubstanceadsorbedon theTenax GC'O was recoveredby thedesorption proceduredescribedabove (exceptthatthe substancewas extractedwith 10, 10, and 5 mL ofacetonitrialned thefiltratweesre transferreidnto25-mL volumetric glassflask,which was then filletdo 25 mL). The concentratioonf the test substancein the finalsolutionwas measured by GC to evaluatedesorption efficiency. The desorptionefficiencwyas determinedtobe 97 %. For measurement ofvapor pressure,the concentratioonf thetestsubstancewas correctedby thisfactor. [Table6.3 and Figure6.3] 6.1.5 Calculatioonf vapor pressure V@por pressureof thetestsubstancewas calculatedas follows: P = (W/M)(RT/V) where P: W: M: R: V: T: vapor pressure(Pa) amount of thetestsubstancetrapped(g) molecularweightof thetestsubstance(g mole-1) gas constant(8.31Pa m3 mole-I K-1) totalvolume of thecarriergas corrected(m3) temperatur(e'K) 6.2 Resultsand Discussion Vapor pressureof thetestsubstancweas 6.0 x 10-3Pa. [Table6.1 toTable 6.2,and Figure 6.4] But the chemicalstructureof the testsubstance(shown in 1.1.2)suggeststhat vapor pressureof each component isdifferentfrom one another. Vapor pressureof benzoic acid (a referencesubstancein the OECD was 0.10 Pa (24 C) under theconditionsemployed in thisstudy. Guideline) Table 2.1 Calculationof concentrationof the testsubstance dissolvedin water. Measurement No. 1 time at 40.OC-,days 2 conc.ibStd,.;.mg/L' Peak area,pV-sec Std. testsolution concentratiofnactor A 1.01 1.01 1.01 B 2589644 2717449 2732276 c 2030738 2414771 2230296 D 1 1 1 conc.of thesubstance,mg/L E 0.84 0.95 0.88 Calculatioenquation E = A(C/B)(I/D)(1/0.94) Measurement No. 2 time at40 OC, days 2 3 conc.in..Stdm.g1l,l, peak area,ijv-sec Std. testsolution concentratiofnactor A i.o'i 1.01 1.01 B 2589644 2717449 2732276 C 2115067 2319263 2293546 D I I I conc.ofthe substance,mg/L E 0.88 0.92 Calculatioenquation E = A(C/B)(l/D)(1/0.94) averagewater solubility:0.89itg/l 0.90 -Table2.2 Calculationof recovery conc.of the substanceadded, mg/L A 0.97 conc.'iSntd.,mg/L B 1.01 peak area,jjv-sec Std: C 2723221 test,solution 0 2456907 concentratiofnactor E I conc.of the substancerecovered,mg/L recovery F calculatioenquation F=B(D/C)(I/E)(IOO/A) 0.91 94 Figure2.1 Calibratiocnurve of the testsubstance Curve Fitting (Least Square Method] Inp-ut Data ----------------------------------------- Concentration Peak Area No X (mg/ I Y iUV -sec) --------------------------------------------- 1 0 0 2 0.2016, 550784 3 1.008 2758732 4 5 .'04 13119948 --------------------------------------------- Y = 5.0483X 104 + r = 0.99995 2.5966X 106 X Xl X ioooooo 14 13 12 11 io 4) tn 9 8 d) 7 6 5 4 3 2 1 Oc 0.00 Calibration Curve 1.00 2.00 3 .00 4.00- Concentration(mg/1) 5.00 6.00 XI U) 33.,4U-i77;8-_ 5. !65 7. C) c@o OC) p 11%110 IME AREA liK 3.767 80396 v 3 4.492 317962 V 4 5.1@fi 1821019 v 5 5.982 539354 S Y TOTAL 2769805 -@KtiC, t 4 5 7.. t-ZZ-" c@9! r+ TIME AREA hK IDNO CONC a 3.783 4.503 5.123 5.992 ;Z C4 co 2 -;Za 4 467507 V 1545826 V 8586993 V 211519622 SV Cl.4073 c@ 3.5436 11.7168 65.0866 19.0979 0. 1478 ------------- TOTAL 13193186 WARMING NO PEAK 2-0 4'. PKIio 1 2 3 4 TIME AREA 3.76 15993 4.4'c.'5 645C.8 5.1 373677 5.97 96547 --------- TOTAL 5 5rj78 4 cn 12 -'3F-itt;'2 9.228 T Ih E C5 c@o co AREA liK 3.792 74532 V 4.518 30913 !6 v 4 5.137 1812582 v 5 0 12 527793 SY TOTAL 2735141 r+ CD 7.0 0 PKI-10 3 4 5 TIHE !3 Z..792 4.517 6.005 TOTAL AREA 11K t t ri5 C, 69258 v Ir-180909 v IE-28500 V 478240 OVV 24679le7l @(@4 54. '53 33 0 T IIlE AREA IIK 2 3.77 24910 V 3 4.473 927738 V 4 5.065 879701 V 5 5.943 198339 v P.S'12 9. TOTAL- 2253382 -,477 CD cn 0 r K 110 2 3 4 5 6 7 T I iiE AREA IIK 62 3 . 7 E.17, 16505 V 4.477 1039350 V 5. 077 8!o97 5 1 V 5.9--13 199461 V 94F- i T 13 C4 1 C4 TOTAL 2303814. ?KliO 2 3 4 5 9@55 77 Cl'-..' T IfiE AREA FL c@ C:) co li 1.11 3.757 4.49 5.087 5.455 TOTAL 67922 y '093103 y 1732292 y 496327 SY --------- 2600681 Figuret.4.2 GC chromatograms of the testsubstance ---for measurement of concentrationin water (after2 days at400C)--- Std. 1.008mg/l 4m c LO N U-) C-i I f.%i ui m k@j 0') - I Z- . ix ID n U") r. -:%I <z a- CU cr, I :o C-J cli 0 co tr) I N I cli C-V' rb,p cfk C's %N*) C-) Cli u-@ f%. r@, I", -i <r k %Vi -r u-i n f-> T cu cl@Ir u-i testsolution n=l ci u-) u-) o r . WA, <r c I :@ C- Li c,, -<j= r klj CIO V tro C- . testsolution n=2 <r Li ix <r > D C-i C-) N cl@ N C-i CN %") Cli CU a% cu Lri cli LAJ -:o 0.- tN O.C. -:0 tri rl% Ir I cu e-i ce@ try L- N 5 4@,41 .437 U;3 0 TIME AREA hK :3.4 OF 9-)4 2 f-.:7' 7 7 29460 V .:3 4 @7@l loi5oo2 V 4 5 . fJ73 965788 V 5 5.947 e 0 ri4 6 eiz 9c47- v c'7 TOTAL 2497030 u ti,:. u 8 Orj7 PKIIO i 2 3 4 5 6 T I liE 3 .4 i 3.797 4.512 5.118 6.003 r:o. A- E4;z AREA HK 24657 V 1008133 V 1020090 y 240666 V 14 C42!9 TOTAL 2463445 -t4 cn K Il0i 2 3 4 5 T 1 ME AREA liK co a ce 3.737 4. 447 Ic. r;5i 3 5.917 OTAL 76145 V :?1 ! 144 V 1819770 V 525^c 17 SY 27e:4 I Table 3 Calculationsof reactionrateconstant and halflifetime of the testsubstance (Table3.1-Table3.3) Table 3.1 (pH 4 at 250C) time, days (t) residualpercent rateconstant, of the'substance,day-1 ryig/(LC) (k) 5 89.7 0.022 8 87.3 0.017 15 77.0 0.017 is 76.8 0.014 26 68.3 0.015 33 62.1 0.014 average rateconstant 0. 017dayhalflifetime(t 1, 2 42 days calculatioenquations k=l/tX In 100/c tl,2=1/kXln 2 Table 3.2,(pH7 at 25*C) time, days (t) residualpercent ofthe substance mg/L (C) rateconstant@ day-1 (k) - 5 88.6 0.024 8 86.9 0.018 is 74.1 0.020 19 69.5 0.019 26 59.9 0.020 33 53.8 0.019 average rateconstant 0.020day tl,2 35 days Table 3. (pH 9-at250C) time, days (t) 2 5 8 12 15 residualpercent rateconstant, ofthesubstance, day-1 mg/L(C) (k) 92.4 78.4 69.0 57.1 50.2 37.7 0.040 0.049 0.046 0.047 0.046 0.051 average rateconstant 0.046day-I tl/2 15 days Table 3.4 Calculationsof residualconcentrationof the testsubstance ---far Further investigatidn(at 2 5.0. :t0.2 0C) --- time, days pH 29 4 57 9 4 87 9 12 9 4 is 7 9 4 19 7 9 4 26 7 4 33 7 peak area,ijv*sec test. Std solutioh B c conc.in residual testsoin.percent 0 2889193 2523765 0.371 92.4 2951466 2951466 2951466 2502799 2473261 2186759 0.361 0.356 0.315 89.7 88.6 78.4 2868192 2968192 2968192 2449526 2439536 1936885 0.351 0.350 0.278 87.3 86.9 69.0 2939623 1585436 0.229 57.1 2871688 2871668 2871668 2089210 2011015 1361482 0.309 0.298 0.202 77.0 74.1 50.2 2871088 2871088 2871088 2084603 1886031 1022942 0.309 0.279 0.152 76.8 69.5 37.7 2962788 1912723 2962788 1676378 0.275 68.3 0.241 59.9 2928913 1718699 0.250 62.1 2928913 1490039 0.216 53.8 concentratioonfthestandardsolution(A): 1.01 mg/L concentratiofnactor(E): 2.5 initicaolncentratioonf thetestsubstance(F)0:.402 mg/L recovery(G). 95 % calculatioenquation: -D-A(C/D)-E--1F-1(100/G)-100 Table 3.5 Calculationsof residualconcentrationof the test substance ---for Preliminarytest(after5 days at 50.0 0.1 OC) --- pit4 PH 7 PH 9 initiacloncentration of thetestsubstance,mg/L A 0.402 0.402 0.402 .;afte5r-days conc. ih@Std..,.mg/L peak area,ijv-sec Std. testsolution concentratiofnactor conc.in..thtee:stsolution.. B 1.00 1.00 1.00 C 3022269 3022269 3022269 D 1867539 1876183 1264465 E 2 2 2 0.306 0.307 0.207 residualpercent of the testsubstance F 76 76 - 52 calculatioenquation F B(D/C)(I/E)(I/A)(1/1.01)100 Table 3.6 Calculationof recovery ---for Preliminarytest conc.ofthetestsubstance added,mg/L conc. ;PLStd.,mg/L peak area,uV*sec Std. testsolution concentratiofnactor A 0.402 a 1.00 C 2977220 D 2420272 E 2 conc.of thetestsubstance recovered,mgll recovery,% F 0.406 101 calculatioenquation F.= B(D./C)(I/E)(I/A)100 Table 3.7 Calculationof recovery ---forFurtherinvestigation conc.ofthetestsubstance added,'mg/L conc. i'n@,Stdm.g@/,L peak area,jjv-sec Std. test-solution concentratiofnactor A 0.402 B 1.01 C 2782217 0 2639636 E 2.5 conc.of thetestsubstance recovered,mg/L recovery,% F calculatioenquation F = B(D/C)(I/E)(I/A)100 0.383 95 START 77 :'K i-0f 4 5 U) 9.817 1+ FL T 1 (iE AREA liK I DUIC, COHC liA 11E A 4 4:7 E3 34 577 4.347 4.9145 68147 V 276040 V 2100565 V 2.2809 9.2393 70.3078 5.77 532469 SV 17.8222 ------------- TOTAL '987667 T A F,T 7' r+ ((Dn-i 9 .E.1-2 r+ i-AIliF- 2. T IliE AREA liK I DNO c ct 2 0 55151 V :3 9 . ctri@4 2 4. 229949 V 70-9537. 3 4 .41 1717273 V 17. cl'667 4 5.773 4 1790rl S..-)- ------------- TOTAL 2420272 i (ICI START T IIIE AFEA K 5'1'3 39049 V 4 35 176830 V 4 '@27 13 1 ! 0 (14 V 4 TOTAL IE,76183 .START 5 94 PK[i@ T 1liE AFEA ['KI co 1 3.578 2 (117 3 2 4 . (IE-7 15877 V 3 4.@l '@'97747 4 ti.772 211,0668 sv 4 C4 TOTAL 12.*34 8 5 C4 .-,TART 9.795 cn FL T Ili[ A izE. A (i Iz1! 00 595 69716 y 4. 3 4 cl*U1Z3IriI y 4 4 1 'z 21 10,252 5 .7 5".@2271? SY -- -- ---- - TOTAL @-Ici 4 5 0 i START 9 17 ?KtiO 1 2 3 4 5 TIME AREA M K --1.592 4 . '14 @@' 4.9!7 5. 79 3 6:32 i-*. 186550 v 1308Z49 335415 V 5,@C4G TOTAL 1 2 8 47 6. 9 r -c 99ii E4 c@l T I liE AREA H K 3.615 7f$688 V 4.38 307633 V 4 4.938 1888329 v 5.813 507568 V v - --- --------- TOTAL 2816536 r+ .CD cn r 9 e. PKHO 4- 4 5 TIIIE "1.572 4.322 4.882 5.757 (4 1 TOTAL AREA MK C4 71301 v 270242 Y 17909O.C- IT, 507107 y v 2677385 eg, cn FL c@ ?KHO TIME AREA H@, 2 3.6',5 *.1198 v 3 4.367 3f$i 785 y 4 4.938 !928231 v 5 5.813 547979 V v TOTAL 2928667 CD 33 24@p.-. tp44, @89 9 p ie1-10 3 4 T flie 3 .6, 3 4.382 4.942 5.822 AREA [iK 69455 V 241338 Y 1738036 V 474936 V v TOTAL 2542849 5. E-98 9'7 P.<ti0 I 2 4 TIME AREA 1-1K 3.535 6992i v 4 . 2 ec7*- 265288 v 4.837 1672584 V 5.6-98 465468 SV -i 4@;z6 --------- TOTAL 24920'c*c' 1 DliO. -7 i.@r,7 Z7 Fl!1".10 2 3 4 TIME AREA hl' IDMO 3 57 4.313 4 8!f,7 5. 7 i 7 9 rl9- 6 14 !9 v 171377 1 610692cl $26543 S V TOTAL ---21-9-7-69-6-- CL "9 's m ri T 1H AREA MK DIIO 3.58 84103 v 3 4. 3c-*2 332077 v 4 4 . C'.? '@. 1973856 V 5 5.742 561432 SV --------- TOTAL 21?63383 a 7 @KMO 2 3 4 Tlt'IE AREA tiK 1 D[iCi 568 4.317 4.87 fl.73 TOTAL 72548 6 43 6 4 1763114 v 402272 S';' --------- !E@.77 6: 7 e@ 77' 0 TIME AREA h K IDNO 7 2 5 7 @j V 4 31 276869 4. '37 1764394 v 4 74 '@'25699 SY TO-t AL '-1*439536 3@C.3 cn 7C, 6.77 i" 9 . Ee. PKNO T 1 ME APEA iiK ID ['(II 2 3.54 84865 v 4.283 34 2 4 2:@ Y 4 4. 811:7 1976176 5 5.7133 5E.4727 V 4 6,. P A- TOTAL 3riBOO70 co d. 83r$ r'KliO 1 2 11 4 5 6 7 TIME AREA MK IDNO 3.537 4.035 4 . 03 4.838 S. 7L45 9 ;L@-: 47846 Y. 34361 V 5637@ 1442243 Y 356058 SY a 4- T TOTAL 199-8049 7 PKIIO 2 3 4 5 T IME AREA 1-1K I DI*10 3.607 4 . 'DE,. 4.92 5.702 TOTAL 70742 Y 22@@64S V 1 6894E,4 V 463678 SV - - - - - - -- - 2460565 33 Z1.04&2 C/) CL li0 T IliE AREA MK Dtio 3 4.358 341391 V 4 4.922 2006286 v 5 5.81;13 506656 sv TOTAL 295229@, 7..E55.7.t7;72 7 7. -E7. 5 tio T!hE AREA MK IDliO 3.557 41384 V 2 4.048 27159 V. 3 4.332 7cici3 8 4 4.86 lll5oci7 v 5 5.727 331843 Y TOTAL 1585436 @;33 c- . 6 7 PKIIO 1 2 3 4 TIME 3.54 4.275 4.833 5. c@87 TOTAL AREA 1111% 1 litio 54941 V 176806 V 1362655 V 416614 SY 2@ii@15 5,2.17 - 4.93 pkiio 2 Z" 4 5 TIME 4.108 4.368 4.93 5.'$17 AREA MK 2 1) 22387 v 35866 v 1010484 V 259$45 SV IDNO TOTAL 1378387 33 1',F;ig 81 .7 77 r+ 6. SPS7 FL PKNO i 2 3 4 5 T I ME 9. 3.582 4.327 4.9 5.77 TOTAL AREA IIK I DtiO 78196 V 329297 V 1883392 v 580784 SV 2884962 12 PKNO 4-2 3 5 TIME i i i; -@.525 4.267 4.812 5.65 .. TOTAL AREA h 1'% IDNO tee43 59779 V 169135 v 1444165 V 4 1 13.1 SV 2099453 PKTIO 1 2 3 4 2 95 qI, TIIIE 57 8 4.322 4.895 5.752 TOTAL AREA MK 52284 Y 155916 V 1248817 V 429014 SV 1886031 IDHO 4.953 ?Klio 1 2 3 4 5 !HE AREA tiK 3.62 22481 4 . IrJ 14349 V 4.417 40911 Y 4.95.'$ 723651 V 5 .83 221549 Y' ----------- Tri TO 1 1022942 I B 11C, PKIIO 6i 7tt' ;-'27 T 1h E AREA liK 3 4.252 328533 V 4 4.827 1879027 V 5 5.68 583525 SY TOTAL 2804227 %v783' 17 PKMO 1 2 3 4 TIME AREA liK ID li0 3.597 55724 Y 4.347 149067 V 4.917 ----1-41-4-1-72-- V .5.783 465641 SV TOTAL 2084603 84 PK[iri 1 2 4 TIME 3.54 4.308 4.842 5.692 TOTAL AREA MK IDNO 54270 'V' 1302 19 v 110024*0 V :2@9 650 V 1676373 PKiiO 33 1 .3 T I ME Cl) AREA I'Kl I D 1-10 2 3.52 86807 v 3 4.245 338231 v 4 4.803 1970817 V 5 5.657 566933 SV TOTAL 2974607 Ic.67 1"23 PKIIO I 2 4 TIME AREA MK !DMO 3.,i'c7 4.28 4.823 5.667 58084 V 152256 V 1349336. V 352547 SY TOTAL 1912723 84 ti0 T I 1-!E AREA liK I DliO 1 3. 533 46752 V 4.297 116978 Y 3 4.84 1032i99 v 4 5.688 294110 SV --------- TOTAL 1490039 ;,Klio cn 7 T I tiE AREA IIK idtio 2 3. 5 1 P, 86430 V 3 4.245 341379' V 4 4.82 1937852 V 5 5.67 563253 Y 6. V TOTAL 29 669:30 9 43 p 110 1 2 3 4 T I I-IE :3.5 4 5 4.3Ci8 4.843 5.69 TOTAL AREA 11K 43236 Y 114465 Y 1193570 11, 362408 SY 1718699 1 DI-10 Figure4.1 UV spectrum of the testsubstance inacidic,neutral,and alkalinesolutions concentratioonfthe testsubstance 0.8mg/ i acidic neutral alkaline ILriC- Figure5.1 Correlationbetween HPLC retentiontime and Pow cpfthe roferpnce,..Compounds: Curve Fitting [Least Square Method3 Input Data --------------------------------------------- log Rt log Pow N-o-.------------------------------------------- 1 0. 355 0.626 2. 12 2..99 2 3 0.813 3 .76 0.981 4 .81 4 1 .123 6 .20 --5------ ------------------------------------ Y = -2.1919X r = 0 . 97258 10-2 + 5.1282 X XI X 7 .00 6.00 5 .00 9= 4 .00 C4 3.00 2.00 0 0 0 0 0 1 .00 0.00 0.0 0.2 0.4 0.6 0.8 1.0 1.2 Xj log Rt Figure 5.2 HPLC chromatograms of the referencecompounds and the testsubstance Li: n Z 10.-1 trji).sn or o-i.oz.o toll. I r3 0 14 C.1 c 0 E tr reference compounds CL Peak& 1 .2 3 4 s G net Time 2.637 4.903 G.eG7 S. i:ss 12. @u04 15.900 T-/PC PV L)V av 6-1 ev vv W; d. h 0:07C 0. 116 e. IS7 0.208 0.274 0.325 a t-i 3LI 40 T Area 11214 1042 633.02 Z78.54 3S3.74 169.83 thiourea Rt-Rto lknsodft 2.266 bromobanzone 4.230 diphanyl 6.502 dibenzyl 9.567 DDT 16.263 log Rt 0.355 0.626 0.813 0.981 1.123 log Pow 2.12 2.99 3.76 4.81 6.20 LC R 2 10. -1 4 fj0 u- ;,U, testsubstance Li In a)(ZCD Qi) OD Peakt I 2 3 4 s 6 7 e 9 le net Time Z. 633 6.144 7.SSS 9.502 I'@.@304 1E .44S 22.SG2 'IS.308 2713ss a .7 2 z 31.87D Type PV vii vv ev ov vv BV eV vv vv WEdth O.CGS 0. '&3G 0.199 0.261 9.2SS 0.377 0.41G 0. 46Z 0.7SI 0.402 0.669 T 7 Area 'L?SS9 53.99 se.es 138.G4 79.43 114.04 67.83 160.lc 3SS.40 37 1.IL3 thiourea (i) (2) RL-Rto 3.511 4.922 6.869 9.671 13.812 19.929 23.675 24.726 26.089 29.245 lQg RL 0.545 0.692 0.837 0.985 1.140 1.299 1.374 1.393 1.416 1.466 log Pow 2.77 3.75 4.27 5.03 5.82 6.64 7.02 7. 12 7.24 7.50 Tabfe6.1 Calculationof-vaporpressure trapped testsubstance (correctedmass), g 1 molecular weight'ofthe testsubstance m gas constant, Pa*M3-.mole-l-K-1 R correctedtotalvolume of the carrier'qas.-M3 v temperature, OK T 1.47 X 10625 8.31 0.973 298 vapor pressure,Pa calculationequations P =(W/K)(RT/V) p B. ox 10-3 Table 6.2 !Calclialtion:of:mass" of the trapped test substance conc...infStd...,,.rng/L peak area,AtV*sec Std. testsolution conc. iKth6 test'sol6tionm,g/L finalvolume, mL dilutionfactor A 1.006 B 2489067 c 1412087 0.571 D 50 E 50 trapped testsubstance as measured mass, g as,correctedmass.,g 1.43X 10-3 w 1.47 X 10-3 calculationequation I A(C/B)E(D/1000)(1/0.97)(1/1000) Table 6.3 Calculationof desorptionefficiency conc. ofthetestsubstance: A added, mg/L conc.ihistd.m,g/L B 0.02515 1.006 peak area,ijv*sec Std. C 2451409 test:solution D 2372240 finalvolume.mL 25 conc. ofthetestsubstance recovered,mg/L E desorptionefficiency%, F 0.02434 97 calculatioenquations E B(D/C)(25/1000) F (E/A)100 FigLire6.1 Calibrationcurve of the testsubstance Curve Fitting CLeast Square Method) Input Data --------------------------------------------- Concentration Peak Area No X (mg/ I Y V -sec) -------------r------------------------------- 1 0 0 2 0.252 602790 3 0.503 1234186 4 1.003 2475687 --------------------------------------------- y = -9.1506X 103 + r = 0.99997 2.4740X 106 X Xl x 1000000 2.6 2.4 2.2 2.0 1 .8 rA > 1 .6 1 .4 I .2 1 .0 0.8 0.6 0.4 0. 2 0.0 0.0 Calibration Curve 0.2 0.4 0.6 0.8 Concentration(mg/1) 1 .0 1 .2 X S.TA F,T F.'T 77 TIME 3.762 4 4e5 5 Ci97 5. ';75 OTAI- 7 K v 17 4 2 V -343 Cl2 3 y rSC,4 2^4 IE*,t I D I-('i U) r+ PL c 0 1*c1 C> C> 1:1. :34 :,I.1.1 5 I C4. 6744 1!@, 670* ------------- I Cie START @RT i @KNO 2 4 5 @7 7..E.;@36 :@@.j@CJ TiliE AREA MK 1 DtiO colic 3.762 62275 V 4. 49 5. i !674132 V 5 . 9E-2 472732 SV - - - ---- - - - TOTAL 2 4 8 5 -?(45 2.51351 10.7224 67.3449 ig.ol6s - - - - - - - -- - - - - 1 CIG U) FL C:3 9 00 t.Ztb F99..4787 6- :,Klio 2 11 4 T 1 ME' AREA MK 3.762 4.497 5. i i 5 .9 9 TOTAL 13433 v 63505 v 4 1729Cj 108563 SV - - -- - -- - - 27 0 DHO colic 69.226 If-.*0 10 - - - - - - -- - - - - 6-1ART 5-5:@;Q.'li5 -3-@7 <!icl 5 A --tA !INC' 75 4. 477 5 0;?7 5 IOT AL E.ci9 e.E. 2 4.2 4 1 t.6 CN9 '1*:" V 4E.4747 S%-' 24 17 i7 crn+ CL c 0 ti.1@ NA[IE Ci 4 2. 4774 (1 6 7 Cl":-t E-7 554 1 I ?, 87'1 1 cli-I S T AF,T 5 @7 7 . < 2 4 T V, 3.752 4. 482 93 T(ITAL A r-A oK 1 DNO 59':-40 V 2 5 3 Cl V IA!2761 V 44i.6'@7 SV CD ri) cn 0. E colic -.A-ifiE !:t 0 2.5225 r-_.l66 st E-7.984' 7 i 8. 8 7 6 1 cl1) Figure6.4- GC chromatograms of the,testsubstance for measurement -of--mass-.ot.hfe..,trappetde.st substance;--.:.@ Std (I.Omg/I test:,qolution C'J LLJ ID N cl: :Cl C-i IT -r 12, (.j I;x r% r4 Li Li U'o vi* C"i cli U:l U:l C-J CIO V U-i Figure6.5 Vapor pressuremeasuring apparatus c E 2 flow meter adsorbent (Tenax GCO) 0 0 M CD N c 0 CD LZ-) CL cp