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BIODEGRADATION (Modified MIT Test) TEST SUBSTANCE Identity: 5)[2-(N-Ethylperfluorooctanesulfonamido) ethyl acrylate; may also be referred to as B1228, D-1, EtFOSEA, or FX-13. (2-Propenoic acid, 2[ethyl[heptadecafluorooctyl)sulfonyljaminojethyl ester, CAS # 423-82- Remarks:byMastpeornisalori.s Tanheampubreirtys/odleindt.itRyepofortthelitsetsstssaumbpslteanacse>c9a9nn%o,tabseprovided substantiated. As presented in the report, the structural formula indicates that "purity" cannot be assigned as "99% or more". Lot number 4. METHOD Method: OECD 301C Type: Aerobic GLP: Yes Year completed: 1995 Inoculum source: Chemical Biotesting Center, Chemical Inspection and Testing Institute, Japan Remarks: It appears that the inoculum was obtained from only one source, rather than the ten sources required in the method. RESULTS Tseusbtstsaunbcsetwanacsed:eAtfetremri2n8e-ddabyyst,htrheee pmeertcheondts:biBodieogcrhaedmaibciallityOxoyfgtehne tDesetmand C(oBnOcDe)n,trDaitsisoonl.veTdhOergreasnuilctsCaarreboans (DOC), follows: and Residual Test Substance Percent Biodegradability (o3f replicates): DBOOCD:: N4,ot-1c,aalncudla6t%ed; test substance not dissolved in solution Test Substance Concentration: 5 and 1% (only two determined) Reference substance: The aniline achieved 68% biodegradability in 28-days. Measurementof test substance: The test substance concentration was only measured at the end of the study. Although this laboratory followed the 000502 procedures determined as outlined rather than rinelOyiEngCDon3n0o1mCi,natlhecosntcaretnitnrgatcioonncse.ntrIantaidodnitsihoon,uld also be subsamples atthe end of were removed from the the study. The sample bioreactors used for pH for pH and DOC determinations determinations and the prreimoaritnoinsgamspalmipnlgefoarfttehrecmenetarsiufurgeamtieonntfoorf DthOeCtewsterseubrsettaunrcneedcotonctehnetrbaitoiroena.ctor These for test samples should have been removed after substance concentration determination. the bioreactors were sampled Calculations of theoretical oxygen possible oxygen consumption due demand should to nitrification. have made which account for CONCLUSIONS "bTihoidsetgersatdinignginidniacearteodbitchaetntvhierotnemsetnstusb.stance is not capableofreadily DATA QUALITY _ Reliability: provided in Kiimisch the study. rTanhkeinqguo3.teTdhepurreitiys/indoenrteiftyeroefntcheettoetsthesusoblsutbialnictye information is not spuebrsftoarnmteidataetd.theMeeansduorfetmheentstoufdyt.estThseubbsitoarnecaectcoornscweentrreatpoitoenntwiaasllyonly contaminated by the determinations. The practice of calculation retuming subsamples of theoretical oxygen after pH demand and DOC should have been performed using the correction for niification. REFERENCES `Study conducted at Safety Institute Ltd., the request Yokohama, of Sumitomo Japan 3M LTD by Mitsubishi Chemical OTHER Submitter: Minnesota, 3M Company, 55133 Environmental Laboratory, P.O. Box 33331, St. Paul, Last changed: 5/22/00 000503 M.S.I. REPORT No.1B363(2)G Ready Blodegradability Test of b-1 submitted to : sunrrono a Lorre Prepared by + Mitsubishi Chemical Safety Institute Ltd. october 31, 1995 Le 000504 BR 0) E CEIVE_, | rae IiTi Ready Biodegradability Test of D-1 (English version) study ve. 1 merc Study Title Ready Biodegradability Test of 5-1 sponser + saartovo aw LawiTeD This reporc was conducted in Chemical Safety Institute Ltd., the Yokohama Laboratory of the Mitsubishi 1000 Kamoshida-cho, Acba-ky, Yokohama 727, separ. This zepors is an English version of the orginal, which was written in Gapanese. The underesigned hereby declare that this version faithfully ceflects the original report to the best of our knowledge. Mitaubiahi Chemical Safety Institute Ltd. bres Kiko Yoshida, Ph.D. vate :_Oetodor 31, (976 Environmental Science Division of Yokohama Laboratory pe---- -- a. Sin sun Toray, 8.5 Head of Yokohama Laboratory oases Lotatin 1/4 976 000505 COMPLIANCE WITH THE GLP STANDARDS Study No. Study Title Sponsor : 18363(2)6 Ready Biodegradability Test of D-1 SUMITOMO 34 LIMITED To the best of our knowledge and belief, the study described in this report was conducted in compliance with the Good Laboratory Practice (GL?) standards applied to industrial chemicals under the Chemical Substances Control Law. Facility management Sakae Koike, B.Sc. Facility management Head of Yokohama Laboratory Sealed date April 28, 1992 Study director Kikuo Yoshida, Ph.D. Study Director Sealed date April 28, 1952 Environmental Science Division 000506 QUALITY ASSURANCE STATEMENT Stuay No. Study Title sponsor + 183630206 : Ready Biodegradability Test of D1 + SUMITOMO 3M LIMITED We hereby certify that the above study has been performed in accordance with the Good Laboratory Practice (GLP) standards applied to industrial chemicals under the Chemical Substances Control Law. Dates of inspections and reportings to the Study Director and the Facility Management are as follows : TmT aInt spece tionor In-progress study : AMparriclh 115,, 11999922 April 16, 1992 Final report heril 2s, 1952 dReapotrteinog t AMparriclh 1350,, 11999922 April 16, 1992 April 28, 1992 Quality Assurance Staff Junko Katoh, Ph.D. Yumiko Miura, B.Sc. Sealed date :April 28, 1992 Sealed date April 28, 1992 000507 We, the undersigned declare that the work was performed according to the procedures herein described, and that this report provides a true and correct record of the results obtained. Experiment Staff Ayumi Okuda, M.Se. Sealed date :April 26, 1952 Environmental Science Division Junko Kogasu, B.Sc. Sealed date :April 28, 1352 Environmental Science Division Yukiko Mizoguchi Sealed date :April 28, 1992 Environmental Science Division Staff for cultivation of activated sludge Yasuo sato Sealed date :April 26, 1992 Environmental Science Division 000508 CONTENTS age EXPLANATORY REMARKS ttteeeeeeeeeiittiitiiiennaeaaaaaeenaainann1n 1 STUDY TITLE. etiieieeeeeeeeeeeeeeeaeeeeeeeniinn eee eaeeeeeaanan8n) 3 TEST GUIDELINES......eeriiseeneeesensienieeesiiiiinnnnaaaaaannnnil 8 4 GLP COMPLIANCE... nennnnnnnnnnnnnnnnnnneneenrenninnnnnnnaaaaaeeni8s 5 STUDY BERIOD....:nnnnsnssnnnsnnnnnnnnnnnnnnnnnennnnnnnnnnaanaaneessd 6 SPONSOR......uvunnnnnnssnnnnssnnntnnntnnnaaaeaaeerennnnnnnaaaaaaass 7 TESTING FACILITY. ...tiiiiiesseneeeesiiesinieiaaeasaaiaiaeiaeeinns 8 STORAGE AND RETENTION OF TEST SUBSTANCE AND RECORDS: .:...--.........10 9 TEST SUBSTANCE......eerresiessesesessesisirrrriiiiueennnnnaaaaaenn1ns1 10 CONFIRMATION OF IDENTITYAND STABILITY................esssseseeeeens12 10.1 IDENTITY toiiiiiiiiaaaaeeeeeeeeeceiiieeiie nena ee aaaeeaeiiann12 20.2 STABILITY tevvviiiiiiiiinieeiaaiaaiaeiaiaainnusninnunnannnn 1n2 IL TEST METHODS......iiiiriiisssssessssissresiueieeennnnnnaaaaeeenee1n3n 11.1 ACTIVATED SLUDGE evvviiiiiiissusseusnnnnaeaeaanenenisnnnn1n3 11.2 EXPOSURE CONDITIONS +rtttrirrrrrrsissnnnnnnnnnanaesseenennn1s3 11.3 BODMEASUREMENT oeiiiiiiiiiiiisiuienensnnnaaaeeneneennnnnns 14 11.4 pHMEASUREMENT eiiiiiiiiiiiiiiiiiuuenennaaaaeneneeiinnn 14 11.5 DOC MEASURE+M..E.iNeTeiiiiiissusuueuesnnnaaaaaeneeeennnn, 14 11.6 MEASUREMEONFTRESIDUALTEST SUBSTANCECONCENTRATION ........ 1S 11.7 EQUATIONS FOR CALCULATION OF DEGRADRBILITY .....rvvvrerrnnss. 16 12 RESULTS evita eeeeen e eee eeecennnnnaaaaa1s7 12.1 OBSERVATION OF TEST BOTTLES AFTER EXPOSURE PERIOD ...........17 12.2 plMEASUREMENT oiiiiiieeiiiiiisiieiiiueennnnnnannneeenennn1n7 12.3 ACTIOFVSLIUDTGEY +nnnnnnnneeeeevveennennnnnaaaaeannnss1n7n 12.4 DEGRADABABSEIDOLNIBOTD Y .............seeeeeeeeeeeennnnn1n7s 12.5 DEGRABASDEDOANBDOCI1.L...I...T...Yuessssneeeeeeeennnnnnn 17 12.6 TDEESGTRASDUABBSITLAINTCYE BCAOSENDCEONNTRTAHTEIORENS.I.D.+U+A+L+++. eesesseuueennnnnnns 18 13 CONCLUSIONS. 11s eeee eaeaeeeenennnnnneaeaaaae1s8s TABLESANDFIGURES........uttuunnnnnnnnnnnnnnnnseernninsnneesnnns19233 000509 EXPLANATORY REMARKS Study No. Study Title + 18363(2)6 : Ready Biodegradability Test of D-1 The ready biodegradability test of D-1 was originally started on January 23, 1992, and the BOD measurement of the test substance was initiated on February 13, 1992(Study No.13363G). However, the degradability value of aniline based on the BOD measurements did not exceed 408. Therefore, the test was terminated on February 20, 1992 and the new test (Study No. 18363(2)G) was started with the same test substance and the same test method after approval of the sponsor. Some of data used in this report (IR chart, calibration curve and recovery) are those obtained in the former study (Study No. 183636). 000510 STUDY TITLE SUMMARY Ready Biodegradability Test of D-1 METHOD The method described in the Order, which prescribes the procedure of testing new chemical substances as required by the Chemical Substances Control Lew(Japanese Law 117, 1973). ("Ready biodegradability, Modified MITI test" in OECD Guidelines for Testing of Chemicals). Test bottle : BBoottttllee 21 BBoottttllees 36~5 5 aslnuidlgiene+ +basslauldgmeed+iubmasal medium ++ tteesstt ssuubbssttaannccee +f wsaltuedrge + basal medium Measurements : BDiioscshoelmviecdalorgoaxnyigcencadrebmoannd (0(08C0)D) Residual test substance concentration RESULTS Measured values (day 29) Een We. ED, me s TIT e Le OF Le 00 TDeocs,t msgu/bLstance, mg/l _ 1001..734 1005..32+ 911.65+ 1026..70 + Value corrected with B55 oF DOC Value of Berle ZL ((aftorda26y d28a)ys) (at day 26) heemstiaal vale m20m.a8 100.0 DegradBaobtitllietiNeos. (8) ST % IJ Tneosct substance NA5S 1NAS wwaare * ++ ToSoottlutcciaaollncc.lullaatteedd because because the the Test test SUBSTANCE substance was Wot dissolved in the test might ba lost accidentally during Condensation of the solution under vacuum. CONCLUSTON Fron the degradability results based on the BOD and the residual test substance concentration , it can be concluded that the test substance is not readily biodegradable and is `mot transformed under the conditions prescribed by the Modified MITI test. 000511 1 stooy moms Ready Biodegradability Test of D-1 2 pumeose This study was conducted to evaluate the ready biodegradability of the test substance for notification under the Chemical Substance Control Law of Japan. 3 mest cumeniNEs The test was conducted according to the method described in the Order, which prescribes the procedure of testing new chemical substances as required by the Chemical Substances Control Law (Japanese Law 117, 1973). This method is also known as the "Ready biodegradability, Modified MITI test" in the OECD Guidelines for the Testing of Chemicals No. 301C. 4 owe compLiancs The test was conducted according to the Good Laboratory Practice (GLE) standards applied to industrial chemicals under the Chemical Substances Control Law. 5 sooy semen Maxch 17, 1992 to April 28, 1992 (80D measurement : March 18, 1852 to April 15, 1952) 6 sponsor SUMITOMO 3M LIMITED 8-8 Minamihashimoto 3-chome, Sagamihara 229, Japan 7 mestmNe macILrTy Yokohama Laboratory Mitsubishi Chemical Safety Institute Ltd. (M.S.I.) 1000 Kamoshida-cho, Aoba-ku, Yokohama 227, Japan Head Office: Mitsubishi Chemical Safety Institute Ltd. (M.S.I.) 1-30 Shiba 2-chome, Minato-ku, Tokyo 105, Japan 000512 8 STORAGE AND RETENTION OF TEST SUBSTANCE AND RECORDS The following records, the raw data, and a small sample of the test substance will be stored in the archives of M.S.I. for 10 years after the subnission of the final report. After this period, the sponsor will be contacted for the approval of the disposal of data. Data can be stored in the archives for a further specified period at the sponsor's request with an additional fee. 1) protocol. 2) Final report. 3) Raw data. 4) Quality assurance reports. 5) Test substance (approx. 2 g). 6) Other documents required under the GLE standards. 000513 5 mst sussmmce tamer -(4.5.7. sdentification No. 183630216) 21chentcal naner +a2mi-no(e3t-hyEltacheyyiia-teN-pezf(Cu=l1o~5r)osuallfoknyyll- Structural formulas: Co FanH2aistiCitSy0--0tim2 cin--c, (nw1~8, man component; ned) 4) Elemental composition i5C7i5218.2805, tHryea1s.u5r5e,d bI;y 52..355,.)0; 9.88, 152.08 5) ehysico-chemical properties Sotubitieys iinnssoolluubbllee iinn wDaMtSeOr 3308 in acetone Melting points 127-42 Boiling points ca. 150C/lnmtg 0) sacens stot No. 4 Spurieys 99% (ne1~77c0.21 (total), nesica. 88) 10 Appeasance camber solid L)0ate of receipe iSeptenber 25, 1991 12)5upplied quancicy 200 g + provided by the sponsor. 000514 10 coNermaTION oF TENTITY AWD STABILITY 10.5 tomer For the identification of the test substance, the infrared absorption spectrum of the test substance was measured. The measured spectrum was consistent with that supplied by the sponsor. 1Appazatus + Perkin-Elmer model 1640 infrared spectrophotoneter. (Figure 1 (2-231) 10.2 smatuny After the exposure period, the infrared absorption spectrum of the test substance that had been stored in a refrigerator during the course of the study was measured. The spectrum was consistent with that shown in Fig. 1, fndscating that the test substance was stable under the storage condition. 1Appazatus : Perkin-Elmer model 1640 infrared spectrophotonster. (rigure 2 (2-20) 000515 11 TEST MemEoDs The test was performed according to the method described in the Order, which prescribes the procedure of testing new chemical substances as equired by the Chemical Substances Control Law (Japanese Law 117, 1973). This method is also known as the "Ready biodegradability, Modified MITE test" in OECD Guidelines for Testing of Chemicals No.301C (1981). The test substance was exposed to the activated sludge in a closed-system oxygen consumption measuring apparatus. The biochemical oxygen demand (30D) was measured over a 20 day period. After this period, the concentrations of the dissolved organic carbon (DOC) and the residual test substance in the test bottles were measured. The biodegradability of the test substance was evaluated from these results. 11.1 ACTIVATED supe 1)Mixed liquor suspended solid (MLSS) : 3300 mg/l 2)Souzce CIhnesmpieccatlionBioantdesTteisntginCgentIenrs,titCuhteem,icaJlapan 3)bate of receipt: January 23, 1992 11.2 expose _cowprrrons 1)Temperature tas ET 2)Exposure period : 28 days 3)Test volume : 300m 4)Concentration : atneisltinseubs(tbaontctele(b1o)ttles 3~6) + + 100 100 mg/1 ng/1 activated sludge (bottles 1~5) : 30 mg/l *t Reference substance : Syowa Chemicals, Lot Ne.SC-2726 S)Test bottle contents Bottle 1 An2i5l4i1n(e30+ magc)tiovfataendilsilnuedgweas+ abdadsead) tmoedtihuem basal mediums, then activated sludge was added. Bottle 2 : AcAtcitviavtaetdedslsuldugdege+ wbaassaalddmeeddituom the basal medium: Bottles 3-5 : Test 30 msgubosftatnhcee t+esatctsiuvbastteadncseluwdagse + basa) added to medium the basal mediums, then activated sludge was added. Bottle & + Test 30 msgubosftatnhcee t+esdteisounbiszteadncweatwears added to 300 ml of deionized water (purified by Milli-Q). *: The total volume of the basal medium and the sludge was held fixed at 300 ml. 000516 11.3 800 weAsuRsMENT The BOD was measured for 28 days. Apparatus: closed Ohkuza system oxygen Electric Co., McoodneslumpOMt-i2o0n01mea(sMu.rSi.In.g IaDppaNor.aBt)u.s 11.4 pM MEASUREMENT After the exposure period, 20 ml of the test solution in each test bottle was transferred into a 20 ml glass beaker for pH measurement. After pH measurement, the test solution was returned to each bottle for measurement of residual test substance concentration. Apparatus : pH meter, Denki Kagaku Keiki, Model COM-10 11.5 DOC MEASUREMENT The concentration of the DOC was measured as follows LiApparatus TOC analyzer, Shimadzu Co., model TOC-5000 2)Conditions : AFturrnafcleowteramtpeerature +: 618500Cm(l1/Cm)in. SIennjseicttiivonityvolume fxs 50 wm 3)calibration cuzve The following standard solutions were injected into the TOC analyzer. The calibration curve was prepared by the data processor of the analyzer. standard solutions TC(total carbon) b2i0phatnhdal5a0tem.g C/1 aqueous solutions of potassium IC(inozganic carbon) 20ndmg1C0/1mgdeC/i1oniazqeudeouwsatesrolu(tpiuornifiofedsboydiMuimllhiy-dgr)ogen carbonate. 4)Measurement of DOC in the bottles Ten ml of the test solution in each bottle was transferred into a 10 ml centrifuge tube and centrifuged at 3000 rpm for 10 minutes. Five ml of supernatant was used for the measurement of DOC. The remaining supernatant and the precipitate were returned to each bottle for the measurement of the residual test substance concentration. 000517 11.6 MEASUREMENT OF RESIDUAL TEST SUBSTANCE CONCENTRATION The concentrazion of the residual test substance was measured by the gas chromatography (GC) as follows: Apparatus sc Integrator :Hewlett-Packazd. sHeulett-packard. Model Model S890A 3392A (No.2) with FID detector 2)CCoolnudmintions +7 4 W. DB-17(503fhenyl SOsmethyl polysiloxane), Temperature 30m X 0.25 mm i.d., 0.25um (film thickness) + [Columnoven] 100C (min. } ~10C2/00mCi(Imnin..) Carrier gas (tIienj3e0ctmolr/)mi2n5.0C, [Detector]280C | AtDteteencutaotriofnlow (iHe:e)40m1l/vmin., air 400 ml/min. Injection volume :1 ul 3)calibration curve The standard solutions of the test substance with 0, 250, 500, and 1000 mg/l in ethyl acetate were analyzed. The total peak azea of test substance was calculated by adding the area of the peaks with retention time of about 5.6, 6.2 and 6.5 min. The total area was plotted against the concentration. The correlation coefficient was calculated to be 1.00 by the method described in Japanese Industrial Standards 29041-1968. 4)Recovery test (Figure 3 (P-25) and Figure 4 (p-26)] Duplicate test bottles (identical to the test bottles 3-5 described in 11.2) were kept in a closed-system oxygen consumption measuring apparatus at 25+1C for 30 minutes. Zach test solution was treated by the procedures described in 11.6.6. The concentration of the test substance was measured by GC. The average recovery of the test substance was 978. (Table 3 (2-21) and Figure 5 (p-27)) The concentration of the residual test substance measured for bottles 3 ~6 were corrected with the average recovery value. S)Detection limit The detection limit of the test substance was calculated to be 3 mg/l based on the minimum detectable peak area of 100000 WV- sec for the largest peak (R.T. 6.2 min.) in the GC chromatogram. (Table 3 (P-21) and Figure 5 (2-27) 6)Measuzement of test substance in test bottles The contents in each bottle was separately transferred into a 500 ml lass separatory funnel, which was then extracted twice with 100 ml of 000518 ethyl acetate. The extract was filtered through a grass funnel containing 50 g sodium sulfate, anhydrous to remove water. Then filtrate was condensed to ca. 20 ml under vacuum (<50C). The solution was brought up 0 50 ml with ethyl acetate. The ethyl acetate solution was then analyzed by GC. The concentration of the residual test substance was determined as described in 11.6.2. 11.7 EQUATIONS FOR CALCULATION OF DEGRADABILITY Equations for calculation of degradability based on BOD, DOC, and residual test substance concentration are as follows: 1)Degradability based on 50D Degradability (8) = (80Ds- BODb)/THOD X 100 where BBOODDsb :: OOxxyyggeenn ccoonnssuummppttiioonn ((mngg)) in in bboottttllees 2.1, 3, 4, and S. TROD: Tthesetorestuibcsatlancoex.ygen demand (mg) of aniline and the hop aniline CBN + 35/40; -- 6CO; + 7/280 + NO, THOD 5 90.2 mg 03/30 mg-aniline test substance : CH0;=COz, H#1/40:1/2H0, N4O;~NO:, +0:+50s, F--F TROD 1 27.7 mg 03/30 mg-test substance 2)Degradability based on DOC Degradability (3) = (1 = (DOCs-DOCH) /DOCc) X 100 wheze DOCs DOCb 5 : DOC DOC ((mmgg//ll)) iinn bboottttllees 2.3, 4, and 5. DOCe : DOC (mg/l) in bottle 6. 3)Degradability based on residual test substance concentration Degradability (8) = (1 - Cs/Ce) X 100 where CCes:: ccoonncceennttrraattiioonn ((mmgg//ll)) iinn bboottttllees6.3, 4, and 5. 000519 12 Resoms There was no specific circumstances which might have affected the relia- bility of the test results. 12.1 OBSERVATION OF TEST BOTTLES AFTER EXPOSURE PERIOD The solutions in the bottles were colorless except the bottle 1, in which the solution was cloudy. Growth of the sludge was observed in bottle 1 in contrast with the bottle 2. No growth was observed in the bottles 3, 4 and 5. 12.2 pH MEASUREMENT After 28 days of exposure, the pH was determined to be 7.4, 7.4, 7.5 and 7.4 for bottles 3, 4, 5 and 6, respectively. 12.3 Activiry oe suupce (Table 1 (p-19)] The degradability value of aniline based on the BOD measurements was 59% after 7 days. The activity of the sludge was thus shown to be satisfactory (Table 1 (P-19) and Figure 6 (p-28~31)] 12.4 DEGRADABILITY BASED ON 80D BOD! in bottles 3, 4, and 5 (as corrected with the value in bottle 2) were 1.0, -0.3 and 1.6 mg, respectively. BOD in bottle 6 was 0.0 mg. (6 Maximum theorstical value = 27.7 mg) The degree of degradability based on the BOD measurements were 4, -1 and 68 for bottles 3, 4, and 5, respectively. (Table 1 (B-19) and Figure 6 (p-20~31)] 12.5 DEGRADABTLITY BASED oN Doc DOC: in bottles 3, 4 and 5 (as corrected with the value in bottle 2) were 0.7, 0.3, and 1.6 ng/1, respectively. DOC in bottle & was 2.0 mg/l. (+% Maximum theoretical value = 59.8 mg/l) Degradability based on the DOC measurements was not calculated because the test substance was not dissolved in the test solution. (Table 1 (B-15) and Table 2 (2-20) 000520 12.6 DEGRADABILITY BASED ON THE RESIDUAL TEST SUBSTANCE CONCENTRATION The test substance) was detected at concentrations of 101.3, 105.2, 51.5 and 106.7 mg/l in bottles 3, 4, 5 and 6, respectively. (+3 Initial concentration = 100 mg/l) The degree of degradability based on the residual test substance concent ration were S and 1 3 for the bottle 3 and 4, respectively. (Table 1 (2-19), Table 4 (22), and Figure 7 (-32~33)) During condensation of the filtrate of the bottle 5 under vacuum, a part of the filtrate spouted from an evaporating flask. The spouted filtrate was recovered and then condensed with the remaining filtrate. However, the residual test substance concentration in the bottle S was obviously lower than those in the bottle 3 and 4 and it would impair reliability of biodegradability of the test substance in the bottle S. Therefore, the degree of degradability based on the residual test substance concentration was not calculated for the bottle 5. 13 coxciusrons From the degradability results based on the BOD and the residual test substance concentration, it can be concluded that the test substance is not readily biodegradable and is not transformed under the conditions prescribed by the Modified MITI test. 000521 Table 1 Sumary of the test results Dearadabilitios wero calculated by the aquations shom fn Sec. 11.7 o2iDdegradsbiTliecss basedonB10 | abate 2pir -- 3 -- er -- ---- =-- TT emsete wrani 2 = wso eitian]4] "so il w 5d |---- S1 STT or Zi ara a "i51 IX) [=== Tere %fogsBhTITL vas aleuloted o be TRGRTIve AIS vole a Shoe in brackets --i [===] 1C) E ri s-- easrk ess-- at [TT a. rs o eeara] CE elr e xcio| rgi aie sseanrbon] eee TCsenTol o3n S TTams ssse a7 ool AE IY Canconraition" [Tgradsbiio] I--Ta e ies----1T----| + tot cC olcTSe Ted T1eco ineoteirnst S1 STaRES STA=R B0]Tost Sciduntally ring condersation of the soton der vacun. 000522 Table 2 Results of DOC measurements Sandan soon Tae 3 Sus TUT aesdeentit, TT axrtessenity, TTY satesesesl, Bottle 2 Te an Te Tn Tn TTT 3B LE Bottle3 CST Te he En TTT rn endl sense Bottled. CTT he Te TTT ppebeiiee CTT ee 1 0 Toles TTT eri dieen rere CT Te TTT Te TTT SEER CTT TR TT TTT 3ER Bottle 5 Bottl6e Tae eT TT a eT De TBBH eee mpti releeenl nene CET ST 2 . wv peetisrrtel neees FE: 2 Ey } FY SPE Tea RK 2 icant RE i Tedmei co" sBootltlees2 ee Be EB e EB Ble "Bote 000523 Table 3 Calculation of recovery and detection limit arpeess | Csoelmuttirona,tisoineTen ] Rocovrad beatciteion | [coer tetT fw wen oe 0 mE m&o wa EE 0 -- -- |-- w=py] = eis asm] aen] aT= oerate r] eer TY g eYStololinmdeeoordoffsofslooultuttiiosonnoofonr GECForoaonmc6eiGrnytaeartmaasitiroins J(H0E: @: r500mo est w0o0o1l |s Eantion BHAT CBXISK DOXK FDSEXIO GC rise dociaal fractionsto wit, 8 cotcutavion of detection limit I| Hn7PT -- --] ---- CC o O 7 7 theThofosllionr1 iinng dueattecitoanblebapseedaCkoonortemhosfcintiheauntesd|tetescutbasbtlaencpeewsarceanlcuolfat1e0d00b0y0 a-sic or the largest pak (rcantion tie 6.7 ain) in GC chrosatogron Minin detectable peiskosreesd= GAaIvS0e9e0d X ~10000~ 0 (av~ -sec) a0 (aves + 130 (uesee) <n Gave Table 4 Residual test substance Tocetration Tcwoncentration in test bottles Arreeeoa | [c[olmruielon, we To 7 w + --co|g fa -- sTssun) 150513 061] @8 Samdard olotion ole of solution for GCCFooaenlnytasrreiantsion (0% G) a5000 wweret e oof . 2a Aovelraegeofretceosetrysolutionfor GC analysis (a: x0w1y Baution BAXDEE CAXDSEXFSGHHXI00 Figure 1 Infrared absorption spectra of the test substance Meabystheuspronseor beg a wet EE '' 1 HEagy wn 2B8E22 ww BmEs3e5 ! !1 1 we vy frEh Whe,y wv ake Ilon | | aa Lon run 000526 bd Figure 2 Infrared absorption spectrum of the test substance stored in a refrigerator > .vo-- sms : + I ) | EEE RE ate $20 SHON a Gf gach 000527 Figure 3 Calibration Curve of the Test Substance Calibration Curve Concentration X(ng/1) 0 250 500 1000 Peak Area (pV+sec) 0 1.92785406 3.89003E+06, 8.12341E+05 Y= 7.552104 + 8.139x103 X r= 0.993707 x108 or SE or 6 = 3| 5oJ & oCed pe | 2 | os wl 0 500 Loncentration(ng/l) | 1000 000525 Figure 4 GC chromatograms of the test substance -- Calibration curve 0 matin atyiacotate 250 man in ettvacatate gage " `i = ! i 2 E! 2 = puss . ses ngs = = tH - pe] FREEEE otal pask axenrisaTesOm-sec 5e0t0hmgy/llian cetate # . E3EtREeNRsR = SazEgzYas r FyReEaE - mE Sggoze ssse . z FiIE Sota peak arenas 1000 mg/l in ethylacetate 3 p2i3egdnesosaFg Fsigagbxe JL . TL iy gd ] EscEpm gaeo tzeesseses a i T Fsx3iEynEasnee [---- 000529 Figure GC chromatograms of the test substanc~e Rocovery and detection li Recovery and detection iit are shown in Table 3. 500 mgt in etyiacetate fink IN . $5058 yrsisiee = gEiEeRsE g 2 gia : = Sota peak axensd1s0siour-se Recoverytest-1 Becoverytest-2 2 Espa Z S8EEanEINSRe Ep2r2s5e2zpeLns s i 3 000830 rps gipemness = TARE Toa Sotalepesnk are ensisusziow-ses EERE =3 = Je SR ss3xggavgd3zEdns yesrzzsz im Sgazaeds RL INEE total peak azearda1soscuv-soc [SR EEE === = Shpa ele m. in = E --E = ewE e oo | Sie mmamaeeeeee Sheps oF e fF SSEhEeRree er w==e Eee == eee See eee == Haa ee] EEE RE eee] EE 000531 Fauros (Continued) Fra prema EE Tae ee Be pe FEmogEs orEma Se p Ee | == == H[roeaom ranom [Percentage dexradabilityofaniline at day 7. EEe I e = e aea Ee E== F El sore = Sam E ==E Siremmeaeees : a wim E = ee EET FE rm EE hee n mr e eeee Bodtotlne g5.\H = EFerem eE ee e ee Pe N meeE e] Si e ee mre eeeae? m eee a i. ee 000532 Figur6e (Continued) Eat e Eeee ee eee ee e ee eee eE --Tee= = Eee eee. EEE ee eaee ree Eee eeeee eer = | EE ee EET Ee--ear mai FEe E b P|eNEE =H aE POLE rrr rrr st i 000533 nes many - EE aa e EE ee = ee E CEEeee == = =ue se a ~ e = = e _ = E -- e --yee EEELe Te 000534 jE gress = SsEppast=ss = T ; x ENse g g moe ess srs Bottle3 g = 3 2 Bottled. ; Tgzaune Eg = s2zsas gms geoaabat goon total pesk area:3149880uv:sec 000535 `total peak area:5349150uv:sec Figur7e (Continued) Bottle5 Bottle 8= E$8o2s2a2oaRl2e1 greszse 2 EgEiEaEmEnNeEyS El I genni Sota oak axons6430O-se E FAW 5 - sSEEz]E pez sro. s=eia F pe o cot peak aressizsne 000536 LL