Document ymnMJmq2X630vZEvdLnrxV7jX

BIODEGRADATION (Modified MITI Test) TEST SUBSTANCE Identity: [2-(N-Ethylperfluorooctanesulfonamido) ethyl acrylate; may also be referred to as B1228, D-1, EtFOSEA, or FX-13. (2-Propenoic acid, 2[ethyl[heptadecafluorooctyl)sulfonyl]amino]ethyl ester, CAS # 423-825) Remarks: Material is an amber solid. Report lists sample as > 99%, as provided by sponsor. The purity/identity of the test substance cannot be 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 Test substance: After 28-days, the percent biodegradability of the test substance was determined by three methods: Biochemical Oxygen Demand (BOD), Dissolved Organic Carbon (DOC), and Residual Test Substance Concentration. The results are as follows: Percent Biodegradability (of 3 replicates): BOD: 4,-1, and 6% DOC: Not calculated; 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. Measurement of test substance: The test substance concentration was only measured at the end of the study. Although this laboratory followed the OQ0502 procedures as outlined in OECD 301C, the starting concentration should also be determined rather than relying on nominal concentrations. In addition, subsamples were removed from the bioreactors for pH and DOC determinations at the end of the study. The sample used for pH determinations and the remaining sample after centrifugation for DOC were returned to the bioreactor prior to sampling for the measurement of the test substance concentration. These samples should have been removed after the bioreactors were sampled for test substance concentration determination. Calculations of theoretical oxygen demand should have made which account for possible oxygen consumption due to nitrification. CONCLUSIONS This testing indicated that the test substance is not capable of readily biodegrading in aerobic environments. DATA QUALITY Reliability: Klimisch ranking 3. There is no reference to the solubility information provided in the study. The quoted purity/identity of the test substance is not substantiated. Measurement of test substance concentration was only performed at the end of the study. The bioreactors were potentially contaminated by the practice of returning subsamples after pH and DOC determinations. The calculation of theoretical oxygen demand should have been performed using the correction for nitrification. REFERENCES_______________________________________________ Study conducted at the request of Sumitomo 3M LTD by Mitsubishi Chemical Safety Institute Ltd., Yokohama, Japan OTHER Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 Last changed: 5/22/00 000503 M.S.I. REPORT N o .1B363(2)G Ready Biodegradability Test of D-l Submitted to : SUMITOMO 3M LIMITED Prepared by : Mitsubishi Chemical Safety Institute Ltd. October 31, 1995 00504 Ready Biodegradability Test of D-l (English version) Study No. Study Title Sponsor : 1B363(2)G : Ready Biodegradability Test of D-l : SUMITOMO 3M LIMITED This report was conducted in the Yokohama Laboratory of the Mitsubishi Chemical Safety Institute Ltd., 1000 Kamoshida-cho, Aoba-ku, Yokohama 227, Japan. This report is an English version of the orginal, which was written in Japanese. The underesigned hereby declare that this version faithfully reflects the original report to the best of our knowledge. Mitsubishi Chemical Safety Institute Ltd. Date Kikuo Yoshida, Ph.D. Environmental Science Division of Yokohama Laboratory Approved by Jun Toriya, B. S e r ^ Head of Yokohama Laboratory Date 000505 COMPLIANCE WITH THE GLP STANDARDS Study No. Study Title Sponsor : 1B363(2)G : Ready Biodegradability Test of D-l : SUMITOMO 3M LIMITED To the best of our knowledge and belief, the study described in this report was conducted in compliance with the Good Laboratory Practice (GLP) 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. Sealed date April 28, 1992 Study Director Environmental Science Division 000506 QUALITY ASSURANCE STATEMENT Study No. Study Title Sponsor : 1B363(2)G : Ready Biodegradability Test of D-l : 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 : In-progress study : Final report : Date of Date of Inspection____________ Reporting March 18, 1992 April 15, 1992 April 16, 1992 March 30, 1992 April 15, 1992 April 16, 1992 April 28, 1992 April 28, 1992 Quality Assurance Staff Junko Katoh, Ph.D. Yumiko Miura, B.Sc. Sealed date April Sealed date .April 1992 1992 N) CO CO 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.Sc. Sealed date :April 28, 1992 Environmental Science Division Junko Koyasu, B.Sc. Sealed date :April 28, 1992 Environmental Science Division Yukiko Mizoguchi Sealed date :April 28, 1992 Environmental Science Division Staff for cultivation of activated sludge Yasuo Sato Sealed date April 28, 1992 Environmental Science Division 000508 CONTENTS Page EXPLANATORY REMARKS .................................................. 7 SUMMARY .............................................................. 8 1 STUDY TITLE.......................................................... 9 2 PURPOSE.............................................................. 9 3 TEST GUIDELINES................ .................................... 9 4 GLP COMPLIANCE....................................................... 9 5 STUDY PERIOD......................................................... 9 6 SPONSOR.............................................................. 9 7 TESTING FACILITY..................................................... 9 8 STORAGE AND RETENTION OFTEST SUBSTANCE AND RECORDS.................. 10 9 TEST SUBSTANCE....................................................... 11 10 CONFIRMATION OF IDENTITY AND STABILITY...............................12 10.1 . IDENTITY..................................................... 12 10.2 STABILITY.................................................... 12 11 TEST 11.1 11.2 11.3 11.4 11.5 11.6 11.7 METHODS........................................................ 13 ACTIVATED SLUDGE ............................................ 13 EXPOSURE CONDITIONS ........................................ 13 BOD MEASUREMENT .............................................. 14 pH MEASUREMENT ............................................... 14 DOC MEASUREMENT .............................................. 14 MEASUREMENT OF RESIDUAL TESTSUBSTANCE CONCENTRATION ......... 15 EQUATIONS FOR CALCULATION OFDEGRADABILITY ................... 16 12 RESULTS............................................................. 17 12.1 OBSERVATION OFTEST BOTTLES AFTER EXPOSURE PERIOD ............ 17 12.2 pH MEASUREMENT ............................................... 17 12.3 ACTIVITY OF SLUDGE ........................................... 17 12.4 DEGRADABILITY BASED ON B O D ................................... 17 12.5 DEGRADABILITY BASED ON D O C ................................... 17 12.6 DEGRADABILITY BASED ON THERESIDUAL TEST SUBSTANCECONCENTRATION................................... 18 13 CONCLUSIONS.......................................................... 18 TABLES AND FIGURES................................................ 19-33 000509 EXPLANATORY REMARKS Study No. Study Title : 1B363(2)G : Ready Biodegradability Test of D-l The ready biodegradability test of D-l was originally started on January 29, 1992, and the BOD measurement of the test' substance was initiated on February 13, 1992(Study No.lB363G). However, the degradability value of aniline based on the BOD measurements did not exceed 40%. Therefore, the test was terminated on February 20, 1992 and the new test (Study No. 1B363(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. 1B363G). 000510 SUMMARY STUDY TITLE Ready Biodegradability Test of D-l METHOD 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). ("Ready biodegradability, Modified MITI test" in OECD Guidelines for Testing of Chemicals). Test bottle : Bottle 1 Bottle 2 Bottles 3~5 Bottle 6 : aniline + sludge + basal medium : sludge + basal medium : test substance + sludge + basal medium : test substance f water Measurements : Biochemical oxygen demand (BOD) Dissolved organic carbon (DOC) Residual test substance concentration (for 28 days) (at day 28) (at day 28) RESULTS Measured values (day 23) Bottle No. 3456 BOD, mg 1.0* -0.3* 1.6* 0.0 DOC, mg/1 0.7* 0.3* 1.6* 2.0 Test substance, mg/1 101.3 105.2 91.5 106.7 * value corrected with BOD or DOC value of Bottle 2 Theoretical value 27.7 28.8 100.0 Degradabilities (%) Bottle No. 345 BOD 4 -1 6 DOC NA* NA* NA* Test substance 5 1 NA** * not calculated because the test substance was not dissolved in the test solution. ** not calculated because the test substance might be lost accidentally during condensation of the solution under vacuum. CONCLUSION 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. 000511 1 STUDY TITLE Ready Biodegradability Test of D-l 2 PURPOSE This study was conducted to evaluate the ready biodegradability of the test substance for notification under the Chemical Substance Control Law of Japan. 3 TEST GUIDELINES 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 GLP COMPLIANCE The test was conducted according to the Good Laboratory Practice (GLP) standards applied to industrial chemicals under the Chemical Substances Control Law. 5 STUDY PERIOD March 17, 1992 to April 28, 1992 (BOD measurement : March 18, 1992 to April 15, 1992) 6 SPONSOR SUMITOMO 3M LIMITED 8-8 Minamihashimoto 3-chome, Sagamihara 229, Japan 7 TESTING FACILITY 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 submission 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 GLP standards. 000S13 9 TEST SUBSTANCE 1)Name* 2)Chemical name* 3)Structural formula* :D-l (M.S.I. identification No. 1B363(2)G) :2-[N-Ethyl-N-perfuloroalkyl(C=l~8)sulfonylamino]ethylacrylate C,Hc 0 1 II C n F2n+lS 2 -- N -- C H 2C H 20 -- C -- CH = C H 2 (n=l~8, main component; n=8) 4)Elemental composition :C;28.8%, H; 1.9%, N; 2.3%, 0; 9.8%, F;52.0% S; 5.2% (Measured by M.S.I.) 5)Physico-chemical properties Solubility* : insoluble in water insoluble in DMSO >50% in acetone Melting point* :27~42C Boiling point* :ca. 150C/lmmHg 8)Batch* :Lot No. 4 9)Purity* :>99% (n=l~7;ca.21 % (total), n=8;ca.78%) 10)Appearance amber solid 11)Date of receipt September 25, 1991 12)Supplied quantity* :100 g * provided by the sponsor 000514 10 CONFIRMATION OF IDENTITY AND STABILITY 10.1 IDENTITY 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. UApparatus : Perkin-Elmer model 1640 infrared spectrophotometer. [Figure 1 (P-23)] 10.2 STABILITY 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, indicating that the test substance was stable under the storage condition. l)Apparatus : Perkin-Elmer model 1640 infrared spectrophotometer. [Figure 2 (P-24)] OSlS 11 TEST METHODS The test was performed 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 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 (BOD) was measured over a 28 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 SLUDGE 1) Mixed liquor suspended solid (MLSS) : 3300 mg/1 2) Source : Chemical Biotesting Center, Chemical Inspection and Testing Institute, Japan 3) Date of receipt: January 23, 1992 11.2 EXPOSURE CONDITIONS 1) Temperature 25 1 C 2) Exposure period28 days 3) Test volume 300 ml 4) Concentration test substance (bottles 3~6) aniline*1(bottle 1) activated sludge (bottles 1-- 5) 100 mg/1 100 mg/1 30 mg/1 *1 Reference substance : Syowa Chemicals, Lot No.SC-2726 5)Test bottle contents : Bottle 1 Aniline + activated sludge + basal medium 29^,1(30 mg) of aniline was added to the basal medium*2, then activated sludge was added. Bottle 2 Activated sludge + basal medium Activated sludge was added to the basal medium*2. Bottles 3~5 Test substance + activated sludge + basal medium 30 mg of the test substance was added to the basal medium*2, then activated sludge was added. Bottle 6 Test substance + deionized water 30 mg of the test substance was added to 300 ml of deionized water (purified by Milli-Q). *2 The total volume of the basal medium and the sludge was held fixed at 300 ml. 000516 11.3 BOD MEASUREMENT The BOD was measured for 28 days. 1)Apparatus : closed system oxygen consumption measuring apparatus Ohkura Electric Co., Model OM-2001 (M.S.I. ID No.B). 11.4 pH 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. lJApparatus : pH meter, Denki Kagaku Keiki, Model COM-10 11.5 DOC MEASUREMENT The concentration of the DOC was measured as follows. lJApparatus : TOC analyzer, Shimadzu Co., model TO.C-5000 2)Conditions : FFuurrnnaaccee tteemmppeerraattuurree Air flow rate Sensitivity Injection volume : 680C(TC) 150 ml/min. x5 50 (xl 3) Calibration curve 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) 20 and 50 mg C/l aqueous solutions of potassium biphthalate. IC(inorganic carbon) 0 mg C/l deionized water (purified by Milli-Q) and 10 mg C/l aqueous solution of sodium hydrogen 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 concentration of the residual test substance was measured by the gas chromatography (GC) as follows: 1)Apparatus GC Integrator Hewlett-Packard. Model 5890A (No.2) with FID detector Hewlett-Packard. Model 3392A 2)Conditions Column Temperature Carrier gas Detector flow Attenuation Injection volume :J & W. DB-17(50%Phenyl 50%methyl polysiloxane), 30 m X 0.25 mm i.d., 0.25|rm (film thickness) :[Column oven] 100C {lmin.)-*10C/min.-*2 00C (lmin. ) [Injector] 250C, [Detector] 280C He 30 ml/min. :H2 40ml/min., air 400 ml/min. :[GC] 1 V :1 fxl 3)Calibration curve The standard solutions of the test substance with 0, 250, 500, and 1000 mg/1 in ethyl acetate were analyzed. The total peak area 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 Z9041-1968. [Figure 3 (P-25) and Figure 4 (P-26)] 4) Recovery test 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 251C for 30 minutes. Each test solution was treated by the procedures described in 11.6.6. The concentration of the test substance was measured by G C . The average recovery of the test substance was 97%. [Table 3 (P-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. 5) Detection limit The detection limit of the test substance was calculated to be 3 mg/1 based on the minimum detectable peak area of 100000 p C V sec for the largest peak (R.T. 6.2 min.) in the GC chromatogram. [Table 3 (P-21) and Figure 5 (P-27)] 6) Measurement of test substance in test bottles The contents in each bottle was separately transferred into a 500 ml glass 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 to 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 BOD Degradability (%) = (BODs- BODb)/ThOD X 100 where BODs BODb ThOD Oxygen consumption (mg) in bottles 1, 3, 4, and 5. Oxygen consumption (mg) in bottle 2. Theoretical oxygen demand (mg) of aniline and the test substance. ThOD aniline : CsHiN + 35/402 -* 6C02 + 7/2H20 + N02 ThOD : 90.2 mg O2/30 mg-aniline test substance : C+02-*C02, H+l/402->l/2H20, N+02^ N 0 2, S+02^-S02, F-^F ThOD : 27.7 mg 02/30 mg-test substance 2)Degradability based on DOC Degradability (%) = (1 - (DOCs-DOCb)/DOCc)X100 where DOCs : DOC (mg/1) in bottles 3, 4, and 5. DOCb : DOC (mg/1) in bottle 2. DOCc : DOC (mg/1) in bottle 6. 3)Degradability based on residual test substance concentration Degradability (%) = (1 - Cs/Cc) X 100 where Cs : concentration (mg/1) in bottles 3, 4, and 5. Cc : concentration (mg/1) in bottle 6. 000519 12 RESULTS 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 ACTIVITY OF SLUDGE [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 BOD BOD*1 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. (*L Maximum theoretical value = 27.7 mg) The degree of degradability based on the BOD measurements were 4, -1 and 6% for bottles 3, 4, and 5, respectively. [Table 1 (P-19) and Figure 6 (P-2 8-- 31) ] 12.5 DEGRADABILITY BASED ON DOC DOC*2 in bottles 3, 4 and 5 (as corrected with the value in bottle 2) were 0.7, 0.3, and 1.6 mg/1, respectively. DOC in bottle 6 was 2.0 mg/1. (*2 Maximum theoretical value = 59.8 mg/1) Degradability based on the DOC measurements was not calculated because the test substance was not dissolved in the test solution. [Table 1 (P-19) and Table 2 (P-20)] 000520 12.6 DEGRADABILITY BASED ON THE RESIDUAL TEST SUBSTANCE CONCENTRATION The test substance*3 was detected at concentrations of 101.3, 105.2, 91.5 and 106.7 mg/1 in bottles 3, 4, 5 and 6, respectively. (*3 Initial concentration = 100 mg/1) The degree of degradability based on the residual test substance concent ration were 5 and 1 % for the bottle 3 and 4, respectively. [Table 1 (P-19), Table 4 (P-22), and Figure 7 (P-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 5 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 5. Therefore, the degree of degradability based on the residual test substance concentration was not calculated for the bottle 5. 13 CONCLUSIONS 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 Summary of the test results Degradabilities were calculated by the equations shown in Sec. 11.7. a) Degradability based on BOD lottle Test ThOD day 7 day 14 day 21 No. substance mg BOD % Degra BOD % Degra BOD % Degra mg dability mg dability mg dability 1 aniline 2-- 3 D-l 90. 2 -- 27. 7 56.2 3. 1 3.0 -- 59 0 63.7 5. 0 5. 0 -- 65 0 66. 3 5. 5 6. 1 -- 67 2 4 D-l 5 D-l 6 D-l 27. 7 27. 7 27. 7 2. 4 ` -3 3.5 0. 0 -- 1 4. 1 5.7 0. 0 -- -3 3 4. 8 6.7 0. 0 -- -3 4 Where % degradability was calculated to be negative, this value is shown in brackets. day 28 BOD % Degra mg dability 67.4 5. 8 -- 68 6.8 4 5. 5 -1 7.4 0. 0 -- 6 b) pH measurement Bottle pH No. day 0 day 28 1-- 8. 3 2 6. 9 7. 4 3 6. 9 7. 4 4 6. 9 7. 4 5 6. 9 7. 5 6 6. 8 7. 4 z) Degradability based on DOC lottle Organic carbon TC No. added, mg/1 mg/1 2 1. 7 3 28. 8 2. 5 4 28. 8 2. 1 5 28. 8 3. 3 6 28.8 2. 0 IC mg/1 0. 0 0. 1 0. 1 0. 0 0. 0 DOC mg/1 1. 7 2. 4 2. 0 3. 3 2. 0 d) Degradability based on residual test substance Bottle Concentration Degradability No. mg/1 2 <3 % -- 3 101. 3 5 4 105. 2 1 5 91. 5 NA* 6 106. 7 -- * not calculated because the test substance might be lost accidentally during condensation of the solution under vacuum. 000522 Table 2 Results of DOC measurements Standard solution TviCal50.0:sr1og/L a arer 312 333444667349705 HN 34690 TviCal20.0:sm2g / L # arer 312 111443109804178 MN 14045 ICvia1!0.,0:raa3g/L # arer 312 77118057 7187 MN 7159 1Cvial0.0:rsa4g/L # arer 312 000 MN 0 Bottle2 TC # arer 231 111111573785 MN 1156 mg/L 1.71 11..6658 1.68 RMK IC # arer 321 0 00 MN 0 mg/L 000...000000 0.00 RMK Bottle3 TC # arer rag/L 321 111777414333 222...554339 MN 1733 2.52 IC # arer m g / L 321 00 166 000...200300 MN 55 0.08 RMK RMK Bottle4 TC # arer rag/L 321 111434177840 2.14 22..0060 MN 1420 2.06 RMK IC # arer 321 157 00 MN 52 mg/L 00..0202 0.00 0.07 RMK Bottle5 TC Bottle6 TC # arer m g / L RMK # arer rag/L RMK 321 222233104502 333...432052 312 111334254642 211...199073 MN 2285 3.32 MN 1374 2.00 IC # arer rog/L 1 0 0.00 32 00 00..0000 MN 0 0.00 RMK IC # arer 10 32 00 MN 0 mg/L 0.00 00..0000 0.00 RMK SPL # TC.mg/L RMK 342 56 32122.....0360508622 IC.mg/L RMK 00000.....0000000807 TOC.mg/L / Bottle2 32121.....3046920489 <-<<---<<---- -v^ ' Bottle3 Bottle4 Bottle5 "Bottle6 000523 Table 3 Calculation of recovery and detection limit Recovery test 1 Recovery test 2 Blank Peak area /iV- sec (A) 4805210 4815940 <132800 Concentrat ion in solution, mg/1 for GC in bottle analysis (B) (0 578.8 96. 5 580. 1 96. 7 <16. 0 <2. 7 Recovered Added mass mass mg mg (D) (E) 29. 0 30. 0 29. 0 -- 30. 0 0. 0 Average recovery Recovery % (F) -- 97 97 Detection limit mg/1 (G) -- -- 3 Standard solution Concentration Peak area Volume of solution for GC analysis Volume of test solution for GC analysis (H): (I): (J): (K): 500 mg/1 4150910 yuV-sec 0. 050 1 0. 300 1 Equation '.B=HXA~PI C=BXJ-PK D= C X K F=D-r-EX100 G=C raise decimal fractions to unit. 000524 Calculation of detection limit Retention time, min 5. 6 6. 2 6. 5 Total 500 mg/1 std. Peak area Rate of peak ytiV- sec area, % 298720 7.2 3127000 75. 3 725190 17. 5 4150910 100. 0 Blank Peak area a V-sec <100000 -- <132800 The minimum detectable peak area of the test substance was calculated by the following equation based on the minimum detectable peak area of 100000 yizV-sec for the largest peak (retention time 6.2 min.) in GC chromatogram. Minimum detectable peak area = (yuV-sec) 100000 UV-sec) 4150910 X ------------------ = ( f i V ' s e c) 3127000 (tV-sec) < 132745 (i V- sec) 132800 (u V-sec) Table 4 Residual test substance concentration in test bottles Bottle No. 2 3 4 5 6 Peak area i V* sec (A) <132800 5149880 5349150 4654900 5428120 Concentration in solution, mg/1 for GC in bottle analysis (B) (0 <15. 2 <3 589. 3 101. 3 612. 1 105. 2 532. 7 91. 5 621. 2 106.7 Standard solution Concentration Peak area Volume of solution for GC analysis Volume of test solution for GC analysis Average recovery (D): (E): (F) : (G) : (H): Equation :B=AXD-PE C=AXD-PEXF-rG-f-HX 100 500 mg/1 4369340 n V-sec 0. 050 1 0. 300 1 97 % 000525 Figure 1 Infrared absorption spectra of the test substance Measure by the sponsor Sain>le: ">Cw Kethod: KB*. i Dte: J Operator: Resa rts: Measure in M.S.I. ;; 4 0 0 0 . 0 , 4 0 0 . no c m - 1; o,. 0 7 . BO . 5 3 %r 4 s c a n s; mode r a t i o ; i*esu 1 4 . 0 0 uni-1; 1 0 3 6 JU 0 -1 L o t N o . 4 th re s h o ld 6 . 00%; b an d cm t X c m t X Chi - 1 2 9 0 0 . 3 2 4 . 17 2 3 6 4 . 1 5 0 . 0 5 1729.3 1G19. \UK\i. 1 BOB . 1 Z H .lt 7 .MM sj . a7 1460.0 UM /..1 /B 4. i 10.77 / .SO 22.25 13U2.0 <1114.3 74 7.4 16 p e a k s f o u n d 0 2 /0 1 /1 3 00. 3 l jp o d atru n y A. Okuda X 4.34 5.27 7. in t B . 34 C H I" 1 1639.4 1202.0 USB . 1 6 5 3 .7 X 19.44 2 . ia 36.16 11.30 000526 C-O slrelch Figure 2 Infrared absorption spectrum of the test substance stored in a refrigerator 1: 4000..0, 400.00 c m - 1 ; 0. 14. 77 .86 xr 4 scans; mode ratio; resol 4 .00 cm-1; 18363 (2) G 0-1 Cot No .4 threshold 5.00X; band cm-1 X cm-i X cm-1 2987.2 51.57 2363.0 68.77 173 l.1 1619.8 54.81 1468.8 40.04 1392.9 1062.3 20.58 1018.0 17.95 985. 1 784.2 46.59 747.2 40.04 735.8 16 p e a k s found 92/04/16 16. 39 apod strong A. Okuda X 3.85 5.38 16.75 43.83 cm-l 1630.Q 1205 t5 808.1 653.6 X 42.99 1.32 25.32 25.4Q C-O stretch 000527 Figure 3 Calibration Curve of the Test Substance Calibration Curve Concentrtion X(mg/1) 0 250 500 1000 Peak Area Y(/iV*sec) 0 1.92785E+0S 3.89009E+0G 8.12341E+06 Y= -7.552x104 + 8.139x103 X r= 0.999707 Peak Area(pV*sec) Figure 4 GC chromatograms of the test substance Calibration curve 0 mq/l in ethvlacetate a UCO>t--JCo CO c LU CL X CC O 250 ma/l in ethvlacetate .V cu oe v in <x LU m Oi r-s ac -- v- L CO oc <L CU O' JV CD CL LO >-- LO CO T OS 3; s O' oe N a> LC CE r*s. LO o<c CE GJ CC UJ a. CL cc co 03 > >0 >- 1-- Q o O d as CD CD CD UJ cu ro & CC <z TT in CD CU ro ** a> CO in ro ao\\ cu CD rv LO T> 1-- Mi *T CD a: in LO total peak areai1927850^vsec 500 ma/l in ethvlacetate ---V LC r s <T 0 0 CD < e r - -- o> LO U J -- CU CO LO -- oc < r -c c u cc cu as tr H - CC CD OD -- x c r o> o> lo rs \ cc cd oe a> cd a: " <x a: CD CD CD CC* LU CL > > > 03 CL CL CL > > > H - C Q O CD O <X OE a s CD CD CD U J oc C CD T x -- cc r \ ro --. -- CU < x -- lo in cd ro <T in co CD -- CL -- CO -- CN cu <3? CO <x> CU 00 k-- *X CO cu r o v CC in in lo LO LO 1000 mq/l in ethvlacetate total peak area :3890090jiv sec 000529 TM1 l_j_ Figure 5 GC chromatograms of the test substance- - Rocovery and detection limit Recovery and detection limit are shown in Table 3. 500 mq/l in ethvlacetate Blank V v cn co cn c <X tsO C ro rs c UJ vx>LO c<cZ -- rr: R-<sOcr>-- rs x CNC cn S cn CD Os'sc>n <T CD CD VU = CL. > > O>-. > > CL > h- O O O O ccCC CP s s uj cn oj ss rs rs s CC IO CO -- rs s cn CD O - -- OJ uos , v h - VO cc<r cc ui in in co cn V s l-- a vu coc CO C <z total peak area:4150910|ivsec STATUE OTTTif_______________________ Recovery test-1 w total peak area :4 8 0 5 2 1 0 n v s e c Recovery test-2 2 .(sE* li^cirn]rvdo ur -- --C DOEC CTO Oil Irtcucj o jr -i ro ro n CDLC (U <X *-- -rr h- TT O' O J l s I CD O' cn cr <sO IS in Ccsss scc s S s X s s OJ UJ CD r>c--l- cl Q Q. q_ d > CL a > > c >>a> QC :> co \o<x lu s cn s -- cd in cr cei sv roeo ss C CD OJ oj rs OJ S ' cn ( E ' T O i in cn OJ in --S Uno3 --cn rs in rojo cros cn ro OJ *\0 ro oe .GVhC- CDIDco ojro rr lu in in in uejvo us Qi total peak area:4815940fivsec Figure 6 BOD chart Biological Oxygen Consumption (mg) j -7 5- -- I- -6 0-4 5-3 0-- -1 5-- Measurement of BOD Test substance : D-l S t u d y n u m b e r : 1 B 3 6 3 ( 2 )G Period : 3/18/1992 to 4/15/1992 Temperature 25 1 1C Apparatus : Ohkura Electric Co., M o d e l O M - 2 0 0 1 ( I .D . N o . : B ) Chart speed : 2mm/hour bottlf Substance C o n e . Sludge cone. Ho. mg / 1 1 Aniline 100 2 _____ 3 D-l 100 4 D-l 100 5 D-l 100 6 D-l 100 mg/1 30 30 30 30 30 _____ Operator : Ayumi Okuda -- |-- - 0-- 0 day -- (0) (hour) 000531 Figure 6 (Continued) 000532 Figure 6 (Continued) 000S33 Figure 6 (Continued) ------- 1.... .....-- -- ....__ LO " ..... ' : "" "... ..-- --- -- ---- ;---- -- :-.-:-1- 1! --- ---- -- ----. '_* _"___ ` 1 , -- ;----- --- ;_:__ ! --- - -- -----1.- - -____________ _ : O 1 '' -- _____! H ; . i- 1 fNJ C J t. SOJJ. - 'Ni : i --1--!!---- -- ___ 1___ -- ----- :-- ----- !-:-- -- :-- j-- !-- -- ^ Bottle 1 , i ' 1 _!_!___^ _ -- :-- --i !-- ... . *' -( __:_ ' ; _1_:_:__ : : ; ! 'i ; :1 ___ ___ .-.... 1: : . ' ; __ 1 * ; ' !1 \ i :; i ' _i_ i < Ili; -- 1-- -- :-- !-- i ;. '1 -- -- ;-- r~ ------1 _ :-! .. _:ij _z_:.t : 1i !----- 1------- ; i !i 1 1 i1 : , : i ~i rTi.vt= i : : !ii Il ; ! |--11 ii ii i'ii -- :1 ; " ;` -- j-!-- i i1 ";.'i. , ; ; . :I 1; !1 1 .-i --H !H1-i1 il' 'i! .i : 1 ii ----!j;i---- iiii!'l'1;i;;--.1.-----~*i1i----7--^it--'----ljLr-- 1 -- Ii r- i _i-.i-j 1 --:-j-;-;-- --:--::--| :.-!r~ ... . j _ . ____ , !!! : '1 1 i ;ii ! , 1 1 j_ . :; ' 1 1 i . ' 1 --;-;--- -.. --j- 1 -- -------;-- i_ .1 _ . :1 .j -- ---- ' - = = ----- ---- 1 ! ,; ----- 1-- :-- -- :-- --- ; ....... Bottle 5 / , Bottle 3 -/ .i Bottle 2 -- -- 1-- rL = I i . . | .....--| , . | .-1 -- 1 --__ :_I !:1, ;i | I. ;;; 11 * . J ....... 1 __ i ; j_ -- :-- | j '1 ! ! i: :! iili --- --- | , * -!__:_:-1 : ' ., 1; Bottle 4 : i ?R Bottle 6 (672) i i ! ; : . i i I m n- 00S34 Figure 7 GC chromatograms of the test substance--Measurement of residual test substance concentration The concentrations of the residual test substance in the bottles are shown in Table 4. 500 mq/l in ethvlacetate <xi Cvj SO r-ym L.<VUx -0r0--v CO r x s----r ro '* in 2: so ro scoo 'tf- so J-- X\ 00cu ccCP CcPdi cu aP, so rx d CD CD LU QCL . >Cl Q >>Q 2> o >CD o <X CD locuc nicrcnxuo CU CO --r x CD Cv U ----w1cu CU CO ro oL<,<VucxX QP--d iscnou os n ccuu so rToT so 0"c3 so total peak area:4369340|wsec Bottle 2 ahoC--Oc to <z 5Tmrij Bo.ttig-3 Bottle 4 s.<O<vu1dx --ovcuj*src--xo r-iSnxOrrn"xon smo saos cN"Xu in ;--c <Cr --CO--m03c--u>crxu >l>--u-L>oC L>> o a>. a >C- a >> m> lDcduC: Tx--co->rh--cxoi. rsCcr>xon3iiCcnnUu 0rino3r- 0Ci0nv33J rc--c n-r<x- rmx* cisnou <in73csCouU in sc total peak area: 5149880|iv sec 000535 total peak area:5349150nvsec Figure 7 (Continued) Bottle 5 .V <x -- h rs OS ro C--U<C"VD rv. rs -- m LU 00 CO OD ro m CD OJ o<cx m oj to ro oj m tD hzc icno awo cd oj CO to CO TP to oj rs \C D -- --' CD CD CD <s <C CD CD CD CD 0 CO CD lau. a>. >1 -0 >> 0 a>. 0 >> Cl.> oo >- m >> oo <x LU CD CD CD CD OD OJ S CO CO 00 CD IS oe OC <r to cu cu ro OS tto-- PO in ro ro oroi co --( ro\j rs TT t"*o OJ OJ OtDo ^ CO ro ro rr>oj CD OJ + CO ,V CE tu hD-C Ps TP tn VO in CTiOJ in to OD to V to D<xC total peak area :4654 90 0.ivsec Bottle 6 .S' CD CD tO in <c m n UJ CO M ro o a! 0 <x in ro rs CO to h- PO PO OJ CO X <S cr o d rs N T CD CD C <r cd CD CD CD CD UJ Cl > > CO > a. a. CL D- > CJ h- o O o Q <X OD CD CD 0 UJ QC cc 00 o- CO CD V OJ -4 - cri rs CO m cu to is m CD m m OJ 0> m "O' OJ <S OJ CD CD ,s* *-- VC CM OJ m <X QC tu ce m l in ' to to c <r total peak area: 5428120|av" sec s t w 'Ijf 000536