Document z2aD2z6d1v9YVL5LKQk9KMQg

f t k 26 - 0 ^ 0 0 BIODEGRADATION (MANOMETRIC RESPIROMETRY - ANAEROBIC) TEST SUBSTANCE___________________________________________ Identity: A mixture containing perfluorooctanesulfonate;, which may also be referred to as PFOS, FC-95, or as a component of FC-3041. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was not noted. The test sample is FC-3041. Current information indicates it is a mixture of 6% PFOS, 36% diethylene glycol butyl ether, 36.05% water, 12% Sultone fbamer, 9% sodium octyl sulfate, 0.55% sodium lauryl sulfate, and 0.4% tolyltriazole. The following summary applies to a mixture with incompletely characterized concentrations o f impurities. Datsm ay not accurately reflect the biodegradability o f the fluorochemical component o f the test sample. METHOD____________________________________________________ Method: Manometric Respirometry Test Test type: Anaerobic biodegradability GLP: No Year Completed: 1993 Analytical monitoring: Determination of the mixture ratio of methane and carbon dioxide released in the gas phase by gas chromatography, dissolved organic carbon (DOC), and the release of fluoride by fluoride selective electrode and pH values. Statistical methods: Results were determined by calculation of the percentage biodegradation. Inoculum source: Anaerobic sludge collected from a community sewage treatment plant (Lennestadt-Maumke, Germany) that was fed almost exclusively with residential wastewater. Test condition: Dilution water: Not noted. Mineral Nutrient Medium: Mineral Basal Medium. Stock and test solution preparation: Solutions of the test and reference materials were prepared at 100 mg C/L and 50 mg C/L, respectively, with mineral basal medium. Test vessels: 118 mL serum flasks which contained a liquid phase volume of 40 mL, closed with butyl septa and the gas phase purged with nitrogen. Incubator Conditions: In the dark. Temperature: 37 2 C Agitation: None Number of concentrations: Test sample, test sample inhibited, sodium benzoate (reference substance) and blank inoculum, all in triplicate. 002885 Inoculum condition on test initiation: 0.79 g/L dry weight present in test samples. Element Basis: Biological mineralization by means of the release of methane and carbon dioxide. Test substance flask conditions: pH: Not noted in the report. RESULTS___________________________________________________ Nominal concentrations: Blank control, test material at 100 mg C/L and sodium benzoate (reference substance) at 50 mg C/L. Element values: 60-day % Degradation by Mineralization (methane and carbon dioxide) in test vessels 7, 8*, and 9 were 78.5, 20.7 and 91 respectively. 60-day % Degradation by Determination of DOC in test vessels 7, 8*, and 9 were 81.5,25.2 and 90.5 respectively. Fluoride release, by selective fluoride electrode, was not detected in any of the test samples (detection limit 2 pg F/L). ` Could not confirm what caused the low % degradation in this test vessel. Remarks: Testing was conducted on a mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone. CONCLUSIONS_______________________________________ _______ The test substance under anaerobic conditions was categorized as biologically degradable. The reference substance obtained 89.5% degradation during the incubation period of 60 days as determined by mineralization (108% by DOC), thus the validation criteria of the test was fulfilled (>60% degradation in sodium benzoate in 60 days). Subm itter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY______________________________________________ Reliability: Klimisch ranking 3. Lacks specific methodology used and documentation. Sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in the solution. REFERENCES_______________________________________________ 002886 The studies were conducted by Fraunhofer Institute for Environmental Chemistry and Ecotoxicology, D-57377 Schmallenberg, Germany at the request of the Federal Bureau for Material Technology and Acquisitions, Germany, Grant No.: E/B31E/P0078/N52275, 1993. OTHER_____________________________________ _________________ L ast changed: 6/26/00 002887 FINAL REPORT ECO-TOXICOLOGICAL EFFECT OF FLUORO-SURFACTANTS G ran t No.: E/B31E/P0078/N5275 Client: Bundesamt fr W erktechnik und Beschaffung, Koblenz (Federal Bureau for Material Technology and Acquisition> Investigating Institute: Fraunhofer Institute for Environmental Chemistry and Eco-toxicology D-57377 Schmallenberg Institute Managen Prof. Dr. W. Klein Project leaden Dr. R. Debus Project authors: Dr. A. Fliedner Dr. K. Hund Dipl. BioL R. Niemann Dr. A. Remde December 1993 Translator Andrea-Ingrid Sdmeidcr. 715-549-5734 (German #1453/1995) 002885 Fraunhofer Institute for Environmental Chemistry and Eco-toxicology D-57377 Schmallenberg Investigation for the Eco-toxicological Evaluation of the Fluoro-surfactant Base FC-3041 Anaerobic Degradation Part g by Dr. A. Remde Translator. Andrea-Ingrid Schneider, 715-549-5734 (German #1453/1995) 8 002889 ANAEROBIC DEGRADATION The determination of the biological degradability o f Base FC-3041 under anaerobic conditions was done in the manometric-respirometry test (9). MATERIAL AND METHODS Test substance: Base FC-3041 (combination o f fluoro-surfactants in aqueous solution; Manufacturer. 3M Company Germany, Neuss). Test criterion: Measurement of the biological mineralization by means of the release of methane and carbon dioxide. Test method: A defined volume (40 ml) o f an inoculated m ineral medium with a known concentration of the test substance (100 mg/L) is incubated in the dark in a closed serum flask (volume o f 118 ml) at a constant temperature (37`C 2). During the incubation period of 60 days the release of methane and carbon dioxide is determined by gas chromatography by extracting regularly 0.5 -1 ml samples from the gas phase o f the test run. . Test procedure: 15 test vessels (serum flasks with a volume o f 118 ml, o f which 40 ml liquid phase) were prepared for the performance o f the test: Vessel 1,2 and 3: Vessel 4 ,5 and 6: Vessel 7,8 and 9: mineral basal medium + inoculum mineral basal medium + reference substance (sodium benzoate 50 mg C/L) + inoculum mineral basal medium + test substance (100 mg C/L) + inoculum (30 mg solid maiter/L) ------ T ranslator A ndrea-Ingrid Schneider. 715-549-5734 (Germ an #1453/1995) 9 002890 Vessel 10,11 and 12: mineral basal medium + test substance (100 mg C/L) + inoculum + mercury chloride solution (0.001% final concentration) The serum flasks were closed with butylsepta and die gas phase purged with nitrogen. Then the determination o f the methane and carbon dioxide content in the gas phase occurred. During the incubation period of 60 days at 37 2'C gas samples of 0.5 - 1 ml were taken out at regular intervals and the mixture ratio of methane and carbon dioxide in the gas phase was determined. The total amount of the gases released (CCH4 and CCO2) w as determ ined through the addition o f the concentrations of the gases in the gas phase and the calculated concentration of the gases dissolved in the liquid phase (10). The total amount of the mineralized carbon (Cnn) at a certain time of measurement (t=n) is calculated from the sum o f the release of methane (CCH4) and carbon dioxide (CCO2) in the duration o f incubation of 60 days, following the equation (a). a) Cmjn= [(CcH4)tm *(CcH4)t=o3 + [(QxttXsn - (Cc02X=ol The bio-degradation was determined according to the following equation (b): C. b) Percentage bio-degradation= - ^ aL x 100 Ss Cmin Cts = Total amount o f the mineralized carbon in mg C = Total amount of the test substance used in mg C Additionally, at the start and end of the experiments the pH-value, the dissolved organic carbon and the fluoride concentration in the aqueous phase of the test runs were determined. Inoculum: An anaerobic sludge from a comm unity sewage treatm ent plant (LennestadtMaumke) which was fed almost exclusively with residential waste water, was used as inoculum . The sludge was therm ostated during the transport 4 ml sludge were pipetted into the test samples, such that a dry weight of 0.79 g/L was present in the test samples. The sample collection occurred on 9/28/1993. Translator: Andrea-Ingrid Schneider, 715-549-5734 (German #1453/1995) 10 002891 A nalysis: The determination of the mixture ratio of methane and carbon dioxide in the gas phase was done in a Shimadzu 8A gas chromatograph with a thermal conductivity detector over a Porapak-N analytical separation column. The analysis of the dissolved organic carbon was done in a TOC (Total Organic Carbon Analysis) 500 o f the Shimadzu Company. The fluoride was determined by means of a fluoride selective electrode (Orion 69-90). Reference substance: sodium benzoate (Merck, reagent grade) Test period: 9/30/1993 - 11/29/1993 RESULTS During the incubation period of 60 days, 89.5% of the reference substance sodium benzoate was degraded (Tab. g l, Fig. g l). With that, the validity criterion o f the test, a degradation of sodium benzoate >60% in 60 days, was fulfilled. The degradation of BASE FC-3041 was very different in the three test samples started . in parallel to each other (Tab g l, Fig. g2). According to the methane and carbon dioxide release, 4 days after starting die test a degradation of 10 -13% could be observed in all three vessels. In vessel 9, the test substance BASE FC-3041 was degraded at a constant rate of degradation. After 33 days, 91% of the test substance was mineralized. In the test vessel 7 the test substance BASE FC-3041 did at first not further degrade from day 4 to day 25. Only after 33 days could a significant degradation be observed again. After an incubation period of 60 days, a degradation of 78.5% was achieved. Parallel to vessel 7, the test substance BASE FC-3041 in vessel 8 did at first not further degrade from day 4 to day 25. Here too, a restart in degradation could be observed after 33 days, however, with a substantially lower rate of degradation in comparison to vessel 84. After 60 days, only 20.7% of the test substance had degraded in this test sample. 4sic.Must be T . Translator. Andrea-Ingrid Sdm dder, 715-549-5734 (German #1453/1995) 11 002892 The cause for this degradative behavior in the test vessels 7 and 8 is not known. The possibility exists that during the sample collection or during incubation oxygen diffused in various amounts into the test samples, through which the anaerobic mineralization was inhibited to different degrees. A confirmatory test for oxygen in the vessels 7 and 8 with a suitable color indicator (resazurin) was not posable due to the dark black coloration of the test samples. However, the different degradation of BASE FC-3041 observed in the test samples may also be traced to the inoculum. Sludge represents a very complex inoculum with a large diversity. Not all organisms are capable of degrading the test substance to the same ex ten t Therefore, the possibility exists that a different distribution of various organisms in the test vessels occurred during the addition of the inoculum, which led to the difference in degradation of the test substance observed. - Tab. g2 shows the results of the determination o f the anaerobic degradation of BASE FC-3041 by means of the mineralization to methane and carbon dioxide in comparison to the determination of the decrease of dissolved organic carbon. A very good agreement between the two methods was found in all test samples for the test substance as well as for the reference substance. A release of fluoride by means of a selective fluoride electrode (detection lim it 2 Mg fluoride) was not detectable in any of the test samples. During the mineralization of fluorinated compounds, fluorine can in principle be released only as fluoride ion. A volatilization of fluoride, which is observed for low pH-values, can be excluded under the given slightly alkaline conditions, measured at the start and the end of the experim ent The possibility exists that released fluoride was bonded to the glass vessels or to the organisms. EVALUATION Based on the present test results, the test substance BASE FC-3041 may be categorized as biologically degradable under anaerobic conditions. Translator. Andrea-Ingrid Schneider, 715-549-5734 (German #1453/1995) 12 002893 Tab. g l: Mineralization of BASE FC-3041 under anaerobic conditions Day Inoculum Reference substance cmin l ) C m l A Z 1 X degradation g C mg C Base FC-3041 + HgCl2 Cmin 1 ) mg C 000 0 4 0,279 0 0 11 0.703 0 0 25 1.206 0 ,4 20 33 1,343 1,337 66,8 - 60 1,392 1,789 89,6 0 nb nb nb 0,04 0,06 Day Test Substance Base FC-3041 Vessel 7 Vessel 8 Vessel 9 Cain 31 mg c X degradation Cai.H 3 ) mg C % degradation Cain 3 ) mg C dgradt] 00 i 4 0,428 11 0,394 29 0,442 33 1,913 60 3,141 0 10,7 9,8 11,1 47,9 78,5 0 0,534 0,533 0,407 0,612 0,831 0 13,3 13,3 10,2 15,3 20,7 0 0,481 1,657 3,412 3,641 0 12,: 41,i 85,: 91 1) Average value from 3 experimental runs 2) Average value from 3 experimental runs minus Cmin o f the inoculum control 3) Cmin of the experimental runs minus Cmin of the inoculum control nb = not determined Translater Andrea-Ingrid Schneider, 715-549-5734 (Gennan #1453/1995) 13 00289^! Tab. g2: Determ ination o f the anaerobic degradation of BASE PC-3041 and sodium benzoate by m eans o f m ineralization and measurement of the dissolved organic carbon (DOC) Ha --b en zo ate 1) M inernliratinn % degradation 89,5 Determination of the DOC % degradation 106 BASE F C -3 0 4 1 V essel 7 V essel 8 V essel 9 78,5 20,7 91 81,5 25,2 90,5 1) Average value from 3 parallel [runs] Translator. Andrea-Ingrid Schneider. 715-549-5734 (German #1453/1995) 14 o 02895 Fig. g l: Anaerobic degradation of sodium benzoate (average value from 3 parallel [runs]) DAYS Translator. A ndrea-Ingrid Schneider, 715-549-5734 (Germ an #1453/1995) 15 002896 Fig. g2: Anaerobic degradation of BASE FC-3041 in 3 parallel samples ' Hi Vessel 7 C Vessel 8 O Vessel 9 Translator Andrea-Ingrid Schneider, 715-549-5734 (German #1453/1995) 16 0 0 2 8 9 ';