Document Gm4NR6BQp9VRL2VvZR78Oq6Vx

A ft 9l8l6 - 077g BIODEGRADATION (301E) TEST SUBSTANCE____________________________________________ Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-206. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was 2330,10/79. The test sample is FC-206 (FM-3824). Current information indicates it is a mixture of 0.67% PFOS, 17.5% diethylene glycol butyl ether, 78.91% water, 1.33% Sultone foamer, 1% sodium octyl sulfate, 0.04% sodium lauryl sulfate, 0.5% polyoxyethylene monooctylphenyl ether, and 0.05% benzotriazole. The following summary applies to a mixture with incompletely characterized concentrations of impurities. Data may not accurately reflect toxicity of the fluorochemical component of the test sample. METHOD____________________________________________________ Method: Modified OECD Screening Test, OECD 301E with DOC Analysis Test type: Ready Biodegradability GLP: No Year Completed: 1980 Analytical monitoring: Dissolved organic carbon (DOC) Statistical methods: Results were determined by calculation of the % DOC removal and graphic interpretation. Test organism source: A 50:50 mix of soil extract and secondary effluent. The secondary effluent was from an activated sludge aeration basin at the St. Paul, MN Metro Wastewater Treatment Plant, while the soil was from the city of White Bear Lake, Ramsey County, MN. Test condition: Dilution water: Deionized water. Mineral Nutrient Medium: Nutrient medium per OECD 301E method (1979). Initial pH 7.0. Test and reference solution preparation: The test material was prepared by dissolving 426 mg FC-206 in one liter of mineral nutrient medium. This solution gives a final test concentration of 40 mg DOC/L. The reference substance, hydroquinone, was prepared by dissolving 30.6 mg in one liter of mineral nutrient medium. This solution gives a final test concentration of 20 mg DOC/L. Test vessels: 500 mL Erlenmeyer flasks containing 250 mL test solution and capped with foam plugs. Incubation conditions: Lighting: Constant dark conditions. Temperature: 23.5 - 24C 001415 Agitation: Continuously Number of concentrations: 1 plus hydroquinone (reference substance) and blank, all in duplicate. Inoculum condition on test initiations: Not given. (No measurements are required per method referenced.) Element Basis: Decrease in dissolved organic carbon compared to the blanks. Test substance flask conditions: Not given. RESULTS____________________________________________________ Nominal concentrations: Blank control, hydroquinone at ~20 mg DOC/L and at ~20 mg DOC/L plus HgCh (inhibited, test material at ~ 40 mg DOC/L and at ~40 mg DOC/L plus HgCb, all in duplicate. Element values: 28-day degradation = 91.4%_duplicate 1 and 92.3% duplicate 2. Mean value = 92% Remarks: Testing was conducted on the 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 % degradation based on the mean DOC removal value was 92% after 28 days. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY _____________________________________________ Reliability: Klimisch ranking 2. This study meets the criteria for quality testing. However, the sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in solution. REFERENCES________________________________________________ The studies were conducted by the 3M Company, Environmental Laboratory, Lab Request number 5541S, 1980. OTHER______________________________________________________ Last changed: 6/27/00 001416 Ti^a-m'miln L(jni,jiAI 11A TECHNICAL REPORT SUMMARY Dato 6/30/tC TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN .m poriant - I f report sprinted on both sides o f paper, send two copies to TCC.) Division Environmental Laboratory (EE & PC) Oept. N um tw 0535 I'rojuct Commercial Chemicals Division fiOpOM Vitto Proloct Nurnbar 9970012600 Report Number Biodegradation of "LIGHT WATER" Products in OECD Test-6/80 To' 040 John A. Pignato - Commercial Chemicals Division - 236-2A (01) Auihor(s) Employee NumberUI Eric A. Reiner - Environmental Lab (EE&PC) - 21-BW (63 47816 Notebook Rtftronc No. of Pages Including Coversheet None - See 3M Environmental Lab Request No. 5541S 31 SECURITY ^ Open D Closed 3M CHEMICAL V. REGISTRY ^ New Chemicals Reported Yes H No KEYWOROS: (Select terms from 3M Thosouruj. Suggest other applicable terms.) Environmental Laboratory (EE & PC) CURRENT OBJECTIVE: To use the internationally recognized OECD Screening Test with DOC analysis to show that "LIGHT WATER" products are highly biodegradable. TP 'roc REPORT ABSTRACT: (200-2S0 words) This abstract information is distributed by the Technical Communications Center to alert 3M`ert to Company R&D. This study used the "Modified OECD Screening Test with DOC Analysis" and supplemental parallel sterile controls to conclusively demonstrate the extensive biodegradability of "LIGHT WATER" Brand APFP products: FC-203, FC-206, and FC-3017. In 14 days, the dissolved organic carbon (DOC) levels of FC-206 degraded by 90% and FC-3017 was 94% degraded. FC-203 showed 93% DOC degradation in 21 days. The parallel sterile controls proved that this DOC loss was not due to chemical or physical processes such as adsorption, volatilization, or precipitation of the parent material. Information Liuanitsoorni Initiale VVnO 001417 2- - BIODEGRAOATION OF "LIGHT WATER" PRODUCTS IN OECD TEST INTRODUCTION Tii<_* JM Environmental Laboratory chose to attempt to demonstrate the biodeyradability of "LIGHT WATER" products using the OECD screening method because the test has wide international acceptance/ and the Environmental Lab has had experience using the OECD test as a participant in an international ring test. These results will complement the existing BOD data that show these products to have a high degree of biodegradability. The OECD test is run at 20-25C and it measures only dissolved organic carbon. For these water-miscible "LIGHT WATER" products, which are not expected to volatilize, precipitate, or be adsorbed during testing, the OECD test is an appropriate method for measuring % total degradation. The TOC analysis method even_measures the perfluorinated portion of these products. METHODS AND MATERIALS Methods and Conditions - Wade A. Scheil performed this testing following the September 19, 1979. version of the Modified OECD Screening Test with DOC Analysis^). A copy of this method is attached (Appendix 1). Reductions in the scale of the test were necessary to adapt it to available shaking equipment. Five hundred-ml Erlenmeyer flasks replaced 2-liter flasks, 250 ml of inoculated nutrient solution replaced 900 ml, and the sample size was 25, not 30 ml. As in the prescribed procedure, the analyst used only the last 10 ml of sample filtrate for DOC analysis. A calibrated chart recorder monitored the temperature which stayed within the 20-25C range during the 28-day test, except for a 3-4 hr. period when it reached 25.5C. The temperature was most frequently near the high end of the range (23.5-24C). Chemicals - All chemicals were reagent grade unless otherwise noted. The 3M "LIGHT WATER" products used were samples of commercial material of Antwerp manufacture. Their label identified them as FC-203 NFP, FC-3017 Lot 2303, and FC-206 (FM 3824) Lot 2330 10/79. The hydroquinone used as the calibration compound was Aldrich reagent grade hydroquinone 98.5%, Lot 100147. Four chemical supply houses were contacted in an attempt to obtain the 99.5% pure hydroquinone prescribed by the Modified OECD Screening Test, but this purity is apparenty not readiy available in the USA. We expect that this- * reduced purity will have no signficant effect on results. The impurities have no apparent inhibitory effect on microorganisms and their presence could make only a trivial change in TOC measurements. 001418 -3- Water used in tne preparation of all solutions/ except the inoculum, was St. Paul city water passed through a carbon bed, macroporous anionic resin bed, and 2 mixed bed deionizers, and then filtered tnrough a 0.2-um filter. This water was also used to make up losses to evaporation. Stocks of water collected at the start of the experiment ensured consistent water quality. In preparing nutrient solutions, 26.4 g of anhydrous Na2HP04 replaced the prescribed 33.4 g of NII2HPO4 21I2Q, 36.3 g of CaCl2*21120 replaced 27.5 g CaCl2. These replacements yielded solutions and concentration! identical to those required by the OECD Method. The optional yeast extract solution was used in place of the vitamin solution (1.6.1.2(f)). The 100 mg of Fe-chelate used in the trace element solution consisted of equimolar amounts of FeCl3 and EDTA (30 mg FeCl; and 70 mg EDTA). Test Concentrations - DOC measurements of sample stock dilutions served as the basis for adjusting initial test concentrations to approximately 40 mg of carbon per liter. "This is the highest concentration in the range recommended by the OECD test. Use of this high initial concentration increased the sensitivity of % DOC removal measurements. The low toxicity of "LIGHT WATER" products to microorganisms made significant inhibition at this concentration unlikely. Adjustment of hydroquinone, the calibration compound, to 20 mg of carbon per liter was done by weight based on its known carbon concentration of 65.4%. Two blank controls, instead of 1, were utilized in making blank DOC corrections. Inoculum - The inoculum was 50-50 mixture prepared according to steps 1.6.2.1 and 1.6.2.2 of the OECD test using fresh soil from E. A. Reiner's garden and the supernatant of a sample from an activated sludge aeration basin at the St. Paul Metro Plant. The filter paper used was Whatman** 54, and the water for soil extraction was chlorine-free well water. sterile Controls - Replicate vessels containing 400 mg/1 IlgCl2 served as sterile controls for each test material and the calibration compound. Handling of these sterile controls was identical to the handling of the viable cultures. They had the same innoculum and sampling frequency, but the analyst only made DOC measurements on the 0- and 28-day samples. sample Preservation - Tne sample preservation method used was that prescriued by the TOC manufacturer!4 ). The method involves adding 1 drop of concentrated HC1 to the 10 ml filtered samples, bringing the sample to <pil 2, and storing the samples under refrigeration in vials with aluminum foil-lined caps. The method is an effective preservation method. The manufacturer indicates that the method can stabilize calibration solutions made from potassium hydrogen phthalate, a readily biodegradable material, for several months. 001419 -4- Since the acidification in this preservation method was also a necessary step in the TOC analysis protocol, its use was the most practical and allowed elimination of the IIgCl2 preservation technique described in the OECD method. All TOC analyses were made within a month of the preservation of the samples. Instruments and equipment - The organic carbon analyzer used was a Dohrman DC-52A. Its sensitivity limit is between 1 and 2 mg of carbon per liter. The stoppers for the reaction flasks were clean, porous, plastic foam plugs. Gelman 0.2 urn membrane filters, (Part Ho. 64814) boiled 3 times and stored in deionized water, served to filter samples for DOC analysis. Glassware cleaning involved soaking in chromic acid cleaning solution (ChromergeR ) followed by 6 deionized water rinses. RESULTS AND DISCUSSION Table 1 summarizes the results of this study. Appendices 1 and 2 contain the actual data sheets and plots of the degradation as a function of time. All 3 "LIGHT WATER" AFFF products degraded nearly completely. TABLE 1 Percent Degradation of Hydroquinone and "LIGHT WATER" AFFF Products FC-203, FC-206, and FC-3017 With Time in Modified OECD Screening Test % Degradation at Day: Product 7 14 21 27 28 FC-203 13 85 93 91 FC-206 23 90 94 93 FC-3017 37 94 96 96 Uyd ro- 88 92 97 94 quinone 93 92 95 93 In the case of the highly water soluble "LIGHT WATER" products, it is very unlikely that physical means such as adsorption, volatilization, or precipitation caused the loss of soluble TOC (DOC). This is substantiated by the relatively high oxygen demand observed in BOD tests of the 3M Environmental Laboratory (FC-203 BOD5/COD * 0 . 5 v FC-206 BOD5/COD = 0.5). The sterile control data in Table 2 provide further substantial evidence that the soluble TOC loss from the test samples containing "LIGHT WATER" products is not due to adsorption, volatilization, or precipitation. These samples were handled identically to the test samples except that they contained HgCl2 to prevent microbial growth. 001420 -5- At the end o the 28-day test period, they still contained nearly all the initial DOC. Thus loss of soluble TOC by these physical modes was not a major factor, at least for nonmetabolized "LIGHT WATER" components. This control doesn't prove that physical processes did not remove "LIGHT WATER" metabolites in the test runs, since no metabolism occurred in these sterile controls, but the catabolic* formation of less water soluble materials is unlikely because catabolic products are usually more polar and smaller. TABLE 2 DOC Loss from Sterile Controls Product % DOC Remaining at Day 28 FC-203 FC-206 FC-3017 Hydroquinone 91.3 94.2 88.1 77.5 An off-white precipitate formed in the hydroquinone sterile control on day 0 prior to filtering for DOC analysis. The precipitate was slightly brownish, as was the solution in the hydroquinone sterile control after 21 days. This suggests that hydroquinone is not stable, at least in the presence of ilgCl2, throughout the course of the 28-day OECD experiment. This apparent tendency of hydroquinone to spontaneously form insoluble humus-like material makes it an inappropriate control compound for the OECD test. Its partial precipitation with HgCl2 also casts some doubts about the appropriateness of using the HgCl2 preservation procedure in the OECD method. CONCLUSION The present study conclusively demonstrates that "LIGHT WATER" products, FC-203, FC-206, and FC-3017, are nearly completely degraded in 21 days under the conditions of the modified OECD Screening Test. REFERENCES (1) Organization for Economic Co-operation and Development Chemicals Testing Program Expert Group on Degradation/Accumulation, Dec., 1979, Test Guideline for the Modified OECD Screening Test with DOC Analysis (Level I) (H. G. Nosier) Revision of Sept. 19, 1979. Dohrman DC-52A Operating Manual, 4th Ed., 1978, p. 3-1. Appendices: 1 - OECD Test Guidelines (Sept. 19, 1979) 2 - Data Sheets 3 - Graphs * Catabolism is biologically facilitated breakdown to less complex molecules. 001421 APPENDIX 1 Tokyo, December ist, 197cj C 118/79/Int. OECD - Chemicals Testing Programme Expert Group C, Degradation/ccumulation Mbdified OECD Screening Test with DOC Analysis 001422 Tost Guideline C 118/79/lnt. Test Guideline for the Modified OECD Screening Test with DOC Analysis Date of last revision: Sept. 19,1979 Level I test for ready biodegradability 1. Prerequ isites It has to be known whether the tost material is soluble in the concentration range employed in the test (corresponding to 5 - 40 rag DOC/1). 2. Guidance Information Knowledge of the bacterial toxicity or inhibitory properties of the test material is not unequivocally required but constitutes useful information for the conduction of the test. 3. Qualifying Statements The method is suited for the measurement of the aerobic ultimate biodegradability of water soluble, non-volatile organic compounds. It is unsuited for the biodegradability evaluation of mixtures. 001423 cl I Q / S Sept. 19. 1979 Modified OECD Screening Test with DOC Analysis Preamble It has to be realized that the following procedure for the modified OECD Screening Test has to be regarded in some points as provisional since se veral important features such as the calibration compound and the new test duration of 28 days are completely untried yet. However, since the test constitutes a modification the official, widely practised and well accepted OECD Screening Test for the biodegradability evaluation of surfac tants there should exist a good prospect for success. 001424 September 19* 1979 OECD Chemicals Testing Programme Test Guideline for the Modified OECD Screening Test with DOC Analysis 0. The test procedure constitutes a modification of the OECD Screening Test (OECD Environment Directorate, Proposed Method for the Determination of the Biode gradability of Surfactants Used in Synthetic Deter gents, Paris 1976, and council directive of Nov, 22, 1973 on the approximation of the laws of the member states relating to methods of testing the biodegrada bility of anionic surfactants (73/1*05/EEC), Official Journal of the European Communities No. 4 3^7/53 of Dec, 17, 1973) for the application of the dissolved organic carbon (DOC )analysis. 1. Method 1.1 Introduction: Purpose, scope, relevance, and applica tion of test and explanation of limits. The purpose of the method is the measurement of the ultimate biodegradability of water soluble, non volatile organic compounds in an aerobic, aqueous medium at a starting test concentration corresponding to 5 - **0 mg DOC/1 ( In order to avoid inhibitory effects it is in the investigator's own interest - 2- 001425 2 to choose as low a starting concentration as his analytical capability permits ) 1.2 Definitions and units 1.2.1 Definition of biodegradability where Dt = degradation in.percent DOC-removal at time t CQ = starting DOC concentration of the culture medium (mg DOC/1) Ct = DOC concentration of the culture medium at time t (mg DOC/1) Cbi= starting DOC concentration of the 0 blank (mg DOC/1) CbX= DOC concentration of the blank at * time t (mg DOC/1) 1.2.2 Units The degradation is stated as the percentage DOC-removal within 28 days with respect to the test material ( % DOC-removal ) 3 00142G 3 1.3 Reference compounds 1.3.1 Calibration compound The calibration compound used in this test is hydroquinone at a concentration corresponding' to 20 mg DOC/1. Hydroquinone has to exhibit a DOC-removal of ^ 6 0 % within 28 days, otherwise the test is regarded as invalid. - 1.A The principle of the method A predetermined amount of the compound is dissol ved in an inorganic medium (mineral nutrient solution, fortified with a trace element and essential vitamin solution), providing a concentration corres ponding to 5 - AO mg DOC/1. The solution is inocu lated with a small number of microorganisms from a mixed population and aerated at 293 ~ 2?8 (20 - 25C) in the dark or at least in diffuse light only. The degradation is followed by DOC analysis over a 28 day period. The procedure is checked by means of a standard (hydroquinone). A control with inoculation but without either test material or standard is run parallel for the deter mination of DOC blanks. 1.5 Quality criteria 1.5*1 Reproducibility The reproducibility of the method is appropriate for a screening test which has solely an accep tance but no rejective function. 001427 1.5.2 Sensitivity The sensitivity or the method is largely deter mined by the sensitivity limit of the organic carbon analysis which is 0.5 mg C/l at the present state of the art. 1.5.3 Specificity, applicability Applicable for the biodegradability evaluation of water soluble, non - volatile organic compounds. 1.5*4 Possibility of standardization The test version-with specific analyses .-for anionic and nonionic surfactants is standardized as "OECD Screening Test". 1.5.5 Possibility of automation Parts of the test, e.g., the analysis,can be automated, although hardly the total procedure. The procedure is, though, well suited for being operated with whole series of test materials. 1.5.6 Costs ( in 1978 Swiss Francs ) 1.5.6.1 Equipment Glassware shaking machine 2000,- 6000 - 20000, Carbon analyzer 4*2000,- Miscellaneous (pH meter, balance provision for air dor.ditioning) 5000,- 5- 001428 1.5.6.2 Person - hours 10 1.5.6 .3 Approximate total coat per test 750,- - 1000, (investments assumed to be amortized) 1.6 Description of the method 1 .6 .1 Reagents and materials 1.6.1.1 Deionized water Deionized or distilled water free of toxic substances (copper in particular), for general use a3 a solvent. Water which has been deioni zed by distillation or ion exchange is suitable. A high purity of this test water is necessary in view of the DOC analyses in the concentra tion range of 0 - *10 rag/1. The contaminations result from inherent impurities but also from the ion exchange resins and microbial devel opments (bacteria, algae under the influence of light etc). Only one water charge must be used for one test series which is to be controlled beforehand by DOC analysis. If necessary, suitable water may be gained by UV irradiation or other means. 1.6.1.2 Nutrient solution Mix l ml each of the following solutions (a) to (f) and make up to a volume of 1 1 with water 1 .6 .1 . 1 - 001429 6 (a) KH2?Oa A.R. k 2h p o 4 A.R. Na2RP01J . 2 H20 NHjjCl A R t A.R. 8.5 g 21.75g 33.M g 20*0 S in 1000 ml of water 1 .6.1 .1 the pH value should be 7.2 (b) 22.5 g of MgSOjj . 7 H20 A.R. dissolved in 1000 ml of water (5.3 .1 ) (c) 27.5 g of CaCl2 A.R. dissolved in 1000 ml of water (3.3.1) (d) 0.25 2 of PeCl^ . 6H20 A.R. dissolved ir. 10C0 ml of water (3 .3 .1 ) This solution is prepared freshly immediately before use. (e) Trace element solution Mr.SOu . 4 H,0 H3S03 ZnSOjj . 7 HjO (NH ,, > 6M o 70 21i 39.9 m g (30.23 mg MnSO,. . H 20) 57,2 mg 1)2.8 m g 3H.7 mg (36.85 mg (NHil)6M o 70 2 4 .HK20) Fe - chelate (FeClj,EDTA) water 1 .6.1 . 1 100 mg 1000 ml Sterilisation of the trace element stock solution at 393 U ) (120C), 2 atm., 20 min. - 7 001430 - 7 (f) Vitamin solution Biotin Nicotinic acid 0.2 mg 2.0 mg Thiamine 1.0 mg p-Arainobenzoic acid Pantothenic acid 1.0 mg 1.0 mg Pyridoxamine 5.0 mg o CM Cyanocobalamine mg Folic acid 5.0 mg water 1 .6.1 .1 100 ml The solution is filtered sterile (0.2 p ia). Instead of solution 1 .6 .1 .2 (f) 15 mg of yeast extraxt may be used per 100 ml of water 1 .6.1 .1 . 1 .6.1.3 Biodegradability standard Hydroquinone 995 % DAB Erg. B. 6 1 .6.1.4 Mercuric chloride solution 1 per cent of HgCl^ in water 1.6.1.5 Shaking machine accomodating 2 ltr. Erlenmeyer flasks either with automatic tempera ture control or used in a conrtant temperature room at 293 - 298(K) (20 - 25C) 1.6.1.6 Narrow neck - 2 ltr. Erlenmeyer flasks. (Creased fluted flasks are recommended) The flask3 must be care fully cleaned with, e.g., alcoholic hydrochloric - 8- 001431 3 / ..e use, rinsed and dried in order to avoid i `.nation with residues from previous tests. -lasks also have to be cleaned before their .rst use since they may be contaminated. 1.6.1.7 Membrane filtration apparatus 1 .6.1.8 Membrane filters 0,2 jam 1.6.1.9 Carton analyzer 1.6.2 Inoculation Either of the following three alternatives may be used as inoculum or a composite sample thereof. 1.6.2.1 Inoculum from secondary el*fluent The inoculum is gained preferentially from a secondary effluent of good quality collected from a treatment plant dealing with a predominantly domestic sewage. The effluent must be kept under aerobic conditions in the period between sampling aad use. To prepare the inoculum the sample is filtered through a coarse filter, the first 200 ml ieing discarded. The filtrate is kept aerobic until sed. The inoculum must be used on the day of collec tion. 1.6.2.2 Inoculum from soil 100 g of soil (fertile, not sterile) are suspended in 1000 ml of chlorine-free drinking water (soils with an extremely large content of clay, sand or organic carbon are unsuited). After stirring the suspension is allowed to settle for 30 minutes. 9 - 001432 ( -9- The supernatant is filtered through a coarse filter paper, the first 200 ml being discarded. The filtrate is aerated immediately and until use. The inoculum must be used on the day of collection. 1.6.2.3 Inoculum from a surface water An inoculum is drawn from a suitable surface water. The sample is filtered through a coarse paper, the first 2oo ml being discarded. The filtrate is kept aerobic until used. The inoculum must be used on the day of collection. 1 .6.2.A Composite inoculum Equal volumes of the 3 inoculum samples are united, mixed well, and the final inoculum drawn from this mixture. The suitability of the inoculum is checked by means of the standard hydroquinone. 1.6.3 Conduction of the test 1.6.3.1 Procedure The test materials are evaluated simultaneously in duplicates together with the standard (1 .6 .1 .3) and a control test with inoculation but without either test or standard material for the determination of 00C blanks. The standard material has to attain - 60 % DOC re moval within 28 days at a starting concentration corresponding to 20 mg DOC/1. If < 60 % DOC removal are achieved the whole series has to be discarded +) ) This limit is based on present experience with the 19 day version of the test. A revision of this limit or even of the standard might have to be considered after the accumulation of experien ce with the new 28 day version of the test. - 10 001433 10 A stock solution of the test material in water (1.6.1 .1 ) is prepared. So much of this stock solution is added to the nutrient solution (1 .6 .1 .2) that a carbon concentration of 5 - ^0 mg DOC/1 is attained. The starting concentration of the standard hydroquinone is, though, 20 mg DOC/1. Two reaction vessels (1.6.1.5) are each filled with 900 ml of the nutrient solution and inocula ted with 0,5 ml/1 of the inoculum (1.6.2). The opening of the vessel is covered with, e.g., alumi num foil in such a wa'y that the exchange of iir between the flask and the surrounding atmosphere is not unduly impeded (Cotton wool is unsuited because of the DOC analysis). The vessels are then inserted in the shaking machine. The tempe rature of 293 -298K ( 20 -25C) must be maintained unchanged during the test, and the vessels should be shielded from light. The air should be free of pollutants and toxic materials (chlorinated sol vents etc). In the course of the biodegradation test the DOC concentrations are determined in duplicate (1 .6 .U.2) at the beginning (day 0), and on the 27. and 28. day. Three additional analyses have to be performed - 11 - 001434 11 in rather regular time intervals ( ~ 7 .> '-2 1 . day) and The analyses are registered in the attached form sheet and evaluated. Only the necessary volumes or culture medium may he drawn for each determination; however, they have to he large enough for the membrane filtration or centri fugation preceeding the carbon determination. The latter requires differing volumes for the different instruments. Evaporation losses of the culture medium are to be made up by adding water (1 .6 .1 .1 ) in the required amounts. The culture medium is to be mixed well before withdrawing a sample. Material adhering to the wall of the vessel has to be dissolved or suspended before sampling. The membrane filtration or centrifugation has to be^done immediately. The filtered or centrifuged samples., have to be analyzed on the same day, otherwise they must be preserved with 0,05 ml of the HgCl2 solution (1 .6 .1.4) for each 10ml of nutrient medium or by storing them at 2 - *l0C, The biodegradability test is valid provided the standard exhibits a degradation rate within the specified range. The test can be finished before the 28. day if complete mineralization is accomplished. All steps require great care and cleanliness of the vessels, pipettes etc. but not sterility. 12 001435 12 1.6.3.2 Calculation of results The degradation at the time t is calculated from the determinations of the DOC concer.tra tions at the beginning (CQ ) and the time t CCt> according to 1 x loo where Dt = degradation in per cent at time t CQ = measured starting DOC concentration of the inoculated culture medium (mg DOC/1) C^ = DOC concentration of the culture medium at time t (mg DOC/1) cbl = Martin DOC blank of the mineral nutrient 0 solution with inoculation but without test material (mg DOC/1) Cbl s D0C blank of the mineral nutrient solution ^ with inoculation but without test material at the time t (mg DOC/1) The degradation rates are calculated to the nearest 0,1 2. The means of the D^ values are calculated and reported to the nearest full per cent. Results 13 001436 13 ending in 0,5 are rounded up to the nearest whole number. The course of the degradation test is followed graphically in a diagram as shown in the attached example. The results are reported on the attached data sheet. The results of the degradation test are valid if the condition is met that in the same test series the standard yields -- 60 % DOC-removal.- 1.6.4 Analytical means 1 .6.4 .1 Membrane filter 0;2 yum, 25 mm 0 (1 .6 .1 .6). Pre paration of the filters: membrane filters are impregnated with surfactants for hydrophilizaticn. Thus each filter contains up to several mg of soluble carbon which would interfere in the biode gradability determinations. Therefore the filters are purified from surfactants and other soluble organic interferences by boiling them 3 times 1 hr each in deionized water. These filters may be stored in water (1 .6 .1 .1 ) for at least one week. Other membrane filters are suitable if it is assured that they neither release carbon nor adsorb the compound in the filtration step. If the samples are centrifuged, this has to be done at 4o 000 m sec ( 4000 g) for 15 minutes, preferably in a refrigerated centrifuge, in any case <40C. (Remark: the differentiation TOC:DOC by centrifuga tion at very low concentrations does not seem to work well since either not all bacteria are removed or carbon as part of the bacterial plasma is redissolved At higher test concentrations ( >10 mg C/l) and the same small inoculation the centrifugation error seems to be comparatively small ). 001437 forra Sheet for the M o d i f i e d OE C D Screening Test Q> Exp. no. : f of start of test: Test / standard material: Theoretical test cone.: Inoculum; Carbon analyzer: ___________________________________ ________________________________________ _____________________________________ mg DOC/1 ' Controls: tock solution of the test material t 1000 mg/l, dilution .../1000 ml of nutrient solution) TOC* 1 DOC** Bg/1 U : Carbon do term Inattons : Culture medium Fluk no. Analysas Mineral nutrient solution with test material and with inoculum i *1 *2 *1 . a2 i a ( ct ) Mineral nutrient -iolution without test material but with inoculum 2 bl *2 b b, 2 * "^"5--( ct ) blank C1 c2 C1 c, "a " ~ i ~ 2 < Cbl> Theor. o<!)C - concentrations after x oays cone. *A mg/1 0 (C0). - 7 . i -14 I- 21 27- 1 28 / l 1 / /1 tf **/ / / / / I C : Evaluation of raw data: DOC - concentrations minus blanks Flask no. ID. * - -**1--*"--1"3-- . 100 % DOC tfsoval altar x days 7 14 21 27 i 28 H>2 - * 100 mean D, + D. d- 3 ^ * tor day x f J W aimI WJ4VMK JOwi .C ' / fh# i O O iH W S 1.6.4.2 The DOC measurement The sample withdrawn from the culture medium (about 30 ml) is centrifuged or membrane filtered immediately in the filtration apparatus (1 .6 .1 ,6) using the membrane filters prepared acc. to 1 .6.4.1. The first 20 ml of the filtrate are discarded*. The DOC concentration is determined twice in the remaining filtrate (about 10 ml) by means of the TOC/DOC instrument (1.6.1.8). If the filtrate cannot be analyzed on the same day it has to be conserved acc. to 1.6.3.1. The DOC measurements (ragC/1) obtained *are registered on the 'attached data sheet and the DOC concentrations of the culture medium and of the blanks calculated for each sampling time. 001439 APPENDIX 2 3m Environmontal Laboratory Data Sheot -results of the Modified OECO Screening Test- lob Request No.; SS*I S BEST COPY AVAILABLE Oate of Start of Test: A -"-flo AnaIyst: Wade A. Schei I 'est HaterIaI:A N T W E R P " -7.0-5 N F P JStandard Material; hydroqulnone. 96.?%. Lot 1001^7. Aldrich fhooretlcal DOC of Test Material: frtQT c.ALCiAt-ArD3 Moosured TOC* of Test Material: 142.jOOP mg TOC/kg Inoculum: soli and non-chlorInated secondary effluent frtwa municipal wastewater treatment plant Cnrbon Analyser: Oohrmann 0C-52A Total Organic Carbon Analyser A. Determination of TOC* of Tost Material; A stock solution of the test materiel was prepared by diluting 117 0 aw to a final volume of 5*00 ml with deionized water* cone, test material TOC TOC In stock solution stock solution test material mg/1 mg/1 mg/kg This result was used to calculate the necessary amount of test material to prepare a culture medium of hO mg/1 DOC'* . 340 33^ 000 0. Carbon Oeterminations: 2 Culture medium Flask no. Mineral nutrient solution wlth tost material and wlth Inoculum 1 2 Mineral nutrient solution without test material but wlth inoculum Blank 1 Blank 2 . Evaluation of Row Data; Cone, of test Calculated DOC1 material In of test material flask, mg/1 In flask, mg/1 tez 40.0 al + *2 an ct,- 202. *J-0.0 b. + b- ... b . -J--- 2-----0- *2 n C. C>1 + C* n I DOC 00C - cone, after x days Incubiitlon, m<l/l Anal 0 (Co) 7 11 21 27 2 yses ai 04 3 .... .at *S 37.0 / "3*577" "2 . 6 6 .0 .z v.S" 4 -.?. <4 .1 JbJ. f-t Z g- 37.4 .4 5-<V 4.1 2 . bi 2/V.0 b,.. a h a hj Af/sl bl br 3 $i.k 32./ 3 Z.O s.r <0 . 0 2 .8 ii.o 3.2. m .o 446 r2. z z- 2>*i H . z Z * Cl H 0./ 0 .X i.i -o.4 0*5^ wl /S' -.Z M d. / _ -0 ./ 0 ./ d A i 0 / 0 . if -4 t-1 hb ho 0 <? 0 .y .R 0 O -0 -r c C>2 -- K-- 0 /V OA A3 -( I 1-4 -o.l o-V b Z O.(o t Flask no. i 2 mean Degrada clon - % 1IOC removil after >tdajs Incubation li 21 27 S S . S v t . s 31.3 3.0 2o. 1 S.J 7.0 il.o 1 3 S C <?3 <13 See attached section 1 .6 .3 .2 fo r the formula used to calculate degradation. Results of blank n o .1 and no. 2 were averaged fo r each day x . The average values were used In the calculations. _ _ ^ b lj * blj (from above) b *ave 2 1. The te s t m aterial Is completely soluble In w ater. Therefore, the TOC DOC. 2. 00C analyses performed more than twice on the same sample wore run fo r q u a lity assurance purposes. 001440 3M Environmental Laboratory Oata Sheet -results of the Modified OECD Screening Test- BEST COPY AVAILABLE Lob Request No.: 55b! S Oate of Start of Test: Analyst: Wade A. Schell ist Mater lei; AtvtuJLh k . r cia-?8f t . '"T i 'iV. t a / f t Standard Material: hydroqulnone. 98,5%, Lot 1001b7, Aldrich theoretical DOC of Test Material: /AloT i" A L d A U A - r i o j Meosured TOC* of Test Material: 93.900 mg TOC/kg Inoculum: soil and non-chlorlnated secondary effluent froiia municipal wastewater treatment plant Carbon Analyzer: Oohrmann0C-52A Total Organic Carbon Analyzer A. Determination of TOC^ of Test Material: A stock solution of the test material was prepared by diluting f i O & O me to a final volume of 5 ~ 0 Q m l with deionized water cone.test material TOC In stock solution stock solution mg/l mg/l TOC test material mg/kg This result was used to calculate the necessary amount of test material to prepare a culture medium of bO mg/l DOC* Z ,!Z O 0. Cnrbon 2 Determinations: Culture medium Flask no Mineral nutrient solution wlth test material and wlth Inoculum i 2 Mlnoral nutrient solution wlthout test material but wlth Inoculum Blank 1 Blank 2 . Evaluation of Row Oata: Cone, of test Calculated DOC* material In of test material flask, mg/l In flask, mg/l */0 . > 0] *4* c t," n H 2 . t> 0 ,0 b, b2 ... bn C t2" n DOC DOC - cone, after X days Incubiitlon, mel/l, Anal 0 (C,,) 7 lb 21 27 2 yses at W2.3 ' i h o 17777" T773 s 6 ac ... r.3 . / 3. 3, i r ... 3.V 3 Z) Y3.0 3A<7 hi _ ho _ V A 3 ho bl br J Jto-7 V a 2 '0 .% C 1 . t 0. / c . L/,C* -o.y r-7 'i- r 3.3 ZA 0.2 o- r 3.6 3. V./ 3* Y <4/ 3.2. 3 2 3. 3. /*S" O. V O? ". / " w c < 1 j n (S' o .9 A 2. . r t /> - O . l ./ /.fe o . v - \ <r>- / 0.4. /.o 0.8 - C*>2 -- IS-- w ! + 0 0 .9 /3 o, a - C b l five. ~ 1-9 -0 > l 0 .9 A2 0,<m Flask no. i 2 mean Oegrada tlon - % tIOC removil a f t e r i: dajg Incubation Ib 21 27 Z 3.3 *4 .9 W .f 9 3 ,/ 2 3 .r 9 2 .6 9Y ,/ 9 i4 ?i3 3 7o 53 12. See attached section 1 .6 .3 .2 fo r the*12 formula used to calculate degradation. Results of blank n o .1 and no 2 .wore averaged fo r each day x . The average values were used In the calculations. r cb lj + cb l2 (from above) ,ave --- 5------ 1. The te s t m aterial is completely soluble In w ater. Therefore, the TOC DOC. 2 . DOC analyses performed more than twice on the same sample were run fo r q u a lity assurance purposes. 001441 3H Environment) Laboratory Data Sheet -results of the Modified OECD Screening Test BEST COPY AVAILABLE Ll> Request No: 55*I S Date of Start of Test: fr- 1>-'ftp \nolyst: Wade A. Schell test Material:AM-rvif-RV. u o r-'i-to T . Standard Materia): hydrogulnone. 98.5%. Lot lOOlb?. Aldrich Theoretical DOC of Test Material; / NOT C/vt-Cui_a V & d !) Measured TOC' of Test Material: 1*17,0OQ me TOC/kg Inoculum: soil and non-chlorlnated secondary effluent fro*a municipal wastewater treatment plant Corbon Anolyzof: Dohrmann OC-S^A Total Organic Carbon Analyzer A. Dotcrmlnation of TOC* of Test Material; A stock solution of the test material was prepared by diluting with deionized water* S" mg to a final volume of ml cone, test material TOC In stock solution stock solution mg/l mg/1 TOC test material *9/kg This result was used to calculate the necessary amount of test material to prepare a culture medium of bO mg/l OOC' 2,V<?o rtjo o o 02 Carbon Determinations: Culture medium Flask no. Mineral nutrient solution wlth tost materlol and wlth 1noculum !! 1 2 Mineral nutrient solution wlthout test material but wlth Inoculum Blank 1 Blank 2 Evaluation of Raw Data: Cone, o f te s t material In fla s k , mg/l Zo% Calculated OOC1 o f test material In fla s k , mg/l J O . DOC Anal OOC - cone, o (C^ 7 afte r x lb days Incubiitlo n , 21 27 a ;l/l2 yses 3 /. 2 a.y 3-7 2 fr -- V T T f r m3 o - Z ~ J.S* X -% i'2 . t Ok__ .2-5- a | + 0 2 ...,, ct|" n 2 0 3` 1. o 1_______________ : b ! + b2 " bn &t2 n r _ C + C * Cb l | ,, ccbl2 -- I j Vi-t bi bo " bfr _ -vbl - K 1 < /3 ' $ o .$ M .*T !% % c, .... t . t ef / 7 ^blo* I.S* , d. r- J.4 d j . ii o. J .y -o./ o. <?" 1-4 1 4 -O .f 2 .4 3.5f 3.0 3,7 / 9 l,( * o .a o. r 2.? 0.9 0 -4 o>1 hX ! /. 4 /.o 0 '4 O .H /. z hx 2 . / 7 2.Z j S. 2 i-- 2 .0 C>,9 0 .1 ***c r o.r yvj* o. a 1 O .(o 0 o .t> Flask no. 1 2 lbDegrada lion - X (IOC removiit a fte r : da^^ Incubation 21 27 J 2 .X 1 C .2 - $ 2 ,2 - ^2,1 S.2 W-L Sea attached section 1.6.3.2 fo r the formula used to calculate degradation. Results of blank no. 1 and n o . 2 we r o averaged for each day x . The average values ware used In the calculations. c. b,avt ,, -Cb-l| -+-C-b-lg (from above) mean 3 7 U 1 The te s t m aterial Is completely soluble in water Therefore, the TOC OOC. 0 0 1 4 4 22 DOC analyses performed more than twice on the sai sample were run fo r d u ality assurance purposes. 3M Environmental Laboratory Data Sheet -results of the Modified OECO Screening Test- Lob Request No.: 5541 S BEST COPY AVAILABLE Oote of Start of Test: A -- 11 -- 0ur> Analyst; Wade A. Schell est Material;______________ ______ Standard Material; hydrogulnone, 98,5%, Lot 100147. Aldrich heoretlcal DOC of Test Material; C (``A l l C A R & O U A S S U M & & ' 1 0 LJC `it-uftLE OKCyAW'C CARfioU \ Moasured TOC* of Test Material: me TOC/ka Inoculum; soil ond non-chlorInated secondary effluent frowa municipal wastewater treatment plant Carbon Analyzer: Dohrmonn 0C-52A Total Organic Carbon Analyzer A. Ootcrmlnot Ion of TOC' of Test Material; A stock solution of the test material was prepared by diluting with deionized water roe to a final volume of ml cone.test material TOC In stock solution stock solution mg/l mg/l TOC test material mg/kg This result was used to calculate the necessary amount of test material to prepare a culture medium of 40 mg/l DOC' ^0. Carbon Determinations M L U A r 6 * J O F oA l iQ A A tm . C O M P U h J -- $ 6 STAAJDARb AfA r e AtA t- A B o v e . Culture medium Flask no. Mineral nutrient solution wlth test material ond wlth Inoculum 1 2 Mineral nutrient solution wlthout test material but wlth Inoculum Blank 1 Blank 2 . Evaluation of Raw Oata: Cone, of test material In Calculated DOC*12 of test material ooc Anal ooc o (CJ cone, 7 after x 14 days Incubaitlon. 21 27 m<|/ 2 flask, mg/l In flask, mg/1 yses 30.6 Z0. .i -- 22-- _* /7t> Z -3 1.7 WliWa 2.0 /.Z. a.V :<.3 / 3 ~ T /.? / 1 /e.fe c tj" n 3 0.6 ZO.o b, b, ... bn *2 n /7.3 2*o / * i . n ), (p hi /. S- o.'jr J F bj /6.2 O-s~ J.2. b] o. a /. 1 b. 1> n . o !.% A 8 0.8 h i h . .c ! /;/ e /I o. O'Zo> r* hf >.`f 0.9 T o a. f O ' , _ e l * C2 hlj n /.S- -O.Z P . V - li di O' / o,.M/ /% 1fc /.o o.r 0.V 0 "-<c cbl2 . /V 0 o*i J,Z O.tL - 1l.Cu.tave i^b,v+i-ws - h i ~o.) O'i h % 0(o t Mask no. 1 2 moan Oegrada tlon - % CIOC remov)il after c da^s Incubation 14 21 27 %C,.% 2 S ?3.7 ?/.2. i 1,7 i h o / 02. 3 ? i% il <?3 See attached section 1.6.3.2 for the formula used to calculate degradation. Results of blank no.1 and no. 2 wore averaged for each day x. The average values were used In the calculations . _ cbl| + cbl2 (from abova) " 've --- J------ 1. The te s t m aterial Is complataly soluble In w ater. Therefore, the TOC DOC. 2 . 00C analyses performed more than twice on the same sample were run fo r q u a lity assurance purposes. 001443 I i_ z-- -f-- 3M Environr.ental Laboratory OECO Modified Screening Test with DOC Analysis Analyst: Wade A. Schell Date of Start of Test: 11 -1980 Lab Request No.: 55^1 S APPENDIX 3 3M Environmental Laboratory OECD Modified Screening Test with DOC Analysis Analyst: Wade A. Schell Date of Start of Test: *f-l 1-1980 Lab Request No.: 55^1 S 3M Envlronrental Laboratory OECD Modified Screening Test with 00C Analysis Analyst: Vade A. Schell Date of Start of Test; *-11-1980 Lab Request Ho,: 55**1 S 001447 F m tyrW n-R-P w n LA3 REQUEST BOOK - 42-5W PRODUCT ECOLOGICAL ASSESSMENT /t V*.' A ? - 0 ( > 3 M 'Action'ZOOPii 9Q( (p /Ref: ENVIRASSES?. INQ # REQUEST FOR LABORATORY WORK ____ J iatoattu*! / C e t i l i ,d iv is io n PRODUCT: /=* C > LAB REQUEST NO.. S 5 V - / S Ie --------------------- . V ............ -----------------------*-- REQUESTER LABORATORY Name: V/y Request Date: tSj /Tfrv PhoneNo. O ' / ? ' Date Received Samoli Analyst: I^ -> project No.: Prob. No. , Date Complte : <*-A3 -P Code No.: Hours Spent: \L____________________ ______ Sample Date: /))/>. K I /ffJ Date Needed: Y BP Description: OC0 , J*t<- - 7<it 6.-> fC i fc - i * fc Jc n Please Call Requester If Delays Are Encountered. Purpose of Tests: 7 * , r f <AY-- () ^>ec a> PHYSICAL & CHEMICAL PROPERTIES Heatof Combustion Cal/G IBTU/Lb.) Density H KJPP A U T w i'T fc 3oi 7 L t <?3o3 APCn w3 -io'Pi . (F M 3 S **/) ln+ 3 3 0 Hyi/rt^trriw. Ash Other - / *f/ > 1 /sT eUX. C i-'Mir'? A* V" lM-C Vji- C //km it, it. ryK- , .f WATER POLLUTION pH COD bod5 BOD, Ultimate TOC GC Oxygen Uptake Dehydrogenase Activity (TTC) Other M a j, {<j 0(~ C0 T a k *- D s lu i (tK-pi-i * . 2 *c A T to ff -* / DO c- H e w Vfti .r-- f ^oe ait ***.A3&- ASK*# ywf O p rz rM jC BIOASSAY 'r ` i 13*? ~ T tWo Specify Organism and Method: LEACHATE FROM LEACHING TEST pH COD boo5 BOD. Ultimate TOC GC Bioassay Other atpauaK B*.pe(itr*&kJT OTHER Bioaccumulation f Persistence Degradation Products H 1 T F 7 T ~ Tk^Jo + Y < c 6 ArTTM H-B p a e s u c -rr 70*1 93>*?o_ ^ 2 7e <n*A UA i l F 4 S Whtt* * Return to R ia u ttta r Carvirv Envlronmantil Enora. Offlea Fila Pink - Environmental Engrg. Lab Qoldanrod - Return to Requester