Document jnOMbVqkZ04NkQxzmv0xjB1N

BIODEGRADATION (301E) TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonawthei,ch may alsobe referredtoas PFOS, FC-95, oras a component of FC-203A orof FC-203 NFP. (1-Octanesulfonaicid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was not noted.The testsample isFC-203 NFP Currentinformatioinndicateistisa mixtureof1.34% PFOS, 25% diethyleneglycolbutylether,67.85% water,2.66 % Sultonefoamer,3% sodium octylsulfate0,.1% sodium laurylsulfatea,nd 0.05% tolyltriazole. The followingsummary appliesto a mixture with incompletely characterizedconcentrationsof impurities.Data may not accurately retlectdegradation potentialof the fluorochemicaicomponent of the testsample. METHOD Method: ModifiedOECD ScreeningTest,OECD 301E, withDOC Analysis, 1981 version. Test type: Ready Biodegradability GLP: No Year Completed: 1982 Analyticalmonitoring:Dissolvedorganiccarbon(DOC) Statisticamlethods: Resultswere determinedby calculationfthe% DOC removal and graphicinterpretation. Test organism source: A 50:50 mix of soilextractand secondaryeffluent. The secondaryeffluenwtas the supernatantfrom an activatedsludge aerationbasinattheMetro Wastewater Treatment Plant,St.Paul,MN, while thesoilwas fromthe CityofWhite Bear Lake,Ramsey County,MN. Test condition: Dilutionwater: Deionizedwater MineralNutrientMedium: Nutrientmedium per OECD 301 E method (1981).InitipaHl 7.0. Reference and testsolutionpreparation:The testmaterialwas preparedby dissolving153 mg intwo literosf mineralnutrienmtedium. This solutiognivesa finaltestconcentratioonf20 mg DOC/L. The referencesubstance,sodium benzoate,was prepared by dissolving 30.3 mg inone liteorfmineralnutrienmtedium. This solutiognivesa finatlestconcentratioonf20 mg DOC/L. Test vessels: Not given. Incubationconditions:Constantdark conditions Temperature: 23.5- 240C Agitation: Continuously Number of concentrations: 1 plussodium benzoate (reference substance)and blank,allinduplicate. lnoculumconditioonn testinitiatiNoontg:ivenElementBasis:Decreaseindissolveodrganicarboncomparedto the blanks. Test substance flaskconditions: Not given. RESULTS Nominal concentrations: Blank controls,odium benzoate at-20 mg DOC/L and at-20 mg DOC/L plusHgCL2 (inhibitedt)e,stmaterialat -20 M DOC/L and at-20 mg DOC/L plusHgC12, allinduplicate. Element values: 27-day Degradation = 76.7% duplicate1 and 81.3% duplicate2. Mean value= 79% Remarks: Testingwas conducted on the mixtureas describedinthe Test Substance Remarks field.The valuesreportedapplyto thatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The FC-203 NFP % degradationbased on the mean DOC removal value was 79 after27 days. Submitter: 3M Company, Environmental Laboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2. This studymeets the cdteha forquality testing.However, the sample puritywas not propedy charactedzed and the study lacksanalyticaclonfirmationofthe amount offluorochemicalproportion inthe solution. REFERENCES The studieswere conducted by the 3M Company, EnvironmentalLaboratory, St.Paul,MN, Lab Request number 8483, 1982. OTHER Last changed: 6/28/00 3m - Form 6747-1 I-A TECHNICAL REPORT SUMMARY D ate 6/30/8C TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN (Important-Ifreportisprintoendbo6 sidesofpapesre,ndt" copietsoTCC.) Division - Environmental Laboratory (EE & PC) Dept Number 0535 Project P-roN--fuemct bar Commercial Report Titi Chemicals Division 9970012600 FteportNumber Biodegradation of "LIGHT WATER" Products in OECD Test-6/80 040 To John A. Pignato Author(s) Commercial Chemicals Division - 236-2A (01) Employw Numbsr(s) Eric A. Reiner Notebook Reference Environmental Lab (EE&PC) - 21-BW (63'1 47816 No. of Pages IncludingCoversheat None - See 3M Environmental Lab Request No. 5541S 31 SECURITY I$. 91 Open KEYWORDS: (Select terms from 3M Thesaurus. S@uggest other applicable terms.) Environmental Laboratory (EE & PC) CURRENT Closed OBJECTIVE: 3M CHEMICAL I REGISTRY To use the internationally recognized Test with DOC analysis to show that products are highly biodegradable. Now ChemicalRseported 0 Yes IN No OECD Screening "LIGHT WATER" AFFF TOC REPORT A13STRACT: (200-250words)Thisabstracitnformatioinsdistributbeyd theTechnicaClommunicationsCenterto alert3M'ersto 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 WATZR" Brand AFFF 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. InformatiLon Initials: -2- BIODEGRADATION OF "L IGHT WATER" IN OECD TEST PRODUCTS INTRODUCTION The 3M Environmental Laboratory chose to attempt to demonstrate the biodegradability 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 20-250C 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(l). 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 20-250C range during the 28-day test, except for a 3-4 hr. period when it reached 25.50C. The temperature was most frequently near the high end of the range (23.5-240C). 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 reage nt 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. -3- Water used in the 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 through 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 9 of NH2HP04-2H20, 36.3 g of CaCl2-2H20 replaced 27.5 g CaCl2. These replacements yielded solutions and concentrations 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 FeCl3 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 o.f 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 WhatmanR 54, and the water for soil extraction was chlorine-free well water. Sterile Controls Replicate vessels containing 400 mg/l HgCl2 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 - The sample preservation method used was that prescribed by the TOC manufacturer(4). The method involves adding 1 drop of concentrated HC1 to the 10 ml filtered samples, bringing the sample to <pH 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. -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 HgCl2 preservation technique described in the OECD method. All TOC analyses were made within a month of the preservation of the samples. Instruments and Dohrman DC-52A. per liter. equipment - The organic carbon analyzer Its sensitivity limit is between 1 and used was a 2 mg of carbon The stoppers for the reaction flasks were clean, porous, plastic foam' plugs. Gelman 0.2 um membrane filters, (Part No. 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 contain the actual function of time. completely. the results of this study. Appendices 1 and 2 data sheets and plots of the degradation as a All 3 "LIGHT WATER" AFFF products degraded nearly. Percent TABLE 1 Degradation of Hydroquinone and "LIGHT WATER" FC-203, FC-206, and FC-3017 With Time in Modified OECD Screening Test AFFF Products % Degradation at Day: Product 7 14 21 27 28 FC-203 13 85 93 91 93 FC-206 23 90 94 93 92 FC-3017 37 94 96 96 95 Hydro- 89 92 97 94 93 quinone 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, 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. At the end of 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 HgCl2, 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. Nosler) Revision of Sept. 19, 1979. (2) 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. APPENDIX I Tokyo, December ist, 197 5@ C 118/79/Int. OECD Expert Chemicals Testing Programme Group C, Degradation/Accumulation Mbdified OECD Screening with DOC Analysis Test Test Guideline C 118/79/In-,.. Test Guideline for the Modifi4d OECD Screening Test with DOC Analysis Date of last revision: Sept. 19,1979 Lev6l I test for ready biodegradability 1. Prercqtiisites It has to be known whether the test material is soluble in the concentration range employed in the test (corresponding to 5 - 40 mg DOC/1). 2. Guidance Information Knowledge of the bacterial toxicity or inhibitory properties of the test material i@snot unequivocally required but constitutes useful information for the conduction of the test. 3. Qualifyina Statements The method is suited for ultima*.e biodegradability organic compounds. It is evaluation of mix@uures. the measurement of the aerobic of water soluble, non-volatile unsuited for the biodegradability Sept. 10. 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 several 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 m'odification the of*&'icial,widely practised and well accepted OECD Screening Test for the biodegradability evaluation of surfactants there should exist a good prospect for success. September 19-s 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 Detergents, Paris 1976, and council directive of Nov. 22, 1973, on the approximat4-on of the laws of the member states relating to methods of testing the b4-odegradability of anionic surfactau@s (73/405/EEC),Official Journal of the European Communities No. 4 347/53 of Dec. 17, 1973) for the application of the dissolved organic carbon(DOC)analysis. 1. Methcd 1.1 Introduction: Pur20se, scope, relevance, and aoulica- tion of test and eXDlanation 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 - 4o mg DOC/1 ( In order to avoid inhibitory effects it is in the investigator'sown interest 2 2 to choose as low a starting concentration as his analytical capability permits 1.2 Definitions and units 1.2.1 Definition of biodegradability Dz t L where Ct - x 100 co - c bl 01 D t = degradation in.pqrcent DOC-removal at time t c 0 = starting DOC concentration of the culture medium (mg DOC/1) c = DOC concentration of the cul4kluremedium at time t (mg DOC/1) c bl= starting DOC concentration . oar the 0 blank (mg DOC/1) Cbl = DOC concentration of the blank a*.. t 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 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/l. Hydroquinone has to exhibit a DOC-removal of@@t6O % within 28 days,, otherwise the test is regarded as invalid. 1.4-The principle of the method A predetermined amount of the compound is disso' ved in an inorganic medium (mineral nutrient solution, fortified with a trace element and essential vitamin solution), providing a concentration corresponding to 5 - 40 mg DOC/l. The solution is inoculated with a small number of microorganisms from a mixed population and aerated at 293 - 2,08. (20 25 0 C) 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 determination of DOC blanks. 1.5 Quality criteria 1.5.1 Reproducibility The reproducibility of the method.is appropriate for a screen4-ng test which has solely an acceptance but no rejective function. Oy-7 4 1.5.2 Sensitivity The sensitivity of the method is largely deter- mined by the sensitivitylimitof.theorganic carbonanalysiswhich is 0.5 mg C/1 at'thepresent state of the art. 1.5.3 Specificity,applicability Applicable for the biodegradability evalua,-,ion of water soluble, non - volatile organic compounds. 1.5.4 Possibilityof standardization The test versiom with specific analyses:.for anionic and nonionic surfactants is standard4-zed as "OECD Screening Test". 1.5.5 Possibilityof automation Parts of the test, e.g., the analys4"s,canbe 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 2000,,- shaking machine 6000 - 200001,m Carbon analyzer 42000,- Miscellaneous (pH meter,.balance, provisionfor air donditioning) 50OOs- 5 1.5.6.2 Person hours 10 1.5.6.3 Approx4-mate total cost per test750s(investments assumed to be amortized) 44.000t- 1.6 Description of the method 1.6.1 Reagents and materials 1.6.1.1 Deionized water Deion4-zedor distilled water free of tcxic substances (copper in particular),,for general use as a solvent. Water which has been de4-onized by distillation or ion exchange is suitable. A high purity of this test water is necessary in view of the DOC analyses in the concentb-ation range of 0 - 40 mg/j. "Llhecontaminations 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 Nutrien*- solution Mix I ml each of the following solutions (a) to (f) and make up to a volume of 1 1 with water 1.6.1.1 6 6 (a) KH 2 PO4 X 2 HP04 N,,32. HP04 NH4cl A.R. A.R. 2 H 20 A.R A*Ro g 21-759 33.4 g 20-0 13 in 1000 ml of water 1.6.1.1 the pH value st-.ouldbe 7.2 (b)22.5 9 of MgS04 - 7 R20 A.R. dissolved in 1000 ml of water (3-3.1) (c)27.5 g of CaCl 2 A.R. dissolved in 1000 ml of water (3-3-1) (d)0.25 g of FeCl 3 * 6H 2 0 A.R. dissolved in I-OCOmi of water (3-3.1) This solution is prepared freshly immed4-ately before ,,.se. (e)T.-ace element solution MnS04 H BO 33 ZnS04 6 4 H20 - 7 H20 (NH4)6Mo 7 0 24 39.9.mg 57,2 mg 42.8 mg 34.7 mg (30.23 mg (36.85 mg MNSO 4 6 H 20) (NH4)6Mo 7 0 24* 4H 20) Fe - chelate (FeCl EDTA) 3,* water 1.6.1.1 100 mg 1000 ml Slk-,erilisatioonf the trace element stock solution 393 (.7,,()"&200c)"2 atn..,,20 min. 7 7 (f) Vitamin solution Biotin 0.2 mg Nicotinic acid 2.0 mg Thiamine 1.0 mg p-Aminobenzoic acid 1.0 mg Pantothenic acid 1.0 mg Pyridoxamine -Cyanocobalamine 5.0 mg 2.0 mg Fol.4L,eacid 5.0 mg water 1.6.1.1 100 ml The solution is filtered sterile (0.2jum). Instead of solution 1.6.1.2 (f) 15 mg of yeast extraxt may be used per 1.00ml of water 1.6.1.1. 1.6-1.3 Biodegradability standard Hydroquinone 99.5 % DAB Erg. B. 6 1.6.1.4 Mercu ric chloride solution 1 per cent of HgCl2 in water 1.6.1.5 Shaking machine accomodating 2 ltr. Erlenmeyer flasks.either with automatic temperature control or used in a conrl.ant temperature room at 293 2.018(K)(20 - 250 C) 1.6.1.6 Narrow neck 2 ltr. Erlerneyer flasks. (Creased flasks are reco=ended) The flasks must be carefully cleaned witha e.g., alcoholic hydrochloric fluted 8 use, rinsed and dried in order to avoid .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 Meribranefilters 0.2 1.6.1.9 Carbon analyzer 1.0'.2Inoculation Eithe-,!of the following three alternatives may- be used ts inoeulum or a composite sample thereof, 1.6.2.1 ino,.uluraf-x,omsecoridary effluent The inoculum is gained preferentially from a seedndary effluent of good quality collected from a t'Ireatmenptlant dealing with a predominantly docestic sewage. The effluen t must be kept under rdbic conditions in the pe'riod between sampling d use. To prepare the inoculum the sample is iltered through a coarse filter, the first 200 ml @eesjing discarded. The filtrate is kept aerobic until sed. The inoculum must be used on the day of collecion. 1.6.2.2'Lnoculumfrom soil 100 g of soil (fertile, not sterile) are suspended in 1000 ml of chlorine-free drinking water (soils ith an extremely large content of clay, sand or rganic carbon are unsuited). After stirring %-,he uspensionis allowedto settlefor 30minutes. 9 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 1.6.2.4 Inoculum f.-om-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. Composite inoculum Equal volumes of the 3 inoculum samples are united, mixed well, and the f4.nal inoculum drawn k*rom this mixture. The suitability of the inoculum is checked by means of the standard hydroquinone. 1.6.3 Conduct4hon of the test 1.6.3.1 Procedure The test materials are evaluated simultaneously 4-n 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 DOC blanks. The standard material has to attain 1- 60 % DOC removal within 28 days at a starting concentration corresponding to 20 mg DOC/l. 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 experience with the new 28 day version of the test. 10 -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) thit a carbon concentration of 5 - 40 mg DOC/1 is attained. The starting concentration of the standard hydroquinone is, though, 20 mg DOC/l. Two reaction vessels (1.6.1.5) are each filled with 900 ml of the nutrient solution and inoculated with 0.5 ml/l of the inoculum (1.6.2). The opening of the vessel is covered with, e.g., aluminum foil in such a wa:ythat the exchange of dir between the flask and the surroundi,-,gatmosphere is not unduly impeded (Cotton wool is unsuited because of the DOC analysis). The vessels are then inserted in the shaking machine. The temperature of 293 -298K ( 20 -250C) 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 solvents etc). In the course of the biodegradation test the DOC concentrations are determined in duplicate (1.6.4.2) at the beginning (day 0). and on the 27. and 28. day. Three additional analyses have to be performed in rather regular time intervals -21. day) 14.,, and The analyses are registered in the attached form sheet and evaluated. Only the necessary volumes of culture medium may be drawn for each determination; however., they have to be 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 biamixed 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..d:oneimmediately. The filtered or centr@ifuged samples..have to be analyzed on the same day, otherwise they must be preserved with 0,05 ml of the HgCl.@ solution (1.6.1.4) for each 10ml of nutrient medium or by storing them at 2 - 40C, The biodegradability test 13 valid provided the standard exhibits a degradation rate within the specified range. The test can be@finish'ed before the 28. day if complete minerali-zation is All steps require great care and cleanliness of the vessels, pipettes etc. but not sterility. - 12 12 1.6-3.2 Calculation of results The degradation at the time t is calculated from the determinations of the DOC concentra- tions at the beginning (C0 (Ct according to and the time t Dt I I c t c bl t x loo co c bl 0 where D t = degradation in per cent at time t c 0 = measured starting DOC concentration of the inoculated culture medium (mg DOC/1) c.t = DOC concentration of the culture medium at time t (mg DCC/1) c blo = starting solution DOC blank of the mineral nutrient with inoculation but without test material (mg DOC/1) c bl t = DOC blank of the mineral nutrient solution with inoculation but without test naterial at the time t (mg DOC/1) The degradation rates are calculated to the nearest 0,1 %. The means of the Dt values are calculated and reported to the nearest full per,c@ent. Results 13 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 ':t60 % DOC-removal. 1.6.4' Analytical means 1.6.4.1 Membrane filter 0;2 pm, 25 mm 0 (1.6.1.6). Pre- paration'of the filters; membrane filters are impregnated with surfactants for hydrophilizat4.on. Thus each filter contains up to several mg of soluble carbon which would interfere in the biodegradability determinations. Therefore the filters are purified from surfactants and other soluble organic '*interferencesby boiling them 3 times 1 hr each in deionize.d 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 40,000 m sec-2 ( 4000 g) for 15 minutes., preferably in a refrigerated centrifuge, in any case <400C. (Remark: the differentiation TOC:DOC by centrifugation 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/1) and the same small inoculationthe centrifugationerror seems to be comparatively small Form Sheet for the Modified OECD Scr*ening Test -xp. no.: ;at@ of start of test; .est / standard material: rheoretical test conc.: Inoculum: .arban analyzer: mit DOC/I A: Controls: tock solution of the test material 1000 mg/l, dilution .../1000 ntl of nutrient solution) TOC DOC mg B: Carbon determinations: Culture medium Flask no. Analys Mineral nutrient solution withtest material and with inoculum Mineral nutrient solution without test material but with inoculum 2 Blank 2 a +a 1 2 ( ct ) b b2 b m- 2 ct + b.) c C2 Th r. - DOC - concentrations after x cays coonoc. at/I mg/l 0 (Co). 7 -14 1-21 27. 2S 3 2 Cbl) C: Evaluation of raw data: DOC - concentrations minus blanks Flask no. M3 D 100 m ii2 '43 2 D2 loo 2 % DOC removal after x days 7 14 21 27 28 mean 0 D I + D2 2 for day x 14 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 ace. to 1.6.3.1. The DOC measurements (mgC/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. APPENDIX 2 3H Envirormantal Laboratory Data Shoot -results of the Modified OECD Screening Test- Lab Request No.: 5541 s Date of Start of Test: 4-%%-So Analyst:_Wade A. Scholl Test Material:XW-rwf-plp FC- 14FP tandard Material:_hydrogulnone, 98*5%. Lot 100147, Aldrlch Theoretleal 70C of Test Material: (t.Jore-RLC-tALATED) Measured TOC of Test MotorIa I: 1,@Z pe4 mg TOC/kg lnoculum: sotI and non-chlorinated ifecondary-Aft-luetnrtcme municloal wastewater treatment plant Carbon An'alyzer: Dahrmann OC-5zA Total Organic Carbon Analyzer A. Determination of TOCI of Test Haterlal.L A stock solutionof the test material was prepared-bydiluting L170 with delonlzedwater. mg to a final volume of 5C)O ml conc.test material TOC In stock solution stock solution mg/l mg/l TOC test material mg/kg This resultwas used to calculate the necessary amount of test materlql to prepare a culture medium of 40 mg/i DOC'- Z'340 1 332, 2 B. Carbon Determinations: 1 142zj 00 Culture medium Flask no. Mineral nutrient I solutionwith test material and with Inoculum Mineral nutrient solutionwithout to$ t material but with inocI=um Blank I Blank 2 C. Evaluation of Raw Data, Conc. of test' Calculated DOC] DOC material In of testmaterial Analflask, mg/l in flask.mg/l yses al tO 7. 40-o al + 82 ...an ti- n -02- zg:z AS- DOC - cone. after x 0 (co) 7 14 I 37,0 1/ 'q/-S' 37-4 C-q days Incubation,mg/i 21 27 1 28 G.Z 44.1 Z-7 u #.I 17':, 5-'Cr 4.1 ?--8 b Alq.0 3).g 3.a 2-9 b 51-il 32.1 L. rm Z b Y. Z. b, + bz b ct2m ql. (o 32.,o (o.o 2.4 14.7- z.:s Ci + C2 Cbll" n d+ I 2 Cbl2' n -.94- 16 c @C 61b0# sr- -6-2, d co -0./ d!) z 0.1 rCbIor 0 0-/ 0.6 o. L C'01 O.,e- /00 3 I-2. 0-9 0. 0.1 0. 0.64 n.R 0-/ -0.1 -o-I 0.6 t)-6 6 Flask no. 1 2 mean Dogrods ton - % DOC removal after d s tncubatlon 7 14 21 27 I 818 (0.5, q ZO. 1 -9 3. 1D 13 See attached Section 1.6.3.2for the formula used to calculatedegradation. Results of blank no.1 and no. 2 were averaged for each day x. The average values were used In the calculations. Cbll + Cbl2 (from above) Cbl.v.' 1. The test material Is completelysoluble In water. Therefore, the TOC - DOC. 2. DOC analyses performed more than twice on the save sample were run for quality assurance purposes. 3M Environmental Laboratory Data Shoot -results of the Modified OECD Screening Test- Lob Reques Mo 5541 S Date of St!rt ;i Test: 1-1-84 Analyst: Wadt k, Schell Test Material: MTVXFKP. Fet- A. m Standard Material: hydrogulnom, 98.5%o Lot Theoretical OC of Test mastrlil:- A# e LCC41.A-rF-bj MeasuredTOC7 of Test Material:-..U.Ioo mg TOC kg Inoculum: soil and o"-chlorin-ted- secondary e?fluent froma munteloal wastewater treatment plant Carbon Analyzer: Dohrmai LA Total Organic Carbon Analyzer 100147, Aldrich I A. Determination of TOC of Test Materlal.L A stock solution of the test material was prepared by dllutina I#OCOO with delonlzed water. to a f Inal volume of 5'00 mi conc.test material In stock solution mg/i TOC stock solution mg/l TOC test material mg/kg This result was used to calculate the necessary amount of test mteril! to prepare a culture medium of 40 "/1 Doc z@ /ZO 199 -3; Do 2 B. Carbon Determinations.*, Culture medium Flask no. Mineral nutrient solution with test material and with In lum 2 Mineral nutrient solution without test material but with inocu=um Blank I Blank 2 C. Evaluation of Raw Data: Conc. of test material In flask. mg/l Calculated DOC I DOC DOC cone. of test material Anel- 0 (CO) 7 In flask. mg/i yses - al - 02 a :a!47- after x 14 6.1 days Incubation. mg/i 21 27 28 3-sr 3.04 al + a2 ...an n 2- Z/o, C) b + b: b c t2 n I Ci + C 2 Cbllm n q.?o. bl_ b b b b- 2.3 /,s cl c 2- C.. i.q 7 30,1 30-7 212M4 3-@ t- o,Y 3..q 47"r 2. 3.z 3,4 1.!r Of 102- I 0./ d+ 1 2 c bl2' n c AVP- *X P(CCbb lo) 1.4 n. f0 1.141-01/ -o.1 1 /,2 7o Flask no. 1 2 mean Degrade ton % DOC removal after 7 21 27 93,1 2 14, 3 741 93 dais Incubation 8 2- See attached section 1.6.3.2 for the formula used to calculate degradation. Resultsof blankno.1andno.2 were .averaged for each day x. The average values ware used In thecalculatlons. Cbl&ve Cbli + Cbl2 (fron above) 1. The test material Is completely soluble In water. Therefore, the TOC DOC. 2. DOC analyses performed more then twice on the same sample were run for quality assurance purpose$. 3M Environmental Laboratory Data Shoot -results of the Modified OECD Screening Test- Lab Request No.- 554) S Date of Start ;i@-Test- Analyst: Wade A. Schat I Test Haterlal:AwvwEE1->.r-,t-3ol-7, Standard haterlal-hydroquinam, 98.5%.Lot Theoretical IOC of Test Material-: T CALCTALAIF-D Measured TOC of Test material: 132,000 ._mg TOC kg lnorulum: soil and non-chlorln ad secondary.Iffluent frcma municipal wastewater treatment plant Carbon Analyzer, Dohrmann DC-5ZA Total OrgarkLc Carbon Analyzer 100147,Aldrich A. Determination of I TOC of Test Material: A stock solutionof the test material was prepared by diluting with delonized water* to a final volume of !5'00 mi. conc.test material In stock solution mg/l TOC stock solution mg/i TOC test material mg/kg This result was used to calculate the necessary amount of test materill to prepare a culture modlum of 40 mg/i DOC 4/qo %71 /'12j000 2 B. Carbon Determinations: Culture madlum Mineral nutrient solution with test material and with Inoculum Flask no. 1 2 Mineral nutrient solution without test material but with Inocu=um Blank I Blank 2 C. Evaluation of Raw Data: Conc. of test material In flask, mg/l Calculated DOC DOC of test material Anal- In flask,'mg/l SOS alL @(0,0 -a2a DOC conc. after x 0 (co) 7 14 I 3 12 ley 30.3 a. r- days Incubation, mg/i 21 27 28 3.7 /.17 2,0 -"7.z -Tz- al + a2 ...an n 2-03 41,9,0 aS_ _Al_ b b 1//./130-8 2-4 .7,3 4/.A 1q.1 A. 30 f? 3,o S. /.91 2,r /o7 212 1,5- A# .2.Y b+ c t2" n F Ci + C 2 Cbli" n d I + d2 Cbl2m n b 5 n d @-61qua L 3.(p 2,.l 2-c.) 0-) Of A 1.r 011 Of/ 0. r 0.9 oil 4>0 0#7 o.q /.2. 0,5' 0,/ 1 0.1 I.q -L).t 0 /02- O#R -as 1 0.(* o. 6 . Flask no. 1 2 mean Degrade Ion DOC removal after dals Incubation 7 1 l@ 11 21 27 1 0 1'4 See attached section 1.6-3.2 for the formula used to calculate degradation. Results of blank no.1 and no. 2 were averaged for each day x. The average values were used In the calculations. bigve Cbll + Cbl2 (from above) 2 1. The test material Is completely soluble In water. Therefore, the TOC - DOC. 2. DOC analyses performed more than twice on the same sample were run for quality assurance purposes. 3M Envlrommntal Laboratory Data Sheet -results of the Modified OECD Screening Test- Lab Request No.: 5541 S Date of Start of Test*. 4-- 11-IFIC An lyst: Wade A. Scherl Te:t Material-. St&Ward matertal.-hydre4ulnons,98.5%. Lot 100147,Aldrich Theoretical70C of Test Material:_ C.S.4-1 r- MLL-L@6REM ASSUMF-D 10 f6C SOLUSLF- ORCANIC CARr6,ow_@- Measured TOC of Test Material- 09 OC/kg Inoculum: soft and pan-chlort4stedsecond;ry-effuent frcme municipalwastewatertreatmentplant Carbon Analyzer: Dohrmann DC-5ZA Total orqonlc Carbon Analyzer A. Determinationof TOCI of Test Material.L A stock solutionof the test materialwas preparedby diluting mg to a finalvolume of ml with dolonizedwater. conc.testmaterial TOC TOC In stocksolution stock solution test material mg/i mg/i mg/kg This resultwas used to calculatethe necessary amountof testmaterlilto preparea culture medium of 40 mg/i DOC 2 S. Carbon Determinations: Culture madlum Flask no. Mineralnutrient I solutionwith test material and with lnoculum 2 Mineralnutrient Blank solutionwithout 1 testmaterial but with lroculum Blank 2 C. Evaluationof Raw Data: eom ftu t4*-rLON/At- SLCLC STAAJDARD A86VE Conc.of test CalculatedDOC] DOC DOC - cone. afterx days Incubation,mg/l materialIn of testmaterial Anal- 0 (CO) 7 14 21 27 1 28 flask,mg/l In flask.mg/l yses - al ).7t1 Z.3 2.0 .2.4 /.3 /-7 a,_ l(D-3 1-7 1* 2. 9. /-9 t) 30.ro zo-O a -JR. :27- IG.7-I I I I al + 02 ...an CtIn n 30.ro b I + bz c . t2- n 20.0 b a- Ci + C2 Cbll" n Ib bl b14. 16.2 2.0 O-S' /7-0 c o-i c -o-4 ./2-.G4t 0.2. 0.r 2,7 1-2. 115, 2. d 1.4 /.S' L*)9 0# 1 10-5' 6.V y o. ia I ot 1 1 6.6 a./0,/ 0-1 -0,/ mi d I + d2 Cbl2 n C-61&ve lbi + 61,. Flask no. 1 2 mean Degrade Ion 7I 96,13 % DOC removal after 21 1 27 7o.(o 11.3,7 lq.1 177 .911 d a Incubation @1,2- qq. I q3 See attachedsection 1.6.3.2f;a1 the formulaused to calculatedogr atton. Results of blank no.1 and no. 2 were averagedfor each day x. The overage valueswere used In the calculations Cbll + cbl2 (from.above) c blavo -2 1. The testmaterialIs completelysoluble In water. Therefore, the TOC DOC. 2. DOC analyses performed more then twice on the same sample were run for qualityassurancepurposes. 9D%: 80%70% 50X. 40X LU - 3 OY.- 20X 10% 0. 3m Environr,.entLaalboratory OECD Modified Screening Test with DOC Analysis Analyst: Wade A. Scheil Date of Start of Test: 4-11-1980 Lab Request No.: 5541S Test Material: At4TWF-R? FC- -2-03 t4FP i At IJF 7' 14 21 NUIISER OF DAYS INCUBATED t:-= X Ot D O'A _j -------------7C%: 60%-s 50%. z 40%30%20%- dOP-1 4 3M Environmental Laboratory OECD Modified Screening Test with DOC Analysis Analyst: Wade A. Schell Date of Start of Test: 4-11-1980 Lab Request No.: 5541 S Test Material:At4rwegL FC-2-06 F-Et4.-36 2+ I-OT - 2.3'3 jolag -T- -14 21 NUMBER OF DAYS INCUBATED 100%- go%- 80%. 70% 5(F/. 40% UJ 20% lox 0-0 20 0 -I--- 3m Environr-entalLaboratory OECD Modified Screening Test with DOC Analysis Analyst: Wade A. Schell Date of Start of Test: 4-11-1980 Lab Request No.: 5541S Test MaterialA:ti,rWeit?.E,c- '3017 L-OT -2301 7 WE ---------- 7' 1 21 -2 NUP.BER OF DAYS INCUBATED '3LI'Z 1 '310 '%4 10 0/.9 (rh 80Y.'70Y6(fk 5 OY.- z 4Wo 3 (P/-. 20Y, 0 3M EnvironrentalLaboratory OECD modified Screening Test with DOC Analysis Analyst:Wade A. Schell Date of Start of Test: 4-ii-ig8o Lab Request No 5541 S Test Material: 14yDRoQiAlOOtIE ........... 14 21 2 NUVISER OF DAYS INCUBATED HO'41 Hdv?il b!@-a =t X M3@@z-L310 sesow.v too 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 tfieDetermination of the Biodegradability of Surfactants Used in Synthetic Detergents, 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 biodegradability of anionic surfactants (73/405/EEC),Official Journal of the European Communities Mo. 4 347/53 of Dec. 17, 1973) for the application of the dissolved organic carbonODOC)analysis. 1. Method 1.2 Introduction: Purpose, scope, relevance, and application of test and explanationof limits. The purpose of the method is the measurement of the ultimate biodegradabilityof water soluble, non volatile organic compounds in an aerobic, aqueous medium at a starting test concentration corresponding to 5-- -40mg DOC/1 ( In order to avoid inhibitory effects it is in the investigator'sown interest 2 3M Envircm otal LaboristaryData shoot -results of the Modified KCO Screening Test- Lab.RequestNo.: $541S Date of Start of Test: Analyst.- We" A. Schell Testmot*rTs-i:AN-r%@%MltP q-&nA Theoretical of T--t hot-pi-I Mossu...- of Testhmtiirlel: Inoculum: soil Ong non-C Carbon Anslyz4kr: Dahroann OC-SU NFP StoWard Material:--tiy4roquirw3of,.5%. Lot lOOi47,Aldrich ED e=fi2--&t, mg TOC/kg uent fross municipal wastewater treaggnt plant ti.;WlilcCarbon Analvzor I A. Determination of-TOC of Test Material* A stock Solution of the test mterlal was prepared by diluting 1170 mgto a final volume of 520 ol with dolonized water. corw-test mst*rlal In stock solution mg/l TOC stock solution 09/1 TOC test material ag/kg This result was used to calculate the necessary amount of test mtertti to prepare a culture medium of 40 mg/i OM Z340 33Z 4@"j000 8. Carbon Determ;notlons-, Culture medium flask no. Mineral nutrient solution with test matirt-ai nd with inoculum Mlnarai nutrient Solution without test mateflelbut with lnocu=um blank I Blank 2 C. Evaluation of Raw Data: Conc. of test material In flask, mg/i Calculated DOCI of test material In flask, mgll 40.0 DOC Analysos DOC - conc. after x days Incubation. mg/i 0 (CO)l 7 14 1 21 27 28 44.1; 137,0 IJO-6 39-1 a$Z S.1 ca . Ars 6.0 I G-Z 1 4.15 41.1 #,I Z -7 41 + 82 ...an utl- n + ct2 n I Cl + c 2 Cbll' n I d I + d2 Cbl2" n o - ar -kj-. .0 b 39-a 37-4 r..9 b4 b qi.,(3o2..o (o.0 Cl 1.) 10./ O-Z @Oe4L 0. !r d d9 q (Cblo) -4e- o To- 4.1 Z .8 2.8 .22, -2.9 51.4- Z..g a.) /.0 /.3 6. Fk 0.6 0.6 o./ -o-i -0. -o. 6 F?ask no. 1 2 mean Degra&Wda lcomn- % 7 1 14 OC rom@mwvI after x d s Incubation 21 27 818 13.0 2-o.1 17,fj J.D ct3.,D q3 91. 13 See attached section 1.6-3.2 for the formula used to calculate degradation. Results of blank no.1 and no. 2 were averaged for each day x. The average values were used In the calculations, Cbll + Cb]2 (from above) 1. The test material Is completely soluble In water. Therefore, the TOC - DOC. 2. DOC analyses performed more then twice on the same sample were run for quality assurance purposes. IOIY/l 901/. 8 (P/-o 7 0'/o w 60:/o-. 0-0 4 (P/o LU 3(r/-o 0 3M Environr.,entaLlaboratory OECD Modified Screening Test with DOC Analysis Analyst: Wade A. Schell Date of Start of Test: 4-11-1980 Lab Request No.: 5541S Test Material:At-1Tw ERP FC- -2-03 OFF T -------- -2 2@- NUP@BER OF DAYS INCUBATED .3 cl -G"E 3H Enviromentat LaboratoryData Shoot -resultsof the ftdlflodOECD ScreeningTest- Lob Request No.: 5541 S Oat* of Startof T*st- -+-it--iE Analyst,-We" A. Scholl Toot Material:AUZUEltp.FL-3ft-7. &@OT-2--bo@s StwAard material.- hydroquinam, TheoreticalIOC of Test hatorialtTNOT r-ALC-IALATCD)-- Measured TOC of Test Material: W4 TOCfkg Inoculum: .. rinat m2aigcendereyffluent frcog municipal CarbonAn I i r: Ochroom K-52A TotalOr2oni CarbonAnalyzer 98.9%. Lot 100147, Aldrich nt slant A. I Determinationof TOC gf I2@t Mteriali A stock solutionof the testwtorlel was preparedby diluting.Ii2t-zq;5t'-mgto a finalvolumeof goo al with 4slantzedwater. conc.test@terial Tot Toc In stocksolution stock solution testmaterial 09/1 09/1 mg/kg This resultwas used to calculatethe necessary amountof testmaterillto pr"aro a culture madlum of 40 mg/i DOC 2-4;q 0 47 9 2 8. Carbon Daterminatlonsz Culture medium Flask noo Mineral nutrient 1 solutionwith test met*-rTa7 and with Inoculum 2 Mineralnutrient Blank solutionwithout test material but with lnocu=um Blank C. Evaluationof Raw Data: I Conc.of test Calculated DOC] ooc DOC - conc. after x days Incubation,69/1 materialIn of testmaterialAnal- 0 (CO) .7 14 21 27 1 28 flask,mg/l__ In flask,mg/l yses - si v 31,Z 2-Y 3.7 /.1? 2,0 2-0!9 22- I0. 0,0 r, -z.z cti. 81 + 02 ...On n 410,0 bi +-bz .. ct2. n 2-4 3,o y 2. -C" :s.et /07 %1 b 3,r -77T- bb- lqt,q 1q.I 3,(p 2-1 22-1-o ;L c. Of 0-/ + c, -.2 Cbll' n dI + 2 Cbl2" n c- ILCLIBOT -olz O.q -0-f d ff;L 10.1 Wb Io)- 0 o.(4 C61 4.je 0,5' O.R -0.L- I.1 a..(* -0 /,2. Flask no. 2 mean Degrade ton - % 7 __14 ;L 9!9"0 2-, 1 lc4 OC rafmwoIv of,ter x dala Incubation 21 27 95-s' 9!;, ctb-S' 94-4. 19@ 9!;, See attached section1.6-3.2f:rI the formula used to calculatedogr stlon. Resultsof blank no.1 and no. 2 were averaged for each day x. The average valueswar* used In the calculations, Cbll + Cbl2 (fromabove) ' 1. The testmaterialIs completelysolubleIn water. Therefore,the TOC - DOC. 2. DOC analysesperformedmore than twice on the same samplewere run for quality assurancepurposes. 90/.8(YI-@ 70'/o 6(1/-, 5 CP/. 4V/o <C 31YIo- ------------ 2 IOY)lc, Lu 0 3M EnvironrentalLaboratory OECD modified Screening Test with DOC Analysis Analyst: Wade A. Schell Date of Start of Test: 4-11-1980 Lab Request No.:5541S Test Mater ia I At4,rWEiZ? Fc-- S017 I-OT -230'S OF OF. t T -------- 21 NUtiBER OF DAYS INCUBATED "I 0.@72 N3!3ZI310 lj3ziv=i 14evIND N@-0=131C) :11-OVC 'Z:@- 3H EnvlrardmntatLaboratoryDate Sheet -resultsof the ModifiedOECD Screening Test- Lob Request wo.: 5541 S Date of Start Analyst- -4& A- Vehal-f-- StandardMaterial:"roquinane. TestMet7ortal-AMI Theoreticaln of Test mot*rlaltr"frf?&Le-&411itrBD3 MeasuredTOC of Test Mmteriall *%-100 T0 lnaculum: soil on4 a"-chlorina-tedsoc SQ( effluentfrcisammicipat Carbon A@-olyzer:Ochrmwm K-* Totai uE2ln_iccarbon Analyzer_ 96,a. Lot 100147,Aldrich nt plant A. Determinationof I Toe If.Tyst #bLarlai-, A stock solutionof ttistestmaterialwas preparedby dilutingI-j)OroOt-o-a" finalvolumoef 5W I with dolonlzedwater* cww. test material TOC TOC In stocksolutionEsktolc"k/sooIlluution testmaterial 09/1 01/1 mg/kg This resultwas used to calculatethe necessary amountof testmterill to pr"aro a culture md Ium of 40 mg/I om Z, /ZO 8. CarbonDatermlnatlonst2 Culture medium Flask no. Mineralnutrient solutionwith testmatirl-al and with lnoculum 2 Mineralnutrient blank solutiownithout I testmaterial butwith lnocu=um elan 2 C. Evaluatioonf Ra4 Data: Conc. of test CalculatedOOCI DOC DOC - conc. after x do" materialIn of testmaterialAn&]- 0 (CO) 7 14 flask. mg/i I In flask,ml/i yses "aCl- Incubation. oVI 21 27 1 28 1 1 3.0 ai + 82 ...an ctim n 2. (o 410,0 I - bi + bn ...bn ct2.. Ci + C2 Cb] I' n m dI+ 2 Cb]2 n q2. 0 W2-a 30,2_ 2.Co b92 ql, 3 l@bbb4 3(3,% 3-@ 3.4 ot 3.11 o.1 0.2. 7 0, 1 cl D,y O.-S, , 0.17 -S.t/- b.5l'o 0,V /,2- --Oo/ 1.(o tcblo) 0 If /12 0.(P Flask no. 1 2 mean Dogrj&Wda ton,@ % 7 OC rorlovI after x dalt Incubation 21 27 94,1193,) el 12.,6 q4,1 12., - 13. 3 10 clq 13 92- Soo attachedsection1.6-3.2for the formulaused to calculatedegradation. Resultosfblankno.)andno.2 were averagedfor eachday x. The average valueswere used In the calculations. Cbll + Cbll (froambove) CbI v. 1. The testmaterialIs completelysolubleInwater. Thereforet,heTOC DOC. 2. DOC analysesperformedmorethantwiceon the samesamplewere run for qualityassurancepurposes. 901/0 8 (f/. 7 0'/o 6TI. 5(F/. 1,030/- UJ 30'/-. 2 (P/, .......... 0 3M Environmental Laboratory OECD modified Screening Test with DOC Analysis Analyst: Wade A. Scheil -4 Date of Startof Test: 4-11-1980 Lab Request No.: 5541 S 2+ Test Material:AN*rwEp,?. Fc-;?-L,6 F 1-i L-OT - 2.33 1OZ39 -T- 7-- 2@ NUT!BER OF DAYS INCUBATED N3!3Zl3lC3 a---=Yd HDVNO %-@--'Z-13!301-0:-C 1 3m Envlronmntal Laboratory Date Shoot -results of the Modified OECD Screening Test- Lob R4quest No.: 5541 S Date of Start oi Yost: 4--1 Analyst-. wade A. 11 Test Material- Theoretical IOC of Test rAte-rl-alt- IR-C%.4 obasur*d TOC of Test Material: Inoculum: $oil on4 -chlorl"t-d second* Carbon An-a-lyzer,-Ochroonn OC Total organ $toward material: bwroqulnom, 96.!I@.Lgl-lo0147Al,drich RBON ASSIAMEJD TO BE SOLUSLE OR6ANW- catmrsow) =O-g!gTTJO-C-OLf9,9 0.ifIU*At_ rcoa wAnlclpal.Mtftmter c Co analyzer treatment slant I A. Determination of TOC of Lost Material-* A stock solution of the test mterial was prepared by diluting ag to a final volume of mi with dolonlaod watera conc.test material In stock solution as/i TOC stock solution 09/1 TOC test material mg/kg This result was used to calculate Ow necessary amount of test mterlil to pr"wro a culture medium of 40 mg/i DOC CAi./)3t4.47PI44 eooMftut)D-- SELT SarAA)DA)RD B. Carton Datorminotionst Culture madlum Flask no. Mineral nutrient solution with test matert-al and with Inoculum 2 Miunnstelortual nutrient sonclcution without test material brbIut with ti0onocu=um Blank I Blank 2 C. Evaluation of Raw Data: Conc. of test material In flask, mg/i Calculated DOC of test material In flask, mg/i 30.6 zo.0 cti- al + 02 ...an n DOC Anstysos a ac DOC - conc. after x days Incubation,mg/i 0 (co) 7 14 21 27 26 J'7.1 z.3 2.0 .2. 1( .@s 1-7- 1.2- /-3 /-7 -/.9 1 /7-3 2.0 1 1,4 1.4 30.G zo.0 b ::g= J-2. bh. bi + bn c 1'7- J.q t2. n Cl 9 Ci + c 2 n d Cbl m A + d2 n C6t ICblo) 11(y di /, ;L 0.1 (Cblo) C bt I + 4:bl, j.(a o.q o t3= -O"t o Fl osk an 2 mean Degrade Ion OC remov I after x do)@s Incubation 7 iz 11 21 27 8 3 2 10,(o13,7 llj2-- 7 7/.o Iii.@ q *7 q 4/ q3 sea attached action 1.6.3.2 for the formula used "to catcu lot* degradation. Results of blank no.1 and no. 2 were averaged for each day x. The average values wore used In the calculations cblays Cbll + Cb12 (from above) 2 1. The test material Is completely soluble In water. Therefore, the TOC - DOC. suVIs were run for quality assurance purposes. 2. DOC analyses performed more than twice on the same 10 V/-9 0:/@ 8 VI. 7TI, 5 (Tfl. 40:/, 3 O@/.2 (r/, 0 3M Environrental Laboratory OECD Modified Screening Test with DOC Analysis Analyst: Wade A. Scheil Date of Start of Test: 4-11-1980 Lab Request No.: 5541 S Test Material: 14'fi)RogiAltJotJIE (CALii3iZATION e-OM?@6LI14b) 4F '7 14 21 2 NUMBER OF DAYS INCUBATED NDI*-%'2!0=!UDZ) N3E)Zi3pa HONI HdVd!3 X ::t N3@@Z.L31(3 ::)t-OT-C =.14 FORM10420-0.FWO SAMPLE DESCRIPTION ENVIRONMENTAL File: Page No. ENGR. LAB - WORK SHEET Date: 4-08-E3c::, Analyst: Hrs.: ,@Ld ija ;4 36.o f wa..2..A".got = FiJ= )97-02 14-7 2. )+,0 612. W-@ -0*-ILA J A r, gi)t-r ,,rA@@a- 4A ;t Reviad by: Fro1m0m420-8-FWO SAMPLE DESCRIPTION ENVIRONMENTAL 7a, Lk IOL File: Pap No. ENGR. LAS - WORK SHEET Date* Analyst: Hrs.: -D ig- Zo) -n AAAA - 1 8o 2M@ AM22&@@@ 1 t 5- OoO LA L -,t@ /i@@ @ ,@t + -A on-.A Reviewed by: froo1m0620-6-rwo File:LP-sPk-No. S SAMPLE DESCRIPTION ENVIRONMENTAL ENGR. LAB - WORK SHEET L Date: Analyst, Hrs.: 23 cf- ,J iA" v v@. -UAR.,60 9g.5vo oq 2/y 4b or A v Ft e- AL-.c A,#A 4L) 70 r)-TA UP-^If @e, I'-t.0, 0 Inn 0224 '4i 44 Reviewbeyd: .@.Pwo SAMPLE DESCRIPTION ,A ENVIRONMENTAL ENGR. LAS - WORK SHEET File: Fa-geNo. @f15- po-'ac *" Date: %, - @ I- 70 Analyst:LJ S. Hrs.: v 0404 r dog A OF @A 0O@e J, 4,r LAA A -4Y .4AIVA-1 IJI.9 14A J/@ 7tzl ok /ii A% -a4 A c leZO4 Revibeyw:ed fromm10620-9-FWO SAMPLE DESCRIPTION ENVIRONMENTAL 9,2 I :ZAJI.4 S-4i i File: Pao No. ENGR. LAB - WORK SHEET Date: 5-2-,!5-go Analyst: Mrs.: 6,@ o A.4v ed-rOU C4..3 A" -14^ DOC 4iL" A@ d42A, 4L u f-o)c rA @)AY DA@ 2.!g 2:1 %Aioo .)(too r"4l@ too 914 (50t APt ;L DOC 4,A t Av4 Reviewebdy: FORM 10620-0-PWO L (Z L( f File: No. SAMPLE DESCRIPTION ENVIRONMENTAL ENGR. LAB - WORK SHEET Date: Analyst: Hrs.: -.Ida 4- ,Ulu 4d4 A) Z:'@A5 p -A4.) 2-0^1.-4 v 4@@, 1 2.50 -FOC L 0-0 Y2 A.A -d in I-# v 4io v c -V I.,T)O ki 'ca-rt T- f% 0.@L.%?- FC-2-03 ------ol A 3oiI LO'r-,aIO3 (IC12too0wi, jib 11"0 a FC.- 3q) 0 /,7q 4 lo e, 14 X o @o Reviewed by: .,o 3 or tL A,L PA-T (,Dt3 COI\APDIKNJI) SftrAP L E -iiI- Aft-L 1-1 (-1. 3-1 boc- Dce- 2.1 910 -77 -ro 4 FORM10620-4-PWO SAMPLE DESCRIPTION Fe -263 D-t L,tL File:)U" No. ENVIRONMENTAL ENGR. LAB - WORK SHEET (late,..4a44 Z)o e- Y 0, rd Date: Is_ ILIAnalyst: LJ Hrs.: 7-'/ '7 ME&O 61. 1367a ,Cc.-2,03 1 v Iq) Fc. 107 rl- I- C-0 L k-/Oo z Z30810 100 13 -XJo L4, 2 qo. 2@3 0 q3 3- Reviewed by: FORM 10410-8.rwo SAMPLE DESCRIPTION ENVIRONMENTAL A-k File: PageNo. ENGR. LAB - WORK SHEET LA j@ '@ Date: Analyst: Hrs.: k loo qj, i,q ------ )ql (-D.1) :@j )C/00 ?>-,7I 3-6 -0,ti 4.3. - v 2- L F:c- 30)7 f- o, XIL7,01 70 v Qo, Reviewebdy: room IOslo-s-pwo SAMPLE DESCRIPTION -,2a(, -7) T)i ENVIRONMENTAL File: Page No. ENGR. LAB - WORK SHEET IC) Date: Analyst: Hrs.: d 3.3 13,5 3 f7o FC - 7-,o )4) Fc - 206 3.q -o41 .q.o...,.q....... Fc -ao %x too 3, Ll- El 31 1-.0 a 0 13 qb Reviewed by: FORM 10420-0-PWO SAMPLE DESCRIPTION File:Page No. Is ENVIRONMENTAL ENGR. LAB - WORK SHEET -/ Date: Eo I Analyst:ZJ, '@7 4r Hrs.: L L 7 J D4 @3.-79o Reviewed by: 110My 2 4 a 6 10 1.t2 4 6 8 10 V 2 4 4 8 10 12 2 4 4 6 10 V 2 4 6 a 10 . 2 4 6 8 10 IV 2 4 5 8 10 2 2 4 6 8 10 10 2 4 6 1 10 122 4 4 1 10 . 2 4 6 6 '0 12 2 4 6 8 10 *1 2 4 f P F FIN m fit 60 4 L 71 ff I 1- L '1--: 2 4 -0' Un 1 4 6 210122 slow 2 4 a 1 10 129 4 4 8 0 10 2 4 6 0 10 2 4 6 1 10 10 2 4 6 1 10 --- - - - - - . ........ - - - . . . . . . . . . .. - - - - - ---- ----- - - - - 2 4 6 1 10 V 2 4 6 1 10 1 4 1 8 10 V 2 4 6 1 10 2 4 0 9 10 r 2 4 6 1 10 2 4 a 14 r----- - - - -- - - - - - l@ 4 6 1 10 2 4 1 1 10 It 2 4 4 B 10 12 2 4 6 8 10 0 2 4 10 IF2 4 6 8 10 r 2 4 4 0 10 12 2 4 4 8 INO NOT2 4 8 10 122 4 4 8 10 IP 2 4 - - - - - - - - - - -- 4 4 6 10 IT 2 4 9 1 10 -- -.-.-. -.-.-.-.-.-.-.-. - - - - - ---- -- -1 L 1-1 1 rt ;8 10 10 2 4 6 1 10 2 4 6 6 2 4 6 1 to . . . . . ... .. . . . . . . . . 4 9 1 10 0 2 4 9 1 10 2 4 4 2 4 6 1 10 1 4 1 310 IP 2 4 6 0 10 2 4 4 1 0 0 t 4 1 1 to - - -- - - - - - --- - - - - .. . . . . . . . . . . ... -------- --- - - - - - - - 4 4 1 le illI 4"'i 14 jr2 . .. --- .. 7+ room I0420-8-PWG SAMPLE DESCRIPTION File:Pap No.. ENVIRONMENTAL ENGR.LAS - WORK SHEET Date: Analyst: Hrs.: dJtIAXi MII&47J Reviewed by: Forin3337 itob, Ki)l)lFlel)c n ANALYSIS FORM DE-SCRI(>T)ot4 SLANI< -TF-ST DCAP-A-TICO)14 t)A--(0 0 E L-kgokA-rOPy -<)C- /A?J-rPJFAP FC-ZO5 14FI, F --(7; H I qo.-Al I>A,-rA Wil C FrD it,3M6/L MTWEAP FC-3ol7i.OTZ303 APJ'RWEAP /FS,(,C-Z0 6 (F m - 3s34)i-@-2.-5so 4) L 1. q2. 1/2.-3yp..S_ 0 11711 14 sio 5-rA 17-10 16.2- 0.1 -0-11 .3lOLIL-9 3 1,2. IN-q M3 3#1,,0.30.17 2 'A .2. -04 0.1 3A3 lc" 3ilo 3a-71 1 -7 D.r. 1 7 9 10 0,1 2-4 31 -.16" 5.3 .3.6' 0 @r 8.1 I-s' 3.6 -4.1 3,3 /.2 12 0.5 0.'ro Co s-.s-l 13 14 - I1176s [!r5>-A0Y2.-R2-o1) O1..5q 1.0 ro.2-12..90 41,?-L :I3.,z41I3I..9 q -3.613.5 34,.o1 12.4 o-5 1.2- 4.2. 20 21 i)AY 2.'l 22 zi @424 25 26 DA-4 2B 29 -0.4 3.2- q-1 5"0 __,q (. -0-/,010.1 I'll 2.-7 ;a.9 3aq )LIS2. (oI*.6 -f,17 l,7 3-@L 3-v li.1 3 119 3r..l I.o ,z.gt3_Gq --310 2;2.. 3'). 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