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AR&9.6 _ OSOO m ACTIVATED SLUDGE RESPIRATION INHIBITION TEST TEST SUBSTANCE________________________________________________ Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-206A. (1Octanesulfonic acid) (CAS # 2795-39-3). Rem arks: The 3M production lot number was 2052. The test sample is FC-206A. Current information indicates it is a mixture of 0.92% PFOS, 18.75% diethylene glycol butyl ether, 75.44% water, 1.83% Hydroxy foamer, 1.75% sodium octyl sulfate, 1.2% poloxyethylene monooctylphenyl ether, 0.06% sodium lauryl sulfate, and 0.05% benzotriazole. The following summary applies to a mixture with incompletely characterized concentrations o f impurities. Data may not accurately reflect die toxicity o f the fluorochemicai component o f the test sample. M E T H O D _________________________________________________________ Method: 3M Environmental Laboratory Protocol for Determining Acute Toxicity of a Compound to Activated Sludge GLP: No Year Com pleted: 1979 Analytical m onitoring: Dissolved oxygen concentrations. S tatistical methods: Not given. Test organism source: Activated sludge mixed liquor collected from the Metropolitan Wastewater Treatment Plant (Pig's Eye Plant), St. Paul, MN. Test condition: Dilution w ater: Deionized water. Reference and test solution preparation: A primary stock solution was created by diluting 0.60 g test sample to 100 mL with dilution water. Test vessels: Not given. Num ber o f concentrations: one plus 1 blank control Tem perature: Not given. Total suspended solids and pH on day o f testing: Not given. Elem ent Basis: Respiration inhibition as determined by oxygen consumption. RESULTS________________________________________________________ Nom inal concentrations: 1 Blank control and 1000 mg/L Elem ent values: No acute inhibitory effects on activated sludge respiration rate at 1000 mg/L with a contact time of 5 minutes. 00162S Element values based on nominal concentrations. Rem arks: Testing was conducted on a mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical component alone. C O N C L U S IO N S ___________________________________________________ The test sample exhibited no inhibitory effects at 1000 mg/L over a 5-minute exposure period. Subm itter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY__________________________ . _ ____________________ Reliability: Klimisch ranking 3. Study lacks documentation. The inoculum is not properly characterized. There is no characterization of the test substance purity and the study lacks analytical confirmation of the amount of fluorochemical proportion in solution. R E F E R E N C E S ____________________________________________________ This study was conducted by the 3M Company, Environmental Laboratory, St Paul, MN, Lab Request number 4970S, 1979. OTHER_______________________________________________ ____________ Last changed: 6/26/00 001630 mm Fit: cJ No. < /i? o s ENVIRONMENTAL EN QR. LAB - W O R K SHEET Reviewed b y : __,__________ 001631 & ** A ' 1 3 . " '3C BEST COPY AVAILABLE10 13 O ICTZO C N M A M P A W N 1 0 X IO PEN INCH DISSOLVED OXYGEN CONCENTRATION - mg/1 r vn O 3M ENVIRONMENTAL L A B O R A T O ^ PROTOCOL FOR DETERMINING ACUTE TOXICITY OF A COIVOUD TO ACTIVATED SLUDGE EAR 1/79 'ROCEDURE MATERIALS AND REAGENTS .l Obtain 4 liters of activated sludge from as aeration basin at the Metro Wastewater Treat ment plant. .2 Use sludge within 4 hours of collection. .0 Prepare a 100-ml, 6,000-mg/l stock solution of the material to be tested in air-saturated deionized (DI) water. .0 Starting with this stock solution, m ake serial dilutions by transferring 10 nl o t the pre ceding solution into 100-al volumetric flasks and bringing to volume with air-saturated water. In this manner, prepare 600-, 00-, 6-, and 0.6-ag/l solutions. .1 Prepare a 1,200 ag/1 nutrient broth solution by dissolving 1.2 g in approxinately 100 ml of DI water at 95 C and diluting to 1 liter with air-saturated water. ,2 Aerate to bring to air saturation. .1 Pass the sludge through a 100-mesh stainless steel sieve. .2 Place 3 liters of the activated sludge in a 4-liter beaker. 3 Place the.beaker on an insulating sheet on v top of a magnetic mixer. .4 Set the mixer speed so that it keeps the sludge completely suspended. .5: Aerate the sludge rapidly to bring it to near air saturation. X Place a 1-in. magnetic stirring bar in a BOO bottle. .2 Plaee tbs BOD bottle on a sheet of foam insulation on top of a magnetic mixer. 3 Add SO ml of nutrient broth soluti and 50 ml of aerated DI water. 4 Start the mixer at a rate that will keep sludge suspended. Transfer enough of the completely dis persed , aerated; activated sludge to fill the BOD bottle using a 500-ml beaker. The following materials and reagents are required for this procedure: 1) Oxygen meter 2V DO probe 3) ASTM thermometer 4) 4 "Liter beaker 5) 500 ml beaker 6) 10 ml pipets 7) 100 ml volumetric flasks 8) Compressed air 9) Air spargers 10) Plastic tubing 11) 300 ml BOD bottles 12) 3-in. Magnetic stirring bar 13) 1-in. Magnetic stirring bar 14) Magnetic mixer 15) 3/4 in. Foam insulation 16) Stop, watch 17) 100-inesh steel sieve 18) Deionized water 19) Nutrient broth 20) Activated sludge 6.6 Start the stopwatch immediately. 6.7 Insert the DO probe into the BOD bottle, and record the time and DO concentration at fre quent intervals for 10 min or until the DO concentration reaches 0.5 mg/1. 6.8 Obtain fresh sludge if the DO is greater than 0.5 mg/1 at 10 min. 6.9 If the DO is depleted to 0.5 mg/1 in less than 5 min., bring the time required for complete 0,, uptake to between 5 and 10 min by diluting the sludge stock with air-saturated DI water. 7.1 Repeat the procedure described in steps 6.1 to 6.7, except substitute 50 ml of the test material solutions or aerated DI water controls for the aerated DI water used in step 6.3. 7.2 Run the 0 assays in the following order: 1) 6,000-mg/l test material, 2) DI control No. 1, 3) 0.1 mg/1, 4) 100 mg/1, 5) 1 mg/1, 6) Control No. 1, 7) 10 mg/1. 8.1 Use a full sheet of graph paper to plot the oxygen concentration as a function of time for the control and each test concentration. 8.2 Label each plot with the final concentration of test material in each BOD bottle. These concentra tions are 1,000, 100, 10, 1, and 0.1 mg/1. C 59TQ P 5 o /-/7 2/7