Document MJO3exyg2Eekw59b5qxZDwMpk

m i-w 5j PROTOCOL DETERMINATION OF THE WATER SOLUBILITY OF PFOS BY THE SHAKE FLASK METHOD OECD Guideline for the Testing of Chemicals, 105 Water Solubility 3M Lab Request No. U2723 Submitted to 3M Corporation Environmental Laboratoiy P.O. Box 33331 St. Paul, Minnesota 55133 Wildlife International ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 December 2,1998 PROTOCOL NO.: 454/120298/830.7840/SUB454 000140 3M LAB REQUEST NO. U2723 W il d l if e In t e r n a t io n a l l t d . -2- DETERMINATION OF THE WATER SOLUBILITY OF PFOS BY THE SHAKE FLASK METHOD SPONSOR: 3M Corporation Environmental Laboratory P.0. Box 33331 St. Paul, Minnesota 55133 SPONSOR'S REPRESENTATIVE: Ms. Susan A. Beach TESTING FACILITY: Wildlife International Ltd. 8598 Commerce Drive Easton, Maryland 21601 STUDY DIRECTOR: Ronda Jacobs foym w vU V fln-ljeikn, P h D. Senior Chemist* ierA is4" LABORATORY MANAGEMENT: Willard B. Nixon, Ph.D. Manager of Chemistry FOR LABORATORY USE ONLY Proposed Dates: Experimental Start Date- Project No : H 6 itfi-io n Test Substance No : ___ ^ fo T 5 ______ Experimental Termination Date- Tnt/Date- fili-lnlff Receipt Date- io toqfafi a/io/n DATE DATE iz h h i DATE PROTOCOL NO.: 454/120298/830.7840/SUB454 000141 3M LAB REQUEST NO. U2723 Wil d l if e In te r n a tio n a l ltd . -3 - INTRODUCTION Wildlife International Ltd. will experimentally determine the water solubility o f PFOS (perfluorooctane sulfonic acid, potassium salt). The study will be conducted at the Wildlife International Ltd. analytical chemistry facility in Easton, Maryland. The study will be performed following the guidance presented in the OECD Guideline for Testing of Chemicals, 105: Water Solubility (1) for shake flask methodology. This study is intended to meet data requirements under this guideline and under EPA Product Properties Test Guidelines, OPPTS 830.7840, Water Solubility: Column Elution Method; Shake Flask Method (2) and TSCA Title 40 of the Federal Code of Regulations, Part 796, Section 1840: Water Solubility (3). The method of analysis for quantitation provided by the Sponsor will be verified at Wildlife International Ltd, Raw data for all work performed at Wildlife International Ltd. and a copy of the final report will be filed by project number in archives located on the Wildlife International Ltd. site or at an alternative location to be specified in the final report. PURPOSE The purpose of this study is to determine the water solubility of PFOS at 20 0.5 C by the shake flask method. Determination of water solubility by the shake flask method is applicable to test substances with water solubilities equal to or exceeding 0.01 gram/Liter (g/L). The methodology may not be applicable to surface active or volatile test substances. EXPERIMENTAL DESIGN The water solubility of PFOS will be determined at a temperature of 20 0.5 C. A preliminary test will be performed to estimate the water solubility at room temperature. The preliminary test consists o f additions of increasingly large amounts of water to a known weight of test substance until solubilization is effected or the solubility is estimated to be less than 0.01 g/L. For substances with an estimated water solubility greater than 0.01 g/L, a definitive test will be performed. The definitive test consists o f equilibration of an excess amount of test substance with water at an elevated temperature, 30C, followed by equilibration at 20C. MATERIALS AND METHODS Test Substance The test substance will be the highest purity available. The test substance should be at least 99% pure by weight. The Sponsor will be asked to provide the following information concerning the test substance, if available: PROTOCOL NO.: 454/120298/830.7840/SUB454 000142 3M LAB REQUEST NO. U2723 Wil d l if e In t e r n a t io n a l l t d . -4 - Chemical Name Empirical Formula and/or Molecular Weight Dissociation Constant in Water Expiration Date Method o f Analysis Lot or Batch Number Chemical Purity: Vapor Pressure Storage Conditions The attached form IDENTIFICATION OF TEST SUBSTANCE BY SPONSOR (Appendix I), along with a Material Safety Data Sheet, will be used to provide information necessary for the handling and testing of the test substance. The Sponsor agrees to accept any unused test substance and/or test substance containers at the end o f the study. Reagents Water that meets ASTM Type II standards (ASTM D 1193-91) will be used (4). Other solvents may be needed for the analytical method or preparation of stock solutions and analytical standards. All solvents will be ACS reagent grade or better, and will be determined to be free of contaminants that interfere with the quantitation of the test substance. Preliminary Test Procedure A preliminary test will be conducted to estimate the solubility of the test substance at room temperature. Approximately 10 mg of the test substance will be placed in a plastic tube. Increasing volumes o f water will be added stepwise as shown below: Total volume of water added (mL) Approximate solubility (g/L) 0.1 0.5 1 2 10 >100 20 10 51 After each addition of water to yield the total volume indicated, the container will be capped and shaken vigorously for approximately 10 minutes and visually examined for undissolved test substance. If the test substance is not dissolved after addition of 10 mL of water, the contents of the container will be transferred to a larger container and water added for a final volume of 100 mL. Following capping and shaking, the container will be visually checked for undissolved material, i.e., undissolved material would indicate a water solubility <0.1 g/L. Preliminary testing of the solubility at lower concentrations (<0.1 g/L) may be performed by transferring the contents of the 100-mL vessel to a 1-L vessel or reinitiating ' the test with a lesser amount o f test substance, e.g., 1 mg. Amounts and volumes noted above are only guides for conduct of the preliminary study; larger amounts of test substance with proportionally larger volumes of water may be used. 000143 PROTOCOL NO.: 454/120298/830.7840/SUB454 3M LAB REQUEST NO. U2723 Wil d l if e In t e r n a t io n a l ltd. -5 - Definitive Test Procedure The quantity of test substance necessary to saturate the desired volume of water is estimated from the preliminary test. The volume of water used will depend on the analytical method and solubility range. Greater than (or equal to) five times the quantity of test substance required to achieve water solubility will be transferred to a minimum of three test vessels (plastic and/or other suitable container material). The desired volume of water will be added to each vessel and the vessels sealed. The closed vessels will be agitated (shaken or stirred) at 30 1.0 C in a water bath. After 1 day, one o f the vessels will be removed and re-equilibrated for 24 hours at 20 0.5 C with occasional shaking or stirring. The contents of the vessel will then be centrifuged in a centrifuge regulated at 20C. The concentration of the test substance in a minimum of two aliquots ofthe aqueous phase will be determined. The remaining two vessels are treated similarly following initial equilibration periods o f 2 and 3 days at 30 1.0C. The test will be terminated if the concentrations of the test substance in the aqueous phase of two consecutive vessels agree within 15%. If greater variation is observed and/or at the request o f the Sponsor, the test may be reinitiated with longer equilibration periods. Method The analytical method for quantitation of the test substance will be based on procedures provided by the Sponsor. The method will be appended to this protocol in Appendix II and referenced in the report. The method may be verified as part of the preliminary test(s) and may be modified to measure the observed test substance concentrations. Major modifications will be approved by the Sponsor and Study Director, and documented in the raw data. Calculations The concentration of the test substance in each vessel will be expressed in g/L, mg/L or |ig/Lin water as appropriate. The average solubility for test vessels with solubilities within 15% of each other will be averaged and reported. Sample Handling and Safety The Sponsor will identify any special handling or safety precautions to be used with the above referenced test substance. All normal precautions with respect to handling and storage will be taken. 000144 PROTOCOL NO.: 454/120298/830.7840/SUB454 3M LAB REQUEST NO. U2723 Wil d l if e In t e r n a t io n a l l td . n 6- - Sample and Test Substance Retention Upon completion of testing, portions of the test substance used as part o f this study will be disposed of in accordance with federal, state and local regulations. Any unused portion of the test substance will be returned to the Sponsor. RECORDS TO BE MAINTAINED Records to be maintained for data generated by Wildlife International Ltd. will include, but not be limited to: 1. A copy o f the signed protocol. 2. Identification and characterization o f the test substance, if provided by the Sponsor. 3. Dates of initiation and completion of the study. 4. Dates o f experimental start and termination. 5. Storage conditions o f the test substance. 6. Test substance use log. 7. Concentration calculations and records of solution preparation. 8. Instrument operating conditions and chromatograms, if applicable. 9. Statistical calculations. i 10. Test conditions. 11. A copy o f the final report. I FINAL REPORT A final report o f the results of the study will be prepared by Wildlife International Ltd. The report will include, but not be limited to the following, when applicable: 1. Name and address of the facility performing the study. 2. Dates upon which the study was initiated and completed. 3. A statement o f compliance signed by the Study Director addressing any exceptions to Good Laboratory Practice Standards. 4. Purpose and procedure, as stated in the approved protocol, including all amendments and deviations to the protocol. 5. The test substance identification, including name or code number, purity, empirical formula, molecular formula, lot or batch number, method of analysis, and any other information provided by the Sponsor. 6. Description o f the test method or reference to the method used along with any m odifications made. 000145 PROTOCOL NO.: 454/120298/830.7840/SUB454 3M LAB REQUEST NO. U2723 Wil d l if e In tern a tio n a l ltd . -7 - 7. The individual concentrations o f each sample. 8. The means and standard deviations of solubility determinations at each interval for each temperature tested. 9. Description of any problems experienced and how they were resolved. 10. A statement prepared by the Quality Assurance Unit listing the dates that study inspections and audits were made and findings reported to the Study Director and Management. CHANGING OF PROTOCOL Planned changes to the protocol will be in the form of written amendments signed by the Study Director and the Sponsor. Amendments will be considered as part of the protocol and will be attached to the final protocol. Any other changes will be in the form of written deviations filed with the raw data. All changes to the protocol will be indicated in the final report. GOOD LABORATORY PRACTICES This study will be conducted according to the Good Laboratory Practices described in OECD (ISBN 92-84-12367-9) and EPA (40 CFR Part 792). Each study conducted by Wildlife International Ltd. is routinely examined by the Wildlife International Ltd. Quality Assurance Unit for compliance with Good Laboratory Practices, Standard Operating Procedures and the specified protocol. A statement of compliance with Good Laboratory Practices will be prepared for all portions of the study conducted by Wildlife International Ltd. The Sponsor will be responsible for compliance with Good Laboratory Practices for procedures performed by other laboratories. Raw data for all work performed at Wildlife International Ltd. and a copy of the final report will be filed by project number in archives located on the Wildlife International Ltd. site or at an alternative location to be specified in the final report PROTOCOL NO.: 454/120298/830.7840/SUB454 0*00146 3M LAB REQUEST NO. U2723 Wil d l if e In te r n a tio n a l ltd. 8- - REFERENCES 1 O rganisation for Economic Cooperation and Development. 1995. Guideline for Testing of Chemicals, 105: Water Solubility. 2 P ro d u ct Properties Test Guidelines. 1996. OPPTS 830.7840. Water Solubility: Column Elution Method; Shake FlaskMethod. 3 TSCA T itle 40 of the Federal Code of Regulations. 1994. Part 796, Section 1840. Water Solubility. 4 American Society for Testing and Materials. 1991. Standard Specification for Reagent Water. D 1193-91, ASTM Section II Water and Environmental Technology, Yol. 11.01: 45-47. PROTOCOL NO.: 454/120298/830.7840/SUB454 000147 3M LAB REQUEST NO. U2723 W il d l if e In t e r n a t io n a l ltd. -9 - APPENDIXI IDENTIFICATION OF TEST SUBSTANCE BY SPONSOR To be Completed by Sponsor I. Test Substance Identity (name to be used in the report): PFOS (Perfluorooctane Sulfonic Acid Potassium Salt Reference Standard (if applicable): Analytical Standard: N/A__________ Internal Standard: 1.1.2.2H.H.H.H Perfluorooctane Sulfonic Acid Test Substance Sample Code or Batch Number: Lot 217___________ ___________________________ Test Substance Purity (% Active Ingredient): 98.9______ Expiration Date: 2008_______________ II. Test Substance Characterization Have the identity, strength, purity and composition or other characteristics which appropriately define the test substance and reference standard been determined prior to its use in this study in accordance with GLP Standards? Y es_____ No X III. Test Substance Storage Conditions Please indicate the recommended storage conditions at Wildlife International Ltd. Ambient room temperature_____________________________________________ _ Has the stability o f the test substance under these storage conditions been determined in accordance with GLP Standards? Y es____ No X - Other pertinent stability information: _______________________________________ ________________ IV. Test Concentrations: Adjust test concentration to 100% a.i. X based upon the purity (%) given above. Do not adjust test concentration to 100% _____a.i. Test the material AS IS. V. Toxicity Information: Mammalian: Rat LD50 251 mg/kg Mouse LD50 N/A Aquatic: Invertebrate Toxicity (EC/LC50) Fish Toxicity (LC50) Daphnia maena: 27 mg/L__________ Rainbow Trout: 11 mg/L Daphnia maena: 50 mg/L__________ Fathead Minnow: 38 mg/L Other Toxicity Information (including findings of chronic and subchronic tests): Please see MSDS___________________________________________________________ PROTOCOL NO.: 454/120298/830.7840/SUB454 000148 3M LAB REQUEST NO. U2723 Wil d l if e In t e r n a t io n a l ltd. - 10- APPENDIX II ANALYTICAL METHOD Analytical Method for the Determination of PFOS in Water PFOS may be determined in water using high performance liquid chromatography with mass spectrometric detection (LCMS). Sample preparation for the fortification samples will involve dilution (using the same matrix water) into the concentration range of instrumental response. Matrix blanks will be directly injected. The following presents approximate chromatographic conditions for LCMS quantitation of PFOS. Chromatographic conditions may be changed to provide conditions more optimized for retention and/or chromatographic separation of PFOS. Instrument: Hewlett-Packard Model 1090A HPLC Column: Keystone Betacil C-18, 100 mm x 2.0 mm, 3/mn Mobile Phase: Methanol: Ammonium Acetate Buffered Water 65:35 (v:v), isocratic. Flow Rate: 250 L/minute Column Temperature: 40C Injection Volume: 100 /L Ion Source: Turbo IonSpray Mass Spectrometer: Perkin-Elmer API 100 Run Time: 6.0 minutes PROTOCOL NO.: 454/120298/830.7840/SUB454 000149 3M LAB REQUEST NO. U2723