Document pmg1qwNqDNjwQdqqXQZeOerOE

28) OECD 105-OPPTS 830.7840, water solubility in nanopure water, 454C-118 SANITIZED PERFLUOROBUTANE SULFONATE, POTASSIUM SALT (PFBS) , BY THE SHAKE FLASK METHOD WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454C-118 I OECD Guideline for the Testing of Chemicals, 105 Water Solubility AUTHORS : Raymond L. VanHoven, Ph.D. Jon A. MacGregor, B.S. Willard B. Nixon, Ph.D. STUDY INITIATION DATE: July 27,2000 STUDY COMPLETION DATE: August 30,2000 Submitted to 3M Corporation Environmental Laboratory 935 Bush Avenue St. Paul, Minnesota 55 144 Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 2 1601 (410) 822-8600 Page 1 of34 WILDLIFE INTERNATIONALL, TD. SANITIZED PROJECT NO : 454C-118 -2- OEC 0 9 2003 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT SPONSOR: 3M Corporation TITLE: Determination of the Water Solubility of Perfluorobutane Sulfonate, Potassium Salt (PFBS) by the Shake Flask Method WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454C-118 3M ENVIRONMENTAL LAB PROJECT NUMBER E00-1429 STUDY COMPLETION':'August 30,2000 This study was conducted in compliance with Good Laboratow PracticeStandards as describedin OECD Principles of Good Laboratory Practice (ENV/MC/CHEM (98) 17), with the following exceptions: The test substance was not characterized in accordance with Good Laboratory Practice Standards. The stability of the test substance under storage conditions at the test site was not determined in accordance with Good Laboratory Practice Standards. STUDY DIRECTOR: Wildlife International, Ltd. SPONSOR APPPROVAL: 530-00 DATE WILDLIFE INTERNATIONALLT, D. -3- PROJECT NO.: 454C-118 QUALITY ASSURANCE STATEMENT T h s study was examined for compliance with Good Laboratory Practice Standards as described in OECD Principles of Good Laboratory Practice (ENVMCKHEM (98) 17). The dates of all inspections and audits and the dates that any findings were reported to the Study Director and Laboratory Management were as follows: ACTMTY Sample Preparation Draft Report and Data Final Report DATE CONDUCTED: July 27,2000 August 1,2000 August 30,2000 . DATE REPORTED TO: STUDY DIRECTOR MANAGEMENT: July 27,2000 July 27,2000 August 1,2000 August 3,2000 August 30,2000 August 30,2000 Susan L. Coleman, B.A. Senior Quality Assurance Representative DATE 8 - 3 0 - 0 0 WILDLIFE INTERNATIONAL,LTD. -4- PROJECT NO.: 454C-118 REPORT APPROVAL SPONSOR: 3M Corporation TITLE: Determination of the Water Solubility of Perfluorobutane Sulfonate, Potassium Salt (PFBS) by the Shake Flask Method WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER 454C-118 3M ENVIRONMENTAL LAB PROJECT NUMBER: E00-1429 STUDY DIRECTOR: Ra&dnt?L. VanHoven, Ph.D. Scientist MANAGEMENT: Willard B. Nixon, Ph.D. ' Manager, Analytical Chemistry DATE DATE ' WI LDLlFE I NTERNATIONAL. LTD. PROJECT .NO. 454C-118 TABLE OF CONTE'NTS TitleKover Page................................................................................................................................................. 1 Good Laboratory Practice Compliance Statement............................................................................................. 2 Quality Assurance Statement............................................................................................................................. 3 Report Approval ....................... .................................................................................................................... 4 Table of Contents ............................................................................................................................................... 5 summary............................................................................................................................................................. 7 Introduction ........................................................................................................................................................ 8 Purpose ............................................................................................................................................................... 8 Experimental Design .......................................................................................................................................... 8 Materials and Methods....................................................................................................................................... 8 Test Substance ........................................................................................................................................... 9 Reagents..................................................................................................................................................... 9 Preliminary Test Procedure ....................................................................................................................... 9 Definitive Test Procedure.......................................................................................................................... 9 Calibration Curve .................................................................................................................................... 10 Reagent Blanks and Limit of Quantitation.............................................................................................. 10 Example Calculations.............................................................................................. :............................... 11 Results ............................................................................................................................................................ 11 Water Solubility....................................................................................................................................... 11 Conclusions ...................................................................................................................................................... 12 References ........................................................................................................................................................ 13 TABLES Table 1. Typical LC/MS Operational Parameters ........................................................................................ 14 Table 2. Reagent Blanks Analyzed Concurrently During Sample Analysis................................................. 15 Table 3. Solubility of PFBS inNANOpure(9 Water ..................................................................................... 16 WILDLIFE INTERNATIONAL. LTD. -6- PROJECT .NO. 454C-118 TABLE OF CONTENTS .Continued . FIGURES Figure 1 .Analytical method flow chart for the analysis of PFBS in NANOpureO Water............................. 17 Figure 2 .A typical calibration curve for PFBS .............................................................................................. 18 Figure 3 .A representative ion chromatogram of a low-level PFBS standard................................................ 19 Figure 4 .A representative ion chromatogram of a high-level PFBS standard .............................................. 20 Figure 5 .A representative ion chromatogram of a reagent blank sample...................................................... 21 Figure 6 .A representative ion chromatogram of a Day 2 solubility sample.................................................. 22 APPENDICES Appendix I -Protocol and Protocol Deviation................................................................................................ 23 Appendix I1 -Certificate of Analysis................................................................................................................ 33 Appendix I11 -Personnel Involved in the Study................................................................................................. 34 WILDLI FE INTERNATIONALLT, D. -7- PROJECT NO. 454C- 118 SUMMARY STUDY TITLE: WILDLIFEINTERNATIONAL, LTD. PROJECT NUMl3ER: SPONSOR: TESTING FACILITY: LOCATION OF STUDY, RAW DATA AND A COPY OF THE FINAL REPORT: Determination of the Water Solubility of Perfluorobutane Sulfonate, Potassium Salt (PFBS) by the Shake Flask Method 454C-118 3M Corporation Wildlife International, Ltd. Easton, Maryland 2 1601 Wildlife International, Ltd. Easton, Maryland 2 1601 TEST SUBSTANCE: TEST DATES: PFBS Experimental Start - July 27,2000 Experimental Termination - July 3 1,2000 TEST SYSTEM: NANOpure@Water 1 SUMMARY: The overall mean solubility concentration of PFBS in NANOpureO water was 46.2 g PFBSL (SD= 0.18; CV = 0.38%; N = 6). WILDLIFE INTERNATIONAL, LTD. -8- PROJECT NO.: 454C-118 INTRODUCTION This study was conducted by Wildlife International, Ltd. for 3M Corporation at the Wildlife International, Ltd. analyticalchemistry facility in Easton, Maryland. A test was conducted to determine the water solubilityof PFBS (perfluorobutane sulfonate, potassium salt). The experimentalportion of thls study was conductedbetween July 27, 2000 and July 3 1, 2000. The orignal raw data generated by Wildlife International, Ltd. and a copy of the final report are filed under Project Number 454C- 118 in archives located on the Wildlife International,Ltd. site. PURPOSE The purpose of this study was to determine the water solubility of PFBS at 20 k 0.5"C by the shake flask method. Determinationof water solubility by the shake flask method is applicable to test substances with water solubilities equal to or exceeding 0.01 gram/Liter (a). EXPERIMENTAL DESIGN The water solubility of PFBS was determined at a temperature of 20 k 03C. A preliminary test was performed to estimate the water solubility. The test consisted of additions of increasingly larger amounts of NANOpure@water to a known weight of test substance at room temperature. The deffitive test consisted of equilibration of an excess amount of test substance with NANOpure@water at 30C followedby equihbrationat 20C and analyzing subsamples by high performance liquid chromatography with mass spectrometric detection (LCNS). MATERIALS AND METHODS This study was conducted accordingto the procedures outlinedinthe protocol, "Determinationof theWater Solubilityof Perfluorobutane Sulfonate, Potassium Salt (PFBS) by the Shake Flask Method." The protocolwas based on procedures outlined in the OECD Guideline for Testing of Chemicals, 105: WaterSolubility (1). This study meets data requirements under this guideline and under U.S. EPA Product Properties Test Guidelines, WILDLIFE INTERNATIONAL, LTD. -9- * PROJECT NO.: 454C-118 SANITIZED Serie~830.7840,Water Solubi1ity:Column ELution Method; Shake Flask Method (2) for shake flask methodology and TSCA Title 40 of the Federal Code of Regulations, Part 796, Section 1840: r a t e r Solubility (3). Test Substance The test substance was received from 3M Environmental Technology and Safety Services on March 27, 2000, assigned Wildlife International, Ltd. Identification number 5216, and stored under ambient conditions. The test substance, a white powder, was identified as: Potassium Perfluorobutane Sulfonate expiration date: March 20 10. The test substance was further identified with the 3M Environmental Laboratory test control and reference number . The test substance had a reported purity of 97.90% (Appendix 11). Reagents All solvents used in the solubility analyseswere of ACS reagent grade. NANOpureO water (equivalentto ASTM Type I1 Designation D1193-91) was used (4). Preliminaw Test Procedure A preliminary test was conducted to estimate the water solubility of PFBS. The test was performed at room temperature by adding approximately 10 mg of the test substance to a 10-mL volumetric flask. Increasing volumes of NANOpure@water were added (0.1 and 0.4 mL) to the flask. After each addition, the flask was shaken vigorously for approximately 1 minute and visually examined for undmolved test substance. After the addition of a total volume of 0.5-mL of NANOpure@water, no particulates were observed. Definitive Test Procedure Approximately 10.0 grams of the test substance was weighed into each of three labeled 8-ounce French square bottles. NANOpure@water, 100mL equilibrated at 20C for -30 minutes, was addedto each bottle. The bottles were sealedwith screwcaps and wrapped with Parafilm@.The bottles were agitated in a shaker-waterbath (LindbergBlue, Model No. S W B 1122A)maintained at 30 k 1.O"C. After one day, one bottle was removed and placed in a second constant temperature bath (LindbergBlue, Model No. SWB1122A, with agitation switched off) maintained at 20 k 0.5"C for 24 hours with occasional shaking.,Triplicate 10-mLsubsampleswere removed WILDLI FE INTERNATIONAL,LTD. PROJECT NO.: 454C-118 from the bottle, placed into 15-mLglass centrifugetubes, and Centrifuged at -2,000 rpm (845g) for -5 minutes. A single-IO-mL subsample of the NANOpureOwater matrix was centrifuged along with the treated samplesfor use as a reagent blank. Because undissolved, suspended material was evident upon completion of this centrifugation, a second and more vigorous centrifugation was incorporated into the methodology. Approximately I-mLaliquots of the supernatant from each subsample in the 15-mL centrifuge tubes were then transferred to plastic microcentrifuge tubes and centrifuged at -13,000 rpm (14,OOOg) for -5 minutes. An appropriate volume from each subsample aliquot was volumetrically removed and diluted into the calibration range of the LCMS methodology with 50% methanol and 50% NANOpure@water dilution solvent. The dilutions were placed into autosampler vials for analysis by reverse-phase high performance liquid chromatography (HPLC) using a Hewlett-PackardModel 1100 High Performance Liquid Chromatographwitha Perkin-Elmer API IOOLC Mass Spectrometer equipped with a Perlun-Elmer Turbo IonSpray ion source. The remaining two bottles were treated in a similar manner following initial equilibration periods of 2 and 3 days at 30.0 k 1.0"C. Instrumental parameters for the analysis of PFBS are summarized in Table 1 and a method flow chart is provided in Figure 1. Calibration Curve Calibration standards of PFBS, ranging in concentration from 1.00mg PFBSL to 5.00 mg PFBSL, were analyzedwith each sample set. A linear regression equation was generated using the peak area responses versus the respective concentrations of the calibration standards. A typical calibration curve is presented in Figure 2. The concentration of PFBS in the samples was determined by substituting the peak area response into the applicable linear regression equation. Representative ion chromatograms of low and high calibration standards are presented in Figures 3 and 4, respectively. Reagent Blanks and Limit of Ouantitation Reagent blanks were analyzed to assess the presence of potential interferences. The reagent blank, consisting of a subsample from the same batch of NANOpure@water as was used for preparation of the 100 g PFBSL (target nominal concentration) test samples, was processed in an identical manner as for the treated samples. The limit of quantitation (LOQ) for these analyses, calculated as the product of the lowest calibration standard (1.00 mg PFBSL) and the dilution factor of the reagent blank samples (12500), was 12500mg PFBSL = 12.5 g PFBSL. No chromatographic interferenceswere observed in the reagent blanks at or abovethe limitof WILDLIFE INTERNATIONALL, TD. - 11 - PROJECT NO. 454C- 118 quantiiation of 12.5 g PFBSL (Table 2). A representative ion chromatogram of a reagent blank sample is presented in Figure 5. Example Calculations Sample number 454C-118-2A, target nominal concentration of 100 g PFBSL in NANOpure@water First Initial Volume: 0.200 mL First Final Volume: 25.0 mL Second Initial Volume: 0.100 mL Second Final Volume: 10.0mL Dilution Factor: 12500 PFBS Peak Area: 17031882 Calibration curve equation. Slope: 3535642.25 Intercept: 4000869 Curve is weighted: (l/x) Instrument Software: MacQuan, version 1.6 PFBS (S/L) in sample = - areaslo'-pinetercept' x dilution factor x unit conversion factor - 17031882-4000869 3535 642.25 l2'Oo Ig 1000 mg = 46.1 RESULTS Water Solubility Triplicate subsamples were removed from the appropriate bottles after one, two and three days of shaking in a water bath maintained at 30 f 1.O"C and following one day of a 20 f.0.5"C equilibration period (Table 3). Analysis of aqueous subsamples after one day had a mean analytical result of 44.1 g PFBSL (SD = 0.40, WILDLIFE INTERNATIONAL,LTD. - 12- PROJECT NO.: 454C-118 CV = 0.92%). For subsamples collected after two and three days, the mean concentrations were 46.1 g PFBSL (SD = 0.15, CV = 0.33%) and 46.3 g PFBSL (SD = 0.15, CV = 0.33%), respectively. A representative ion chromatogram of a Day 2 solubility sample is presented in Figure 6. CONCLUSIONS The Day 2 and Day 3 mean solubility concentrations were well withinthe 15% agreement criterion of the protocol and were averaged to obtain the overall mean solubility concentration. The overall mean solubility concentration of PFBS in NANOpure@water was 46.2 g PFBSL (SD = 0.18; CV = 0.38%; N = 6). WILDLIFE INTERNATIONAL, LTD. - 13- PROJECT NO.: 454C-118 REFERENCES 1. Organisation for Economic Cooperation and Development. 1995. Guideline for Testing of Chemicals, 105: Water Solubility. 2. Product Properties Test Guidelines. 1996. OPPTS 830.7840. Water Solubility: Column Elution Method; Shake Flask Method. 3. TSCA Title 40 of the Federal Code of Regulations. 1994. Part 796, Section 1840: WaterSolubility. 4. American Society ,for Testing and Materials. 1991. Standard Specification for Reagent Water. D 1193-91, ASTM Section I1 Water and Environmental Technology,Vol. 11.O 1:45-47. WILDLIFE INTERNATIONAL, LTD. - 14- PROJECT NO. 454C- 118 Table 1 Typical LC/MS Operational Parameters INSTRUMENT: Hewlett-PackardModel 1100High Performance Liquid Chromatograph with a Perkin-Elmer API lOOLC Mass Spectrometer operated in Selective Ion Monitoring (SIM) Mode SOURCE: Perkin-Elmer TurboIonSpray ANALYTICAL COLUMN: Keystone PRISM Rp (30 mm x 1.5 mm, 3-pmparticle size) GUARD COLUMN Keystone Javelin C18 cartridge (20 x 2 mm) OVEN TEMPERATURE: 40C STOP TIME: 3.00 min FLOW RATE: 200 pL/& MOBILE PHASE: 25% NANOpure@Water with 0.1%AmmoniumFormate:75%Methanol INJECTION VOLUME: 5.0 pL PFBS PEAK RETENTION TIME: Approximately 1.5 minutes PFBS MONITORED MASS: 299.0 m u WILDLIFE INTERNATIONALLT, D. - 15 - PROJECT NO.: 454C- 118 Table 2 Reagent Blanks Analyzed Concurrently During Sample Analysis Number (454C-118-) REB- 1 Sample T_yp- e Reagent Blank Measured Concentration of PFBS (g PFBSL)' < LOQ REB-2 Reagent Blank < LOQ ' REB-3 Reagent Blank < LOQ The limit of quantitation (LOQ) of 12500 mg PFBSL = 12.5g PFBSL was based upon the product of the lowest calibration standard analyzed (1.00 mg PFBSL) and the dilution factor of the respective blank sample (12500). WILDLI FE INTERNATIONAL, LTD. -16- PROJECT NO.: 454C-118 Table 3 Solubility of PFBS in NANOpure@Water Sample Concentration of PFBS Mean > Number Day of Fortified Measured SD (454C-1 IS-) Test ( PFBSL) cv 1A 1 100 1B 1 100 44.3 Mean = 44.1 SD = 0.40 43.6 CV = 0.92% 1c 1 100 44.3 2A 2 100 2B 2 100 46.1 Mean = 46.1 SD = 0.15 45.9 CV = 0.33% 2c 2 100 46.2 3A 3 100 3B 3 100 Mean=46.3 46.3 SD = 0.15 cv = 0.33% 46.4 3c 3 100 46.1 Totals: Mean (Days 2 and 3)= 46.2 SD = cv = N= 0.18 0.38% 6 Note: Results generated using MacQuan version 1.6 software and manual calculations. Values have been rounded for reporting purposes. WILDLIFE INTERNATIONALL, TD. - 17- PROJECT NO.: 454C-118 METHOD OUTLINE FOR THE PROCESSING OF PFBS IN NANOPURE~WATER Prepare calibration standards in a solution of 50% methanol and 50% NANOpure@water using volumetric flasks and gas-tight synnges. 1 Prepare solubility samples by weighing approximately 10 grams of PFBS into three 8-ounce French square bottles and adding 100 mL of NANOpureQwater to each. After the appropriate equilibration periods, centrifuge triplicate subsamples at approximately 2,000 rpm for approximately 5 minutes 1 Centrihge an aliquot of each subsample at approximately 13000 rpm for approximately 5 minutes 1 Dilute subsample aliquots into the range of calibration With a solution of 50% methanol and 50% NANOpure@water I Transfer samples and standards to autosampler vials for analysis by LCMS. Figure 1 . Analytical method flow chart for the analysis of PFBS in NANOpure@water. WILDLIFE INTERNATIONALLT, D. - 18- PROJECT NO.:454C-118 PWS 299.0 No Internal Standard Weighted ( l / x ) Intercept = 3103767.75000 Slope = 2979999.00000 Correlation Coeff. = 0.9965 ~ ~~ ~~ Area 18000000- 16000000- 14000000- 12000000- 10000000- 8 0 00 0 00- 6000000- 4000000 20 00000- 01 1 1 I I 8 I I I I I I I I I 8 I I 8 6 1 4 1 8 I I I Figure 2. A typical calibration curve for PFBS. WILDLIFE INTERNATIONALL, TD. - 19- PROJECT NO.:454C- 118 intensity: 5000000 cps *01 91 1 20 I 2I1 35 I 4I1 57 I 6I1 1 811 l 10, 1 I 121 14,1 I 161 I Scan 0.35 0.69 1.02 1.36 1.N 203 2.36 2.70 Time Figure 3. A,representative ion chromatogram of a low-level (1.OO mg/L) PFBS standard. WILDLIFE INTERNATIONAL, LTD. -20- PROJECT NO.:454C-118 inlensily: 5000000 Cps 90 2 1 21 4 1 61 81 101 - l i t - 141 ' 1st ' scan 0.35 0.69 1.02 1.36 1.69 2.03 2.36 2.70 Time Figure 4. A representative ion chromatogram of a high-level (5.00 m a ) PFBS standard. WI LD LIFE I NTER NATIoNAL , LTD. -21 - PROJECT NO.: 454C-118 intensity: 5oooOOO cps -1 1 3 ' 2 10. 22 37 49 6 7 90 132 151 170 Figure 5. A representative ion chromatogram of a reagent blank sample (454C-118-REB-1).The arrow indicates the retention time of PFBS. WILD LIFE INTERNATIoNAL, LTD. - 22 - PROJECT NO.: 454C-118 intensity: 5000000 cps :E 89 4 3 2 1 0 1927 21 0.35 41 49 65 1 4'1 6'1 0.69 1.02 81 1.36 101 1.69 121 2.03 141 2.36 16SI1 2.70 scyl Time Figure 6 . A representative ion chromatogram of a Day 2 solubility sample, 454C-118-2A (nominal concentration of 100 g PFBSL. Sample diluted 12500~). WI LDLIFE INTERNATIoNAL , LTD. - 23 - APPENDIX I Protocol and Protocol Amendment PROJECT NO.: 454C-118 WILDLIFE INTERNATIONAL,LTD. - 24 - PROJECT NO.:454C-118 PROTOCOL DETERMINATION OF THE WATER SOLUBILITY OF PERFLUOROBUTANE SULFONATE, POTASSIUM SALT (PFBS) BY THE SHAKE FLASK METHOD OECD Guidelinefor the Testing of Chemicals, 105 Water Solubility 3M Lab Project No. E00-1429 Submitted to 3M Corporation Environmental Laboratory 935 Bush Avenue St. PauL Minnesota 55144 Wildlife International7Ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 July 17,2000 WILDLI FE I NTERNATI ONAL, LTD. SANITIZED - 25 - DEC 0 9 2003 PROJECT NO.: 454C-118 wiIdlife InternationaI, Ltd. DETERMINATION OF THE WATER SOLUBILITY OF PEWLUOROBUTANE SULFONATE, POTASSWM SALT (PFBS) BY THE SHAKE FLASK METHOD SPONSOR: 3M Corporation Environmental Laboratory 935 Bush Avenue St. Paul, Minnesota 55144 SPONSORS REPRESENTATIVE: TESTING FACILITY: Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 STUDY DIRECTOR Raymond L. Van Hoven, PbD. Scientist LABORATORY MANAGEMENT: Willard B. Nixon. Ph.D Manager of Chemistry FOR LABORATORY USE ONLY Start Date: Project No.: 7137 (00 454C-118 Experimental Termination Date: SJ 97 j 0 PROTOCOL APPROVAL LAWRATORY h4ANAGEMENT PROTOCOL NO.: 454/071700/105/SUJ3454 3M LAB PROJECT NO. E00-1429 WILDLI FE INTERNATIO N A L , LTD. - 26 - PROJECT NO.: 454C-118 Wi1d1ije Intemationa1, Ltd. -3 - INTRODUCr'ION Wildlife International, Ltd. will experimentally determine the water solubility of Perfluorobutane Sulfonate,Potassium Salt (PFBS). The study will be conducted at the Wildlife International,Ltd. analyt~cal chemistry facility in Easton, Maryland. The study will be performed based on guidance presented in the OECD Guideline for Testing of Chemicals, 105: WuferSolubility ( I ) 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, Wuter Solubility: Column Elution Method; Shake FIaskMethod ( 2 ) and TSCA Title 40 of the Federal Code of Regulations,Part 796, Section 1840: WuterSolubility (3). Raw data for all work performed at Wildlife International, Ltd. and a copy of the final report will be filed by project number in archiveslocated on theWildlife International,Ltd. site or at an alternative location to be speczed in the final report. OBJECTIVE The objectiveof this study is to determinethe water solubilityof Peduorobutane Sulfo~teP, otassium Salt, hereafter referred to as PFBS, at 20 f 0.5"C by the shake flask method. EXPERIMENTAL DESIGN The water solubility of PFBS will be determined at a temperatureof 20 f 0.5"C. A preluninarytestwill be performed to estimate the water solubility at room temperature. The preliminary test consists of additions of increasingly large amounts of water to a known weight of test substance until solubilization is effected. The definitivetest consists of equilibrationof an excess amount of test substance with water at an elevated temperature,30"C,followed by equilibration at 20C. MATERIALS AND METHODS Test Substance The test and reference items (hereinafter referred to as test and reference substances) will be PFBS. Informationon the characterizationof test and reference substances is r q u i r d by Good Laboratorj~Practice Standards (GLP). T h e Sponsor is responsiblefor providingWildlifeInternational,Ltd. written verification that the test and reference substances have been characterized according to GLPs prior to initiation of the study, If written verificationof GLP test and reference substancecharacterizationis not provided to Wddlife International,Ltd.. it will be. noted in the compliance statement of the final report. PROTOCOL NO.: 454/071700/1OS/SUB454 3M LAB PROJECT NO. E00-1429 WILDLIFE INTERNATIONALLT, D. PROJECT NO.: 454C- 118 Wildlife International, Ltd. -4- Reagents Water that meets ASTM Type I1 standards (ASTM D 1193-9I) will be used (4). Other solventsmay be needed for the analyticalmethod or preparationof stock solutions and analyticalstandards. All solventswill 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 preliminarytest will be conducted to estimatethe solubilityof the testsubstance at room temperature. Approximately 10mg of the test substancewill be placed in a 10-mLglass-stoppered graduatedcylinder 01 flask. Increasingvolumes ofwater will be added stepwise as shown below: Total volume of water added (mL) Approximate solubility (gn) 0.1 0.5 1 >loo 20 10 2 10 5 1 After each addition of water to yield the total volume indicated,the containerwill be shaken vigorously for approximately IOminutes and visually examined for undissolvedtest substance. Lfthe test substanceis not dissolved after addition of 10 mL of water, the contents of the container will be transferred to a 100-mL cylinder or flask and water added for a final volume of 100mL. Followingshaking, the flask or cylinderwiu be visuallychecked for un&ssolved material,i.e., undissolvedmaterialwould indicatea water solubility4 . 1 g5. Preliminary testing of the solubility at lower concentrations (<0.1 g5)may be performed by bansferring the contents of the 100-mL cylinder or flask to a 1-L vessel or reinitiating the test with a lesser amount of test substance, e.g., 1mg. Definitive Test Procedure The quantity of test substance necessary to saturate the desiredvolume of water is estimatedfrom the preliminary test. T h e volume of water used will depend on the analytical method and solubility range. Greater than (or equal to) five times the quantityof test substancerequired to achieve water solubilitywill be transferred to a minimumof three glass or Teflon@test vessels. 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 f 1.O"Cin a water bath. After 1 day, one of the vessels will be removed and re-equilibrated for 24 hours at 20 * 0.5"C PROTOCOL NO.: 454107 1700/105/SUB454 3M LAB PROJECT NO. E00-1429 WIL D L l FE INTERNATIONAL, LTD. PROJECT NO.:454C- 118 WiLirife InternationaI,L,td. -5- with occasionalshakingor stirring. The contentsof the vessel will then be centrifuged. The concentrationof the test substancein a minimum of two aliquotsofthe aqueousphase will be determined.Theremaining two vessels are treated similarly following initial equilibration periods of 2 and 3 days at 30 f 1.0"C. If the concentrationresults from the last hvo vessels agree to within 15%, the test is satisfactory.Iftheresults &om the three vessels show a trend of increasing values, andor at the request of the Sponsor, the test may be reinitiated with longer equilibration periods. Method The analytical method to be used for quantitation of the test substance will be LClMS based on procedures developed at Wildlife International, Ltd. The method used was developed at Wildlife International, Ltd. and entitled "Analytical Method Validation for the Determination of Perfluorobutane Sulfonate, Potassium Salt (PFBS) in Freshwater" (Wildlife International,Ltd. Project No. 454C-115). Calculations The concenbation of the test substance in each vessel will be expressed in g5,m g 5 or p g L inwateras 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. Sample and Test Substance Retention Upon completionof testing, portions of the test substanceused as part of this study will be dqosedof in accordancewith 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 of the signed protocol. PROTOCOL NO.: 454/07 1700/105/SUB454 3M LAB PROJECT NO. E00-1429 WILD LIFE INTERNATIoNAL , LTD. - 29 - PROJECT NO.: 454C- 118 WildliJeInternational, Ltd. 2. Identification and characterization of the test substance, if provided by the Sponsor. 3. Dates of initiation and completion of the study. 4. Dates of experimental start and termination. 5. Storage conditions of 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. IO. Test conditions. 11. A copy of the final report. FINAL REPORT Wildlife International, Ltd will prepare a final report of the results of the study. 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 statementof compliancesigned by the StudyDirector addressingany exceptionsto GoodLaboratoxy Practice Standards. 4. Purpose and procedure,as stated in the approved protocol,includingall amendmentsand deviationsto the protocol. 5 . The test substanceidentification, includmg name OKcodenumber,purity,empkal fomulqmolecular formula, lot or batch number, method of analysis,and any other informationprovidedby the Sponsor. 6. Description of the test method or reference to the method used along with any molfications made. 7. The individualconcentrationsof each sample. 8. The means and standard deviations of solubility determinationsat each interval for each temperature tested. 9. Description of any problems experienced and how thq. were resolved. 10. A statementprepared by the Quality AssuranceUnit listing the dates that study inspectionsand audits were made and findings reported to the Study Director and Management. PROTOCOL NO.: 454/071700/105/SUB454 3M LAB PROJECT NO. E00-1429 WILDLIFE INTERNATIONALL, TD. - 30 - PROJECT NO.:454C- 118 WiId 1ije InternationaI, Lid. CHANGING OF PROTOCOL Planned changes to the protocol will be in the form of written amendmentssignedby the StudyDuector 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 LABORATORYPRACTICES This study will be conducted according to the Good Laboratory Practices described in OECD (ENVRVICICHEM(98) 17). Each studyconducted by Wildlife International, Ltd. is routinely examined by the Wildlife International; Ltd. Quality Assurance Unit for compliance with Good Laboratory Practices, StandardOperatingProcedures and the specifiedprotocol. A statementof compliancewithGood Laboratory Practiceswill beprepared for all portions of the study conductedby WildlifeInternational,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 PROTOCOLNO.: 454/07 1700/105/SUB454 3M LAB PROJECT NO. E00-1429 WILDLIFE INTERNATIONAL, LTD. - ^. 31 - PROJECT NO.: 454C-118 WiZdlfe International, Ltd. -8REFERENCES 1 Organisation lor Economic Cooperation and Development, 1995. Guideline for Testing of Chemicals, 105: Wuter Solubility. 2 Product Properties Test Guidelines. 1996. OPPTS 830.7840. Wurer Solubility: ColumnElution Method; Shake FlaskMethod. 3 TSCA Title 40 of the Federal Code of Regulations. 1994. Part 796, Section 1840. Wuter Solubility. 4 American Society for Testing and Materials. 1991. Standard Specification for Reagent Water. D1193-91, ASTM Section I1 Water and EnvironmentalTechnology,Vol. 11.01: 45-47. PROTOCOL NO.: 454/071700/105/SUB454 3M LAB PROJECT NO. EOO-1429 WILD LIFE I NTERNATIoNAL , LTD. - 32 - PROJECT NO.: 454C-118 Wildlife International, Ltd. WLI Project No.: 454C-118 Page 1 of 1 DEVIATION TO STUDY PROTOCOL STUDY TITLE: Determination of the Water Solubility of Perfluorobutane Sulfonate, Potassium Salt (PFBS) by the Shake Flask Method PROTOCOL NO.: 454/07 1700/105/SuB454 DEVIATION NO.: 1 SPONSOR: 3M Corporation PROJECT NO.: 454C-118 DATE OF DEFACTO DEVIATION: July 27,2000 DEVIATION: Page 4, Preliminary Test Procedure The protocol states that the container will be shaken vigorously for approximately 10 minutes and visually examined for undissolved test substance upon each addition of water. In the experiment, the container was shaken for only approximately one minute. REASON: Analyst oversight. IMPACT: In the opinion of the Study Director, the cited deviation during the preliminary test had no impact on the design and conduct of the definitivetest. Raymond L. Van Hoven, Ph.D. Willard B. Nixon, Ph.D WILDLIFE INTERNATIONAL,LTD. SANITIZED PROJECT NO.:454C-118 APPENDIX I1 Certificate of Analysis C ~ F ~ S O ~ - K + Lot 1; PFBS) Lot 2 (1999) April 17,2000 (Supcrdcs ccrtifigtc of Ardysis dated April 11,ZOoO) This sample was analyzed using and elemental analyses techniques. The results of these tests show the sample to contain the follolvlng weight percent composition: 0.13 % c4Fs.so,x+ 97.900/. I I 1.96 % Additionally, the isomer distribution of the sample was determined using "F-NMR techniques and found to contain the following weight percent composition: I (Normal chain) I Analytical Study Number: 60488 WILDLIFEINTERNATIONALL, TD. PROJECT NO.: 454C-118 APPENDIX 111 Personnel Involved in the Study The following key Wildlife International, Ltd. personnel were involved in the conduct or management of this study: 1. Willard B. Nixon, Ph.D., Manager, Analytical Chemistry 2. Raymond L. VanHoven, PbD., Scientist 3. Jon A. MacGregor, B.S.,Scientist 4. Frank J. Lezotte, B.S., Chemist