Document DGvV9Yd7mVZ4dQ7pZa548kemO

BACK TO MAIN Study Title Solubility Study of Perfluorooctanesulfonate(PFOS), SD-018 and Perfluorooctanesulfonamide(FOSA), SE-027 in Male/Female Human Urine Data Requirement 40 CFR 160.105(b) Author Mark E. Ellefson Phase Completion Date Date of signing Performing Laboratory 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Environmental Laboratorty Project# E01-0768 3M LIMS#: 25160 (PFOS SD-018), and #25161 (FOSA SE-027) Total Number of Pages 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 TABLEOF CONTENTS Study Information ......................................................................................... ................................. 4 Summary ........ ................................................................................... 5 Purpose ...................................................................................................................... Test Substance ............................................................................... .................................... 5 Reference Substance......................................................................................................................... 6 Test System. .............................................................................................................................. 6 Method Summaries .................................................................................................. ...................7 Preparatory Methods........................................ ... .................................................... 7 Sample Collection and Analysis ........................................................................................ 8 Analytical Method .................................................................. .............................. 10 Analytical Results ............. .............................................................................. 10 Data Summary .......................................................................................................... Statistical Methods ............................................................................................................................. 14 Statement of Conclusion ....... ................................................................. References ..... ........................................................................................................................... 15 List of Attachments ................................................................ .............................. 15 Signature Page.................................................... ....................................................................... 16 Page 2 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 LISTOF TABLES Table l a . Summary Table of Solubility of PFOS SD-018 in Urine. .................................................... 5 Table 1b. Summary Table of Solubility of FOSA SE-027 in Urine. .................................................... 5 Table 2. Characterizationof the Test Substance.............................................................................. 6 Table 3. Characterizationof the Analytical Reference Substances ....................................... ........... 6 Table 4. Description of the test system used in this study ...................................... Table 5. Solubility Screen Test Sample Preparation ............ ................................. 7 Table 6. Shake Flask Method Preparation for Study ........................................................................ 8 Table 7. Sample Collection and Preparation for Solubility Study .......................................... .... 9 Table 8a. PFOS Calibration Standard Reprocessing Summary ...... .................................. 11 Table 8b. FOSA Calibration Standard Reprocessing Summary .......................... . .... 12 Table 9a. PFOS SD-018 Solubility in Urine Data Summary .. ............................................ 13 Table 9b. FOSA SE-027 Solubility in Urine Data Summary ............................................................. 13 Table loa. Summary Table of Solubility of PFOS SD-018 in Urine. ....................... Table 1Ob. Summary Table of Solubility of FOSA SE-027 in Urine ................................ 14 Page 3 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 STUDYINFORMATION Analytical Chemistry Laboratories Solubility Jesting 3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory Bidg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Dr. William K. Reagen, Laboratory Manager Mark E. Ellefson, Principal Analytical Investigator Kristin L. Terrell, Analytical Chemist Mark L. Anderson, Analytical Chemist Cindy M. Carlson, Analytical Chemist Sponsor 3M Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, MN 55106 Dr. William K. Reagen Experimental Dates Experimental Start Date: 23 April 2001 Experimental Termination Date: 01 May 2001 Arc hives Reserve samples of the reference standards will be maintained by the 3M Environmental Laboratory. All study data and a copy of the final report are archived by the 3M Environmental Laboratory. Page 4 of 171 BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 SUMMARY The approximate solubility of Perfluorooctanesulfonate(PFOS) and Perfluorooctanesulfonamide FOSA in human urine was determined by performing a solubility screen test, followed by a quantitative solubility determination via Shake Flask Method. Results are presented in table l a and Ib: Table l a . Summary Table of Solubility of PFOS SD-018 in Urine. Day 1 302pgImL 12 4% Day 2 325 pghL 40 12% Day 3 287 pg/mL 30 10% Average Value 305 pglmL 14 5% The solubility of PFOS SD-018 in urine was determined to be 305 pg/mL at 23-24 "C. Table 1b. Summary Table of Solubility of FOSA SE-027 in Urine. I Day 1 I Day 2 Day 3 Average Value 2.56 uo/mL 2.88 pg/mL 2.61 udmL 2.68 pglmL I 0.38 I 0.12 0.05 0.18 15% I 4% 2% 7% PURPOSE The purpose of this solubility study was to determine the solubility of PFOS and FOSA in urine. TESTSUBSTANCE PFOS, Test Control Reference #SD-018 and FOSA, Test Control Reference # SE-027 Page 5 of 171 BACK TO MAIN 3M Environmental Laboratory Project# E01-0768 Table 2. Characterizationof the Test Substance ~ source 98-021 1-0888-5 Expiration Date 08/31101 Stora-ge Conditions Frozen 3M Laboratory Identification Number SD-018 I Physical Description Purity I White powder 1 86.9% REFERENCSEUBSTANCE R. Buckanin, 3M Not Available Frozen SE-027 1 I Whitepowder/chunks ~ To Be Determined Table 3. Characterizationof the Analytical ReferenceSubstances Source Expiration Date Re-crvstallized bv George Moore, 3M Specialty Materials, BldQ. 236-1B-I0 01-01-2010 II yiBuckanin' II SynQuestLabs I I Not Available 1 I Not Available Storage Conditions 3M Laboratory Identification Number Physical Description Purity Frozen TCR-00017-46 White Powder 97.9% 1 Frozen SE-027 I ?i:er/chunks To Be Determined 1 Frozen TCR-OOO17-047 Lot #Q75-30 II Whitepowder * The purity of T H fF( analykal findings since it is used as a reference substance only. I Pat Savu, 3M 1 I Not Available Frozen Primary Standard, TCR99030-23 (L-7038, I F-8442): Lot 101 WhitePowder TESTSYSTEM Table 4. Descriptionof the test system used in this study Expiration Date I I 03-16-05 I Storage Conditions Frozen 3M Laboratory Identification Number Purity/Grade TN-A 4822 ~ Normal MaleFernaleUrine Page 6 of 171 BACK TO MAIN 3M EnvironmentalLaboratory Project# E014768 METHODSUMMARIES The solubility determination of PFOS SD-018 and FOSA SE-027 in urine was performed according to 3M Environmental Laboratory methods ETS-8-170.1 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents;" and ETS-8-172.1 "Shake Flask Method; Solubility Determination of a Test Substance in Various Solvents" as adapted from United States Environmental Protection Agency OPPTS 830.7840, "Water Solubility: Column Elution Method; Shake Flask Method" and OECD 105, "Water Solubility" Guidelines. All analyses were performed according to the 3M Environmental Laboratory method ETS-8-155.0 "Analysis of Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-Electrospray/MassSpectrometry". The Solubility Screen Test Method (ETS-8-170.1) was used to determine the solubility range of PFOS SD-018 and FOSA SE-027 in urine. Incremental amounts of urine were added to PFOS and FOSA neat material and a qualitative determination of the solubility point was made. The screen test indicated a solubility of PFOS in urine of less than 200 pg/mL, and FOSA in urine of less than 200 pg/mL at 23 "C. Therefore the Shake Flask Method was used for the quantitative determinationof PFOS and FOSA solubility in urine. Detailed descriptionsof the Solubility Screen Test Method and Shake Flask methods used in this study are located in Attachment A. PREPARATORMYETHODS ETS-8-170.1 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents." This method is a prerequisite to the shake flask method and gives an estimate of the solubility point of the test substance in the solvent of choice. Approximately ten milligrams of PFOS/FOSAwas weighed into glass vials, and varying amounts of urine were added in a step-wise fashion. After each urine addition, the vials were shaken, sonicated, and observed for particulate. When particulatewas observed in a final volume of 2 mL of urine (approximately 5000 pg/mL) for both PFOS and FOSA, the final step of the screen test was initiated. Approximately 10 mg PFOS/FOSAwas placed into a 100 mL volumetric flask and urine was added in a step-wise fashion up to 50 mL (with shaking, vortex-mixing, sonicating, and observations for particulate in between). Due to fine particulates present in the urine, it was difficult to tell the difference between urine that did and did not have the test substance in it. This indistinguishable difference between FOSA and PFOS in urine and a urine blank sample was determined to be the stopping point for the screen test. The approximate solubility of PFOS and FOSA in urine at 23 "C was 200 pg/mL. This concentration estimation was used as a starting point for the shake flask method, and as a guide to determine the amount of dilution necessary to bring the final sample concentration into the analytical range of method ETS-8-155.0 (approximately 2.5 - 1000 ng test substance/mL). Table 5. Solubility Screen Test Sample Preparation I 1 I 10 mg test substance weighed into tared 4.0 mL glass screw-capped vial I I 2 I 0.1 mL of urine added to glass screw-capped vial I 3 Followingthe additionof urine,vial was capped,shaken vigorously and vortexed,and sonicated. Mixing and sonication times recorded on the worksheet. Page 7 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Table 5. Solubility Screen Test Sample Preparation 14 5 6 7 8 Sample checked visually for undissolved particles of the test substance. (If all of the test substance is dissolved,the solubility is determinedto be greaterthan 10% (mass/volume) and no additional solubility testing is required. If a portion of the test substance remains undissolved, additional dilutions are performed using the appropriate solvent.) Obselvation made: undissolved particlespresent... continueto step 5. Step 2 repeatedwith total volume of urine of 0.5,1, and 2 mL, respectively. Following urine addition, vial was shaken, sonicated, and observed for particulate. Followingthe final urine additionto bringthe volume of urine to 2 mL, there was still undissolved PFOS and FOSA. The screentest methodwould therefore be continued to step 7. Approximately 10 mg PFOSlFOSA were weighed into 100 mL volumetric flasks and urine was added stepwise up to 50 mL. Followingeach addition of urine the flask was capped, vortex-mixed,sonicated, and allowedto settle. The appearanceof urinewithout test substance looked about the same as urine with PFOS or FOSA at 50 mL. The concentrationof urinewas approximately200 pg/mL and there was still particulate. The shake flask method shall be usedfor the quantitativedeterminationof PFOS/FOSA solubility in urine. ETS-8-172.1 "Shake Flask Method: Solubility Determination of a Test Substance in Various Solvents." Approximately 10 mg test substance was weighed into a tared 15 mL plastic centrifuge tube. Urine was added gravimetrically to approximately 4.7-5 g (approximately 5 mL if density of urine is assumed to be approximately 1 g/mL). For each test substance study, nine centrifuge tubes were prepared, along with three method blanks (tubes with 5 g f 0.5 g urine only). The centrifuge tubes were placed in a temperature- controlled orbital shaker set at approximately 30 OC. See the following table documenting the shake flask method preparation: Table 6. Shake Flask Method Preparationfor Study 9 FOSA 2 Urine added to centrifuge tube such that total weight of PFOSlFOSAand urine is approximately5 g. 9 PFOS/ Q FOSA 04-23-01 3 Method Blank Preparation-Approximately5 g urine addedto a 3 PFOS/ tared centrifuge tube gravimetrically. 3 FOSA 04-23-01 4 Samples sealed with tape, vortex-mixed, and placed in a 12 PFOS 04-23-01 temperature-controlledorbital shaker set at approximately30.0 OC, /I2 FOSA rotationspeed set at approximately150 rpm. SAMPLE COLLECTION AND ANALYSIS On each of Days 1, 2, and 3 (approximately 24, 48 and 72 hours after initial sample preparation) three centrifuge tubes (3 containing PFOS and 3 containing FOSA) and a method blank were removed for analysis. The centrifuge tubes with PFOS and FOSA in urine, and method blanks (urine only) were allowed to equilibrate at 23-24 OC for 24 hours, centrifuged, and aliquots of Page 8 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 supernatant were collected. For the PFOS samples, aliquots were first diluted IOOX (990 pL methanol: 10 pL sample), and then diluted further to 1,OOOX (900 pL methanol: 100 pL sample). For the FOSA samples, aliquots were diluted 1OX (900 pL methanol: 100 pL sample). Final dilutions were spiked with internal standard (17 pL of 15 pg THPFOS and PFBS/mL) and every fourth sample replicate was spiked with 4 pL test analyte (25.05pg/mL PFOS, 24 pg/mL FOSA). Samples were analyzed via HPLC/ES/MS according to analytical method ETS-8-155.0. Four injections (three samples and one sample spike) of each sample were performed. The following table describes the sample collection and preparation regimen: Table 7. Sample Collection and Preparationfor Solubili 1 At 24,48, and 72 hours, 3 test substance+ urine, and 1 urine (method blank) centrifuge tubes per PFOS and FOSA are removedfrom &IStudy 4 centrifuge tubes per day for the incubator and equilibrated at 23-24 OC for approximately 24 hours. PFOS and FOSA 2 Aliquots of the supernatant (urine containing 4 per dissolved test substance or urine only from the method blanks) were collected and dispensed into autovials. tube per FOSA and 04-24-01 04-26-01 PFOS 3a II II PFOS The PFOS aliquots were diluted 1OOX with methanol (using a dual syringe diluter: 990 pL methanol, I O pLsample), followed by a Day 1 Day 2 04-26-01 0501-01 second dilution of 1OX (900 pL methanol, 100 pLsample) for a final dilution factor of 1,OOOX. Day 3 04-26-01 17 pL THPFOS and PFBS solution at 15 pg/mL in methanol were then added to the final dilutions, and every fourth sample was spiked with 4 pL 25.05 pg/mL PFOS 3b FOSA 4 The FOSA aliquots were diluted 1OX with methanol (using a dual syringe diluter: 900 pL methanol, 100 pLsample), 17 pL THPFOS and PFBS solution at 15 pg/mL in methanolwere then added to the final dilutions, and every fourth samplewas sDiked with 4 UL24 w/mL FOSA. Sampleswere analyzedvia HPLCESNS on HP 1100 LCMSD Day 1 Day 2 Day 3 04-26-01 04-26-01 05-01-01 PFOS: 0501-01/FOSA: 04- Day 3 PFOS:04-30-011FOSA: 0 5 01-01 Page 9 of 171 BACK TO MAIN 3M Environmental Laboratory Project# E01-0768 ~ ANALYTICAML ETHOD 0 ETS-8-155.0 "Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLG-Electrospray/MassSpectrometry". Diluted samples (i.e. diluted supernatants) were analyzed using HPLCIESIMS in the negative ion mode. PFOS levels were evaluated versus calibration standards ranging in concentration from 2.5-1000 ng PFOS/mL. FOSA levels were evaluated versus calibration standards ranging in concentrationfrom 2.5-1000 ng FOSNmL. Internal Standard quantificationwas used to normalize the data. PFBS was chosen as the internal reference standard to quantify the data, THPFOS was added as an optional internal standard during preparationfor analysis, but was not used to quantify the data. Target ions were 499 m/z (PFOS anion), 498 m/z (FOSA anion), 427 m/z (THPFOS anion), and 299 m/z (PFBS anion). Analytical Equipment Liquid Chromatograph: Hewlett-Packard@Series 1100 Liquid Chromatograph system Analytical column: Keystone@BetasilTMC182x50mm, 5vm particle size Column temperature: Ambient Cycle Time: 10 minutes Flow rate: 300pL/min Injection volume: 1OpL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in water Solvent C: Methanol Solvent Gradient: Time Yo c 0.00 15 % 0.50 15 % 0.80 60 % 3.50 100 % 6.50 100 % 7.00 15 % 10.00 stop Mass Spectrometer: Hewlett-Packard- Series 1 100 APllMass Spectrometer Detector Software: HP ChemStation" 6.0 FragmentorVoltage: m/z 299=100 V; m/z 427=100 V; m/z 499= 140 V; mlz 498=80 V Capillary Voltage: 3500 V Gain = 2.0 EMV Mode: Electrospray Negative Gas Temperature: 350 *C Drying Gas: 10.0 L /min. Nebulizer Pressure: 25 psig Analysis Type: Single Ion Monitoring (SIM) ANALYTICARL ESULTS Data quality objectives outlined in the 3M Environmental Laboratory method were met (see Appendix A). Page 10 Of 171 BACK TO MAIN 3M Environmental Laboratory Project # EOi-0768 Regressions. Quadratic curve fit was applied to calibration standards and sample data to improve quantitation over the concentration range appropriate to the data. All calibration curves had least-square fits of 0.980 or greater (R2 values ranged from 0.9993-0.9998). Calibration Standards. Twelve standards ranging in concentration from approximately 2.5 to 1000 ng/mL PFOS/FOSA in methanol were used for the calibration curves. Calibration curves were run before and after every analytical sequence. Calibration standards were also evaluated for accuracy, and were consistently I 30% difference from the expected value. Lab Control Spikes. 1 mL methanol was spiked with 4 pL 25.05 pg/mL PFOS spike solution, and another 1 mL was spiked with 4 pL 24 pg/mL FOSA spike solution, (the same 25 ppm spike solutions spiked into every fourth replicate for sample recoveries). The lab control sample was used to evaluate the recovery of the test substance in methanol only. Recoveries for PFOS ranged from 89-90%, and recoveries for FOSA ranged from 103-111% for Days 1,2, and 3. Matrix Spikes. Matrix spikes were prepared to demonstrate any possible matrix affects (i.e. signal suppression or enhancement) from the urine. Matrix spike recoveries for PFOS and FOSA samples were as follows: k 30% with the following PFOS sample exceptions: 042301-PFOS3.1.4MS (65%),042301-PFOS-2.1.4(52%),042301-PFOS-2.2.4 (135%), 042301-PFOS-2.3.4 (53%). The MS samples from day 2 were done with a 5 ULsyringe that seemed to have a loose plunger, therefore giving erratic results. Excluding the Day 2 PFOS spike recoveries may be disregarded. Overall, there appears to be no significant difference between Lab Control Spikes (test analyte spiked into methanol only) and Method Blank (test analyte spiked into urine only), or Sample Spikes (test analyte spiked into urine samples containing test analyte). Sample Replicates. Samples were within the 15% precision requirement. Continuing Calibration Verification. For quantitative determinations, a mid-level matrix calibration check was analyzed every ten samples to monitor instrumental drift, with a limit of k30% deviation of the target concentrations. Limit of Quantitation (LOQ). The LOQ is equal to the lowest standard in the calibration curve. See tables 8a and 8b for the calibration curve ranges used in the study. Table 8a. PFOS Calibration Standard ReprocessingSummary Day 1 data: Day 2 data: Day 3 data: I PFOS: 5501 ng/mL I PFOS: 10-501 ng/mL PFOS: 5501 ng/mL 2.5 8 1000 ng/mL since only 10 calibration levels may be reprocessed using Target data analysis software. 2.5 & 1000ng/mL since only 10calibrationlevels may be reprocessed in Target. 5 ng/mL std. was disabled because the 5 ng/mL CCVs did not meet the 30% accuracy requirement. (Data at the hgh end of the curve passed all criteria). 2.5 & 1000ng/mLsince only 10 calibrationlevels may be reprocessed in Target. Page 11 of 171 BACK TO MAIN 3M Environmental Laboratory Project # EO1-0768 Table 8b. FOSA CalibrationStandard ReprocessingSummary Day 1and 2 data: IIDay 3 data: 0 Blanks. FOSA: 10499 FOSA: 10499 2.5, & 1000 ng/mLwere not used as calibrantssince Target allows 10 levels only. The 5 ng/mL standard was disabled due to an elevatedlevelof FOSA in some blank samdes. ~_______ 2.5,5, & 1000 ng/mL standards were excluded/disabledto remain consistentwith Day 1 and 2 data. _______~ 0 Method blanks (urine taken through the entire sample preparation, dilution, and analysis process) provided a measure of laboratory contamination. Acceptable values for the blanks were less than 50% of the limit of quantitation (LOQ). Method blank samples for Day 3 FOSA had a level of FOSA (20,18,and 15 ng FOSNmL on column) that was most likely due to FOSA dust in the analytical balance on the day of initial prep of the centrifuge tubes. Method blank samples from Days 1 and 2,a diluter blank, and internal standard blank ruled out all other possible forms of contamination. Therefore since it was determined that the FOSA was not in the urine prior to the sample preparation, this finding (Le. FOSA in the day 3 method blank samples) is inconsequential. 0 Solvent blanks (methanol injections) provided a measure of instrument contamination. Acceptable solvent blanks must contain levels of target analyte less than 50% of the limit of quantitation (LOQ). Acceptable solvent blanks were analyzed before each calibration curve. 0 Specificity (according to OPPTS 830.7840): The solubility determination as stated in the EPA Guidelines follows. This method should be applied to: 0 Pure substance. Purity of PFOS SD-018has been determined to be 86.9%. The purity of FOSA SE-027has yet to be determined. Since it is possiblethat impurities may exist in the FOSA, and the PFOS SD-018is 86.9% pure, the reported solubility may be greater than that of a pure substance. 0 Substances that are stable in water. Hydrolytic studies have been conducted for PFOS TCR-0001746 (primary standard material, Report # Wl878) and FOSA (Project #EL-I 132) at the 3M Environmental Laboratory showing stability. DATASUMMARY Tables 9a and 9b summarize individual sample data. Representative chromatograms are presented in Attachment C. The table displays PFOS and FOSA concentrations for individual injections. Also included are the average concentrations for each Centrifuge Tube replicate, standard deviation, and % coefficient of variation. Page 12 of 171 Table 9a. PFOS SD-018 Solubility in Urine Data Summaw 042301-PFOS-1.1.1 Smp 286 Day Tube #1 042301-PFOS-1.I.2 Dup 325 042301-PFOS-l. 1.3 Trip 323 042301-PFOS-1.2.1 Smp 292 Tube#C2entrifuge 042301-PFOS-1.2.2 Dup 289 ' 042301-PFOS-1.3.1 Smp 314 TDuabye#C3entrifuge 042301-PFOS-1.3.2 Dup 301 042301-PFOS-1.3.3 Trip 302 042301-PFOS-2.1.ISmp 305 Tube #1 042301-PFOS-2.1.2 Dup 412 042301-PF( ~~ ~ 042301-PFOS-2.2.1 Smp 289 Tube #2 042301-PFOS-2.2.2 Dup 288 042301-PFOS-2.2.3 Trip 263 Day2 Centrifuge 0o442301-PFOS-2.3.1 Smp 332 Tube#3 nAA2.2330011--PPFFOOSS--22..33..32DTrUipD 33~~~ 3485 042301-PFOS-3.1.ISmp 262 TDIahya%1C e n ~ g e 042301-PFOS-3.1.2 Dup I""TZ7rI 287 042301-PFOS-3.1.3 Trip 259 042301-PFOS-3.2.1 Smp 302 TDuabye3#C2entrifuge 042301-PFOS-3.2.2 Dup 352 042301-PFOS-3.2.3 Trip 31 2 Day 3 Centrifuge 042301-PFOS-3.3.1 Smp 286 Tube #3 042301-PFOS-3.3.2 DUP 257 042301-PFOS-3.3.3 Trip 270 Table 9b. FOSA SE-027 Solubility in Urine Data Summary TDuabye#C1entrifuge -Davl ' ' _ Tubej tLLenunuge Tube #C3 entrifuge berlulluye Tube#, 14I ". DayZL- enunuge TubeLn I Day2 c-enm.-ge 1- 042301-FOSA-1.1.1 Smp 042301-FOSA-1.I.2 Dup 042301-FOSA-1.I.3 Trip 042301-FOSA-1.2.1 Srnp 042301-FOSA-1.2.2 Dub 042301-FOSA-1.2.3 Trip 042301-FOSA-1.3.1 Srnp 042301-FOSA-1.3.2 Dup nA7?nl-FnSA-l.3.3 Trip U~LJUI-ruSA-2.1.1Smp 042301-FOSA-2.1.2 DUP 042301-FOSA-2.1.3 Trip 042301-FOSA-2.2.1 Srnp 042301-FOSA-2.2.2 DUP 042301-FOSA-2.2.3 Trip 042301-FOSA-2.3.1 SmD 042301-FOSA-2.3.2 DUD 042301-FOSA-2.3.3 Trip 3d3 2.23 2.13 3.09 3.13 3.12 2.48 2.33 2.30 3.17 2.96 2.91 2.98 2.80 2.69 2.89 2.79 2.75 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 31 1 22.0 7% 288 4.0 306 7.2 2% 358 54 280 15 5% 338 7 2 Yo 269 15.4 6% 322 26.5 8% 271 14.5 5% 2.18 4.6 % RPD (Rep 1 excluded from calc.) 3.1 1 0.0 1% 2.37 0.1 4% 3.02 0.1 5% 2.82 0.1 5% 2.81 ~~ 0.1 3% Page 13 of 171 BACK TO MAIN 3 M Environmental Laboratory Project # E01-0768 -7 Day3 Centrifuge 042301-FOSA-3.1.IS,mp 2.69 2.60 Tube #I 042301-FOSA-3.1.2, DUP 2.51 0.1 042301-FOSA-3.1.3, Trip 2.60 3% Day3 Centrifuge 042301-FOSA-3.2.1, Smp 2.57 042301-FOSA-3.2.2. DUD 2.61 042301-FOSA-3.2.3, Trip 2.51 Day3 Centrifuge 042301-FOSA-3.3.1, Smp 2.63 2.66 T u k #3- 042301-FOSA-3.3.2, DUD 2.74 042301-FOSA-3.3.3, Trip 2.60 3 % The average PFOS and FOSA concentrations and standard deviations for each day, as well as the final overall average for all non-excluded replicatesand time points were as follows: Table loa. Summary Table of Solubility of PFOS SD-018in Urine. Day 1 302 pg/mL 12 4% Day 2 325 pg/mL 40 12% Day 3 287 pg/mL 30 10% Average Value 305 pglmL 14 5% Table lob. Summary Table of Solubility of FOSA SE-027 in Urine. I Day 1 I Day 2 Day 3 Average Value 2.56 pg/mL 2.88 pg/mL 2.61 pg/mL 2.68 pglmL 0.12 0.047 0.2 15% I 4% 2% 7% Attachment B contains data summary tables. STATISTICAMLETHODS Statistical methods included calculating means and standard deviations. Outlier data points were excluded using Dixon's Q-Test. Refer to Attachment E for formulas and example calculations. STATEMENT OF CONCLUSION Under the conditions of the present study, the solubility of PFOS SD-018 in urine is 305 pg/mL at 23-24"C. The solubility of FOSA SE-027 at 23-24"Cis 2.68pg/rnL. Page 14 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 REFERENCES 1. Fate, Transport and Transformation Test Guidelines Office of Prevention, Pesticides and Toxic Substances (OPPTS) 830.7840 Water Solubility: Column Elution Method; Shake Flask Method. EPA 712-C-96-041, August 1996. 2. OECD 105: Water Solubility. Adopted 27July, 1995. LISTOF ATTACHMENTS Attachment A: Extraction and Analytical Methods 0 Attachment B: Data Summary Tables Attachment C: Sample Chromatograms Attachment D: Deviations from the Protocol Attachment E: Sample Calculations Page 15 of 171 SIGNATURPEAGE ~ _ _ _ _~~ BACK TO MAIN 3M Environmental Laboratory Project #EI01-0768 We certify that this report is a true and complete representation of the data for this phase of the study: Principal Analytical Investigator - William K. Reagen Laboratory Manager Date Page 16 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 ATTACHMENAT: EXTRACTIOANND ANALYTICAML ETHODS Page 17 of 171 3M ENVIRONMENTLAALBORATORY BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 SOLUBILITSYCREENTEST:APPROXIMATSEOLUBILITDYETERMINATIOONF A TESTSUBSTANCE IN VARIOUS SOLVENTS Method Number: ETS-8-170.1 Adoption Date: 09/08/00 Effective Date: 03/14/01 Approved By: Wil6am K. Reagcn, Laboratory Manager Date 1.0 SCOPE AND APPLICATION 1.1 Purpose. According to methods set forth by the United States Environmental Protection Agency (US EPA) and the Organization for the Economic Cooperation and Development (OECD) a preliminary study, which will be outlined in this method, serves as a prerequisite to performing solubility testing via the Column Elution Method and Shake Flask Methods. The EPA and OECD solubility testing guidelines state that if the preliminary solubility determinationtest indicates a solubility of > 10-2g/L (10 ppm), the Shake Flask Method (ETS-8-172.0) is to be used. If preliminary testing indicates a solubility of < g/L (10 ppm) the Column Elution Method (ETS-8-171.0) will be used to determine the solubility in the solvent of interest. Additionally, due to the tendency of certain compounds to be highly soluble in particular solvents, no further solubility determinationwill be necessary for compounds having solubility greater than 10% (mass/volume). 1.2 Compatible Analytes. Test substance and degradation products for solubility testing. ETS-8-170.1 Solubility Determination: Screen Test Method Page 1 o f 9 Page 18 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 1.3 Acceptable Matrices. Aqueous (e.g. Milli-Q water, 0.01M CaCl,), Acetone, Methanol, or other solvent(s) of interest. 2.0 SUMMARY OF METHOD 2.1 Preliminary Solubility Determination in solvent. Weigh approximately 10 mg of test substance into a 2.5 mL-4 mL glass screw-capped vial. Add test solvent in varying increments to the vial. After each solvent addition, vortex mix for approximately 15 sec., and sonicate 2-5 minutes. Make observations of the solution and visually confirm whether or not particles are present; it may be necessary to allow the solution to sit for up to 5 minutes. Add test solvent in a stepwise fashion up to 2 mL. If the substance hlly dissolves after the first solvent addition of 100 pL, the test substance shall be considered highly soluble, and no further testing is required. If there are still undissolved particles in the vial, weigh 10 mg of test substance into a 100 mL glass stoppered volumetric flask or graduated cylinder. Add test solvent until visual confirmation of the substance dissolution has been achieved. Record all observationson a standardized preparation sheet or logbook. Once 100 mL solvent has been reached, the approximateconcentration is at 100 pg/mL. If the solubility limit has not yet been achieved, weigh approximately 10 mg of test substance and transfer to a 1 L graduated cylindedvolumetric flask (concentration is approximately 10 ppm). The container should be sonicated and allowed to sit overnight to allow for maximal dissolution. If the undissolved particles are still observed after sitting > 12 hours, the column elution method will be utilized (ETS-8-171.0). If the test substance dissolves in 1 L or less of solvent, the shake flask method (ETS-8-172.0) will be used to determine the solubility. 3.0 DEFINITIONS 3.1 Test substance. A liquid or solid material for which the relative solubility in a specified solvent will be determined. 3.2 Test solvent. The matrix to which the test substance of interest is introduced. The test solvent may include but is not limited to aqueous matrices, including ASTM Type I Water and various salt matrices (e.g. 0.01M CaCl,,,,,); and organic solvent matrices, including methanol, and acetone. 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: . 4.1.1 Wear the proper lab attire for all parts of these procedures. Wear gloves and eye protection at all times. 4.1.2 Handle all solvents in a hood for all parts of the described sample preparation procedure. 4.1.3 For potential hazards of each chemical used, refer to material safety data sheets, packing materials, and 3M Environmental Laboratory's Chemical Hazard Review 4.1.4 No mouth pipetting is allowed. ETS-8-170.1 Solubility Determination: Screen Test Method Page 2 of 9 Page 19 of 171 BACK TO MAIN 3M Environmental Laboratory Project ## E01-0768 4.2 Cautions: 4.2.1 Glassware in which standards are prepared should be triple rinsed with acetone and methanol to reduce the possibility of accidental contamination. 5.0 INTERFERENCES 5.1 Impurities may significantly affect the solubility of the test substance. The purity of the test substance should be known and documented prior to starting the screening procedure. 6.0 EQUIPMENT 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Vortex mixer. 6.3 Sonicating device. 7.0 SUPPLIES AND MATERIALS 7.1 Disposable glass graduated pipettes, 1mL to 100mL. 7.2 Disposable glass Pasteur pipettes and rubber bulbs. 7.3 Glass beakers, various sizes. 7.4 2.5 mL-4 mL glass screw-top vial. 7.5 Glass volumetric flasks, 10 mL to 1000 mL. 7.6 10 pL-1000 pL PipettemanTMmanual pipettor and plastic pipette tips, or equivalent. 8.0 REAGENTS AND STANDARDS 8.1 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science, or equivalent. 8.2 Acetone, HPLC/SPEC/GC grade from EM Science or equivalent. 8.3 Water, ASTM Type I, or equivalent. 8.4 Calcium Chloride Dihydrate, Approximately 99%or better, from Sigma or equivalent. 8.5 0.01 M CaCI,, A 0.01 M CaCl, stock solution is prepared by weighing 1.5 g CaCI, Dihydrate in a weigh boat and transferring to a 1 L volumetric flask and diluting to the mark with Milli-QTMwater. 8.6 Test substance of known purity. 9.0 SAMPLEHANDLING 9.1 Record times of initial preparation and dilution on a sample preparation sheet or logbook. 9.2 Once the preliminary testing has been completed and the appropriate information is extracted, the screen test samples should be disposed into the proper waste stream. ETS-8-170.1 Solubility Determination: Screen Test Method Page 3 of 9 Page 20 of 171 10.0 QUALITCYONTROL 10.1 Not applicable. 11.0 CALIBRATIOANND STANDARDIZATION BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 4 mL vial or equivalent Total volume solvent added (mL)... Approximate Solubility (pg/mL)... Solvent addition 1 0.1 100000 Solvent addition 2 0.5 20000 Solvent Solvent addition 3 addition 4 1 2 10000 5000 12.1.3 Following addition of solvent, vortex mix approximately 15 seconds, sonicate 2-5 minutes. Visually check for undissolved particles. If undissolved test substance remains, continue adding solvent according to the above chart. It may be necessary to allow the solution to settle for up to 5 minutes before making observations. Record observations on the standardized prep sheet. 12.1.4 If all particles dissolve, then estimate the approximate solubility and document the concentration. Because the concentration is >10 pg/mL, the shake flask method (ETS-8-172.0) will be used to determine the accurate solubility point. However, if the test substance dissolves after t h e j r s t solvent addition of 100 pL the solubility is determined to be >lo% (mass/volume). The test substance shall be regarded as "highly soluble," and no further testing is required. 12.1.5 If the test substance did not dissolve in the 2 mL of solvent, weigh 10 mg f 1 mg of test substance into a 100 mL glass volumetric flask or equivalent. Record the weight (refer to attachment A for an example of a standardized prep sheet). 12.1.6 Add solvent according to the table below to the test substance to deliver the appropriate amount of solvent. As described in 12.1.3,vortex mix, sonicate, and allow the test solution to settle. Record all observations and procedures on the preparation sheet. ETS-8- 170.1 Solubility Determination: Screen Test Method Page 4 of 9 Page 21 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 100 mL volumetric Solvent addition 1 Solvent addition 2 Solvent addition 3 flask or equivalent Total volume solvent 10 50 100 added (mL)... Approximate Solubility 1000 200 100 (pg/mL)... 12.1.7 If the solubility point is reached at or prior to the complete addition of 100 mL, the concentrationestimation at which the substance is soluble should be documented and shall be used as a starting point for the shake flask method (ETS-8-172.0). 12.1.8 If the 100mL solvent level has been reached without complete dissolution of test substance, weigh out 10 mg k 1 mg test substance and transfer to a 1 L volumetric flask, graduated cylinder, or equivalent. Dilute to 1 L with test solvent. The approximate concentration of test substance is at 10 pg/mL. The flask should be allowed to sit 12-24 hours to allow for maximal dissolution. If the undissolved particles are still observed, the column elution method will be utilized (ETS-8171.O). If no visible particulates are observed, the shake flask method (ETS-8172.0) shall be used to determine the solubility. 13.0 DATAANALYSIASND CALCULATIONS 13.1 The solubility determination in this method is qualitativelsemi-quantitative. 13.2 The point to which the substance dissolves in solvent is confirmed visually. The solubility point is therefore a qualitative determination. 13.3 The concentration estimation is semi-quantitative in that the approximate concentration is calculated by the following equation: c=a/b Where: c= the semi-quantitativeconcentration pg/mL, a= amount of substance weighed out (pg), and b= the approximate amount of solvent added (mL). 14.0 METHODPERFORMANCE 14.1 Limitation of data. The accuracy to which the solubility is determined is subject to a large margin of error due to the way in which the solvent is added to the solute. Since large and varying increments of solvent are being added to the solute, this error margin must be considered when reporting the concentration estimation. 14.2 The data obtained through this study is an estimation only and should be treated as a qualitative estimation of the solubility of test substance in a given solvent. ETS-8-170.1 Solubility Determination: Screen Test Method Page 5 of 9 Page 22 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 14.3 For an accurate measurement of the substance solubility concentration, the shake flask method, or column elution method will be used. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 16.0 RECORDS 16.1 Sign and date all observations and calculations. 17.0 ATTACHMENTS 17.1 Attachment'A-Example of a Standardized Sample Preparation Sheet: Solubility Screen Prep Sheet. 18.0 REFERENCES 18.1 Organization for Economic Cooperation and Development. OECD Guideline for Testing of Chemicals. Water Solubility -0ECD Guideline 105:pp. 1-7, Adopted 1995 18.2 United States Environmental Protection Agency. OPPTS 830.7860 Water Solubility (Generator Column Method). Prevention, Pesticides and Toxic Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-042: pp. 1-17, 1996 18.3 United States Environmental Protection Agency. OPPTS 830.7840 Water Solubility: Column Elution Method; Shake Flask Method. Prevention, Pesticides and Toxic Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-041: pp. 1-12,1996 18.4 3M Environmental Laboratory Method ETS-8-171.O,"Column Elution Method: Solubility Determination of Test Substance in Various Solvents.'' 18.5 3M Environmental Laboratory Method ETS-8-172.0, "Shake Flask Method: Solubility Determination of Test Substance in Various Solvents.'' 19.0 AFFECTEDDOCUMENTS 19.1 ETS-8-171.0, "Column Elution Method: Solubility Determination of Test Substance in Various Solvents." 19.2 ETS-8-172.0, "Shake Flask Method: Solubility Determination of Test Substance in Various Solvents." ETS-8-170.1 Solubility Determination: Screen Test Method Page 6 of 9 Page 23 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 20.0 REVISIONS Revision Number Reason For Revision 1 It was desirable to make the method more universally applicable by removing references to specific test substances. Revision - Date 03/13/01 ETS-8-170.1 Solubility Determination: Screen Test Method Page 7 of 9 Page 24 of 171 1 Attac/inierit A: Example Preparation Worksheet,page 1 of 2 I BACK TO MAIN GLP Study Number: Test Substance: m Source: II I N SCRFFN Sample preparation worksheet Date: Solvent: ID# Source: Step 1 Weigh approximately 10mg f 1mg of test substance into a 2.5 mL-4 mL glass screw-top vial. Weight of test substance: ! n g Balance ID: Datellnitials: 2nd replicate optional: mg Thermometer ID: I I Analyst ___ 2 Add test solvent accordin-gto the table below. following each addition of solvent, shake vigorously/vortex, and sonicate the mixture and visually check for undissolvedp a des. Datenim- Ivolume added (rnL)/ Iamrox conc. after I Sample Prep I Step 2-1 Flaskvortex-mixed? Y ~ ~ R/~~N~ T~~ Q OBSERVATIONS~NOTES I Is there solute still presentxescontinue to step 2-2 Total volume solvent added (mL) (1.1 Approximate Solubility (uglmL) 100,000 Flask Flask allowed t0 Settle? YeslNoT-i NO. The solution is at IO%, and is considered"infinitelysoluble." h3 further solubility testing isrequired, Analyst - Analyst __ Analyst __ Datemim- Step 2-2 Total volume solvent added (mL) (1.5 Approximate Solubilitv (ualmL) 20,000 Datenim- Steo 2-3 Total volume solvent added (mL) 1. Approximate Solubilitv (udmL) 1.0.000 Datenim- Step 2-4 Total volume solvent added (mL) 2 Approximate Solubilih, (uqlmL) 5,000 Flask vortex-mixed?YesMo RoomT e d Is there solute still present? Yes/No Flask sonicated? YeslNo T-i Flask allowed to settle? YeslNo T-i Flask vortex-mixed?YeslNo Room Te@ Flask sonicated?YeslNo T-i Flask allowed to settle? Yes/No T-i ls there solute still present? Yes/No Flask vortex-mixed?Yes/No Room T e d ls there solute still present? YedNo Flask sonicated? YeslNo -Ti Flask allowed to settle? YeslNo T-i Did the substance dissolve in 2 mL or less of solvent? Yes -Approximate concentration at which the test substanceis soluble in solvent: - d g h l The solubility of the test substance is to be determinedvia the shake flask method. 9 or No Continue to page two for further testing. m ro 01 s Notes/ additional comments: ETS-8-170.1 Solubility Determination: Screen Test Method Page 8 of 9 BACK TO MAIN I Attnclinient A Cont.: Examule PreDaratioii Worksheet.Dape 2 o f 2 ~~~~~ GLP Study Number: Test Substance: 3M Environmental Laboratow SOLUBlLlN SCREEN Sample preparation worksheet Iwy: Source: Date: Solvent: IWI: Source: Step "Did test. (continued from page I) the test substance dissolve in 2 mL? YeslNo If YES, the test Is complete, and there Is no need to continue with the If NO, continue to step 3 to determine the approxlrnate solubility of the substance. 3 Weigh approximately 10 mg f I mg of test substance into a 100 mL graduated stopperedcylinderholumetricflask. Weight of test substance: mq Balance ID: 2nd rep/iwle optional: mg Thermometer ID: 4 Add test solvent according to the table below. following each addition of solvent, shake vigorouslyhortex, and sonicate the mixture and visuallv check for undissolved oarticles. screen Datellnitials: IAnalvst Analyst - Analyst - volume added (mL)I approx conc. after Solubility data addwon (WmL) Daterrime I Step 4-1 DvIIsIIIouusu (mL)... 10 e Solubility(ug/mL)... 1,000 Daterrime I Step 4-2 Total volume solvent added (mL)... 50 Approximate Solubility (uglmL)... 200 Datmme I Step 4-3 Total volume solvent added (mL)... 100 Approximate Solubility (ug/mL)... 100 Sample Prep Flask vortex-mixed?YedNo Room Temp Flask sonicated? YeslNo Time Flask allowed to settle? YesMo Time OBSERVATIONSINOTES 'C Is there solute still present? YeslNo min min Flask vortexmixed? YeslNo ROM Temp Flask sonicated? Y&No Time Flask allowed to settle? YeslNo l i m e "C Is there solute still present? YeslNo min min Flask vortexmixed? YeslNo Room T e m p D C Is there solute still present? YedNo Flask sonicated? YeslNo Time min Flask allowed to settle? YeslNo Time min Weight of test substance: mq Balance ID: Date/lnitials: IAnalvst - Dalerrime I .. , . . .. Approximate aoiuoiiiry (,ug,mLj. Flask vortex-mixed? YeslNo Room flask sonicated? Y&No Time Flask allowed to settle? YeslNo l i m e Is there solute still present? YeslNo min ~~ Conclusions: Did the substance dissolve in 1000 mL or less o f solvent? Yes -Approximate concentration at which the test substance is soluble in solvent: ualml The solubility of the test substance is to be determinedvia the shake flask method. or No -The solubility of the test substance is to be determinedvia the column elution method. Notes1additional comments: ETS-8- 170.1 Solubility Determination: Screen Test Method Page 9 of 9 3M ENVIRONMENTLAALBORATORY BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 SHAKE FLASKMETHODS:OLUBILITYDETERMINATIOONF A TESTSUBSTANCE IN Vmous SOLVENTS Method Number: ETS-8-172.1 Adoption Date: 09/08/00 Effective Date: 03/27/01 Authors: Kristin L. Terrell, Mark L. Anderson, and Mark E. Ellefson Approved By: - William K. Reagen, Laboratory Manager OJ$&/, Date 6ate ' 1.0 Scope and Application 1.1 Purpose. According to the United States Environmental Protection Agency (U.S. EPA) and Organization for the Economic Cooperation and Development (OECD) guidelines, solubility determination of substances that have solubilities in a given test matrix (e.g. water/acetone/methanol) of greater than 100 pg/mL must be analyzed via the shake flask method (OECD Guideline 105, and OPPTS Guideline 830.7840). The prerequisite to this' . method is a preliminary screen of the test compound for its approximate solubility level. Refer to ETS-8-170.0 for the preliminary solubility screen test procedures. ETS-8- 172.1 Solubility Test: Shake Flask Method Page 1 of 11 Page 27 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 1.2 Compatible Analytes. Test substance and degradation products for solubility testing. 1.3 Acceptable Matrices. Water, Acetone, and Methanol, or other solvents of interest. 2.0 SUMMARY OF METHOD 2.1 Using the qualitativehemi-quantitative data obtained from the preliminary solubility screen test, weigh out more than five times the estimated soluble concentration of test substance into each of twelve screw-capped vessels (three time points with four test vessels each-sample, duplicate, triplicate, and matrix blank). Add the appropriate amount of test solvent gravimetrically. Cap the tubes, seal the caps with tape, and place horizontally on an incubatorhhaker set to 30 "C k 2 "C, shaking for approximately 24 hours. At approximately 24 hours, the first time point will be pulled and equilibrated at room temperature (about 20 'C-26 "C)for approximately.24 hours. The equilibrated , samples are centrifbged and aliquoted into autovials. Samples are then diluted 1:10 or higher with a suitable solvent for analysis via LCMS. At approximately 48 hours and 72 hours, the second and third sets, respectively, will be pulled, equilibrated, centrifuged, aliquoted and diluted with a suitable analytical solvent for LCMS analysis. Depending on the analytical range, dilutions may be made using a diluter or syringe to make successive serial dilutions using a suitable analytical solvent to reach the desired concentration (e.g. 1:10 or higher samp1e:methanoVacetone serial dilutions), and an appropriate internal standard (e.g. lH,1H,2H,2Htetrahydroperfluorooctane sulphonic acid (THPFOS)) will be added to the final dilution prior to analysis. Samples are to be analyzed via LCMS against a standard curve containing the test substance and an appropriate internal standard. 3.0 DEFINITIONS 3.1 Method blank: An analyte-free matrix (e.g. methanol, water, or acetone) to which all reagents agree are added in the same volumes or proportions as used in the sample processing. The method blank is used to document contaminationresulting from the entire sample treatment and analytical process. The method blank is carried through the complete sample preparation, treatment, and analytical procedure. 3.2 Solvent blank: A sample of analyte-freemedium (e.g. methanol/water/acetone solution) that is not taken through the sample treatment process. This blank is used to evaluate instrument and reagent contamination. 3.3 Shake Flask Sample Triplicates: Three test vessels taken from and representative of the same sample source carried through all steps of the treatment, extraction, and analytical procedures in an identical manner. 3.4 Sample replicates: Replicate samples (two or more) taken from and representative of the same sample source (e.g. test vessel containing test analyte and solvent) and separately carried through analytical procedures in an identical manner. 3.5 Internal Standard (IS): A known amount of a compound similar in analytical behavior to the compound(s) of interest, added to all samples and standards, and carried through the entire measurement process. ETS-8-172.1 Soltibility Test: Shake Flask Metliod Page 2 of 11 Page 28 of 171 BACK TO MAIN 3M Environmental Laboratoly Project # E01-0768 3.6 Calibration Standard: A dilution of various amounts of a stock, intermediate or purchased standard to achieve standard solutions in a concentrationrange of interest. 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Wear the proper lab attire for all parts of these procedures. Wear gloves and eye protection at all times. 4.1.2 Handle all solvents in a hood for all parts of the described sample preparation procedure. 4.1.3 For potential hazards of each chemical used, refer to material safety data sheets, packing materials, and 3M Environmental Laboratory's Chemical Hazard Review. 4.1.4 No mouth pipetting is allowed. 4.2 Cautions: 4.2.1 Glassware in which standards are prepared are to be triple rinsed with acetone and methanol to reduce the possibility of accidental contamination. 4.2.2 All test vessels (e.g. centrihge tubes) are to be sufficientlyrinsed with solvent prior to their usage. 5.0 INTERFERENCES 5.1 Impurities may significantly affect the solubility of the test substance. The purity of the test substance should be known and documented prior to starting the preliminary solubility screening procedure. 5.2 Contaminants in solvents, reagents, glassware and other sample processing or analysis hardware may cause interference. The routine analysis of laboratory method blanks must ' be used to demonstratethat there is no interference under the conditions of the analysis. 6.0 EQUIPMENT 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Centrifuge capable of holding 15 mL centrifuge tubes or equivalent. 6.3 Incubator with heatingkooling capabilities. 6.4 Diluter, Hamilton Microlab@ 500 Series, or equivalent. 6.5 Vortex-mixer. 7.0 SUPPLIES AND MATERIALS 7.1 Thermometer capable of reading at least 15 'C-40 'C. 7.2 5-250 mL polypropylene centrifuge tubes, or equivalent. 7.3 Disposable glass graduated pipettes, 1mL to 10 mL. 7.4 Disposable glass Pasteur pipettes and rubber bulbs. 7.5 Glass beakers, various sizes. 7.6 Crimp cap autovials-1.5 mL, caps, crimper, and decapper. 7.7 Hamilton GastightB syringes (precision f 1% of the total volume), 5 pL to 1000 pL. 7.8 Pipette-man manual pipettor. ETS-8-172.1 Solubility Test: Shake Flask Method Page 3 of 11 Page 29 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 8.0 SUPPLIES AND MATERIALS 8.1 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science, or equivalent. 8.2 Acetone, HPLC/SPEC/GC grade from EM Science or equivalent. 8.3 Water, ASTM Type 1. 8.4 Test substance of known purity. . 8.5 Calcium Chloride Dihydrate, Approximately 99% or better, from Sigma. 8.6 0.01 M CaC12 solution, Example: A 0.01 M CaC12 stock solution is prepared by weighing 1.4 g CaC12 in a weigh boat and transferring to a 1 L volumetric flask and diluting to the mark with Milli-QTMwater. 9.0 SAMPLHEANDLING 9.1 Record times of initial preparation, set-up, and sample aliquotinghalysis on a sample preparation sheet or logbook. 9.2 Once the samples have been diluted, they may be analyzed via LCMS. Alternatively, the diluted samples may be kept in cold storage (e.g. approximately 1-5 "C) in crimp-capped autovials until the time of analysis. 10.0 QUALITYCONTROL 10.1 Shake Flask Sample Triplicates. Set up each test vessel in triplicate to provide a measure of the precision on sample preparation. 10.2 Sample Replicates. Prepare and analyze all samples (from each test vessel for each time point) in multiple replicates (2 or more) to provide a measure of the precision on analysis. 10.3 Quality Control Blank Samples. 10.3.1 Method blank. Set up a fourth test vessel without test substance to measure any contamination accrued throughout the sample preparation process. The method blank is carried through the same sample preparation procedures as the three test vessels with test substance, only no test substance is added throughout the entire procedure. 10.3.2 Solvent Blank. An aliquot of the dilution solvent (Le. methanol) directly analyzed for possible contaminants. The solvent blank is used to document any possible contamination of the solvent(s) used during the sample preparation process, and to detect any instrumental contamination or background interferences. 11.0 CALIBRATION AND STANDARIZATION 11.1 The compounds of interest must be standardized according to laboratory specifications. 11.2 All equipment used, such as the analytical balance and automated diluter, should be calibrated prior to use (daily, weekly, etc.) as specified in its standard operating procedure. ETS-8-172.1 Solubility Test: Shake Flask Method Page 4 of 11 Page 30 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 11.3 All samples analyzed will be run against a standard curve containing varying amounts of test substance, and a fixed amount of internal standard. 12.0 CALIBRATION AND STANDARIZATION 12.1 Preparation of Test Vessel. Record all data and observations on the example sample preparation worksheet (Attachment A) or equivalent. ' 12.1.1 Prepare a solution at least five times more concentrated than the concentration at which the substance was known to be dissolved at in the preliminary solubility screen test (ETS-8- 170.0). 12.1.2 Weigh out test substance into a tared test vessel (e.g. 15 mL polypropylene centrifuge tube). 12.1.3 Add the test solvent to the test vessel gravimetricallyuntil the desired volume has been obtained (note: solvents have differing densities, therefore the weight needed to achieve a specijk volume will be dependent on the density.). Record all weights on a standardized preparation sheet or logbook. 12.1.4 Visually confirm that there are undissolved particles. If no particles are present, repeat 12.1.2 with more test substance or add more test substance and record the additional weight. 12.1.5 Prepare the solution described in 12.1.1-12.1.4 nine times and label (at minimum) the test vessels as Day 1, -Rep 1, -Rep 2, and -Rep 3; Day 2, -Rep 1, -Rep 2, and - Rep 3; and Day 3, -Rep 1, -Rep 2, and -Rep 3. Also include the day of initial sample prep, the person(s) responsible for the sample, the test compound, nature of the study (e.g. water solubility), the solvent utilized, and the study number. 12.1.6 Prepare three test vessels as described in 12.1.3 without the test substance and label (at minimum) as Dayl,Rep4; Day2,Rep4; and Day3,Rep4 along with the information described under 12.1.5. These three test vessels are the "method blank" samples. 12.1.7 Seal the cap to the test vessel. Midshake test vessel to ensure contact between the test substance and the solvent. And wrap the cap with tape. 12.2 Equilibration of samples. 12.2.1 Place all of the test vessels on their side (horizontal) in an orbital incubator set to approximately 30 OC k 2 O C and rotating at a considerable speed to ensure sufficient contact/mixing between the test substance and solvent. 12.2.2 At approximately 24 hours, four test vessels are removed: three containing test substance labeled "Day 1,Repl", "Day1,Rep2", "Dayl ,Rep3", and "Dayl ,Rep4". 12.2.3 The samples are shaken to ensure no test substance is stuck to the side of the tube. 12.2.4 The tubes are then placed in a stable temperature environment of approximately 20- 26 "C (record the actual temperature) for about a 24 hour period. 12.2.5 After about 24 hours of equilibration at 20 "C-26 "C, centrifuge the samples until the solution is visibly clear. If micelle formation is suspected, an additional highspeed centrifugation step may be added (20,000 RCF for approximately 1 hour). The test vessels are now ready for dilution. See section 12.3 for further sample preparation. ETS-8-172.1 Solubility Test: Slinke Flnsk Method Page 5 of 11 Page 31 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 12.2.6 At approximately 48 hours of incubation time, the second set of test vessels may be pulled (labeled "Day2,Repl", "Day2,Rep2","Day2,Rep3", and "Day2,Rep4") and equilibrated as described in 12.2.3-12.2.5. 12.2.7 At approximately 72 hours, the final set of samples is pulled (labeled "Day3,Rep1", "Day3,Rep2", "Day3,Rep3", and "Day3,Rep4") and equilibrated as described in 12.2.3- 12.2.5. 12.3 Sample preparation for analysis. 12.3.1 Observe the test solution. Due to the possibility of emulsions/or a concentrated layer of solute at the surface of the solution, it may be necessary to pipette off the top layer of solution (or an aliquot of sample may be centrifuged as stated in 12.2.5). The use of a Pasteur pipette or equivalent is recommended. 12.3.2 Aliquot the test solution into each of four autovials (e.g. 1.5 mL polypropylene or glass crimp-cap vials). 12.3.2.1 Sample aliquots may be taken by submerging the pipette or syringe tip below the surface of the test solution. Note: when selecting the type of pipette to be used, take into consideration the test substance's tendency to adsorb to certain materials (i.e. adsorption of test substance to glass or plastic), 12.3.2.2 Prior to aliquoting sample to the autovial, the pipette tip must be equilibratedsaturated with the test solution by drawing and gently expelling the test solution in and out of the pipette tip, taking care not to disturb the solid particulate. 12.3.3 Label the four autovials and record the sample id. 12.3.4 Using a diluter, make a 1:lO or higher dilution of samp1e:extractionsolvent (i.e. methanol, acetone) into a labeled autovial. 12.3.5 Using the solubility information obtained in the preliminary screen test, estimate the appropriate dilutions, if any, required to bring the concentration of sample into the appropriate analytical range (e.g. approximately 3 ng/mL to 3000 ng/mL test substance). Utilize the diluter to make serial dilutions of the sample using a suitable analytical solvent. 12.3.6 To the final analytical sample, add internal standard at a concentration suitable to the analytical method. 12.3.7 Samples will be analyzed via LCMS against a standard curve of varied, known concentrations of test substance with a constant concentration of internal standard equal to the concentration in the samples. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Samples will be analyzed via a calibration curve. The amount of test substance in the sample will be quantified against a standard curve. 13.2 Means will be calculated by adding the individual entities and dividing the resultant sum by the number of individual entities. 13.3 Standard deviations will be calculated using either Microsoft Excel@ or Microsoft Access@to calculate standard deviation. The built in function contains the following equation which is based on the individual entities (n) being less than 30: - 1 ETS-8-172.1 Soltibility Test: Shake Flask Method Page 6 of 11 Page 32 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 13.4 Sample precision will be reported as %RSD (or %CV). Sample precision will be calculated using the following equation: A/B X 100 = Sample %RSD where: A= standard deviation of averaged samples B= average of samples I I Sample RPD will be calculated using the following equation: A-B /((A+B)/2)= Sample RPD where: A= concentration of first replicate B= concentrationof second replicate 13.5 Exclusion of an outlier data point may be performed by utilizing Dixon's Q-Test. The questionable data point may err to the high or low end of the data set. Calculate the variable "Qobserved." If Qobserved > Qtabulated, then the data point may be rejected with 90% confidence (see table 1below for Qtabulatedvalues). Qobserved= gap/range where: Gap= the difference between the questionablepoint and the nearest value. Range= total spread of the data. Table 1 Criteria for Rejection of Outlier Values: I 1 Number of observations I Qtabulated, 90% Confidence Range 0.886 0.679 0.557 14.0 DATAANALYSIASND CALCULATIONS 14.1 Precision of data. Sample data must have a percent relative standard deviation (%RSD) (or relative percent difference) of < 15%. Non-compliant data must be evaluated for obvious outliers. The Q-Test may-be applied to exclude questionable data points. If an outlier value exists, sample average and precision is re-calculated and reported without the questionable data point. Document the non-compliant data on data summary sheets, and include results of the statistical analysis with the final results. 14.1.1 14.1.2 Sample replicate %RSD's should be 4 5 % . If the average of the sample triplicate data is >15% RSD, evaluate the threevalues for an outlier value. To questionable data points, apply the Dixon's Q-Test. If the outlier value is rejected with 90% confidence, exclude the sample from further data calculations and calculate the average and RPD for the remaining two values. If no data points can be excluded and the data does not meet the criteria, then the data set may not be used in the final reported data. The remaining shake flasks for the timepoint shall be used to report the data. The %RSD between shake flasks should agree within 15%. If this criteria is not met, apply the Q-test to outlier datum. If the questionable data point is discarded ETS-8-172.1 Solubility Test: Shake Flask Metliod Page 7 of 11 Page 33 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 via the Q-Test, then the non-compliant shake flask data is not to be used for the timepoint. Includejustification for dropping the shake flask (e.g. q-test results). 14.1.3 And finally, the %RSD between the three days should be <15 % for the shake flask method. If this criteria is not met, it may be necessary to re-evaluate the data for possible outliers. Three agreeable time intervals are required to report the final solubility. If three consecutive timepoints do not agree within 15%, it may be necessary to repeat the entire test, or re-dilutehe-shoot samples to check for error. 14.2 Limit of Quantification. For this study, the LOQ will be equal to the lowest calibration standard used in the calibration curve containing more than twice the area counts of the highest Quality Control Blank. 14.3 Quality Control Blanks: Method and Solvent Blank samples. The level of analyte (test analyte OR internal standard analyte) shall be less than 50% of the area counts of the LOQ. If background levels of analyte exist in the initial method blanks, but subsequent . blanks prior to the calibration standard curve are clean, the data may be accepted (provided that there are no other indicators that either the samples or the instrument contain significant background levels of analyte). 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 16.0 RECORDS 16.1 Sign and date all observations and calculations. 16.2 Fill out all appropriate sample preparation worksheets. 17.0 ATTACHMENTS 17.1 Attachment A-Example Sample Preparation Worksheet. 18.0 REFERENCES 18.1 Organization for Economic Cooperation and Development. OECD Guideline for Testing of Chemicals. Water Solubility -0ECD Guideline 105: pp. 1-7, Adopted 1995 18.2 United States Environmental Protection Agency. OPPTS 830.7860 Water Solubility (Generator Column Method). Prevention, Pesticides and Toxic Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-042: pp. 1-1 7, 1996 18.3 United States Environmental Protection Agency. OPPTS 830.7840 Water Solubility: Column Elution Method; Shake Flask Method. Prevention, Pesticides and Toxic ETS-8-172.1 Solubility Test: Shake Flask Method Page 8 of 11 Page 34 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-041: pp. 1-12,1996 18.4 ETS-8-170.0, Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents. 18.5 Harris, Daniel C. Statistics. Quantitative Chemical Analysis. 4`h ed.; W.H. Freeman and Company: New York, 1982; p 70-71. . 18.6 Natrella, Mary Gibbons. The Treatment of Outliers. Experimental Statistics, National ' Bureau of Standards Handbook 91; U.S. Government Printing Office: Washington, D.C., 1963; Pages 17-3, and T-27. 19.0 AFFECTEDDOCUMENTS 19.1 ETS-8-170.0, "Solubility Screen Test: Approximate SolubilityDetermination of a Test Substance in Various Solvents." 20.0 REVISIONS Revision Number 1 Reason For Revision Section 1.2, references to specific test substances were removed. Section 3.6, removed "internal standard blank" from definition section. Section 12.2.5, an optional high-speed centrifugation step was added. Fixed the incorrect numbering of sections 13.5 through 14.3. Section 14.3, more clearly defined the acceptance criterion for method and solvent blanks. Revision Date 03-0 1 ETS-8-172.1 Soliibility Test: Shake Flmk Method Page 9 of 11 Page 35 of 171 BACK TO MAIN ATTACHMENTA: Example Sample Preparation Sheet @age 1 of 2). Was the solubiliy screen test petformedprior to sfart? Y/N Iso, what is the approximate concenfration?-&nl Balance ID Thermometer I D . RoomTemp: . SFM vials I D # 'samplelD" = Sample Description weight test substance added: Total mass (solute + solvent) added: 'Internal Standard addition uu ds.sdn IS added to dilution factor: samplelD-1.1.1 thru -1.1.4 Dayl. Smpl I: samplelD-1.2.1 thru -1.2.4 Dayl, Smp2 1: samplelD-1.3.1 thru -1.3.4 Dayl. Smp3 1: samplelD-1.4.1 thru -1.4.4 Dayl, Smp4 blank ' none added 1: samplelD-2.1.1 thru -2.1.4 Day2, Smpl 1: samplelD-2.2.1 thru -2.2.4 DayP, Smp2 1: samplelD-2.3.1 thru -2.3.4 Day2, Smp3 1: samplelD-2.4.1 thru -2.4.4 Day2, Smp4 blank none added 1: samplelD-3.1.1 thru -3.1.4 Day3. Smpl 1: samplelD-3.2.1 thru -3.2.4 Day3, Smp2 ,I: samplelD-3.3.1 thru -3.3.4 Day3, Smp3 1: samplelD-3.4.1 thru -3.4.4 Day3. Smp4 blank none added 1: DateKime: Initials: Test Vessels sealed wl tape? YiN Test Vessels shakedvortex-mixed? Y/N Test vessels placed on orbital incubatoR Y/N IncubatorID: Speed: 'Internal Standard: int. Std.Conc.: rpm InitialTemperature: ID#: ug/ml 0 "C DatelTimdlnitials: ObSeNatlOnS/COmmentS: Location of samples: Temperature of environment: D r ID: Visual observations: I I I ETS-8- 172.1 Solubility Test:Shake Flask Method Page 10 of 11 BACK TO MAIN ATTACHMENTA CONTINUED (page2 of 2). Day 2: Four "Day 2" samples are pulled and allowed to be placed in a temperature controlled environment for approx. 24 hours with no agitation. Location of samples: Temperature of environment 0 r ID: Visual observations: I Date/T&ndInitials: "Day 1" Test Vessels Centrifuged: Cent. ID: RPM or RCF readout: duration: mi& DaIe/Time/lnllisir: i Visual observations: -Four aliquots from each of the four samples are to be made into glasdplastic autovials (also, check autovials for correct labeling). YesMo -Immediately dilute the solution -:1 with methanol. MethanolTN-A: Date/TunellnlliaiS' Additional dilutions: Dilution factor (test so1ution:methanol): 1: X's how many dilutions: =final dilution factor: I: Diluter ID used: Amounts used: uL methanol, ul samples -Store samples and extracts in a cooler at untiltime of analysis. Cooler ID: Cooler Temp: 0 Date/Timellnitials: Day 3: Four "Day 3"samples are pulled and allowed to be placed in a temperature controlled environment for approx. 24 hours with no agitation. Dal.9Timellni(lals: Location of samples: Temperature of environment c Thermometer- Visual observations: "Day 2" Test Vessels Centrifuged: Cent. ID: RPM or RCF readout: duration: mia l l Visual observations: I each of the four replicates are to be made into glasdplastic autovials (also, check autovials for correct labeling). Yes/No Datefllmellnllials: the solution -1: with methanol. MethanolTN-A: Dateffimelinitialo: Additional dilutions: Dilution factor (test so1ution:methanol): 1: X's how many dilutions: =final dilution factor: I: Diluter ID used: Amounts used: uL methanol, uI samples -Store samples and extracts in a cooler at untiltime of analysis. Cooler ID: Cooler Temp: 0 DatwThelinitials: At approx. 24 hours of equilibration. the 'Day 3. samples are to be aliquotedlextracted. EnvironmentTemp.: 'G Therm. ID: Test Vessels Centrifuged: Cent. ID: Visual observations: 11 I RPMor RCF readout duration: mio, Dateffime/lnlUak II-Four aliquots from each of the four samples are to be made into glasdplastic autovials (also, check autovials for correct labeling). YesMo -0 -Immediately dilute the solution 1:- with methanol. MethanolTN-A. DateCTunelnitiair: m Additional dilutions: Dilution factor (test so1ution:methanol): 1: X's how many dilutions: =final dilution factor: 1: 0 (D Diluter ID used: Amounts used: uL methanol, ul samples I-Store samples and extracts in a cooler at until time of analysis. Cooler ID Cooler Temp: 0 Oatefrimellnilials: ETS-8-172.1 Solubility Test: Shake Flask Method Page 11 of 11 I I I I I 0 0T.(Di 8s *r n rm 3M ENVIRONMENTLAALBORATORY BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 METHOD ANALYSIS OF POTASSIUM PERnUOROOCTANESULFONATE OR OTHER FLUOROCHEMCAINLWS ASTE STREAM OR WATER EXTRACTUSSING HPLC-ELECTROSPRAYLMASS SPECTROMETRY Method Number: ETS-8-155.0 Author: Mark L. Anderson, Mark E. Ellefson Approved By: Adoption Date: (//p/UU Revision Date: Laboratory Manager Date Word 97 ETS-S- 155.0 Page 1 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayNass Spectrometry Page 38 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 1.0 SCOPE AND APPLICATION 1 1.1 Scope: This method describes the analysis of waste stream or water extracts using HPLC- electrospray/mass spectrometry. 1.2 Applicable Compounds: Fluorochemicals or other electrosprayionizable compounds. 1.3 Matrices: Tap water, ground water, wastewater and other aqueous solutions. 2.0 SUMMARY OF METHOD 2.1 This method describes the analysis of fluorochemicals or other electrospray ionizable compounds extracted from water, using HPLC-electrospraymass spectrometry (HPLCES/MS). The analysis is performed by the mass selection of a single ion characteristic of a particular compound, such as the perfluorooctanesulfonate (PFOS) anion, m/z = 499 or perfluorooctanoate(PFOA), d z = 413. 3.0 DEFINITIONS 3.1 Atmospheric Pressure Ionization (MI):The Micromass Platform LCZ single quadrupole system and other commercially availableLCMS systems allow for various methods of ionization by utilizing a variety of sources, probes, and interfaces. These include but are not limited to: Electrospray Ionization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization in these processes occurs at atmosphericpressure (Le., not under a vacuum). 3.2 Electrospray Ionization (ES, ESI): A method of ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These droplets are produced by the application of a strong electrical field. 3.3 Mass Spectrometer (MS): The Platform LCZ and other commerciallymanufactured ESMS systems are equipped with a single quadrupolemass selectivedetector. Ions are selectivelydiscriminated by mass to charge ratio (dz)and subsequently detected. 3.4 Conventional vs. Z-spray probe interface: The Micromass Platform LCZ system utilizes a "Z-spray" conformation. The spray emitted from the probe is orthogonal to the cone aperture. In the conventional conformation it is aimed directly at the cone aperture, after passing through a tortuous pathway in the counter electrode. Though the configuration is different, the methods of operation, cleaning, and maintenance are the same. However, Zspray components and conventionalcomponents are not compatiblewith one another, but only with similar systems (Le., Z-spray components are compatible with some other Z-spray systems, etc.). Other commerciallymanufactured ESMS systems may have similar features. 3.5 Mass Lynx Software: System software designed for the specific operation of Micromass LCZ Mass spectrometer. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details see the manual specific to the instrument (MassLynx NT User's Guide or Micromass Platform LCZ User's Guide). ETS-8-155.0 Page 2 of9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry Page 39 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage of approximately 5000 Volts. 4.1.2 When handling samples or solvents wear appropriateprotective clothing, gloves, and eyewear. 4.2 Cautions: 4.2.1 Operate solvent pumps below a backpressure of 400 bar (5800 psi). If the backpressure exceeds 400 bar, the HP1100 will initiate automatic shutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 Interferences 5.1 To minimize interferences when analyzing samples, Teflon should not be used for sample storage or any part of instrumentation that comes in contact with the sample or extract. 6.0 EQUIPMENT 6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications in the raw data as method deviations. 6.1.1 Micromass Platform LCZ Mass Spectrometer equipped with an electrospray ionization source. 6.1.2 HPl 100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler. 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.1.1 High purity grade nitrogen gas regulated to approximately 100 psi (or house air system.). 7.1.2 HPLC analytical column, such as a Betasil C18 column (5Ox2mm, 5 pm particle size) or equivalent. 7.1.3 Capped autovials or capped 15 mL centrihge tubes. ETS-8-155.0 Page 3 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry BACK TO MAIN 3M Environmental Laboratory Project# E01-0768 8.0 REAGENTS AND STANDARDS d 8.1 Reagents t 8.1.1 Methanol, HPLC grade or equivalent. 8.1.2 Milli-QTMwater (ASTM type I), all water used in this method should be Milli-QTM water or equivalent, and may be provided by a Milli-Q TOC Plus system or other vendor. 8.1.3 Ammonium acetate, reagent grade or equivalent. 8.1.3.1 When preparing different amounts than those listed, adjust accordingly. 8.1.3.2 2.0 mMammonium acetate solution: Weigh approximately0.300 g ammonium acetate. Pour into a 2000 L volumetric flask, add the appropriate volume of Milli-Q water, mix until all solids are dissolved. Store at room temperature. 8.2 Calibration Standards 8.2.1 Typicallytwo method blanks (Milli-Q water), two matrix blanks, and solvent standards are prepared during the sample extraction procedure. 9.0 SAMPLHEANDLING 9.1 Standards and sample extracts are stored in capped autovials or capped 15 mL centrifuge tubes until analysis. 9.2 If analysis will be delayed, standards and sample extracts may be refrigerated at approximately4"C until analyses can be performed. 10.0 QUALITY CONTROL 10.1 Solvent Blanks, Method Blanks and Matrix Blanks 10.1.1 Solvent blanks, method blanks, and matrix blanks are prepared and analyzed with each sample set to determine contamination or carryover. 10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve. 10.2 Matrix Spikes 10.2.1 Matrix spikes are prepared for each sample set and analyzed to determine the matrix effect on the recovery efficiency. 10.2.2 Matrix spike duplicates are prepared periodically to measure the precision associated with the analysis. 10.2.3 Analyze the matrix spike and matrix spike duplicate (if prepared) in the same run as the original sample. 10.2.4 Matrix spike and matrix spike duplicate concentrations should fall in the mid-range of the initial calibration curve or should be prepared at 1.5-5 times the endogenous ETS-8-155.0 Page 4 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry Page 41 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 concentration of the analyte. Spike concentrations should fall in the low-range of the initial calibration curve if extremely low levels are expected. C; 10.3 Continuing CalibrationVerifications 10.3.1 Continuing calibration verifications (CCV) are analyzed to verify the continued accuracy of the calibration curve. 10.3.2 Analyze a mid-range calibration standard after every tenth sample, with a minimum of one per sample set. 10.4 Internal Standardsurrogate Standard 10.4.1 An internal standard (IS) may be used to quantify the target analytes by establishing a relationship between the ratio of analyte response to IS response and a known concentration of the analyte of interest. The IS should be spiked at an amount that will fall within the mid-range of the calibration curve. The IS should be added after the extraction process and before analysis. 10.4.2 A surrogate standard may be used for qualitycontrol. The surrogateis used to quantitatively evaluate the entire analytical procedure including sample preparation and analysis. The surrogate should be spiked to fall within the low to mid-range of the calibration curve. 11.o CALIBRATION AND STANDARDIZATION 11.1 Analyze the standard curves prior to and following each set of extracts. The average of two standard curves may be plotted by linear regression (y = mx + b) weighted l/x, or quadratic fit (y = ax' + bx + c) using MassLynx or other suitable software. The calibration curves should not be forced through zero. 11.2 If the calibration curve does not meet acceptance criteria perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. 11.3 For purposes of accuracy when quantitating low levels of analyte, it may be necessary to use the low end of the calibration curve rather than the full range. Example: when attempting to quantitate approximately 10 ppb of analyte, generate a calibration curve consisting of the standards from 5 ppb to 100 ppb rather than the full range of the curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear regression weighting of high concentration standards. 12.0 PROCEDURES 12.1 Acquisition Set up 12.1.1 Set up the sample list. 12.1.1.1 Assign a sample list filename using the first letter of the name of the instrument (T for Tucker), the year (00 for 2000), the month (04 for April), and the day (TO01012 for October 12,2000). If more than one list is made on the same day, use increasing letters of the alphabet starting with A at the end of the list. ETS-8-155.0 Page 5 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrosprayhfass Spectrometry Page 42 of 171 BACK TO MAIN 3M EnvironmentalLaboratory Project # E014768 12.1.1.2 Assign a method (MS) file. 12.1.1.3 Assign an HPLC program (Inlet file). c 12.1.1.4 Type in sample descriptions and vial position numbers. 12.1.2 To create a method, click on method in the Acquisition control panel then mass spectrometerheadings and select SIR Set ionization mode as appropriate and mass to 499 or other appropriate masses. A full scan is usually collected in addition to the SIRS. Save acquisition method. See the Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information. 12.1.3 Typicallythe analytical batch run sequence begins and ends with a set of solvent standards. 12.1.4 Samples are analyzed with a continuing calibration verification (CCV) injected after every tenth sample. Solvent blanks should be analyzed periodically to monitor for possible analyte carryover. 12.2 Using the AutosamplerKolumnHeater 12.2.1 Place sample vials into the sample tray according to the sample list prepared in Section 12.1.1. 12.2.2 Attach the proper analytical column in the column heater compartment. If using the switching valve, make sure that the tubing is run to the appropriate ports. 12.3 Using the Inlet Editor 12.1.1 Set-up the HP1100 using the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook 12.1.1.1 Sample size = 10 pL injection 12.1.1.2 Flow rate = 300 pL/min. 12.1.1.3 Cycle time = 10.0minutes 12.1.1.4 Mobile phase components: Solvent A: 2.0 mM Ammonium Acetate Solvent B: Methanol (MeOH) Solvent Gradient: Time (min.) 0.00 1.oo 4.50 8.00 8.50 10.0 yJ l 5.00 Yo 5.00 Yo 95.0 Yo 95.0 Yo 5.00 Yo stop ETS-8-155.0 Page 6 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrosprayhlass Spectrometry Page 43 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 12.4 Instrument Set-up 12.4.1 Refer to the Platform LCZ User's Guide, the MassLynx NT User's Guide or ETS9-36, "Operation and Maintenance of the Micromass Platform LCZ Electrospray/Mass Spectrometer". 12.4.2 Check the solvent level in reservoirs and refill if necessary. 12.4.3 Check the tip of the stainless steel capillary at the end of the probe with an eyepiece. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary. 12.4.4 Turn on the nebulizing gas. 12.4.5 Open the tune page. Click on `Operate'to initiate the desolvation heaters. 12.4.6 Open the Inlet Editor. 12.4.5.1 Set HPLC pump to "On". 12.4.5.2 Set the solvent flow to the desired flow rate. 12.4.5.3 Observe droplets coming out of the tip of the probe. A fine mist should be expelled with no nebulizing gas leaking around the tip of the probe. Readjust the tip of the probe if no mist is observed. 12.4.5.4 Allow to equilibrate for at least 10 minutes. 12.4.6 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response: 12.4.6.1 Drying gas 250-425 litershour 12.4.6.2 ESI nebulizing gas 10-15 litershour 12.4.6.3 HPLC constant flow mode, flow rate 10 - 500 pUmh 12.4.6.4 Pressure <400bar (This parameter is not set, it is a guide to ensure the HPLC is operating correctly.) 12.4.6.5 Source Block temperature 150". 12.4.6.6 Desolvation temperature 250". 12.4.7 Print the tune page with its parameters, the Inlet page, sample list, mass spec information, and all other applicable information and store it in the study binder with copies taped into the instrument run logbook. 12.4.7.1 All copies must be initialed and dated. 12.4.8 Click on start button on the MassLynx toolbar. Ensure start and end sample numbers include all samples to be analyzed. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.1 Calculate matrix spike percent recoveries using the following equation: ETS-8- 155.0 Page 7 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry Page 44 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 YORecovery = Observed Result - Backmound Result x 100 Expected Result 13.1.2 Calculate percent difference using the following equation: - % Difference = Expected Conc. Calculated Conc. x 100 Expected Conc. 13.1.3 Calculate actual concentration of analyte in matrix (p.g/mL): On-Column Concentration (pg/mL) x Dilution Factors = Calculated Concentration 14.0 METHODPERFORMANCE 14.1 The Limit of Quantitation (LOQ) is method, analyte, and matrix specific. For many analytes, the LOQ concentration is selected as the lowest acceptablenon-zero standard in the calibration curve. 14.2 Solvent and method blank values must be < '/z that of the lowest standard used in the calibration curve. 14.3 The coefficient of determination (r2)value for the calibration curve must be greater than or equal to 0.980. 14.4 Continuing Calibration Verification (CCV) percent recoveries must be k 30% of the standard concentration. 14.5 Internal Standard recoveries should be within k 50% of the spiked concentration. 14.6 If criteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the raw data. 14.7 If data is to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 RECORDS 16.1 Each page generated for a study must have the following information included either in the header or hand written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. 16.2 Print the tune page, sample list, and acquisition method from MassLynx and other applicable information to include in the appropriate study folder. Copy these pages and tape into the instrument runlog. 16.3 Plot the calibration curve then print these graphs and store in the study folder. ETS-8-155.0 Page 8 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayMass Spectrometry Page 45 of 171 BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 16.4 Print data integration summary, integration method, and chromatograms, ftom MassLynx, and store in the study folder. *; 16.5 Summarize data using suitable software and store in the study folder. 16.6 Back up electronic data to appropriate medium. Record in study notebook the file name and location of backup electronic data. 17.0 ATTACHMENTS 17.1 None 18.0 REFERENCES 18.1 Platform LCZ User's Guide, Micromass UK Limited, Tudor Road, Altrincham, WA14 5RZ;or Floats Road, Wythenshawe M23 9LZ; United Kingdom. 18.2 MassLynx NT User's Guide, Micromass UK Limited, Tudor Road, Altrincham, WA14 5Rz;or Floats Road, Wythenshawe M23 9LZ; United Kingdom. 18.3 MassLynx NT Guide To Data Acquisition, Micromass UK Limited, Tudor Road, Altrincham, WA14 5RZ;or Floats Road, WythenshaweM23 9LZ; United Kingdom. 18.4 ETS-9-36.0, "Operation and Maintenance of the Micromass Platform LCZ Electrospray/Mass Sprectrometer". 19.0 AFFECTEDDOCUMENTS 19.1 None 20.0 REVISIONS Revision Number. Reason For Revision Revision Date ETS-8-155.0 Page 9 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrosprayhfass Spectrometry Page 46 of 171 BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 ATTACHMENBT: DATASUMMARTYABLES Page 47 of 171 BACK TO MAIN Solubilitv Studv Test Substance: PFOS, SD-018 (purity = 86.9%); and FOSA, SE-027 (purity TBD) Test Matrix: Male/Female Human Urine Test Temperature: 23-24 OC Method(s) used: The solubility determinationwas performed according to United States Environmental ProtectionAgency OPPTS 830.7840, "Water Solubility: Column Elution Method; Shake Flask Method" and OECD 105, "Water Solubility" Guidelines, as implemented by 3M Laboratory in methods ETS-8-170.1 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents;" and ETS-8-172.1 "Shake Flask Method; Solubility Determination of a Test Substance in Various Solvents". All analyses were performedvia LC/MS according to the method ETS-8-155.0 "Analysis of PerfluorooctaneSulfonate or Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-Electrospray/MassSpectrometry". Results: PFOS Averane ualmL Day I 302 Day 2 325 Day 3 287 Total 305 Std Dev 12 40 30 14.2 Precision (%RSD) 4% 12% 10% 5% 9 ln (D 3A0% L4E 4 FOSA Day 1 Day 2 Day 3 Total Average ualmL 2.56 2.88 2.61 2.68 Std Dev 0.38 0.12 0.047 0.2 Precision (%RSD) 15% 4% 2% 7% Results for PFOS were corrected for standard purity. Purity for FOSA is unknown, therefore no correctionswere made. ..--I Results Summary 3M Confidential E Page 1 BACK TO MAIN FOSA SE-027Urine Solubility Results SarnpTvpe &f Sample Name Sample Desc. Compound Name Amount lnalmLL SAMPLE HILL0126.D W 042301-FOSA-1.1.1 Smp JasA 5sa&sCwLl 343 SAMPLE HlLLOl27.D FOSA-I.I.2 042301-FOSA-1.1.2 DUp FOSA 5.89 6180725 223 SAMPLE HILL0128.D FOSA-1.I.3 042301-FOSA-I.I.3 Trip FOSA 5.886 5838914 213 SAMPLE HILLO130.D FOSA-1.2.1 042301-FOSA-1.2.1 Smp FOSA 5.889 8385467 309 SAMPLE HILL0135.D FOSA-1.2.2 042301-FOSA-1.2.2 DUp FOSA 5.885 8541720 313 SAMPLE HILL0136.D FOSA-1.2.3 042301-FOSA-1.2.3 Trip FOSA 5.888 8450504 312 SAMPLE HlLLOl 30.D FOSA-1.3.1 042301-FOSA-1.3.1 Smp FOSA 5.802 6003507 248 SAMPLE HILL0139.D FOSA-1.3.2 ' 042301-FOSA-1.3.2 DUp FOSA 5.886 6433005 233 SAMPLE HILL0141.D FOSA-1.3.3 042301-FOSA-1.3.3 Trip FOSA 5.886 6255010 230 from HPLC/MSD"Hillaiy" analytical sequence H010426.s, reprocessedwith Target Sofhvare by KLT SAMPLE HILL0152.D FOSA-2.1.1 042301-FOSA-2. I.1 Smp FOSA 5.879 8103858 317 SAMPLE HlLLOl 53.D FOSA-2.1.2 042301-FOSA-2.1.2 DUp FOSA 5.879 8151376 296 SAMPLE HILL0154.D FOSA-2.1.3 042301-FOSA-2.1.3 Trip FOSA 5.88 7853370 291 SAMPLE HILL0157.D FOSA-2.2.1 042301-FOSA-2.2.1 Smp FOSA 5.887 7993752 298 SAMPLE HlLLOl 58.0 FOSA-2.2.2 042301-FOSA-2.2.2 DUp FOSA 5.886 7697039 280 SAMPLE HILL0159.D FOSA-2.2.3 042301-FOSA-2.2.3 Trip FOSA 5.887 7443513 269 SAMPLE HILL0165.D FOSA-2.3.1 042301-FOSA-2.3.1 Smp FOSA 5.879 7850050 289 SAMPLE HILL0166.D FOSA-2.3.2 042301-FOSA-2.3.2 DUp FOSA 5.882 7536620 279 SAMPLE HlLLOl67.D FOSA-2.3.3 042301-FOSA-2.3.3 Trip FOSA 5.88 7535404 275 from HPLCMSD "Hillaiy" analytical sequence H010426.s, reprocessedwith Target Sobare by KLT Corrected for 1:lO dilution pglmL FOSA ,343 2.23 2.13 3.09 3.13 3.12 2.48 2.33 2.30 Caluculations Average (pg/mL) Std Dev % RSD Spike % Recov. 2.60 2.18 0.7 4.4% 28% Rep2.3 ave&RPD 3.1 1 Day 1results, pglmL 0.0 average 2.56 1% stddev 0.38 2.37 %RSD: 15% 0.1 4% 3.17 3.02 2.96 0.1 2.91 5% 2.98 2.82 2.80 0.1 2.69 5% 2.89 2.81 2.79 0.1 2.75 3% Day 2 results, pglrnL average 2.88 std dev 0.12 %RSD: 4% SAMPLE HILL0097.D FOSA-3.1.1 042301-FOSA-3.1.1, Smp FOSA 5.909 7201703 269 SAMPLE HILLO099.D FOSA-3.1.2 042301-FOSA-3.1.2. DUp FOSA 5.901 6731194 251 SAMPLE HILLO1OO.D FOSA-3.1.3 042301-FOSA-3.1.3, Trip FOSA 5.908 6998562 260 SAMPLE HILLOIOI.D FOSA-3.2.1 042301-FOSA-3.2.1, Smp FOSA 5.908 6990791 257 SAMPLE HILLOI02.D FOSA-3.2.2 042301-FOSA-3.2.2. DUP FOSA 5.922 7009317 261 SAMPLE HILL0103.D FOSA-3.2.3 042301-FOSA-3.2.3, Trip FOSA 5.895 6836606 251 SAMPLE HILLOI08.D FOSA-3.3.1 042301-FOSA-3.3.1, Smp FOSA 5.916 7065688 263 SAMPLE HlLLOl 09.D FOSA-3.3.2 042301-FOSA-3.3.2, DUP FOSA 5.906 7377514 274 2 SAMPLE HlLLOl 10.D FOSA-3.3.3 042301-FOSA-3.3.3, Trip FOSA 5.9 7030743 260 (P from HPLClMSD "Hillary" analytical sequence H010501.s,reprocessedwith Target Software by KLT 2.89 2.60 2.51 0.1 2.60 3% 2.57 2.56 2.61 0.1 2.51 2% 2.63 2.66 2.74 0.1 2.60 3% Day 3 results, pglmL 2 average 2.61 std dev 0.05 Y %RSD: 2% <a-. 7; 8.5Srn -.;4 3M Confidential Page 1 BACK TO MAIN *-.- PFOS SD-018 Urine Solubility Results Sample Name Misc Info ComDound Name Corrected for 1:1,OOQ dilution Average pglmL Corrected for std - RT Area AmtlnslmL] puritvnslrnL Std Dev %RSD SAMPLE HILL0037.D PFOS-1.1.1 042301-PFOS. 1.1 Smp PFOS 5.271 7570273 292.6 286 311 SAMPLE HILL0038.D PFOS-1.1.2 042301-PFOS-1.1.2 DUP PFOS 5.277 8619490 331.8 325 22.0 SAMPLE HILL0039.D PFOS-1.1.3 042301-PFOS-1.1.3 Trip PFOS 5.255 8589480 329.8 323 7% SAMPLE HILL0041.D PFOS-1.2.1 042301-PFOS-1.2.1 Smp PFOS 5.251 7600633 298.7 292 288 SAMPLE HILL0046.D PFOS-1.2.2 042301-PFOS-1.2.2 DUp PFOS 5,263 7571396 294.9 289 4.0 SAMPLE HILL0047.D PFOS-1.2.3 042301-PFOS-1.2.3 Trip PFOS 5.264 7594271 290.2 284 1% SAMPLE HILL0049.D PFOS-1.3.1 042301-PFOS-1.3.1 Smp PFOS 5.27 8267658 320.6 314 306 SAMPLE HILL0051.D PFOS-1.3.2 042301-PFOS-1.3.2 DUp PFOS 5.263 8045600 307.6 301 7.2 SAMPLE HILL0052.D PFOS-1.3.3 042301-PFOS-1.3.3 Trio PFOS 5.272 8019673 308.5 302 2% ~ from HPLClMSD "Hillary" analytical sequence HO10426.s, keprocessedwith Target Software by KLT Day 1results, pglmL average 302 std dev %RSD: 12 4.0% SAMPLE HILL0037.D PFOS-2.1.1 042301-PFOS-2.1.1 Smp PFOS 5.248 7420988 312 305 SAMPLE HILL0038.D PFOS-2.1.2 042301-PFOS-2.1.2 DUP PFOS 5.248 9885661 421 412 SAMPLE HILL0039.D PFOSZ.1.3 042301-PFOS-2.1.3 Trip PFOS 5.256 8832844 363 356 358 54 15% SAMPLE HILL0045.D PFOS-2.2.1 042301-PFOS-2.2.1 Smp PFOS 5.263 7021826 295 289 SAMPLE HILL0046.D PFOS-2.2.2 042301-PFOS-2.2.2 D u ~PFOS 5.255 7094280 294 288 SAMPLE HILL0047.D PFOS-2.2.3 042301-PFOS-2.2.3 Trip PFOS 5.25 6410858 269 263 280 15 . 5% Day 2 results, pglmL average 325 std dev 40 SAMPLE HlLL0050.D PFOS-2.3.1 042301-PFOS-2.3.1 Smp PFOS 5.257 8177762 339 332 338 SAMPLE HILL0051.D PFOS-2.3.2 042301-PFOS-2.3.2 DUP PFOS 5.249 8319423 345 338 7 SAMPLE HILL0052.D PFOS-2.3.3 042301-PFOS-2.3.3 Trip PFOS 5.249 8547947 352 345 2% from HPLC/MSD "Hillary" analytical sequence HO10501.s, reprocessed with Target Software by KLT %RSD: 12% SAMPLE HILL0064.D PFOS-3.1.1 042301-PFOS-3.1.1 Smp PFOS 5.237 6815087 267.6 262 SAMPLE HILL0065.D PFOS-3.1.2 042301-PFOS-3.1.2 Dup PFOS 5.242 7544482 292.7 287 SAMPLE HILL0066.D PFOS-3.1.3 042301-PFOS-3.1.3 Trip PFOS 5.242 6794540 265.0 259 SAMPLE HILL0072.D PFOS-3.2.1 042301-PFOS-3.2.1 Smp PFOS 5.'245 8117378 308.4 302 SAMPLE HILL0073.D PFOS-3.2.2 042301-PFOS-3.2.2 DUP PFOS 5.249 8786445 359.3 352 -0 SAMPLE HILL0074.D PFOS-3.2.3 042301-PFOS-3.2.3 Trip PFOS 5.251 9095599 319.1 312 InP m SAMPLE HILL0077.D PFOS-3.3.1 042301-PFOS-3.3.1 Smp PFOS 5.238 7558687 292.4 286 VI 0 SAMPLE HILL0078.D PFOS-3.3.2 042301-PFOS-3.3.2 Dup PFOS 5.243 6689145 262.7 257 A r? SAMPLE HILL0079.D PFOS-3.3.3 042301-PFOS-3.3.3 Trip PFOS 5.234 7103605 275.7 270 2 from HPLClMSD "Hillary" analytical sequence H010430.s, reprocessed with Target Software by KLT z 269 Day 3 results, pg/mL 15.4 average 287 6% std dev 30 rn 322 26.5 8% $ gsx 271 14.5 5% 222 CXAGt Copy- or OriQis35.1 -.:J 3M Confidential BACK TO MAIN Solubility Study, Shake Flask Method: Test Compound P F O S Test Matrix: MaleFernale Normal Human Urine; Timepoint(s): Days Iand 2 Analytical Instrument: HPLCIMSD "Hilfary"; AnalyticalSequencelMethod H010426.dPFOS0425.m (ChemStation):Reprocessing Method: H010426t.m (Target) Batch Sam T e 4/26/01 1957 D:khemWiWO10426.b SA&: 4/26/01 19:48 D WemWillary iW010428b SAMPLE 4/25/01 1959 D \chemWdlaryiW010426b CAUE-1 4/26/01 2O:lO D khemwdlary RHO10426 b CALIE-2 4/26/012021 D khemWdlary iW010426b CALIE-3 4/26/01 2033 D khemWillary RHO10426b CALIE-4 4/26/01 20.44 D khem\Hillary tW010426 b CAUE-5 4/26/0120:55 OkhemWillary lW010426b CALIE-6 4/26/0121:06 D khemWillary iW010426b CALIE-7 4/25/01 21:17 D khemWdlaryRHO10426b CALlE-8 4/26/01 21:28 D khemwdlary iW010426b CALlE-9 4/26/0121:40 D khemWillary iW010426b CALIB-10 4/26/012151 D khemWillary iW010426b SAMPLE 4/27/01 0 2 8 D khemWillary iW010426b SAMPLE 4/27/01039 D khemWullaryiW010426b SAMPLE 4/27/013 0 5 D \chem\HllaryiWOlM26 b SAMPLE 4/271013:17 D khemWillary iW010426b SAMPLE 4/27/01 5 5 3 D khem\Hllary RHO10426 b SAMPLE 4/27/016.04 D \chemUiollaryhH010426b SAMPLE 4/27/01734 D WemWillary iWO10426 b SAMPLE 4/27/01745 D khemWdlaryiWOlM26 b SAMPLE 4/27/01 8'19 D WemWillary MOlMZ8 b SAMPLE 4/27/018 3 0 D khemWllary iW010426b SAMPLE 4/27/01 8:41 D WemWWdry iWOl0426b CALlE-11 4/27/01 8:52 D khem\HdlaryiW010426b CALlE-12 4/27/019:03 D WemWillary RHO10426 b CALIE-13 4/27/01 9:15 D khemWillary iw010426 b WE-14 4/27/019 2 6 D khemWillary i!HOI0426 b CALIE-15 4/27/019x37 OkhemWillary iWO10426 b CALlE-16 4/27/01948 D khemwillary RHO10426b CALIE-17 4/27/01 9:59 D WemWdlary iWOlM28 b CALIE-I8 4/27/011O:lO D \chcmWdlaryiw010426b CAUE-19 4/27/01 10:22 D khemWillarv lW010426b CALIE 20 4/27/01 10:33 D:khem\Hifl~~.iWOlO426.b SAM< Two curves were averagedusingTargels o M m lo qunnlify dab. File Srrnsle Name Samele Derc. H 1 L L ~ i e . o 01003-07-01 0 nglmL PFOS HILL0017.D 01003-07-02 2.5 ndmL PFOS HILLW18.D 01003-0743 5 n&L PFOS HILLW19.D 01003-07-04 10 ngmL PFOS HlLLW20.D 01003-07-05 25 nglmL PFOS HILL0021.D 01W3-0746 40 n#mL PFOS HILLW22.D HILLOM3.D 01003-07-07 01003-07-08 50.1 ng4nL PFOS . 75.1 nglmL PFOS HILLW24.D 01003-07-09 100.2 nglrnL PFOS HILLW25.D 01003-07-10 250.5 ndmL PFOS HILLOO26.D HILLW27.D 01003-07-11 01003-07-12 400.8 ndmL PFOS , 501 nglrnL PFOS HILL0028.D 01003-07-13 1002 n@nL PFOS HILLW42.D 01003-07-03 5 nglmLPFOS HILL0043.D 01003-07-09 100.2 nglmL PFOS HILL0056.D 01W3-07-03 5 ndmL PFOS HILLW57.D 0100507-09 1002 nglrnLPFOS HILL0071.D 01W3-07-03 5 ndmL PFOS HILLW72.D 01003-07-09 100.2nglrni PFOS HILLW8O.D 01003-07-03 5 nglmLPFOS HILLW81.D OlW3-07-09 100.2ndmL PFOS HILLO084.D 01003-07-01 0 ngJrnL PFOS HILLOO85.D 01003-0742 2.5 nglmL PFOS HILLW88.D 010050743 5 n g h L PFOS HILL0087.D 01005.07-04 10ngmL PFOS HILL0088.D 01003-07-05 25 nplmL PFOS HILLW89.D 0100507-06 40 ndmL PFOS HILLW9O.D 01003-07-07 50.1 ngrnL PFOS HILLW91.D 010050748 75.1 ngrmL PFOS HILLOO92.D 01003-0749 1W.2 n#mL PFOS HILLW93.D 01003-07-10 250.5 nglmLPFOS HlLL0094.D 01003-07-11 400.8 n@mLPFOS HILL0095.D 01003-07-12 501 ndmL PFOS HlUW98.D 01003-07-13 1002 n&L PFOS A standardc v r a rangeofonly 10levelsis allowsd. 5 5 W n # m L m r THPFOS THPFOS THPFOS THPFDS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFDS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS llie rangeshoren. !?%I 5.115 5.1 1 5.125 5.112 5437 5.115 5.094 5.131 5.097 5.117 5.11 5.11 5.109 5.111 5.116 3.141 5.125 5.122 5.133 5.133 5.13 5.127 5.13 5.123 5.13 5.123 5.124 5.13 5.124 5.122 5.123 5.123 5.132 5.123 &?a 1238088 1251680 1273628 1280488 1289080 1294694 1319187 1320686 1316830 1333532 1326197 1324859 1323984 1428715 I441029 1487926 1456794 1499176 1477095 1504963 1469125 1491338 1484350 1517040 1493288 1476860 1492539 1489446 1438972 1481229 1455720 1457008 1455144 1426062 1406375 88296 6.3% &?.Wa 254 254 254 254 ,254 254 254 .254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 ' SirnuTwe File Sample Name Sample Doas. ComDound Narnm 4/26/01 2224 SAMPLE HlLLT3I.D 042301-PFOSLCS PFOS Solvent Blank Spike THPFOS 5.105 1356608 254 4/26/01 22:36 D khemWillary iW010426b SAMPLE HILLW32.D PFOS1.4.1Mk 042301-PFOS1.4.1 Mtd Elk THPFOS 5.112 1357657 254 4/26/01 2247 D khemWillaiy iW010426b SAMPLE HILLOO33.D P F O S l l Z b l k 042301-PFOS1.4.2 Mld BIk THPFDS 5.111 1371641 254 4/26/012258 0 WemWillary iW010426b SAMPLE HILL0034.D PFOS1.4.3Mk 042301-PFOS1.4.3 MM Blk THPFOS 5.117 1347413 254 4/26/0123:W D khcmWdlary iW010426b SAMPLE HILLW35.D PFOS1.4.4blkMS 042301-PFOS1.4.4 Mld Elk Spke THPFOS 5.137 1362229 254 4/26/012332 D khemWdlaryiW010426b SAMPLE HILL0037.D PFOS1.l.l 042301-PFOS1.l.l Smp THPFOS 5.123 1239933 254 4/26/0123:43 D khemWillrry RHO10426b SAMPLE HILLOO38.D PFOSl.l.2 042301-PFOS1.12 Dup THPFOS 5.123 1242749 254 4/26/01 23:54 D khemWdlary AH010426 b SAMPLE HILL0039.D PFOS1.1.3 042301-PFOS1.1.3 Trip THPFOS 5.115 1244758 254 4/27/01 0:05 D WemWillary W010426 b SAMPLE HILLO04O.D PFOSl.1.4MS 042301-PFOS1.1.4 Spike THPFOS 5.117 1243279 254 -0 4/27/010:16 D khcmWllary iW010426b SAMPLE HILL0041.D PFOS1.2.1 042301-PFOS1.2.1 Smp THPFOS 5.111 1262274 254 a, ul 4/27/01 1:12 D khemWillary iW010426b SAMPLE HlUW46.D PFOS1.2.2 042301-PFOSl22 Dup THPFOS 5.123 1279599 ' 254 (D 4/27/01124 D khemWdiary iW010426b SAMPLE HILL0047.D PFOSi.2.3 042301-PFOS1.2.3 Trip THPFOS 5.124 1288849 254 2 4/27/01 135 D khemWillary iW010426b SAMPLE HILLW48.D PFOSt24MS 042301-PFOS1.2.4 Spike THPFOS 5.115 1283580 254 0, A 2 Page 1 Of 5 PFES PFBS PFES PFES PFBS PFBS PFES PFES PFES PFES PFES PFES PFES PFBS PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES PFES 4.383 4.39 4.383 4.38 4.38 4.367 4.376 4.375 4.382 4.389 4.374 4.397 4.401 4.405 4.397 4.407 4.411 4.411 4.408 4.412 4.409 4.408 4.409 4.409 4.41 4.418 4.409 4.407 4.409 4402 4.41 4.402 ARI 1933641 1933710 1932682 1943358 1949841 194 1643 1930302 1953486 1971932 1956172 1987259 1977603 1983865 2021892 2000887 2033053 2045778 2055427 20449% 2044088 2044316 2052700 2038788 2035215 2007995 2020150 2027822 2029826 I952364 2023371 1982961 2031354 2025464 1994320 1997302 41289 2 1% Amount 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 2596492 1398111 +30% response -30% response CornuoundNama PFBS PFBS PFBS PFES PFES PFES PFES PFES PFBS PFES PFBS PFBS PFES 4.377 4.37 4.369 4.374 4.388 4.388 4.374 4.393 4.395 4.397 4.388 4.395 4.394 &g 1932182 1900574 1910506 1921068 1925821 2179012 2222026 2225836 2165910 2148762 2164574 2202302 2190236 AM 254 254 254 254 254 254 254 254 254 254 254 254 254 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Tesl Matrix: MatelFernale Normal Human Urine; Tlrnepolnt(s): Days Iand 2 Analylical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod HOl0426.r/PFOS0425.m (ChemStation); Reprocessing M e t h o d H010426t.m 4/27/01 1:47 D:khem\Hiliary.iWOl0426.b SAMPLE HILLW49.D PFOS1.3.1 042301-PFOS-1.3.1 Smp THPFOS 5.13 1269547 4/27/01 209 O:WemVlillary.i~O10426.b SAMPLE HlLL0051.D PFOS1.3.2 042301-PFOS1.3.2 Dup THPFOS 5.118 1282405 4/27/012 2 0 4/27/012 3 1 DkhemVlillary.iWOlO426.b O:Wem\Hillary.~H010426.b SAMPLE HILL0052.D SAMPLE HILL0053.D PFOS1.3.3 PFOS1.3.4MS 042301-PFOSl.3.3 Trip 042301-PFOS1.3.4 Spike THPFOS THPFOS 5.132 5.116 1283479 1277667 4/27/012 4 3 4/27/01 254 4/27/01 3:50 4/27/01 4:Ol 4/27/01 4:13 4r27mi 424 4/27/01 435 4/27/01 457 4/27/01508 4/27/01 5 2 0 4/27/01 531 4/27/01 5:42 4/27mi 6 : 4/27/01 6:49 4/27/01 7 0 0 4/27/01 7 1 1 4/27/01 7 2 3 O:Wem\Hillary.iW010426.b DWemWillary.iWO10426.b D khemWillaG iW010426b 0 khem\HdlaryiW010426b D WemWillary iWO10426 b D khemWillary iv1010426b D WemWillary iW010428b D \chem\Hillary iWO10426 b D khem\Hillary iW010426b 0 khcm\H4laryiW010426b D khemWillaryiW010426b D Wem\Hillary i\H010426b ~ D khemVlllary 1W010426b D khrmWJlaryiW010426b D WemWllary M010426 b DWemW.1lary iW010426b D Wem\Hillary iW010426b SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILLW54.D HILL0055.D HlLLO06O.D HlLL0061.D HILL0062.D HILLCW3.D HILL0064.D HILL0066.D HILL0067.D HILL0068.D HILL0069.D HILLW70.D HILL0075.D HILL007S.D HILL0077.D HILL0078.D HlLL0079.D 042301-PFOSLCS PF062.4.lblk PFOS2.4.2blk PFOS2.4.3blk PFOS-2.4.4MkMS PFOS2.1. I PFOS2.12 PFOS2.1.3 PFOS2.1.4MS PFOS-2.2.1 PFOS-2.2.2 PFOS2.2.3 PFOS2.2.4MS PFOS2.3.1 PFOS2.32 PFOS2.3.3 PFOS2.3.4MS PFOS Solvent Blank Spike 042301-WOS-2.4.1 Mld Blk 042301-PFOS2.4 2 Mld Blk 042301-PFOS24.3 Mld Blk 042301-PFOS24 4 Mtd Blk Spike 042301-PFOS2.1 1 Smp 042301-PFOS2.12 Oup 042301-PFOSZ 1 3 Tnp 042301-PFOS2.1 4 Spike 042301-PFOS22 1 Smp 042301-PFOS2 2 2 Dup 042301-PFOSZ2.3 Trip 042301-PFOSZ 2 4 Spike 042301-PFOS2.3.1 Smp 0423Ol-PFOSZ 3 2 Dup 042301-PFOS2.33 Trip 042301-PFOS2.3.4 Spike THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS 5.123 3.13 5.131 5.119 5.13 5,122 5.136 5.123 5.13 5.136 5.137 5.131 5.137 5.129 5.123 5.133 5.137 1423459 1433506 1413614 1421004 1403989 1287849 1278347 1294319 1272974 1434509 1421082 1441500 1430472 1423581 1417186 1411658 1451012 (Target) 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 me 4/26/01 1914 4/26/01 1926 4/26/01 2 2 0 2 4/26/01 2213 4/26/01 2320 4/27/01 0:50 4/27/01 1:Ol 4/27/01 1:58 4/27/01 3 2 8 4127/01 339 4/27/01 4:46 4/27/01 6:15 4/27/01 6-27 4/27/01 7:56 4/27/01 8:08 4/27/01 1044 4 ~ 7 1 0 1io:^ Batch Dkhem\Hillary.i\WOlO426.b D khem\HdlaGiW010426.b D WemWdlary nH010426 b D khemWillary iW010426b D khemWinaryN1010426b D khemWillary iW010426b D khemWillary iW010426b 0 khem\Hillary AH010426 b D khern\H#llsryRHO10426b D khcm\Hillary iW010426b D WemWdlary iW010426b D khemWtllary iW010426b D\shemW4laryiW010428b D khem\Hillary iW010426b D khemWillary iW010426b 0 khemWillary iW010426b 0.khemWdlary iW010426b Sam ",:SA: SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0014.D HILL0015 D HILL0029 D HILLW30 D HILL0036 D HILL0044 D HILL0045 D HILL0050 D HILLW.56 D HILLW59 D HILLGC65 D HILL0073 D HILL0074 D HILL0082 D HILL0083 D HILLW97 D HILL0098.D S8mole ame TN-A480:MeOH TN-A 4802 MeOH TN-A4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH T K A 4802 MeOH TN-A 4802 MeOH TN.A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MaOH TN-A4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN.A 4802 MeOH TN-A 4802 MeOH Samdr Dele, Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank SolvenlBlank Solvent Blank Solvent Blank Sdvenl Blank Solvenl Blank Solvenl Blank Solvent Blank Solved Blank Solvenl Blank Solvenl Blank Solvenl Blank Solvent Blank ComDoundNamq THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS BI 0 0 0 0 0 5.255 0 0 0 0 5258 '0 0 0 0 0 0 1342769 73848 6 6%._ 0 0 0 0 0 7825 0 0 0 0 7710 0 0 0 0 0 0 Amount 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 PFBS 4.394 2195975 254 PFBS 4.394 2216090 254 PFBS 4.396 2203291 254 PFBS 4.394 2220887 254 PFBS 4.394 1977507 254 PFBS 4.395 1931697 254 PFBS 4.409 1944188 254 PFBS 4.398 1913426 254 PFBS 4.408 1698991 254 PFBS 4.4 2201997 254 PFBS 4.415 2231938 254 PFBS 4.409 2202521 254 PFBS 4.409 2203090 254 PFBS 4.401 1973173 254 PFBS 4.409 1956138 254 PFBS 4.41 1940430 254 PFBS 4.423 1930607 254 PFBS 4.415 1936770 254 PFBS , 4.416 1899523 254 PFBS 4.419 1917144 254 PFBS 4.408 1951546 254 2056957 2674044 ComDoundNamQ E 137331 e.. f-.7 . a 1439870 &y+yJj PFBS 4.387 8824 0 PFBS 0 0 0 PFBS 4.39 9125 0 PFBS 0 0 0 PFBS 4.402 8664 0 PFBS 4.407 8480 0 PFBS 0 0 0 PFBS 4.408 8742 0 PFBS 4.423 8244 0 PFBS 0 0 0 PFBS 4.423 9199 0 PFBS 4.429 8487 0 z PFBS 0 0 0 w PFBS 4.436 8442 0 PFBS 0 0 0 PFBS 4.431 9918 0 PFBS 0 0 0 ... J Page 2 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MaldFemala Normal Human Urine; Timepolnt(s): Days 1and 2 h . Analytical Instrument: HPLClMSD "Hillary'; Analytical SequencelMethod H010426.slPFOS0425.m (ChemStatlon); R e p r o c e r r l n g Method: H010426t.m (Tahet) Batch Sam T e File SamoleNann Samota Dess, Comoound Nanm XTheomtlca( , 4/26/01 1937 D:khem\HiiHO10426.b SAIP; HILLx16.D 01003-07-01 0 n@mLPFOS PFOS 0 4/26/01 19:48 D khem\HiB& iW010426b SAMPLE HILL0017.0 01003-07-02 2.5 ngmL PFOS PFOS 5.32 81289 2.8 4/26/01 1959 D WlemWdhry MOlM26 b CALIE-1 HILL0018.D 01003-07-03 5 nglmL PFOS PFOS 5.335 138659 5.0 99.5% 4/26/012010 D khem\HdlaryRHO10426 b CALIE-2 HILLW19.D 01003-07-04 10 nglmL PFOS PFOS 5.329 254205 8.3 93.4% 4/26/01 2021 D khem\Hillary iW010426b CALIE-3 HILLW2O.D 0100507-05 25 ngmL PFOS PFOS 5.315 660047 24.9 99.5% 4/26/01 20.33 DkhemWillary iW010426b CALIE-4 HILLW21.D 01WSO7-06 40 ngmL PFOS PFOS 5.332 1055533 40.5 1012% 4/26/01 20:44 D khemWillary iWOlM26 b CALlE-5 HlLLW22.D 0100507-07 50.1 ng'mL PFOS PFOS 5.318 1291097 50.1 100.1% 4~610120:55 OkhemWiilary iWO10426 b CALIE-6 HILLOO23.D 0100507-08 75.1 nglmL PFOS PFOS 5.348 1939424 75.6 1w.7x 4/26/01 21:06 0 khemWillaryRHO10426 b CALIE-7 HILL0024.D 01003-07-09 100.2 nglmLPFOS PFOS 5.322 2537734 99.3 99.1% 4/26/0121:17 D khemWllary iW010426b CALM-8 HILL0025.D 01003-07-10 250.5 nglmL PFOS PFOS 5.335 6004211 254.8 101.6% 4/26/0121:28 D Wern\HillacyRHO10426b CALIE-9 HlLL0026.D 01003-07-lt 400.8 nglmL PFOS PFOS 5.327 8957654 394.8 98.4% 4/26/0121:40 D khemWllary tW01M26 b CALlE-10 HILL0027.D 01003-07-1 2 501 ngmL PFOS PFOS 5.328 10985Lu5 504.8 1W.8% 4/26/012151 D khem\HIIarf iW010426b SAMPLE HILL0028.D 01003-07-13 1002 ngmL PFOS PFOS 5.326 18864507 983.7 4/27/010 2 8 D khemWdlaryhH010426b SAMPLE HILL0042.D 01003-07-03 5 n@mLPFOS PFOS 5.329 176475 6.1 122.5% 4/27/01 039 D !chemWllary Mi010426 b SAMPLE HILL0043.D 0100307-09 100.2 nglmL PFOS PFDS 5.333 2506427 99.7 99.5% m 7 m i 3.05 D khem\Hillary hHOlM26 b SAMPLE HILLO056.D 01003-07-03 5 ndmL PFOS PFOS 5.351 150106 5.1 102.6% 4/27/01 3:17 D khemWdlaryRH010426b SAMPLE HILLOO57.D 0100507-09 100.2 ngmL PFOS PFOS 5.335 2627085 99.0 98.8% 4/27/01553 D IchemW.llary iW010426b SAMPLE HILLW71.D 01003-07-03 5 nglmL PFOS PFOS 5.325 151691 5.1 99.5% 4/27/01 6:04 D khemWillary N-1010426 b SAMPLE HlLL0072.D 01W3-07-09 100.2 n g h L PFOS PFOS 5.336 2666281 100.6 100.4% 4/27/01 734 D khemWillary iWOlM28b SAMPLE HILLOO8O.D 01w307-03 5 ngmL PFOS PFOS 5.324 148887 5.1 101.1% 4/27/01 7:45 0 khem\Hillafy iWOlM26 b SAMPLE HILLW81.D 0100307-09 100.2 ngmL PFOS PFOS 5.333 2633934 98.4 99.2% 4/27/018:19 D khemWillaryRHO10428b SAMPLE HILL0084.D 01003-07-01 0 ngmL PFOS PFOS 0 0 0.0 4/27/018 : ~ o D khemWillary iWOlMZ6 b SAMPLE HILL0085.D 01003-07-02 2.5 ngmL PFOS PFOS 5.333 83865 2.7 4/27/018:41 DLhemWillsry iW010426b CALlB-11 HILL0086.D 01003-07-03 5 nglmL PFOS PFOS 5.333 142999 4.9 97.4% 4/27/01852 Dkhem\Hillary iW01M26 b CALIB-12 HILL0087.D 01003-07-04 10 nglmL PFOS PFOS 5.333 254810 9.1 90.5% 4/27/019:03 D khemWdlaryiWOlM26 b CALIB-13 HILL0088.D 01003-07-05 25 nglmL PFOS PFDS 5.334 687053 25.0 100.0% 4/27/01 915 D khemWillary iW01M26 b CALlE-14 HILLOO89.D 01W501d6 40 nglmLPFOS PFOS 5.327 1065908 39.1 97.8% 4/27/01 9:m 0 khemWillary lW01M26 b CALIE-15 HILLOO9O.D 01003-07-07 50.1 ngmL PFOS PFOS 5.34 1380299 502 100.3% 4/27/01937 0 WemWdlary 1W01M26b CALlE-16 HILLW9l.D 01003-07-08 75.1 ngmL PFOS PFOS 5.327 1965188 76.7 102.1% 4/27/019:48 D khem\Hillary RHO10426b CALIE-17 HILLW92.D 010050749 100.2 ndmL PFOS PFOS 5.332 2630530 100.3 100.1% 4/27/01959 D khem\HdlaryRHO10426b CALlE-18 HILL0093.D 01003-07-10 250.5 n#mL PFOS PFOS 5.334 6077465 254.1 101.5% 4/27/01 i o 1 0 D khemWillary M010428 b CALlE-19 HILL0094.D 01003-07-11 400.8 ndmL PFOS PFOS 5.326 9143568 393.9 98.3% , 4/27/01 10:22 D khemWillary iWO10426b CALIE-20 HILL0095.D 01W.?-07-12 501 ndmL PFOS PFOS 5.335 11226796 503.5 100.5% 4/27/01 1033 D khemWillaryiW01M26 b SAMPLE HILL0096.D 0100507-13 1 w 2 n3mL PFOS PFOS 5.326 19703670 1033.7 Two curyeswere avenged usingTargel soltwara 10 quantifydata. A standardwfvennge D l only 10levelsIs allowed. 5-500 n@mLmslhs rangechosen. Page 3 of5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MilelFernale Normal Human Urine; Tlrnepolnt(s): Daya 1and 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod H010426.slPFOS0425.rn (ChernStation); Reprocessing Method: HOfO426t.m (Target] 4/26/01 2224 4/26/01 2236 4/26/01 22:47 4/26/012258 4/26/01 23:og D:\chemWillary.iWO10426.b D:\chemWillary.iWOl0426.b D:\chemWillary.NiO10426.b D.WlemWillaly.iWO10426.b D:khemV(ilIary.NiOlMZ6.b SamDTvDe SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE @ HILLOO31.D HILLW32.D HIlL0033.D HILL0034.D HILLW35.D Sarnolc Nany 042301-PFOSLCS PFOS1.4.lblk PFOS1.4.2Mk PFOS1.4.3blk PFOSlIIbIkMS SarnDlc Derc. PFOS Solvent Blank Spike 042301-PFOS1.4.1 MldBlk 042301-PFOS1.42 Mld Blk 042.301-PFOS1.4.3 Mld Blk 042301-PFOS1.4.4 Mtd Blk Spike PFOS PFOS PFOS PFOS PFOS 5.322 0 0 0 5.354 2260230 0 0 0 2520637 Amoun( 89.8 0.0 0.0 0.0 101.1 Average r@mL Std Dev Purity %RSD corrected nplrnL Spike Recovery 88 90% 99 101% 4n6mi 2332 4/26/01 23:43 4/26/01 23:54 4/27/01 0.05 D:\chemWillary.NiO10426.b D:khemlHi~ary.iWO10426.b D:khemWillary.iWO0l 426.b D:khemWillary.iWOl0426.b SAMPLE SAMPLE SAMPLE SAMPLE HILLO037.D HILLW38.D HILL0039.D HILLW40.D PFOSl.l.l PFOS1.12 PFOS1.1.3 PFOSl.1.4MS 042301-PF051.1.1 Smp 042301-PFOS1.1.2 Dup 042301-PFOS1.1.3 Trip 042301-PFOS1.1.4 Spike PFOS 5.271 7510273 292.6 286 PFOS 5.277 8619490 331.8 325 PFOS 5.255 8589480 329.8 323 PFOS 5.264 9810483 396.8 388 311 22.0 70.6 78% 4/27/01 0:16 4/27/01 1:12 4/27/01 124 4/27/01 1:35 D:khem\Hillary.iWO10426.b D:khemWillary.NiO10426.b D:\chemWillary.NiOl0426.b D:khemWillaly.iWO10426.b SAMPLE SAMPLE SAMPLE SAMPLE HILLW41.D HILLW6.D HILLO047.D HILLDM&D PFOS1.2.1 PFOS1.2.2 PFOS1.2.3 PFOS-1.2.4MS 042301-PFOS-1.2.1 Smp 042301-PFOSl.2.2 Dup 042301-PFOS1.2.3 Trip 042301-PF051.2.4 Spike PFOS 5251 7600633 298.7 292 PFOS 5.263 7571396 294.9 289 PFOS 5.264 7594271 2902 284 PFOS 5.2% ~ ~ 6 7 8 9 9 385.1 377 288 4.0 1% 90% average rlddev XRSD: Day 1results 302 12 4.0% 4/27/01 1:47 4/27/01 2 0 9 4/27/01 2 2 0 4/27/01 2:3i 4/27/01 2 4 3 D:khemWillary.NiO10426.b D:\chemWillary.RHO10426.b D:\chemWillaty.NiOI 0428.b D.khemWillary.i\HO10426.b D:\chemVlillary.iW010426.b SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0049.D HILLOO51.D PFOS1.3.1 PFOS-1.3.2 HILL0052.D PFOS1.3.3 HILL0053.D PFOS1.3.4MS HILL0054.D 042301-PFOSLCS 042301-PFOS1.3.1 Smp 042301-PFOSl.3.2 Ch~p 042301-PFOS1.3.3 Trip 042301-PFOs1.3.4 Spike PFOSSolvsnlBlank Spike PFOS 5.27 8267658 320.6 314 PFOS 5.263 80456W 307.6 301 PFOS 5.272 8019673 308.5 302 PFOS 5.263 10242365 405.2 397 PFOS 5.333 2342701 91.0 89 306 7.2 2% 93% 91% 4/27/012 5 4 D:\chem\Hillafy.iWOl0426.b SAMPLE HILL0055.D PFOS2A.lblk 042301-PFOS2.4.1 Mld Blk PFOS 0 0 0.0 4/27/013 5 0 D:themVlillary.NiO10426.b SAMPLE HILL0060.D PFOS2.4.2Mk 042301-PFOS2.4.2 Mld Elk PFOS 0 0 0.0 4/27/01 4:Ol DWlemWillary.NiO10426.b SAMPLE HILLOOG1.D PFOS2.4.3blk 042301-PFOS2.4.3 Mld Blk PFOS 0 0 0.0 4/27/01 4:13 D\chemWillary.i\HOlM26.b SAMPLE HILLOOG2.D PFOS2.4.4blkMS 042301-PFOS2.4.4 Mld Elk Spike PFOS 5.333 2571930 104.8 103 105% 4/27/01 4 2 4 4/27/01 4 3 5 4/27/01 4 5 7 ~ 7 m5.0i 8 D:WlemWillary.Ni010426.b D:WemWillary.NiOl0426.b D:\chemWillaly.NiO10426.b DkhcmWillary.iWO10426.b SAMPLE SAMPLE SAMPLE SAMPLE HlLLOC63.D HILL0064.D HlLLO066.D HlLL0067.D PFOS2.1.1 PFOS2.1.2 PFOS2.1.3 PFOS2.1.4MS 042301-PFOS2.1.1 Smp 042301-PFOS2.12 Dup 042301-PFOS2.1.3 Trip 042301-PFOS2.1.4 Spike PFOS 5.262 7822147 300.1 294 PFOS 5.276 8758474 3362 329 PFOS 5257 8011616 308.3 302 PFOS 5.27 10379987 415.5 407 308 18.3 6% 701% 4/27/01520 4/27/015 3 1 4/27/01 5:42 DkhemlHillary iW010426.b D.ishernWillary.NiOl0426.b D:\chemWillary.NiO1M26.b SAMPLE SAMPLE SAMPLE HILLGC66.D HILLO069.D HILL0070.D PFOS2.2.1 PFOS2.2.2 PFOS2.2.3 042301-PFOS2.2.1 Smp 042301-PFOS-2.22 Dup 042301-PFOS2.2.3 Trip PFOS 5.36 48170 1.4 PFOS 0 0 0.0 PFOS 0 0 0.0 4/27/016 3 8 DkhemWillary.iWO1MZ6.b SAMPLE HILL0075.D PFOS2.2.4MS 042301-PFOS2.2.4 Spike PFOS 5.34 2618681 104.9 103 105% 1 w 4/27/016:49 D\chemWillary.iWO10426.b SAMPLE HILLW76.D PFOS2.3.1 042301-PFOS2.3.1 Smp PFOS 0 0 0.0 4/27/01 7:W D:\chemWiUary.ilHO10426.b SAMPLE HILL0077.D PFOS23.2 042301-PFOS2.32 Dup PFOS 0 0 0.0 4/27/01 7:11 D:khemWillary.iWO10426.b SAMPLE HILL0078.D PFOS2.3.3 042301-PFOS2.3.3 Trip PFOS 0 0 0.0 4/27/01 72.7 D:WemWillary.NiO 10426.b SAMPLE HILL0079.D PFOS2.3AMS 042301-PFOS2.3.4 Spike PFOS 5.34 2483685 98.1 96 98% Paps4 of 5 BACK TO MAIN Solubility Study, Shake Flask Method: Test Compound: PFOS: Test Matrix: MaleIFemale Normal Human Urine: Timepoint(s): Days 1 and 2 Analytical Instrument: HPLCIMSD "Hillary"; AnalyticalSequencflethod H010426.s/PFOS0425.m (ChemStation): ReprocessingMethod: H010426t.m (Target) lniDatc 4/26/01 1914 SatnuTvm & Samul*Nam SAMPLE HILLW14.D TKA4802 MeOH Sample Desc, Solvent Blank Comuound Name EI PFOS 5.424 Eel 58585 Amount Below LOQ 4/26/oi i9:26 4/26/01 2 2 0 2 4/26/01 2213 4/26/01 2320 SAMPLE SAMPLE SAMPLE SAMPLE HILLWi5.D HILUW19.D HILL0030.D HILLW38.D TN-A4802 MeOH TN-A4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH Solvent Blank Solvent Blank Solvent Blank Solvenl Blank PFOS PFOS PFOS PFOS 0 5.419 5.431 0 0 65012 22312 0 0 B S ~ WLOQ Below LOQ 0 4/27/010 5 0 4/z7/oi i : o i 4/27/01 153 SAMPLE SAMPLE SAMPLE HiLLW44.D TN-A 4802 MeOH HILLW45.D TN-A4802 MeOH HiLL0050.D TN-A 4802 MeOH Solvent Blank Solvent Blank Solvent Blank PFOS PFOS PFOS 0 0 5.367 0 0 31709 0 0 Below L W 4/27/01 328 SAMPLE HILLW58.D TN-A4802 MeOH Solvent Blank PFOS 0 0 0 4/27/01 339 4/27/01 4:46 4/27/016 3 5 SAMPLE SAMPLE SAMPLE HILL0059.D TN-A4802 MeOH HILLOW5.D TN-A 4802 MeOH HILL0073.D TNA4802 MeOH Solvent Blank Solvent Blank Solvent Blank PFOS PFOS PFOS 0 0 0 5.34 33691 BDIOWL o a 0 0 0 4mmt 6:n SAMPLE HtLLW74.0 TN-A 4802 MeOH Sdvenl Blank PFOS 0 0 0 4/27/01 756 4/27/01 8:08 SAMPLE HILLW82.D TN-A 4802 MsOH SAMPLE HILL0083.D TN-A 4802 MeOH Solvent Blank Solvent Blank PFOS 0 0 0 PFOS . 0 0 0 4/27/01 10:44 4/27/01 1055 SAMPLE HlLL0097.D TN-A4802 MeOH SAMPLE HILLOW8.D TN-A 4802 MeOH Solvent Blank Solvent Blank PFOS PFOS 5.433 5.431 50495 14767 Below L o a 0 U a P In Page 5 of 5 BACK TO MAIN I Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Day 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SeqoencemMethod: H010501.slPFOS0425.m (ChemStation); ReprocessingMethod: H010501t.m (Target) Quantrange: 10-501ng/mL Day 2 samples were re-aliquotted from the initial centrifuge tube on 05-01-01 into 1.5 mL centrifuge tubes, centrifuged, and then diluted 1:lOO and 1:lO for a final dilution factor of 1:l.OOO prior to analysis. SampType File SamDle Name Misc Info CompoundName RT Area Amount Compound Name RT Area Amount SAMPLE HILL0016.D 01003-07-01 0 nglmL PFOS THPFOS 5.119 1282312 254 PFBS 4.39 1737872 254 SAMPLE HILLO017.D 01003-07-02 2.5 ng/mL PFOS THPFOS 5.116 1300843 254 PFBS 4.388 1742704 254 CALIB-1 Disabled HILL0018.D 01003-07-03 5 nglmL PFOS THPFOS 5.123 1310415 254 PFBS 4.409 1726536 254 CALIB-2 HILLOOl9.D 01003-07-04 10 nglmL PFOS THPFOS 5.122 1318634 254 PFBS 4.408 1720895 254 CALIB-3 HILL0020.D 01003-07-05 25 nglmL PFOS THPFOS 5.117 1324328 254 PFBS 4.41 1770020 254 CALIB-4 HILL0021.D 01003-07-06 40 ng/mL PFOS THPFOS 5.117 1329731 254 PFBS 4.409 I787802 254 CALIB-5 HlLL0022.D 01003-07-07 50.1 nglrnL PFOS THPFOS 5.124 1346360 254 PFBS 4.416 1786195 254 CALIB-6 MlLL0023.D 01003-07-08 75.1 nglrnL PFOS THPFOS 5.118 1335866 254 PFBS 4.41 1 1789034 254 CALIB-7 HILL0024.D 0 1003-07-09 100.2 nglrnL PFOS THPFOS 5.125 1352724 254 PFBS 4.404 1791645 254 CALIB-8 HILL0025.D 01003-07-10 250.5 nglrnL PFOS THPFOS 5.124 1343468 254 PFBS 4.417 1774591 254 CALlB-9 HILL0026.D 01003-07-11 400.8 ng/rnL PFOS THPFOS 5.119 1338624 254 PFBS 4.41 1 1789155 254 CALlB-IO HILL0027.D 01003-07-12 501 nglmL PFOS THPFOS 5.115 1349352 254 PFBS ' 4.408 1796642 254 SAMPLE HILL0028.D 01003-07-13 1002 nglrnL PFOS THPFOS 5.116 1311155 254 PFBS 4.408 1764193 254 SAMPLE HILL0041.D 01003-07-03 5 nglmL PFOS THPFOS 5.129 1385797 254 PFBS 4.400 1769946 254 SAMPLE HILL0042.D 01003-07-09 100.2 ng/rnL PFOS THPFOS 5.117 1376301 254 PFBS 4.41 1764551 254 SAMPLE HILL0054.D 01003-07-03 5 nglmL PFOS THPFOS 5.122 1375001 254 PFBS 4.401 1783815 254 SAMPLE HlLL0055.D 01003-07-09 100.2 nglrnL PFOS THPFOS 5.117 1372102 254 PFBS 4.402 1769874 254 SAMPLE HILL0058.D 01003-07-01 0 nglmL PFOS THPFOS 5.131 1373368 254 PFBS 4.409 1780686 254 SAMPLE HILL0059.D 01003-07-02 2.5 nglmL PFOS THPFOS 5.108 1387395 254 PFBS 4.407 1788657 254 CALlB-I 1 Disabled HILL0060.D 01003-07-03 5 nglmL PFOS THPFOS 5.115 1396556 254 PFBS 4.401 1769985 254 CALI8-1 2 HILL0061.D 01003-07-04 10 nglmL PFOS THPFOS 5.117 1407101 254 PFBS 4.417 1750426 254 CALIB-I3 HILL0062.D 01003-07-05 25 nglmL PFOS THPFOS 5.117 1388273 254 PFBS 4.416 1770162 254 CALIB-14 HILL0063.D 01003-07-06 40 nglmL PFOS THPFOS 5.131 1399058 254 PFBS 4.416 1793510 254 CALIB-I5 HiLL0064.D 01003-07-07 50.1 nglrnL PFOS THPFOS 5.111 1399532 254 PFBS 4.411 1763775 254 CALIB-16 HILL0065.D 01003-07-08 75.1 nglrnL PFOS THPFOS 5.115 1382061 254 PFBS 4.401 1748667 254 CALIB-17 HILL0066.D 01003-07-09 100.2 nglrnL PFOS THPFOS 5.122 1384218 254 PFBS 4.415 1777945 254 CALIB-18 HILL0067.D 01003-07-10 250.5 nglrnL PFOS THPFOS 5.117 1375930 254 PFBS 4.402 1745338 254 CALlB-19 HILL0068.D 01003-07-11 400.8 nglmL PFOS THPFOS 5.103 1364296 254 PFBS 4.403 1751400 254 CALIB-20 HILL0069.D 01003-07-I 2 501 ng/mL PFOS THPFOS 5.11 1378457 254 PFBS 4.402 1799585 254 ' SAMPLE HILL0070.D 01003-07-13 1002 nglrnL PFOS THPFOS 5.115 1357091 254 PFBS 4.407 1769921 254 m U (P (D Page 1o f 4 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MaleIFemale Normal Human Urine; Tirnepoint(s): Analytical Instrument: HPLC/MSD "Hillary"; Analytical SequenceIMethod: HO10501.s/PFOS0425.m (ChemStation); ReprocessingMethod: H010501t.m (Target) Quant range: 10-501 nglmL Day 2 samples were re-aliquotted from the initialcentrifuge tube on 05-01-01 into 1.5 mL centrifuge tubes, centrifuged, and then diluted 1:lOO and 1 : l O for a final dilution factor of 1:l.OOO prior to analysis. SAMPLE HILL0031.D 042301-PFOS-LCS PFOS Solvent Blank Spike THPFOS 5.122 1322947 254 PFBS SAMPLE HILL0032.D PFOS-2.4.lblk 042301-PFOS-2.4.1 Mtd Blk THPFOS 5.115 1317272 254 PFBS SAMPLE HILL0033.D PFOS-2.4.2blk 042301-PFOS-2.4.2 Mtd Blk THPFOS 5.109 1319308 254 PFBS SAMPLE HILL0034.D PFOS-2.4.3blk 042301-PFOS-2.4.3 Mtd Blk THPFOS 5.122 1316462 254 PFBS SAMPLE HILL0035.D PFOS2.4.4blkMS 042301-PFOS-2.4.4 Mtd Blk Spike THPFOS 5.1 11 1318338 254 PFBS SAMPLE HILL0037.D PFOS-2.1.1 042301-PFOS-2.1.1 Smp THPFOS 5.115 1215865 254 PFBS SAMPLE HILL0038.D PFOS2.1.2 042301-PFOS-2.1.2 DUP THPFOS 5.108 1161890 254 PFBS SAMPLE HILL0039.D PFOS-2.1.3 042301-PFOS-2.1.3 Trip THPFOS 5.116 1218774 254 PFBS SAMPLE HILL0040.D PFOS-2.1.4MS 042301-PFOS-2.1.4 Spike THPFOS 5.116 1224336 254 PFBS SAMPLE HILL0045.D PFOS-2.2.1 042301-PFOS-2.2.1 Smp THPFOS 5.123 1256724 254 PFBS SAMPLE HILL0046 D PFOS-2.2.2 042301-PFOS-2.2.2 DUP THPFOS 5.122 1246843 254 PFBS SAMPLE HILL0047.D PFOS-2.2.3 042301-PFOS-2.2.3 Trip THPFOS 5.11 1245602 254 PFBS SAMPLE HILL0048.D PFOS-2.2.4MS 042301-PFOS-2.2.4 Spike THPFOS 5.117 1219461 254 PFBS SAMPLE HILL0050.D PFOS-2.3.1 042301-PFOS-2.3.1 Smp THPFOS 5.117 1211070 254 PFBS SAMPLE HILL0051.D PFOS-2.3.2 042301-PFOS-2.3.2 DUP THPFOS 5.118 1227470 254 PFBS SAMPLE HILL0052.D PFOS2.3.3 042301-PFOS2.3.3 Trip THPFOS 5.116 1222931 254 PFBS SAMPLE HILL0053.D PFOS-2.3.4MS 042301-PFOS-2.3.4 Spike THPFOS 5.111 1223446 254 PFBS Day 2 4.414 4.407 4.408 4.394 4.403 4.408 4.401 4.409 4.402 4.408 4.401 4.409 4.402 4.41 4.408 4.409 4.397 1715710 254 1718184 254 1695951 254 1698236 254 1686696 254 1912250 254 1969789 254 1995209 254 1951822 254 1900616 254 1926427 254 1881971 254 1935761 254 1958520 254 1962518 254 1984447 254 1937135 254 Average: 1319470 Std Deviation: 65017 %RSD 4.9% 1806527 86617 4.8% SAMPLE HILL0014.D TN-A 4802 MeOH Solvent Blank THPFOS 5.189 7753 0 PFBS 4.418 7561 0 SAMPLE HILL0015.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0029.D TN-A 4802 MeOH Solvent Blank THPFOS 5.195 8699 0 PFBS 0 0 0 SAMPLE HILL0030.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0036.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0043.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0044.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILLO049.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0056.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0057.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0071.D TN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 SAMPLE HILL0072.D IN-A 4802 MeOH Solvent Blank THPFOS 0 0 0 PFBS 0 0 0 w z Page 2 of 4 BACK TO MAIN Solubility Study;Shake Flask Method; Test Compound: PFOS; Test Matrix: MalelFemaleNormal Human Urine; Timepoint(s): Day 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: HOI 0501.slPFOS0425.m (ChemStation): Reprocessing Method: H010501t.m (Target) Qoant range: 10-501 IIglmL Day 2 samples were re-aliquottedfrom the initial centrifuge tube on 05-01-01into 1.5 mL centrifuge tubes, centrifuged, and then diluted . . 1 : l O O and 1 : l O for a final dilution factor of 1:l.OOO Drior to analvsis. SampType File Sample Name Misc Info CompoundName RT Area Amount $ Recovew SAMPLE HILL0016.D 01003-07-01 0 ng/mL PFOS PFOS 0 0 0 SAMPLE HILL0017.D 01003-07-02 2.8 nglmL PFOS PFOS 5.326 71773 4 CALIB-1Disabled HILL0018.D 01003-07-03 5 nglmL PFOS PFOS 5.326 129494 6 CALIB-2 HILL0019.D 01003-07-04 10 nglrnL PFOS PFOS 5.318 230148 10 105% CALIB-3 HILL0020.D 01003-07-05 25 nglrnL PFOS PFOS 5.32 628525 26 103% CALIB-4 HILL0021.D 01003-07-06 40 nglrnL PFOS PFOS 5.32 994126 40 100% CALI8-5 HILL0022. D CALIB-6 . HILL0023.D 01003-07-07 01003-07-08 50.1 nglmL PFOS 75.1 nglmL PFOS PFOS 5.32 1255424 50 PFOS 5.321 1866886 75 101% 100% CALIB-7 HILL0024.D 01003-07-09 100.2 nglmL PFOS PFOS 5.342 2434000 99 99% CALI6-8 HILL0025.D 01003-07-10 250.5 nglmL PFOS PFOS 5.32 5704519 252 100% CALlB-9 HILL0026.D 01003-07-1 1 400.8 nglmL PFOS PFOS 5.322 8544962 397 99% CALlB-10 HILL0027.D 01003-07-12 501 ng/mL PFOS PFOS 5.318 10504197 506 101% SAMPLE HILL0028.D 01003-07-13 1002 n g h L PFOS PFOS 5.319 18199294 1041 SAMPLE HILL0041.D 01003-07-03 5 nglmL PFOS PFOS 5.332 152981 7 SAMPLE HILL0042.D 01003-07-09 100.2 nglmL PFOS PFOS 5.32 2399376 99 99% SAMPLE HILL0054.D 01003-07-03 5 ng/mL PFOS PFOS 5.326 158654 7 SAMPLE HILL0055.D 01003-07-09 100.2 nglmL PFOS PFOS 5.32 2404840 99 99% SAMPLE HILL0058.D 01003-07-01 0 nglmL PFOS PFOS 0 0 0 SAMPLE HILL0059.D 01003-07-02 2.5 nglmL PFOS PFOS 5.311 72552 4 CALlB-11 Disabled HILL0060.D 01003-07-03 5 nglmL PFOS PFOS 5.326 130239 6 CALlB-I 2 HILL0061.D 01003-07-04 10 nglrnL PFOS PFOS 5.32 233273 10 104% CALIB-I3 HILL0062.D 01003-07-05 25 nglmL PFOS PFOS 5.32 621213 25 102% CALIB-14 HILL0063.D 01003-07-06 40 nglrnL PFOS PFOS 5.334 994794 40 100% CALlB-15 HILL0064.D 01003-07-07 50.1 nglmL PFOS PFOS 5.314 1220007 50 99% CALIB-16 HILL0065.D 01003-07-08 75.1 nglmL PFOS PFOS 5.325 1796101 74 99% CALIB-17 HILL0066.D 01003-07-09 100.2 nglmL PFOS PFOS 5.325 2409666 99 99% CALIB-18 HILL0067.D 01003-07-10 250.5 nglmL PFOS PFOS 5.327 5602770 251 100% CALIB-19 HILL0068.D 01003-07-11 400.8 nglmL PFOS PFOS 5.313 8515207 406. 101% CALIB-20 HILLO069.D 01003-07-12 501 nglmL PFOS PFOS 5.313 10313769 494 99% SAMPLE HILL0070.D 01003-07-13 1002 nglmL PFOS PFOS 5.325 17653606 995 TWOwrves were averaged using Targel sohare lo quanWy data. A rlandard a w e range of only 10 levels is allowed. 5500 nglmLwas the initial range chosen. However. Ihe 5 ng/mL sld did not meelacceptancecriteria. therefore the 5 n@mL$Id was disabled and the data was re-quantifiedusingthe sld curve range 10-501 nghL Page 3 of 4 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Day 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical Sequencemethod: H010501.s/PFOS0425.m (ChemStation); Reprocessing Method: H010501t.m (Target) Quant range: 10-501 nglmL Day 2 samples were re-aliquotledfrom the initialcentrifuge tube on 0501-01 into 1.5 mL centrifuge tubes, centrifuged, and then diluted 1:lOO and 1:lO for a final dilutionfactor of 1:1,WO prior l o analysts. SarnpTvoe - File Sample Name Mlsc Info ComooundName Average pg/mL, dil factor corrected S~ amDle - Purity Std Dev %RSD &gg Amt [nalrnll corrected nalrnL Spike Recovery SAMPLE HILL0031.D 042301-PFOS-LCS PFOS Solvent Blank Spike PFOS 5.325 2110116 89 87 89% SAMPLE HILL0032.D PFOS-2.4.1 blk 042301-PFOS-2.4.1 Mtd Elk PFOS 0 0 0 SAMPLE HILL0033.D PFOS-2.4.2blk 042301-PFOS-2.4.2 Mtd Elk PFOS 0 0 0 SAMPLE, HILL0034.D PFOS2.4.3blk 042301-PFOS-2.4.3 Mtd Elk PFOS 0 0 0 SAMPLE HILLW35.D PFOS-2.4.4blkMS 042301-PFOS2.4.4 Mtd Blk Spike PFOS 5.314 2188222 94 92 94% SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0037.D HILL0038.D HILL0039.D HlLLW40.D PFOS-2.1 .I PFOS-2.1.2 PFOS-2.1.3 PFOS-2.1.4MS HILL0045.D HILL0046.D HILL0047.D HILL0048.D . PFOS-2.2.1 PFOS-2.2.2 PFOS-2.2.3 PFOS-2.2.4MS 042301-PFOS-2.1.1 Srnp 042301-PFOS-2.1.2 DUP 042301-PFOS-2.1.3 Trip 042301-PFOS-2.1.4 Spike 042301-PFOS-2.2.1 Srnp 042301-PFOS-2.2.2 DUp 042301-PFOS-2.2.3 Trip 042301-PFOS-2.2.4 Spike PFOS 5.240 7420988 312 PFOS 5.248 9885661 421 PFOS 5.256 8832844 363 PFOS 5.249 9719956 417 PFOS 5.263 7021826 295 PFOS 5.255 7094280 294 PFOS 5.25 6410858 269 PFOS 5.257 9712496 421 305 358 412 54 356 15% 409 52% 289 280 288 15 263 5% 412 135% Day 2 results, pglmL average 325 rtddev 40 %RSD 12% SAMPLE SAMPLE SAMPLE SAMPLE HILL0050.D HILL0051.D HILL0052.D HILL0053.D PFOS-2.3.1 PFOS-2.3.2 PFOS-2.3.3 PFOS-2.3.4MS 042301-PFOS-2.3.1 Srnp 042301-PFOS-2.3.2 DUP 042301-PFOS-2.3.3 Trip 042301PFOS-2.3.4 Spike PFOS 5.257 a i m 6 2 339 PFOS 5.249 8319423 345 PFOS 5.249 8547947 352 PFOS 5.251 9269162 398 332 330 338 7 345 2% 390 53% SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HlLL0014.D HILL0015.D HILL0029.D HILL0030.D HILL0036.D HILL0043.0 HlLL0044.D HILL0049.D HILL0056.D HILL0057.D HILL0071.D HILL0072.D TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH Solvent Blank Solvent Blank Solvent Blank Solvent Blank Soivent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS 5.413 0 5.419 5.403 0 0 0 5.34 0 0 5.412 5.404 24002 0 40197 16788 0 0 0 23584 0 0 38870 14734 below LOQ 0 below LOQ below LOQ 0 0 0 below LOQ 0 0 below LOQ below LOO Page4 of 4 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MalelFemala Normal Human Urine; Timepoint(s): Days 2 and 3; Analytical Instrument: HPLC/MSD "Hillary"; Analytical Sequence/Method: HOi0430.s/PFOS0425.m(ChemStation);ReprocessingMethod: H010430tm (Target) SampTvoe & SamDle Name Mise Info Fomoound NamQ E I A l a l Amou SAMPLE HILL0016.D 01003-07-01 0 nglrnL PFOS THPFOS 5.105 1240800 254 SAMPLE HILL0017.D 01003-07-02 2.5 ng/mL PFOS THPFOS 5.101 1259512 254 CALIB-1 CALIB-2 CALIB-3 CALIB-4 CALIB-5 CALIB-6 CALIB-7 HILL0018.D HILL0019.D HILL0020.D HILL0021.D HILL0022.0 HILL0023.0 HILL0024.D 01003-07-03 01003-07-04 01003-0705 01003-07-06 01003-07-07 C1003-07-08 01003-07-09 5 nglmL PFOS 10 nglmL PFOS 25 nglrnL PFOS 40 ng/rnL PFOS 50.1n g h L PFOS 75.1 nglrnL PFOS 100.2 nglrnLPFOS THPFOS 5.098 1268358 254 THPFOS 5.104 1273859 254 THPFOS 5.103 1287880 254 THPFOS 5.104 1293445 254 THPFOS 5.105 1299052 254 THPFOS 5.101 1297549 254 THPFOS 5.105 1310823 254 CALlB-8 HILL0025.D 01003-07-10 250.5 nglrnLPFOS THPFOS 5.103 1305549 254 CALIB-9 HILL0026.D 01003-07-11 400.8 nglrnL PFOS THPFOS 5.103 1299241 254 CALIB-10 HILL0027.0 01003-07-12 501 nglrnL PFOS THPFOS 5.095 1306826 254 SAMPLE HILL0028.D 0100507-13 1002 nglrnL PFOS THPFOS 5.109 1302797 254 SAMPLE HILL0041.D SAMPLE HILL0042.0 01003-07-03 01003-07-09 5 nglrnL PFOS 100.2 nglrnLPFOS THPFOS 5.1 1390044 254 THPFOS 5.103 1386912 254 SAMPLE HILL0054.D 01003-07-03 5 nglrnL PFOS THPFOS 5.103 1417554 254 SAMPLE HILL0055.D 01003-07-09 100.2 n g h L PFOS THPFOS 5.101 1416339 254 SAMPLE HILL0068.D 01003-0703 5 nglrnL PFOS THPFOS 5.11 1463633 254 SAMPLE HILL0069.D SAMPLE HILL0081.D 01003-07-09 0100307-03 100.2 nglrnL PFOS 5 nglrnL PFOS THPFOS 5.103 1453681 254 THPFOS 5.098 1482897 254 SAMPLE HILL0082.D SAMPLE HILL0085.D 01003-07-09 01003-07-01 100.2 ng/rnL PFOS 0 nglrnL PFOS THPFOS 5.101 1463161 254 THPFOS 5.101 1470609 254 SAMPLE CALIB-11 CALIB-12 CALIB-13 CALIB-14 CALIB-15 CALIB-16 CALIB-17 CALlB-18 CALIB-19 CAL18-20 SAMPLE HILL0086.D HILL0087.D HILL0088.D HILL0089.D HILL0090.D HILL0091.D HILL0092.D HILL0093.D HILL0094.D HILL0095.D HILL0096.D HILL0097.D 01003-07-02 01003-07-03 01003-07-04 01003-07-05 01003-0706 01003-0707 01003-0748 01003-07-09 01003-07-10 0100307-1 1 01003-07-12 01003-07-13 2.5 nglrnL PFOS 5 nglrnL PFOS 10 nglrnL PFOS 25 ng/rnL PFOS 40 ngImL PFOS 50.1 nglrnL PFOS 75.1 n g h L PFOS 100.2 ng/rnL PFOS 250.5 nglrnL PFOS 400.8 nglrnLPFOS 501 nglrnL PFOS 1002 nglmL PFOS THPFOS 5.095 1481303 254 THPFOS 5.097 1492012 254 THPFOS 5.095 1477732 254 THPFOS 5.112 1481121 254 THPFOS 5.095 1479643 254 THPFOS 5.095 1475404 254 THPFOS 5.094 1469508 254 THPFOS 5.096 1496019 254 THPFOS 5.109 1476250 254 THPFOS 5.097 1466888 254 THPFOS 5.109 1475286 254 THPFOS 5.103 1440061 254 f(l-i D5&@-';' Comoound NPmQ PFBS PFES PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFES PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFES PFBS - RT 4.384 4.38 4.382 4.382 4.382 4.376 4.376 4.38 4.376 4.375 4.375 4.374 4.381 4.386 4.381 4.381 4.373 4.381 4.382 4.391 4.401 4.394 4.394 4.396 4.394 4.397 4.394 4.388 4.394 4.395 4.395 4.396 4.38 4.382 Arer 1897371 1883485 1898698 1894001 1891481 1895316 1887750 1894332 1899388 1877817 1884885 1906259 1898513 1873917 1895596 1870203 1863988 1941991 1934618 1898558 1912978 1882104 1898530 1905370 1894008 1892338 1891942 1897650 1911398 1928086 1869148 1898244 1911397 1902178 Amount 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 SAMPLE HILL0031.D 042301-PFOS-LCS PFOS Solvent Blank Spike THPFOS 5.095 1325456 254 SAMPLE HlLLW32.D PFDS-2.4.lblk 042301-PFOS-2.4.1 Mld Blk THPFOS 5.096 1344481 254 SAMPLE HILL0033.D PFOS-2.4.2blk 042301-PFOS-2.4.2 Mtd Blk THPFOS 5.101 1347423 254 SAMPLE HILL0034.D PFOS-2.4.3blk 042301-PFOS-2.4.3 Mld Blk THPFOS 5.103 1347097 254 SAMPLE HILL0035.D PFOS-2.4.4blkMS 042301-PFOS-2.4.4 Mtd Blk Spike THPFOS 5.095 1324124 254 SAMPLE HILL0037.D PFOS-2.1.1 042301-PFOS-2.1.1 Srnp THPFOS 5.101 1163460 254 SAMPLE HILL0038.D PFOS-2.1.2 042301-PFOSd.1.2 Dup THPFOS 5.102 1199081 254 SAMPLE HILL0039.D PFOS-2.1.3 042301-PFOS-2.1.3 Trip THPFOS 5.102 1228695 254 SAMPLE HILL0040.D PFOS-2.1.4MS 042301-PFOS-2.1.4 Spike THPFOS 5.104 1219919 254 SAMPLE HILL0045.D PFOS-2.2.1 042301-PFOS-2.2.1 Smp THPFOS 5.101 1364440 254 SAMPLE HILL0046.0 PFOS-2.2.2 042301-PFOS-2.2.2 Dup THPFOS 5.096 1385823 254 SAMPLE HILL0047.D PFOS-2.2.3 042301-PFDS-2.2.3 Tlrp THPFOS 5.102 1392690 254 SAMPLE HILL0048.D PFOS-2.2.4MS 042301-PFOS-2.2.4 Spike THPFOS 5.102 1376257 254 P U SAMPLE HILLW50.D PFOS-2.3.1 042301-PFOS-2.3.1 Smp THPFOS 5.103 1367578 254 (mP SSAAMMPPLLEE HHIILLLL00005521..DD PPFFOOSS--22..33..23 004422330011--PPFFOOSS--22.3.3.2.3 DTruipp TTHHPPFFOOSS 55..009967 11337832410020 225544 0 0 SAMPLE HILL0053.D PFOS-2.3.4MS 042301-PFOS-2.3.4 Spike THPFOS 5.096 1373073 254 % -.L 2 PFBS PFBS PFBS PFBS PFBS PFBS ' PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFES, PFBS PFBS PFBS 4.381 4.375 4.379 4.389 4.38 4.387 4.388 4.387 4.383 4.387 4.382 4.381 4.374 4.374 4.378 4.374 4.374 1823070 254 1834828 254 1867721 254 1835565 254 1847790 254 1968845 254 2112105 254 2098049 254 2057424 254 1826163 254 1818895 254 1822627 254 1820132 254 1841611 254 1797985 254 1816031 254 1843565 254 Page 1 of 5 w != 8;?! -aC. U= P-.3 a- rnK 00- e4 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: Maleffernale Normal Human Urine; Timep.oint(.s1.: Da-ys 2 and 3: Analytlcal Instrument: HPLC/MSD "Hlllary"; SAMPLE HILL0058.D 042301$FOS-L& PFOS Solvent Blank Splke THPFOS 5.095 1394750 254 SAMPLE HILL0059.D PFOS3.4.lblk 042301-PFOS3.4.1 Mld Elk THPFOS 5.098 1419875 . 254 SAMPLE HILLW6O.D PFOS3.4.2blk 042301-PFOS3.4.2 Mld Elk THPFOS 5.096 1420804 254 SAMPLE HILL0061.D PFOS-3.4.3blk 042301-PFOS3.4.3 Mtd Elk THPFOS 5.098 1433374 254 SAMPLE HILL0062.D PFOS-3.4.4blkMS 042301-PFOS3.4.4 Mtd Elk Spike THPFOS 5.103 1406537 254 SAMPLE HILLOO64.D PFOS3.1.1 042301-PFOS-3.1.1 Smp THPFOS 5.097 1310521 254 SAMPLE HlLL0065.D PFOS3.1.2 042301-PFOS-3.1.2 Dup THPFOS 5.109 1294952 254 SAMPLE HILL0066.D PFOS-3.1.3 042301-PFOS3.1.3 Trip THPFOS 5.102 1296261 254 SAMPLE HILL0067.D PFOS-3.1.4MS 042301-PFOSJ.1.4 Spike THPFOS 5.107 1334559 254 SAMPLE HILL0072.D PFOS-3.2.1 042301-PFOS-3.2.1 Smp THPFOS 5.105 1305210 254 SAMPLE HILL0073.D PFOS-3.2.2 042301-PFOS3.2.2 DUp THPFOS 5.109 1308424 254 SAMPLE HILL0074.D PFOS3.2.3 042301-PFOS-3.2.3 Trip THPFOS 5.111 1312470 254 SAMPLE HILL0075.D PFOS-3.2.4MS 042301-PFOS-3.2.4 Spike THPFOS 5.104 1289916 254 SAMPLE HILLW77.D PFOS3.3.1 042301-PFOS-3.3.1 Smp THPFOS 5.104 1325618 254 SAMPLE HILL0078.D PFOS-3.3.2 042301-PFOS3.3.2 Dup THPFOS 5.1 1 ' 1332228 254 SAMPLE HILL0079.D PFOS3.3.3 042301-PFOS3.3.3 Trip THPFOS 5.101 1339789 254 SAMPLE HILLO080.D PFOS-3.3.4MS 042301-PFOS3.3.4 Spike THPFOS 5.098 1327428 254 Avenge: 1364258.28 Std Deviation: 82080 YmRSD: 6.0% SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0014.D HILLOO15.D HILL0029.D HILLW30.D HILL0036 D HILL0043.D HILL0044.D HILL0049.D HILLO05S.D HILL0057.D HILL0063.D HILL0070.D HILLW71.D HILL0076.D HILL0083.D HILLOO84.D HILL0098.D HILLW99.D TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH Solvent Blank Solvenl Blank Solvent Blank Solvenl Blank Solvent Blank Solvent Blank Solvent Blank Solvenl Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 5.178 7624 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 5.172 8030 0 THPFOS 0 0 0 THPFOS 5.2 7970 0 THPFOS 0 0 0 PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS. PFBS ' PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS 4.367 4.363 4.368 4.369 4.375 4.376 4.38 4.38 4.379 4.384 4.388 4.39 4.397 4.397 4.389 4.386 4.383 0 0 0 0 0 4.409 0 0 0 0 0 0 0 4.423 0 0 0 0 1828732 1870275 1875725 1883710 1858704 2034585 2079222 2046016 2057849 2128725 2170045 2160742 2146697 . 2084860 2030571 2064868 2068536 1925085.4 95672 5.0% 0 0 0 0 0 7786 0 0 0 0 0 0 0 7887 0 0 0 0 254 254 254 254 254 254 254 254 254 254 254 254 254 254 , 254 254 254 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 m-0 (P (D Page 2 of 5 BACK TO MAIN *s- Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MalelFemale NormalHuman Urine; Timepoint(s): Days 2 and 3; Analytical Instrument: HPLClMSD "Hillary"; SampTvue Flle Sample Name Mlsc info Comvound Name E Arer Amt lnalmL1 SAMPLE HILLzl6.D 01003-07-01 0 nglmL PFOS PFOS 0 0 0.0 SAMPLE HILLOO17.D 01003-07-02 2.5 n g h L PFOS PFOS 5.31 1 77068 3.4 CALlE-1 HILLOO18.D 01003-07-03 5 nglrnL PFOS PFOS 5.306 140922 5.7 CALIB-2 HlLL0019.D OlM13-07-04 10 nglmL PFOS PFOS 5.314 250842 9.8 CALlB-3 HILL0020.D 01003-07-05 25 ngImL PFOS PFOS 5.313 644213 24.7 CALIE-4 HILL0021.D 01003-0706 40 nglmL PFOS PFOS 5.307 1035507 39.7 CALIE-5 HILL0022.D 01003-07-07 50.1 n g h L PFOS PFOS 5.308 1297981 50.1 CALIB-6 HILL0023.D 01003-07-08 75.1 n g h L PFOS PFOS 5.311 1923999 74.8 CALIE-7 HILL0024.D 01003-07-09 100.2 nglmL PFOS PFOS 5.308 2507092 98.2 CALlE-8 HiLL0025.D 01003-07-10 250.5 ng/mL PFOS PFOS 5.306 5879693 248.2 CALIE-9 HILL0026.D 01003-07-11 400.8 ng/mL PFOS PFOS 5.306 8875074 394.1 CALIE-10 HILLOO27.D 01003-07-12 501 nglmL PFOS PFOS 5.305 10870332 493.2 SAMPLE HILLW28.D 01003-07-13 1002 ngImL PFOS ' PFOS 5.312 18717748 968.5 SAMPLE HILL0041.D 01003-07-03 5 nglmL PFOS PFOS 5.31 163873 6.7 SAMPLE HILL0042.D 01003-07-09 100.2 ng/mL PFOS PFOS 5.306 2519548 98.9 SAMPLE HILL0054.D 01003-07-03 5 ng/rnL PFOS PFOS 5.313 136445 5.6 SAMPLE HiLL0055.D 01003-07-09 100.2 nglmL PFOS PFOS 5.312 2484938 99.2 SAMPLE HILL0068.D 01003-07-03 5 nglmL PFOS PFOS 5.32 166490 6.5 SAMPLE HILL0069.D 0100307-09 100.2 nglrnL PFOS PFOS 5.306 2588211 99.5 SAMPLE HILL0081.D 01003-07-03 5 nglmL PFOS PFOS 5.301 162640 6.5 SAMPLE HILLO062.D 01003-07-09 100.2 ng/rnL PFOS PFOS 5.304 2575229 100.2 SAMPLE HILL0085.D 01003-07-01 0 nglmL PFOS PFOS 0 0 0.0 SAMPLE HIL1.0086.D 01003-07-02 2.5 ng/mC PFOS PFOS 5.298 74899 3.3 CALlB-11 HILLOO87.D 01003-0703 5 nglmL PFOS PFOS 5.3 130171 5.6 CALIE-12 HILLW88.D 01003-07-04 10 nglmL PFOS PFOS 5.298 248359 9.7 CALIE-13 HILL0089.D 01003-07-05 25 nglrnL PFOS PFOS 5.322 648497 24.9 CALIB-14 HILLO090.D 01003-0706 40 ng/mL PFOS PFOS 5.298 1030084 39.6 CALIE-15 HILLO091.D 01003-07-07 50.1 nglmL PFOS PFOS 5.298 1323882 50.9 CALIB-16 HILL0092.D 01003-07-08 75.1 nglmL PFOS PFOS 5.304 1963056 75.6 CALIE-17 HILL0093.D 01003-07-09 1002 n g h L PFOS PFOS 5.299 2594831 100.2 CALlE-18 HlLL0094.D 01003-07-10 250.5 nglmL PFOS PFOS 5.312 6013719 255.9 CALIE-19 HILLM195.D 01003-07-11 400.8 nglmL PFOS PFOS 5.3 9177083 406.4 CALIE-20 HILL0096.D S-AMP_L_E HI_L_LW97.D 01003-07-12 01003-07-13 501 ngmL PFOS 1002 nalmL PFOS PFOS PFOS 5.319 5.313 11174689 18959651 508.1 982.2 Two curves were averaged usingTarget sofhvara lo quantify data. A standard cum rangeof only 10 levels is allowed. 5500 n g h L was the rangechosen Recovery 114% 98% 99% 99% 100% 100% 98% 99% 98% 98% 133% 99% 113% 99% 100% 99% 130% 100% 112% 97% 100% 99% 102% 101% 100% 102% 101% 101% U lnnl m Page 3 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: 'MalelFemaleNormal Human Urine; Timepoint(s): Days 2 and 3; Analytical Instrument: HPLClMSD "Hillary"; SampTvoe File Samole Name SAMPLE HILL0031.D 042301-PFOS-LCS SAMPLE HILL0032.D SAMPLE HILLW33.D SAMPLE HlLL0034.D PFOS-2.4.lblk PFOS-2.4.2blk PFOS-2.4.3blk SAMPLE HILL0035.D PFOS-2.4.4blkMS Misc Info 042301-PFOS-2.4.1 Mtd Blk 042301-PFOS-2.4.2 Mtd Blk 042301-PFOS-2.4.3 Mtd Elk 042301-PFOS-2.4.4 Mtd Blk Spike SAMPLE HILL0037.D SAMPLE HILL0038.D SAMPLE. HILL0039.D SAMPLE HILL0040.D PFOS-2.1.1 PFOS-2.1.2 PFOS-2.1.3 PFOS-2.1.4MS 042301-PFOS-2.1.1 Smp 042301-PFOS-2.1.2 Dup 042301-PFOS-2.1.3 Trip 042301-PFOS-2.1.4 Spike SAMPLE SAMPLE SAMPLE SAMPLE HILL0045.D HlLLOO46.D HILL0047.D HILL0048.D PFOS-2.2.1 PFOS-2.22 PFOS-2.2.3 PFOS-2.2.4MS 042301-PFOS-2.2.1 Smp 042301PFOS-2.2.2 Dup 042301-PFOS2.2.3 Trip 042301-PFOS-2.2.4 Spike SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0050.D HILL0051.D HILL0052.D PFOS-2.3.1 PFOS-2.3.2 PFOS-2.3.3 HILL0053.D PFOS-2.3.4MS HILL0058.D 042301-PFOS-LCS 042301-PFOS-2.3.1 Smp 042301-PFOS-2.3.2 Dup 042301-PFDSQ.3.3 Ttlp 042301-PFOS-2.3.4 Spike PFOS Solvent Blank Spike SAMPLE SAMPLE SAMPLE SAMPLE HILL0059.D HILLM160.D HILLM161.D HILL0062.0 PFOS-3.4.lblk PFOS-3.4.2blk PFOS-3.4.3blk PFOS-3.4.4blkMS 042301-PFOS3.4.1 Mtd Bik 042301-PFOS3.4.2 Mtd Blk 042301-PF053.4.3 Mtd Blk 042301-PFOS-3.4.4 Mtd Blk Spike SAMPLE SAMPLE SAMPLE SAMPLE HILL0064.D HILL0065.D HILL0066.D HILL0067.D PFOS-3.1.1 PFOS-3.1.2 PFOS-3.1.3 PFOS-3.1.4MS 042301PFOS-3.1.1 Smp 042301-PFOS3.1.2 Cup 042301-PFOS3.1.3 Trip 042301-PFOS-3. 1.4 Spike SAMPLE SAMPLE SAMPLE SAMPLE HILLW72.D HiLL0073.D HlLL0074.D HILLW75.D PFOS-3.2.1 PFOS3.2.2 PFOS-3.2.3 PFOS-3.2.4MS 042301-PFOS3.2.1 Smp 042301-PFOS3.2.2 Dup 042301-PFOS32.3 Trip 042301-PFOS3.2.4 Spike SAMPLE SAMPLE SAMPLE SAMPLE HILL0077.0 HILL0078.0 HILL0079.D HILL0080.D PFOS3.3.1 PFOS-3.3.2 PFOS3.3.3 PFOS-3.3.4MS 042301-PFOS3.3.1 Smp 042301-PFOS-3.3.2 Dup 042301-PFOS3.3.3 Trip 042301-PFOS3.3.4 Spike Comoound Name PFOS PFOS PFOS PFOS PFOS m 5.305 0 0 0 5.305 m 2186915 0 0 0 2461943 PFOS PFOS PFOS PFOS 5.235 5.242 5.242 5.237 7481346 9178901 8215875 9849022 PFOS PFOS PFOS PFOS 0 0 0 5.305 0 0 0 2441252 PFOS PFOS PFOS PFOS PFOS 0 0 0 5.306 5.305 0 0 0 2368154 2208565 PFOS PFOS PFOS PFOS . PFOS PFOS PFOS PFOS 0 0 0 5.313 5.237 5.242 5.242 5.247 0 0 0 2459314 6815087 7544482 6794540 6532934 PFOS PFOS PFOS PFOS 5.245 5149 5.251 5.237 8117378 8786445 9095599 11117925 PFOS PFOS PFOS PFOS 5.238 5.243 5.234 5.238 7558687 6689145 7103605 9203477 Amt tnfa/mL) 69.3 0.0 0.0 0.0 99.1 308.4 359.3 319.1 401.7 0.0 0.0 0.0 99.8 0.0 0.0 0.0 95.4 89.5 0.0 0.0 0.0 98.4 267.6 292.7 265.0 340.6 308.4 359.3 319.1 401.7 292.4 262.7 275.7 369.2 Purity corrected nalmL 87 Average pglmL Std Dev %RSD Spike Recovery 89% 67 99% 302 322 352 26.5 312 8% 393 72% 98 100% 93 95% 88 90% 96 98% 262 269 287 15.4 259 6% 333 gssa. 302 322 352 26.5 312 8% 393 72% 286 271 257 14.5 270 5% 361 92% average std dev %RSD: Day 3 results 267 30 10.4% Page 4 of 5 Solubility Study, Shake Flask Method; Test Compound: PFOS; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Days 2 and 3; Analytical Instrument: HPLClMSD "Hillary"; SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0014.D HILL0015.D HILL0029.D HILL0030.D HILL0036.D HILL0043.D HILL0044.D HILL0049.D HILL0056.D HILL0057.D HILLOO63.D HILL0070.D HILLOO71.D HILL0076.D HILL0083.D HILLO084.D HILLMM8.D HILL0099.D TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH T N A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A4802 MeOH TN-A 4802 MeOH Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank Solvent Blank PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS PFOS 5.398 0 0 0 0 0 0 0 0 0 0 0 0 5.333 0 0 5.424 5.404 27635 0 0 0 0 0 0 0 0 0 0 0 0 30130 0 0 47439 15154 Below L o a 0 0 0 0 0 0 0 0 0 0 0 0 Below L o a 0 0 Below LOQ Below LOQ BACK TO MAIN Page 5 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Days 1 and 2 AnalyticalInstrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010426.slFOSA0425.m(ChemStation); ReprocessingMethod: H010426a.m (Target) SampType File Sample Name Mlsc Info Compound Name RT Area Amount Compound Name RT SAMPLE HILLO105.D 01003-25-01 SAMPLE HILLOIO6.D 01003-25-02 CALIB-1 XX HILL0107.D 01003-2503 CALIB-2 HILLO108.D 01003-25-04 CALIB-3 HlLLOl09.D 01003-2505 CALIB-4 HlLLOl10.D 01003-25-06 CALIB-5 HILLO1lI.D 01003-25-07 CALIB-6 HILLO112.D 01003-25-08 CALIB-7 HILL0113.D 01003-25-09 CALIB-8 HILL0114.D 01003-25-10 CALIB-9 HlLLOl15.D 01003-25-11 CALlB-10 HILLO116.D 01003-25-12 SAMPLE HILL0117.D 01003-25-13 SAMPLE HILL0131.D 01003-25-03 SAMPLE HILL0132.D 01003-25-09 SAMPLE HILL0146.D 01003-25-03 SAMPLE HILL0147.D 01003-25-09 SAMPLE HILLO161.D 01003-25-03 ' SAMPLE HILL0162.D 01003-25-09 SAMPLE HILL0171.D 01003-2501 SAMPLE HILL0172.D 01003-2502 CALlB-I 1 XX HILL0173.D 01003-25-03 CALIB-12 HKL0174.D 01003-2504 CALIB-13 HILL0175.D 01003-25-05 CALIB-I4 HILL0176.D 01003-25-06 CALIB-I5 HILL0177.D 01003-25-07 CALIB-I 6 HlLLOl78.D 01003-2508 CALIB-17 HILL0179.D 01003-2509 CALIB-18 HILL0180.D 01003-25-10 CALIB-I 9 HILL0181.D 01003-25-11 CALIB-20 HILL0182.D 01003-25-12 SAMPLE HILL0183.D 01003-25-1 3 SAMPLE HlLL0120.D 042301-FOSA-LCS SAMPLE SAMPLE SAMPLE SAMPLE HILL0121.D HILL0122.D HILLOl23.D HILL0124.D FOSA-1.4.1blk FOSA-1.4.2blk FOSA-I .4.3blk FOSA-1.4.4blkMS 0 Std 2.5 ng/mL FOSA 5.3 nglrnL FOSA 10.1 ng/mL FOSA 25 nglmL FOSA 40.3 nglmL FOSA 49.9 ng/mL FOSA 74.9 nglmL FOSA 100.8 nglmL FOSA 249.6 nglmL FOSA 403.2 nglmL FOSA 499.2 nglmL FOSA 1008 ng/mL FOSA 5.3 nglrnL FOSA 100.8 nglmL FOSA 5.3 nglmL FOSA 100.8 nglmL FOSA 5.3 n g h L FOSA 100.8 nglmL FOSA 0 Std 2.5 nglmL FOSA 5.3 nglmL FOSA 10.1 nglmL FOSA 25 ng/mL FOSA 40.3 n g h L FOSA 49.9 nglmL FOSA 74.9 nglmL FOSA 100.8 nglmL FOSA 249.6 nglmL FOSA 403.2 nglmL FOSA 499.2 nglmL FOSA 1008 nglmL FOSA THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS THPFOS 5.123 1465515 254 5.111 1447576 254 5.116 1439545 254 5.116 1445222 254 5.109 1441747 254 5.118 1427400 254 5.115 1441987 254 5.116 1447199 254 5.112 1441812 254 5.115 1459549 254 5.116 1432834 254 5.112 1428416 254 5.116 1447369 254 5.1 1 1265387 254 5.119 1298469 254 5.11 1170043 254 5.111 1177767 254 5.111 1145578 254 5.115 1120490 254 5.115 1122982 254 5.1 16 1107259 254 5.118 1096459 254 5.109 1125849 254 5.115 1101993 254 5.11 1104242 254 5.109 1111672 254 5.107 1107520 254 5.112 1113145 254 5.124 1124167 254 5.1 17 1114186 254 5.123 1126570 254 5.119 1146487 254 5.109 1387790 254 5.108 1379938 254 5.115 1385495 254 5.117 1413232 254 5.116 1388787 254 PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS 4.394 4.39 4.395 4.401 4.387 4.389 4.386 4.395 4.39 4.394 4.395 4.39 4.387 4.381 4.39 4.382 4.389 4.309 4.393 4.394 4.388 4.383 4.388 4.387 4.389 4.388 4.386 4.391 4.395 4.389 4.387 4.383 4.381 4.38 4.387 4.381 4.387 SAMPLE SAMPLE SAMPLE SAMPLE HILLO126.D HILL0127.D HILL0128.D HILL0129.D FOSA-1.1.1 FOSA-1.1.2 FOSA-1.1.3 FOSA-1.I.4MS THPFOS THPFOS THPFOS THPFOS 5.112 1223896 254 5.112 1207970 254 5.1 15 1198241 254 5.11 I184556 254 PFBS PFBS PFBS PFBS 4.391 4.384 4.387 4.381 SAMPLE SAMPLE SAMPLE SAMPLE HILL0130.D HILL0135.D HILL0136.D HILL0137.D FOSA-1.2.1 FOSA-1.2.2 FOSA-1.2.3 FOSA-1.2.4MS THPFOS THPFOS THPFOS THPFOS 5.118 1223960 254 5.108 1227152 254 5.11 1210108 254 5.11 I202734 254 PFBS PFBS PFBS PFBS 4.383 4.379 4.375 4.382 .;~ . I Page 1 of5 Area 2007446 2027800 2026487 2021996 201 5250 2007486 2028225 2012838 1997221 1999512 2008100 1999395 1988333 1993211 1945472 1974246 1924449 1984007 1933728 1989610 1975741 1962904 1986462 1965181 1958152 1969738 1941113 I940454 1955186 I940675 1933249 1964405 1875014 1893699 1872319 1876934 1861408 1863247 1889163 1858991 1860013 1887871 1901597 I890574 1875132 Amount 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 BACK TO MAIN *I- Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Days 1 and 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010426.slFOSA0425.m (ChemStation); Reprocessing Method: H010426a.m (Target) SAMPLE SAMPLE SAMPLE SAMPLE HILL0138.D HILL0139.D HILL0141.D HILL0142.D FOSA-1.3.1 FOSA-1.3.2 FOSA-1.3.3 FOSA-1.3.4MS THPFOS THPFOS THPFOS THPFOS 5.112 1177514 254 5.109 1188089 254 5.116 1173989 254 5.116 1179158 254 PFBS PFBS PFBS PFBS 4.39 1900215 254 4.381 1882956 254 4.381 1853824 254 4.388 1873732 254 SAMPLE HILL0143.D 042301-FOSA-LCS THPFOS 5.116 1152061 254 PFBS 4.395 1836551 254 SAMPLE SAMPLE SAMPLE SAMPLE HILL0144.D HlLL0145.D HILL0150.D HILL0151.D FOSA-2.4.1 blk FOSA-2.4.2blk FOSA-2.4.3blk FOSA-2.4.4blkMS THPFOS THPFOS THPFOS THPFOS 5.112 1308838 254 5.115 1332634 254 5.109 1307643 254 5.116 1312788 254 PFBS PFBS PFBS PFBS 4.383 1875879 254 4.386 1889627 254 4.38 1868213 254 4.387 1876885 254 SAMPLE SAMPLE SAMPLE SAMPLE HILL0 152.D HILL0153.D HlLL0154.D HILL0156.D FOSA-2.1.1 FOSA-2.1.2 FOSA-2.1.3 FOSA-2.1.4MS THPFOS THPFOS THPFOS THPFOS 5.116 1110585 254 5.109 1194990 254 5.116 1181747 254 5.1 16 1165222 254 PFBS PFBS PFBS PFBS 4.387 1783250 254 4.388 1910331 254 4.388 1870518 254 4.388 1853384 254 - SAMPLE SAMPLE SAMPLE HILL0157.D HILL0158.D HILL0159.D FOSA-2.2.1 FOSA-2.2.2 FOSA-2.2.3 SAMPLE HILL0160.D FOSA-2.2.4MS THPFOS THPFOS THPFOS THPFOS 5.116 1202436 254 5.115 1233448 254 5.109 1228256 254 5.11 1224603 254 PFBS PFBS PFBS PFBS 4.388 1863768 254 4.387 I900272 254 4.388 I907383 254 4.389 1899528 254 SAMPLE SAMPLE SAMPLE SAMPLE HILL0165.D HILLOl66.D HILL0167.D HILL0168.D FOSA-2.3.1 FOSA-2.3.2 FOSA-2.3.3 FOSA-2.3.4MS THPFOS THPFOS THPFOS THPFOS 5.116 1158902 254 5.111 1162327 254 5.117 1177491 254 5.116 1181915 254 PFBS PFBS PFBS PFBS 4.395 1883593 254 4.39 1866181 254 4.388 1891600 254 4.307 1875392 254 Avenge: Std Devlatlon: %RSD 1249014 125832 10.1% 1926471 60064 3.1% SAMPLE HlLLOl03.D TN-A 4802 MeOH SAMPLE HILLOIW.D TN-A 4802 MeOH SAMPLE HILL0118.D TN-A 4802 MeOH SAMPLE HlLLOl 19.D TN-A 4802 MeOH SAMPLE HILL0125.D TN-A 4802 MeOH SAMPLE HILL0133.D TN-A 4802 MeOH SAMPLE HILL0134.D TN-A 4802 MeOH SAMPLE HILL0140.D TN-A 4802 MeOH SAMPLE HILL0148.D TN-A 4802 MeOH SAMPLE HILL0149.D TN-A 4802 MeOH T SAMPLE HILL0155.D TN-A 4802 MeOH In!Al SAMPLE HILL0163.D TN-A 4802 MeOH (D SAMPLE HILL0164.D TN-A 4802 MeOH SAMPLE HILL0169.D TN-A 4802 MeOH SAMPLE HILL0170.D TN-A 4802 MeOH SAMPLE HlLL0184.D TN-A 4802 MeOH SAMPLE HILL0185.D TN-A 4802 MeOH THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 5.187 8097 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 THPFOS 0 0 0 PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS PFBS 4.426 10752 0 0 0 0 4.403 9817 0 0 0 0 4.41 10811 0 4.409 10138 0 4.408 7466 0 4.403 11533 0 4.41 10126 0 0 0 0 4.41 10883 0 4.417 9378 0 0 0 0 4.403 9038 0 0 0 0 4.415 9491 0 0 0 0 Page 2 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepolnt(s): Days 1 and 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010426.slFOSA0425.m (ChemStation); Reprocessing Method: H010426a.m (Target) SampType File Sample Name Mlsc Info Compound Name RT Area Amount % Recovew SAMPLE HILLO1OS.D 01003-2501 0 Std FOSA 0 0 0.0 Excluded SAMPLE HILL0106.D 01003-2502 2.5 n g h L FOSA FOSA 5.973 92513 1.8 Excluded CALIB-1 XX HILL0107.D 01003-25-03 5.3 ng/mL FOSA FOSA 5.977 175825 4.5 Excluded CALIB-2 HILLO108.D 01003-25-04 10.1 ng/mL FOSA FOSA 5.977 333744 9.5 94.1% CALlB-3 HILLO109.D 01003-25-05 25 n g h L FOSA FOSA 5.977 832878 25.6 102.4% CALlB-4 HlLLOl10.D 01003-25-06 40.3 n g h L FOSA FOSA 5.979 1346447 42.5 105.4% CALlB-5 HILL0111.D 01003-25-07 49.9 nglmL FOSA FOSA 5.976 1622197 51.0 102.2% CALIB-6 HILL0112.D 01003-2508 74.9 nglrnL FOSA FOSA 5.978 2412994 77.6 103.6% CALIB-7 HILL0113.D 01003-25-09 100.8 nglrnL FOSA FOSA 5.98 3249134 106.6 105.8% CALlB-8 HlLLOl 14.D 01003-25-10 1 249.6 nglmL FOSA FOSA 5.976 7439185 255.5 102.4% CALlB-9 HlLLOl 15.D 01003-25-1 1 403.2 nglrnL FOSA FOSA 5.970 11510014 409.4 101.5% CALlB-10 HILL0116.D 01003-2512 499.2 ng/rnL FOSA FOSA 5.973 13881088 507.2 101.6% SAMPLE HILL0117.D 01003-2513 1008 ng/mL FOSA FOSA 5.977 24702641 998.1 Excluded SAMPLE HILLO131.D 01003-25-03 5.3 nglmL FOSA FOSA 5.971 200577 5.3 Excluded SAMPLE HILL0132.D 01003-2509 100.8 nglmL FOSA FOSA 5.973 3024244 101.7 100.9% SAMPLE HILL0146.D 01003-2503 5.3 ng/mL FOSA FOSA 5.965 157381 4.0 Excluded SAMPLE HILL0147.D 01003-2509 100.8 nglmL FOSA FOSA 5.965 2919745 99.1 98.4% SAMPLE HILL0161.D 01003-25-03 5.3 n g h L FOSA FOSA 5.972 190625 5.1 Excluded SAMPLE HILL0162.D 01003-2509 100.8 nglmL FOSA FOSA 5.969 2792405 94.2 93.4% SAMPLE HILL0171.D 01003-25-01 0 Std FOSA 0 0 0.0 Excluded SAMPLE HILL0172.D 01003-2502 2.5 nghnL FOSA FOSA 5.964 75933 1.4 Excluded CALlB-I 1 XX HILL0173.D 01003-25-03 5.3 nglmL FOSA FOSA 5.973 I52433 3.9 Excluded CALIB-12 HILL0174.D 01003-25-04 10.1 nglmL FOSA FOSA 5.971 297834 8.5 84.4% CALIB-I3 HILL0175.D 01003-2505 25 nglmL FOSA FOSA 5.969 743034 23.3 93.2% CALIB-14 HlLLOl76.D 01003-25-06 40.3 nglmL FOSA FOSA 5.958 1216120 39.2 97.3% CALIB-15 CALIB-I6 HILL0177.D HILL0178.D 01003-25-07 01003-2508 49.9 ng/mL FOSA 74.9 nglmL FOSA FOSA FOSA 5.964 1485755 48.0 5.969 2183173 72.6 96.1% 97.0% CALIB-17 HlLLOl79.D 01003-2509 100.8 nglmL FOSA FOSA 5.966 2971186 100.1 99.3% CALlB-18 HILL0180.D 01003-25-10 249.6 ng/rnL FOSA FOSA 5.971 6959290 243.7 97.6% CALlB-19 HILL0181.D 01003-25-1 1 403.2 ng/mL FOSA FOSA 5.972 10720207 393.0 97.5% CALIB-20 HILL0182.D 01003-2512 499.2 n g h L FOSA FOSA 5.977 13077641 492.6 98.7% SAMPLE HILL0183.D 01003-2513 1008 na/mL FOSA FOSA 5.973 23820299 969.4 Excluded ** Only 10 levels may be quantified due to software limitations,lherefore5-500 ng/mL was chosen. However, since not all blank samples had a FOSA response of less than half the 5 ng/mL response. the 5 ng/mL standard was disabled and the LOQ becam Page 3 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MatelFernale Normal Human Urine; Timepoint(s): Days 1 and 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010426.slFOSA0425.m (ChemStation); Reprocessing Method: HOI0426a.m (Target) Caluculatlons Average (pg/mL) Std Dev oncolumn dil factor corrected % RSD SampType ~ SAMPLE Flle Sample Name HILL0120.D 042301-FOSA-LCS Mlsc Info FOSASolvent Blank Spike Compound Name FOSA RT 5.971 Area 3056177 Amount 107 pg/mL FOSA Spike % Recov. 111% SAMPLE HILLOl21.D FOSA-1.4.1 blk 042301-FOSA-1.4.1 Mtd Blk FOSA 5.893 106755 3 0.00 SAMPLE HlLLOl22.0 FOSA-1.4.2blk 042301-FOSA-1.4.2 Mtd Blk FOSA 0 0 0 0.00 SAMPLE HILL0123.D FOSA-1.4.3blk 042301-FOSA-1.4.3 Mtd BIk FOSA 5.894 135380 '4 0.04 SAMPLE HlLL0124.D FOSA-1.4.4blkMS 42301-FOSA-1.4.4 Mtd Blk Spike FOSA 5.977 2921968 103 107% SAMPLE SAMPLE SAMPLE SAMPLE HILL0126.D HILL0127.D HILL0128.D HILL0129.D mSA44.4 FOSA-1.1.2 FOSA-1.1.3 FOSA-1.1.4MS 042301-FOSA-1.1.1 Smp 042301-FOSA-1.1.2 DUp 042301-FOSA-1.1.3 Trip 042301-FOSA-1.1.4 Spike ms.4 FOSA FOSA FOSA 589 9o90620 343 5.89 6180725 223 5.886 5838914 213 5.887 8313598 312 3A3 2.60 2.18 2.23 0.7 4.4% Rep2,3 RPD 2.13 28% 3.12 98% -based on reps 263 only SAMPLE SAMPLE SAMPLE SAMPLE HILLOl3O.D HILL0135.D HILL0136.D HILL0137.D FOSA-1.2.1 FOSA-1.2.2 FOSA-1.2.3 FOSA-1.2.4MS 042301-FOSA-1.2.1 Smp ' 042301-FOSA-1.2.2 DUP 042301-FOSA-1.2.3 Trip 042301-FOSA-1.2.4 Spike FOSA FOSA FOSA FOSA 5.889 8385467 309 5.885 8541720 313 5.888 8450504 312 5.887 10811323 412 3.09 3.11 3.13 0.0 3.12 1 K 4.12 105% Day Iresults, pglmL average 2.56 SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0138.D HILL0139.D HILL0141.D HILL0142.D HILL0143.D FOSA-1.3.1 FOSA-1.3.2 FOSA-I 3.3 FOSA-1.3.4MS 042301-FOSA-LCS 042301-FOSA-1.3.1 Smp 042301-FOSA-I.3.2 DUp 042301-FOSA-I.3.3 Trip 042301-FOSA-1.3.4 Spike FOSA Solvent Blank Spike FOSA FOSA FOSA FOSA FOSA 5.862 6883587 248 5.886 6433005 233 5.886 6255010 230 5.886 9067750 340 5.97 2756837 98 2.48 2.37 2.33 0.1 2.30 4% 3.40 107% 102% stddev 0.38 %RSD 15% SAMPLE SAMPLE SAMPLE SAMPLE HlLLOl44.D HILL0145.D HILLO15O.D HILL0151.D FOSA-2.4.1 blk FOSA-2.4.2blk FOSA-2.4.3blk FOSA-2.4.4blkMS 042301-FOSA-2.4.1 Mtd Blk 042301-FOSA-2.4.2 Mtd Blk 042301-FOSA-2.4.3 Mtd Elk 42301-FOSA-2.4.4 Mtd Blk Spike FOSA FOSA FOSA FOSA 0 0 0 0 0 0 0 0 0 5.97 2811726 98 102% SAMPLE SAMPLE SAMPLE SAMPLE HILL0152.D HILL0153.D HILL0154.D HILL0156.D FOSA-2.1.1 FOSA-2.1.2 FOSA-2.1.3 FOSA-2.1.4MS 042301-FOSA-2.1.1 Smp 042301-FOSA-2.1.2 DUP 042301-FOSA-2.1.3 Trip 042301-FOSA-21.4 Spike FOSA FOSA FOSA FOSA 5.879 8103858 317 5.879 8151376 296 5.88 7853370 291 5.887 10247004 393 3.17 3.02 2.96 0.1 2.91 5% 3.93 96% SAMPLE HILL0157.D FOSA-2.2.1 042301-FOSA-2.2.1 Smp FOSA 5.887 7993752 298 2.98 2.82 SAMPLE HILL0158.D FOSA-2.2.2 042301-FOSA-2.2.2 DUP FOSA 5.886 7697039 280 2.80 0.1 SAMPLE HILL0159.D FOSA-2.2.3 042301-FOSA-2.2.3 Trip FOSA 5.887 7443513 269 2.69 5% 9 SAMPLE HILLOl6O.D FOSA-2.2.4MS 042301-FOSA-2.2.4 Spike FOSA 5.888 9662387 359 3.59 80% (P (D SAMPLE HILLO165.D FOSA-2.3.1 042301-FOSA-2.3.1 Smp FOSA 5.679 7850050 289 2.89 2.81 UI SAMPLE HILL0166.D FOSA-2.3.2 042301-FOSA-2.3.2 Dup FOSA 5.882 7536620 279 2.79 0.1 W SAMPLE HILL0167.D FOSA-2.3.3 042301-FOSA-2.3.3 Trip FOSA 5.88 7535404 275 2.75 3% 4A SAMPLE HILL0168.D FOSA-2.3.4MS 042301-FOSA-2.3.4 Spike FOSA 5.886 9615944 362 3.62 84% 2 rn Day 2 results, pglmL average 2.88 stddev 0.12 %RSD: 4% 7 $ %' J sp r?nt1-7 00- f4 28 z2 Page 4 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepoint@): Days 1 and 2 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010426.slFOSA0425.m (ChemStation); Reprocessing- Method: HO10426a.m (Target) SampType File Sample Name SAMPLE HILLO103.D TN-A 4802 MeOH SAMPLE HILL0104.D TN-A 4802 MeOH SAMPLE HILLOl18.D TN-A 4802 MeOH SAMPLE HILL0119.D TN-A 4802 MeOH SAMPLE HILL0125.D TN-A 4802 MeOH SAMPLE HILL0133.D TN-A 4802 MeOH SAMPLE HILL0134.D TN-A 4802 MeOH SAMPLE HILL0140.D TN-A 4802 MeOH SAMPLE HILL0148.D TN-A 4802 MeOH SAMPLE HILL0149.D TN-A 4802 MeOH SAMPLE HILL0155.D TN-A 4802 MeOH SAMPLE HILL0163.D TN-A 4802 MeOH SAMPLE HILL0164.D TN-A 4802 MeOH SAMPLE HILL0169.D TN-A 4802 MeOH SAMPLE HILL0170.D TN-A 4802 MeOH SAMPLE HILL0184.D TN-A 4802 MeOH SAMPLE HILL0185.D TN-A 4802 MeOH Mlsc Info Compound Name FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA RT 5.981 5.976 5.979 5.978 0 5.978 0 5.972 0 0 5.972 0 0 5.986 5.984 5.977 5.971 Area 143948 47424 129092 53533 0 24760 0 40470 0 0 38428 0 0 48316 17972 136896 49865 Amount G I 2 LOQ resp -412 LOQ resp 412 LOP resp 4 1 2 LOQ resp 4 1 2 LOQ resp <112 LOQ resp 4 1 2 LOQ resp 4 1 2 LOQ resp 4 1 2 LOQ resp <1/2 LOP resp 4 1 2 LOQ resp 4 1 2 LOO resp 4 1 2 LOQ resp 4 1 2 LOQ resp 4 1 2 LOQ resp W2 LOQ resp 4 1 2 LOQ resp 3 ul (0 Page 5 of 5 BACK TO MAIN n Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Day 3 Analytical Instrument: HPLC/MSD "Hillary"; Analytical Sequence/Method: H010501.s/FOSA0425.m (ChemStation); Reprocessing Method: H010501a.m (Target) SampType File SAMPLE HILL0078.D SAMPLE HILL0079.D Sample Name 01003-25-01 01003-25-02 .Misc Info 0 Std 2.5 nglmL FOSA Compound Name THPFOS THPFOS RT 5.115 5.116 Area 1381461 1374933 Amount 254 254 Compound Name PFBS PFBS CALlB-I HILL0080.D 01003-25-03 5.3 nglmL FOSA THPFOS 5.117 1393833 254 PFBS CALIB-2 HILL0081.D 01003-25-04 10.1 ng/mL FOSA THPFOS 5.117 1385169 254 PFBS CALlB-3 HILL0082.D 01003-25-05 25 ng/mL FOSA THPFOS 5.1 13 1380163 254 PFBS CALlB-4 HILL0083.D 01003-25-06 40.3 nglmL FOSA THPFOS 5.116 1376007 254 PFBS CALlB-5 HILL0084.D 01003-25-07 49.9 nglmL FOSA THPFOS 5.129 1374674 254 PFBS CALIB-6 HILL0085.D 01003-25-08 74.9 nglmL FOSA THPFOS 5.11 1374202 254 PFBS CALIB-7 HILL0086.D 01003-25-09 , 100.8 nglmL FOSA THPFOS 5.123 1380970 254 PFBS CALIB-8 HILL0087.D 01003-25-10 249.6 ng/mL FOSA THPFOS 5.111 1382975 254 PFBS CALIB-9 HILL0088.D 01003-25-11 403.2 ng/mL FOSA THPFOS 5.115 1394460 254 PFBS CALlB-IO HlLL0089.D 01003-25-12 499.2 nglmL FOSA THPFOS 5.115 1388280 254 PFBS SAMPLE HILL0090.D 01003-25-13 I008 nglmL FOSA THPFOS 5.122 1389397 254 PFBS SAMPLE SAMPLE SAMPLE HILL0104.D HILLO116.D HILL0119.D 01003-25-09 01003-25-09 01003-25-01 100.8 nglmL FOSA 100.8 nglmL FOSA 0 SM THPFOS THPFOS THPFOS 5.117 5.125 5.115 1244044 254 I247230 254 1266027 254 PFBS PFBS PFBS SAMPLE CALIB-11 CALIB-12 CALIB-I 3 HILL0120.D HILL0121.D HILL0122.D HILLO123.D 01003-25-02 01003-25-03 01003-25-04 01003-25-05 2.5 nglmL FOSA 5.3 nglmL FOSA 10.1 nglmL FOSA 25 nglmL FOSA THPFOS THPFOS THPFOS THPFOS 5.123 5.126 5.114 5.122 1262927 1263723 1282166 1280362 254 254 ,254 .254 PFBS PFBS PFBS PFBS CALIB-14 HILL0124.D CALIB-15 HlLLOl25.D 01003-25-06 01003-25-07 40.3 n g h L FOSA 49.9 ng/mL FOSA THPFOS THPFOS 5.115 5.131 1275643 254 I276350 254 PFBS PFBS CALlB-16 CALIB-17 CALlB-18 CALlB-19 HILLO126.D HILL0127.D HILL0128.D HILL0129.D 01003-25-08 01003-25-09 01003-25-10 01003-25-11 74.9 nglmL FOSA 100.8 ng/mL FOSA 249.6 ng/mL FOSA 403.2 ng/mL FOSA THPFOS THPFOS THPFOS THPFOS 5.124 5.12 5.119 5.117 1274208 254 1269909 254 1287101 254 1292619 254 PFBS PFBS PFBS PFBS CALIB-20 HILL0130.D SAMPLE HlLLOl31.D 01003-25-12 01003-25-13 499.2 nglmL FQSA 1008 ng/mL FOSA THPFOS THPFOS 5.118. 5.115 1288469 254 1293221 254 PFBS PFBS RT 4.408 4.401 4.41 4.416 4.42 4.408 4.407 4.41 4.423 4.411 4.401 4.414 4.414 4.402 4.41 4.4 15 4.415 4.418 4.421 4.414 4.41 5 4.416 4.424 4.419 4.412 4.416 4.41 1 4.422 Area 1798374 1777751 1801694 1810322 1800005 1780106 1812881 1766585 1798232 1773614 1805937 1776350 1788258 1778858 1778060 1789621 1775266 1808173 1804070 1774497 1778220 1785753 1775648 1757613 1767480 1778677 1776855 1781355 Amount 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 254 Page 1 of 5 enr lia! BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemaleNormal Human Urine; Timepoint(s): Day 3 Analytical Instrument: HPLClMSD "Hillary"; Analytical Sequencernethod: H010501.s/FOSA0425.m (ChemStation); SAMPLE HILL0093.D SAMPLE HILL0094.D FOSA-3.4.lblk FOSA-3.4.2blk 042301-FOSA-3.4.1, Mtd Blk 042301-FOSA-3.4.2, Mtd Blk THPFOS THPFOS 5.118 5.123 1236810 254 1299549 254 PFBS PFBS 4.396 1629186 254 4.402 1708821 254 SAMPLE SAMPLE SAMPLE SAMPLE HILL0095.D HILL0096.D HILL0097.D HILL0099.D FOSA-3.4.3blk FOSA-3.4.4bIkMS FOSA-3.1.1 FOSA-3.1.2 042301-FOSA-3.4.3, Mtd Blk 042301-FOSA-3.4.4, Mtd Blk Spike 042301-FOSA-3.1.1, Srnp 042301-FOSA-3.1.2, DUp THPFOS THPFOS THPFOS THPFOS 5.124 5.123 5.118 5.117 1331679 254 1313977 254 1186398 254 I160945 254 PFBS PFBS PFBS PFBS 4.41 1727981 254 4.416 1714291 254 4.403 1735419 254 4.41 1725206 254 SAMPLE HILLO1OO.D FOSA-3.1.3 042301-FOSA-3.1.3, Trip THPFOS 5.124 1174802 254 PFBS 4.409 1738131 254 SAMPLE HILLO1O1.D FOSA-3.2.1 042301-FOSA-3.2.1, Smp THPFOS 5.1 17 1170460 254 PFBS 4.409 1757667 254 SAMPLE HILL0102.D FOSA-3.2.2 042301-FOSA-3.2.2, DUP THPFOS 5.117 1157352 254 PFBS 4.402 1733154 254 SAMPLE HILLO103.D FOSA-3.2.3 042301-FOSA-3.2.3, Trip THPFOS 5.118 I169370 254 PFBS 4.418 1755402 254 SAMPLE HILLO107.D FOSA-3.2.4MS 042301-FOSA-3.2.4, Spike THPFOS 5.116 1142996 254 PFBS 4.408 1710176 254 SAMPLE HILL0108.D FOSA-3.3.1 042301-FOSA-3.3.1, Smp THPFOS 5.124 1120852 254 PFBS 4.403 1739019 254 SAMPLE HILLO109.D FOSA-3.3.2 042301-FOSA-3.3.2, DUp THPFOS 5.128 1115271 254 PFBS 4.407 1748703 254 SAMPLE HlLLOl 10.D FOSA-3.3.3 042301-FOSA-3.3.3, Trip THPFOS 5.123 1120639 254 PFBS 4.415 1743629 254 SAMPLE HILL0112.D FOSA-3.3.4MS 042301-FOSA-3.3.4, Spike THPFOS 5.1 11 1102414 254 PFBS 4.404 1709063 254 SAMPLE HlLLOl 13.13 042301-FOSA-LCS FOSA Solvent Blankspike THPFOS 5.13 1168358 254 PFBS 4.415 1691538 254 SAMPLE HILLO114.D Diluter Blk Diluter blank with IS mix THPFOS 5.122 1184946 254 PFBS 4.415 1645960 254 SAMPLE HILLO115.D MeOH blkw/ IS Methanol with IS mix THPFOS 5.115 809160 254 PFBS 4.414 1075206 254 Average: Std Deviation: %RSD: 1262315 112912 8.9% 1745413 108834 6.2% SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0077.D HILL0091.D HILL0092.D HILL0098.D HILLO105.D HILLO106.D HILL01l1.D HILL0117.D HILLO116.D HILL0132.D HILL0133.D TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH w-0 ln (D 2 s 2 2 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 THPFOS 0 0 0 PFBS 0 0 0 Page 2 of 5 z Y 5a . -0= e-(.D3I sp BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemaleNormalHuman Urine; Timepoint@): Day 3 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequenceMethod: H010501.slFOSA0425.m (ChemStation); SampType File Sample Name Misc Info Compound Name RT Area Amount %Recovery SAMPLE HILL0078.D 01003-25-01 0 Std FOSA 0 0 0 SAMPLE CALlB-I CALIB-2 CALIB-3 CALlB-4 CALIB-5 CALIB-6 CALIB-7 CALI8-8 CALlB-9 CALlB-IO SAMPLE HILL0079.D HILL0080.D HILL0081.D HILL0082.D HILL0083.D HILL0084.D HILL0085.D HILLOO86.D HILL0087.D HILL0088.D HILL0089.D HILL0090.D 01003-25-02 01003-25-03 01003-25-04 01003-25-05 01003-25-06 01003-25-07 01003-25-08 01003-25-09 01003-25-10 01003-25-1I 01003-25-12 01003-25-13 2.5 n g h L FOSA 5.3 nglrnL FOSA 10.1 nglrnL FOSA 25 nglrnL FOSA 40.3 nglrnL FOSA 49.9 ng/rnL FOSA 74.9 nglrnL FOSA 100.8 nglrnL FOSA 249.6 nglrnL FOSA 403.2 ng/rnL FOSA 499.2 nglrnL FOSA 1008 nglrnL FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA 5.991 5.993 5.985 5.982 5.984 5.997 5.985 5.985 5.98 6.004 5.983 5.983 84044 160849 307301 768189 1231657 1519482 2227975 3036314 6971036 10819514 12885941 23137645 3 5 10 25 41 50 76 103 253 405 505 1018 97% 100% 102% 100% 101% 102% 101% 101% 101% SAMPLE SAMPLE SAMPLE SAMPLE CALlB-11 CALlB-12 CALIB-13 CALlB-14 CALIB-15 CALIB-16 CALIB-17 CALlB-18 CALlB-I 9 CALIB-20 SAMPLE HILL0104.D HILLO116.D HILLO119.D HILL0120.D HILLO121.D HILL0122.D HlLL0123.D HILL0124.D HILL0125.D HILLOlL6.D HILL0127.D HILL0128.D HILL0129.D HILL0130.D HILL0131.D 01003-25-09 01003-25-09 01003-25-01 01003-25-02 01003-25-03 01003-25-04 01003-25-05 01003-25-06 01003-25-07 01003-25-08 01003-25-09 01003-25-10 01003-25-11 01003-25-12 01003-25-13 100.8 nglrnL FOSA 100.8 nglrnL FOSA 0 Sld 2.5 ng/mL FOSA 5.3 nglrnL FOSA 10.1 nglrnL FOSA 25 ng/rnL FOSA 40.3 nglrnL FOSA 49.9 nglmL FOSA 74.9 ng/mL FOSA 100.8 nglrnL FOSA . 249.6 n g h L FOSA 403.2 nglrnL FOSA 499.2 nglmL FOSA 1008 ng/rnL FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA 5.999 6 0 5.984 5.987 5.983 5.99 5.984 5.999 5.986 5.988 5.988 5.985 5.994 5.984 2915113 100 2902871 99 0 0 72740 2 152363 5 298779 10 736231 24 1203821 40 1473022 49 2181078 74 2931206 101 6818207 248 10436918 396 12664603 495 22575515 992 102% 102% 95% 97% 99% 98% 99% 101% 99% 98% 99% Page 3 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepoint(s): Day 3 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010501.slFOSA0425.m (ChemStation); Caluculations Average (pg/mL) Std Dev on-column dil factor corrected % RSD SampType File Sample Name Misc Info Compound Name RT Area Amount pglmL FOSA Spike % Recov. SAMPLE HILL0097.D FOSA-3.1.1 042301-FOSA-3.1.1, Smp FOSA 5.909 7201703 269 2.69 2.60 SAMPLE HILL0099.D FOSA-3.1.2 042301-FOSA-3.1.2, DUP FOSA 5.901 6731194 251 2.51 0.1 SAMPLE HILL0100.D FOSA-3.1.3 042301-FOSA-3.1.3, T i p FOSA 5.908 6998562 280 2.60 3% Not enough sample was available to preparethe spike replicate for sample#l. Spike results from samples two and three shall suffice to demonstrate spike recoveries. SAMPLE SAMPLE SAMPLE SAMPLE HILLOlO1.D HILL0102.D HILLOlO3.D HILLO107.D FOSA-3.2.1 FOSA-3.2.2 FOSA-3.2.3 FOSA-3.2.4MS 042301-FOSA-3.2.1, Smp 042301-FOSA-3.2.2, DUP 042301-FOSA-3.2.3, T i p 042301-FOSA3.2.4, Spike FOSA FOSA FOSA FOSA 5.908 5.922 5.895 5.9 6990791 257 7009317 261 6836606 251 9016954 351 2.57 2.56 2.61 0.1 2.51 2% 3.51 99% SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0108.D FOSA-3.3.1 HILLO109.D HILLO1lO.D FOSA-3.3.2 FOSA-3.3.3 HILLO112.D FOSA-3.3.4MS HILL0113.D 042301-FOSA-LCS 042301-FOSA-3.3.1, Smp 042301-FOSA-3.3.2, DuP 042301-FOSA-3.3.3. Trip 042301-FOSA-3.3.4, Spike FOSA Solvent Blank Spike FOSA FOSA FOSA FOSA . FOSA 5.916 5.906 5.9 5.909 5.991 7065688 263 7377514 274 7030743 260 9073879 354 2750436 99 2.63 2.66 2.74 0.1 2.60 3% 3.54 92% 103% Day 3 results, pg/mL average 2.61 std dev 0.05 %RSD: 2% z 9 1. -u nl (m0 Page 4 of 5 BACK TO MAIN Solubility Study, Shake Flask Method; Test Compound: FOSA; Test Matrix: MalelFemale Normal Human Urine; Timepoint@): Day 3 Analytical Instrument: HPLClMSD "Hillary"; Analytical SequencelMethod: H010501.slFOSA0425.m (ChemStation); dil factor corrected SampType SAMPLE SAMPLE SAMPLE File HILL0093.D HILL0094.D HILL0095.D Sample Name FOSA-3.4.1 blk FOSA-3.4.2blk FOSA-3.4.3blk Mlsc Info 042301-FOSA-3.4.1, Mtd Blk 042301-FOSA-3.4 2, Mtd Blk 042301-FOSA-3.4.3, Mtd Blk Compound Name FOSA OSA OSA RT 6.007 5.992 5.986 Area Amt 553440 20 450065 pg/mL FOSA 0.20 0.1 8 0.15 SAMPLE HILL0096.D FOSA-3.4.4blkMS 042301-FOSA-3.4.4, Mtd Blk Spike SAMPLE HILLOI14.D Diluter Elk Diluter blank with IS mix FOSA FOSA 5.984 3129563 Ill 0 0 0 ** SAMPLE HlLLOl15.D MeOH blk w/ IS Methanolwith IS mix FOSA 0 0 0 116% SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE SAMPLE HILL0077.D HILL0091.D HILL0092.D HILL0098.D HILLO105.D HILLOlO6.D HILLO1ll.D HILLO117.D HILL0118.D HlLLOl 32.D HILL0133.D TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH TN-A 4802 MeOH FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA FOSA 5.988 5.991 5.992 5.99 5.992 5.991 5.916 0 0 5.984 5.986 7951 1 0 166363 0 63070 0 40614 0 30961 0 18624 0 46114 0 0 0 0 0 150108 0 56599 0 "Day 3 samples were initially diluted on 04-25-01 and analyzed for FOSA in sequence H010430.s Method blank samples demonstrated a significant amount of FOSA. The Day 3 samples were again diluted 1OX with methanolalong with a diluter blank to check for FOSA backgroundin the diluter, and a methanol blank spiked with IS mix to check for FOSA backgroundlcontamination in the internalstandard mix. Method blank samples analyzed in seq H010501.sstill had a background of FOSA, but it was determined that the FOSA was somehow introducedto the urine during the prep procedureprior to dilution since the diluter blank and IS blank came out "clean." Days 1 and 2 method blank samples demonstrate that the urine used for the study had no initial background of FOSA. and so the levelof FOSA in Day 3 mtd blks was due to a small amt of FOSA introducedto the test system during sample preparation. This backgroundof FOSA in the Day 3 mtd blks may be disregarded since it was only incurred on the day of test analyte in addition, and was a random, inadvertent error that does not affect the solubility results for the other three Day 3 samples. Page 5 of 5 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 ATTACHMENCT: SAMPLECHROMATOGRAMS CHROMATOGRAMS INCLUDED FOR BOTH PFOS AND FOSA: -3 SOLVENTBLANKCHROMATOGRAMS -3 METHODBLANKCHROMATOGRAMS -9 SAMPLE CHROMATOGRAMS (3 CHROM PER DAY) Page 75 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426 .b\HILL0029.D 3MEnvironmentalLaboratogage 1 Report Date: 01-May-200111:16 Project # E01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOO29.D Lab Smp Id: TN-A 4802 MeOH Inj Date : 26-APR-2001 22:02 Operator : KLT Inst ID: hil1ary.i Smp Info : TN-A 4802 MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD rrHillary"H,010426.s Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426t.m Meth Date.: 01-May-200111:12 terrell Quant Type: ISTD Cal Date : 26-APR-200121:40 Cal File: HILL0027.D Als bottle: 92 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 75 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable 1: Page 76 of 171 BACK TO MAIN d Data File: D:\chem\Hillary.i\HOlO426.b\HILLOO29+D 3M Environmental Laboratory Project # E01-0768 Page 7.5: 7.21 6+9: 6.61 6.3: 6.0; 5.7; 5,4{ 5.1: 4,8{ 431 ' 4.21 5, 3.9: - 3.6: 3.3: 3,O: 2.7: 2.4: 2.1- 1.8: 1.5; 1.21 0.9; D:\chem\Hillary.i\HO10426.b\HILL0029.D )t 1 THPFOS (Undetected) I Ion M 7 PFBS 1.61 1.4; 1.2; A IM, 1.0: 4 5 0.8; >- I on 0 rn 1 d .I ...,...,...,...,.. 4,OO 4.20 4,40 4.60 4,80 Hin Page 77 of 171 Data File: D:\chem\Hillary,i\HOlO426.b\HILLOO29.D 2 PFOS BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page ' Page 78 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010501.b\HILL0036.D 3MEnvironmentalLaboratoQage 1 Report Date: 03-May-2001 09:17 Project# E01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO36.D Lab Smp Id: TN-A 4802 MeOH In] Date : 01-MAY-200122:59 Operator : KLT Inst ID: hil1ary.i Smp Info : TN-A 4802 MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylHl0,10501.s Method : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Olt.m Meth Date : 03-May-200108:17 terrell Quant Type: ISTD Cal Date : 01-MAY-2001 21:19 Cal File: HILL0027.D Als bottle: 93 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT S I G MASS ---_-- 427 299 499 CONCENTRATIONS ON-COLUMN FINAL RT BXP RT REL RT RESPONSE (ng/mL) ---- --_---_---____ _--_______ _ _ __-_-___-__---======= (ng/mL) ======= Compound Not Detected. Compound Not Detected. Compound Not Detected. Page 79 of 171 Data File: D:\chem\Hillary,i\HOlO5Ol+b\HILLOO36.D D:\chem\Hillary,i\HOlO5Ol+b\HILLOO36.D 4.6: 4.4; 4.21 4.01 3.8; 3.61 3.41 3.21 3.0: ; 2.6; 2.4; 2.21 2.0- 1.8; 1,6{ 1.4: 1.21 l,O{ 0.8- BACK TO MAIN 3M Environmental Laboratory Page Project # E01-0768 M 1 THPFOS (Undetected) 1.10: 1 1.00; 0,YO-i 0.80; 7 n 0.70; Txi 0.60; v > 0.50; Y 2 PFBS (Undetected) 1+3- 1 1,2; 1.1: Lo: h 0.9; m 0.8; 0 1.I 0,7: " 0.6; >. Page 80 of 171 Data File: D:\chem\Hillary,i\HOlO5Ol.b\HILLOO36,D 3 PFOS 1,20: (Undetected) Ion 0".30:' " 4.4 O4.8 k. 5,2 Min .. ,. ..,.. . 5,6 6.0 BACK TO MAIN 3M EnvironmentalLaboratory page 3 Project # E01-0768 Page 81 of 171 ~~ BACK TO MAIN Data File: D:\chem\Hillary.i\H010430.b\HILL0057.D 3rd EnvironmentalLaboratogage 1 Report Date: 01-May-200113:29 Project # E01 -0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO43O.b\HILLOO57.D Lab Smp Id: TN-A 4802 MeOH Inj Date : 30-APR-2001 18:57 Operator : KLT Inst ID: hil1ary.i Smp Info : TN-A 4802 MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylHl0,10430.s Method D:\chem\Hillary.i\HOlO43O.b\HOlOO430t.m Meth Date : 01-May-2001 12:59 terrell Quant Type: ISTD Cal Date : 30-APR-200113:21 Cal File: HILL0027.D Als bottle: 93 Dil Factor: 1.00000 Inteqrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds _____ _____ _____ ___ __ ___ __- _- - -_- =_= _= =_= -=- - - * 1 THPFOS * 7 PFBS 2 PFOS QUANT SIG MASS ==== 427 299 499 _ _ _ _ RT EXP RT RBL RT RESPONSE r___---- ======== ======E= Compound Not Detected. Compound Not Detected. Compound Not Detected. CONCENTRATIONS ON-COLUMN FINAL (ng/mL) ======= (ng/mL) ======= ,--- -- - - -____- Exaef copy of 0rPglwd Page 82 of 171 Data File: D:\chern\Hi 1 lary. i\H010430,b\HILL0057.~ 6+0: 5.8; 5.61 5.4; 5.2; 5.04 4.8; 4.6; 4,4; 4.21 4+0{ 3.84 3+6{ i 3,4; 3.2; ' 3+0{ 2.81 2.6; 2.4; 2.2; 2.0; 1+8; 1.6; 1,4{ 1.2; 1.0: D:\chem\Hillary,i\HOlO43O+b\HILLOO57,D BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Y 1 THPFOS (Undetected) 600; ; 560: 520 480; 440; 400 ; 360 320 Y 7 PFBS <Undetected) ; 750 ; 700 650; 600 550; 500: 450; 400; ' 350 I on 2 Page 83 of 171 Data File: D:\chem\Hillary,i\HOlO43O.b\HILLOO57,D 2 PFOS (Undetected) I on I 720 680; 640; 600: G 560: 520: > 480; 440; 4001 360; 320: ,. .., .. ., ... ,. .. ,... 4.4 4.8 5.2 5.6 6,O Hin BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 84 of 171 BACK TO MAIN Data File : D:\chem\Hillary.i\H010426.b\HILL0032.D 3M EnvironmentalLaboratoBage 1 Report Date: 01-May-200111:16 Project # E01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOO32.D Lab Smp Id: PFOS-1.4.lblk In] Date : 26-APR-200122:36 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylHl0,10426.s Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426t.m Meth Date : 01-May-2001 11:12 terrell Quant Type: ISTD Cal Date : 26-APR-2001 21:40 Cal File: HILL0027.D Als bottle: 22 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS _-_-___- 427 299 499 RT EXP RT REL RT RESPONSE _--_-_-_ -_-_-_-_-_-_-_-_ _-_-_--_-_-_------___--_- 5.111 5.131 (1.000) 1357667 4.369 4.410 (1.000) 1900574 Compound Not Detected. Page 85 of 171 Data File: D:\chem\Hillary,i\HOlO426.b\HILLOO32.D BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 2 H 1 THPFOS 1 1.61 Lo 1.41 1.21 y)CI 1.0: 0 0.8: Y >. 0.6: 0.4: H 7 PFBS 2.4; r-. 1 2.2; d 2.0; 1.8: 1.6; 1.4: Page 86 of 171 Data File: D:\chem\Hillary,i\HOlO426.b\HILLOO32.D 2 PFOS (Undetected) rI - 1.0: 0.8: 0.6: 0*4i-+L-4 4.4 4.8 5.2 . , . . . , . . . 5.6 6,O I Hin BACK TO MAIN 3M Environmental Laboratory Page Project # E01-0768 Page 87 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010501.b\HILL0032.D 3MEnvironmenta'Laborato age 1 Report Date: 03-May-2001 09:16 Project # Eo1-078 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO32.D Lab Smp Id: PFOS-2.4.lblk Inj Date : 01-MAY-2001 22:15 Operator : KLT Inst ID: hi1lary.i Smp Info ' : 1,OOOX dil 042301-PFOS-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD "HillaryiiH,010501.s Method : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Olt.m Meth Date : 03-May-2001 08:17 terrell Quant Type: ISTD Cal Date : 01-MAY-2001 21:19 Cal File: HILL0027.D Als bottle: 22 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS __ _-_-__ 427 299 499 RT BXP RT REL RT RESPONSE _--_ -_ -_ -_ _____ ___ ___ _--_----_-------_-- ---_-_-------- 5.114 5.109 (1.000) 1317272 4.407 4.402 (1.000) 1718184 Compound Not Detected. CONCENTRATIONS ON-COLUMN FINAL (ng/mL) -------------- (ng/mL) _- _______- _- __ 254.000 254.000 Page 88 of 171 BACK TO MAIN Data File: 'D:\chem\Hillary.i\HOlO5Ol.b\HILLOO32.D 2,s: 2.4; 2.3; 2.24 2.1; 2.0; 1,s; 1.8; 1.7: 1.6; 1.5; 1.4; 1.3; 2 1.2; -' 1,1; 1.0; 0.9; 0,8; 0,7: 0.6; 0.5; 0,4: 0.3; 0,2; D:\chem\Hi 1 la1 i\H010501.b\HILL0032.D 3M Environmental Laboratory Project # E01-0768 Page 2 , . . . . , . . . ..................... 0.5 1.0 1.5 2,O 2,5 3.0 3.5 4,O 4,5 5.0 5,5 6,O 6,5 7,O 7.5 8.0 8,5 9.0 9.5 Min X 1 THPFOS 2 PFBS 2.4; 2.24 2.0; 1.8; i+6; 1+4{ bo 4 d Page 89 of 171 Data File: D:\chemWillary,i\HOlO5Ol.b\HILLOO32.D 3 PFOS (Undetected) I I on BACK TO MAIN 3M Environmental Laboratory Project # E01 -0768 BACK TO MAIN Data File: D:\chem\Hillary.i\H010430.b\HILL0059.D 3MEnvironmentalLaborato~age 1 Report Date: 01-May-200113:29 Project # E01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO43O.b\HILLOO59.D Lab Smp Id: PFOS-3.4.lblk Inj Date ' : 30-APR-2001 19:19 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-3 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylHl0,10430.s Method : D:\chem\Hillary.i\HOlO43O.b\HOlOO430t.m Meth Date : 01-May-200112:59 terrell Quant Type: ISTD Cal Date : 30-APR-2001 13:21 Cal File: HILL0027.D Als bottle: 22 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS CONCENTRATIONS ON-COLUMN FINAL RT EXP RT RBL RT RESPONSE (ng/mL) (ng/mL) Page 91 of 171 Data File: D:\chem\Hillary.i\HOlO43O.b\HILLOO59,D 2.6; 2.5; 2.44 2,3; 2.2; 24 2.0; 1.9; 1.8; 1.7; 1.C 1.5; ; 1.4: ; 1.3; -i 1.2; LA; 1.0: 0.9; I 1 0.8; 0.7; 0+6j 0.54 0,44 0.3; 0,2; 0.1; ........................................ '. 0.5 1,O 1.5 2.0 2.5 3+0 3,s 4.0 W 1 THPFOS 2.0; QI 1,81 1.6; 1.4: W 7 PFBS BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 Page 92 of 171 Data File: D:\chen\Hillary,i\HO1043O+b\HILL0059.D 2 PFOS (Undetected) I on 1.20: 1.10; 1.00; 0.90; 0.80; 20 0,704 Y ~ 0.60; iI iJ 0.50; 0,40; 4.4, . . .4.8, . . .5.2, . . .5.,6 . . .6.,0 . . . BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 93 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426 .b\HILL0037.D 3MEnvironrnenta'Laborato age 1 Report Date: 01-May-2001 11:16 Project# E01- 0 2 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOO37.D Lab Smp Id: PFOS-1.1.1 In] Date : 26-APR-2001 23:32 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylHl0,10426.s Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426t.m Meth Date.: 01-May-2001 11:12 terrell Quant Type: ISTD Cal Date : 26-APR-2001 21:40 Cal File: HILL0027.D Als bottle: 26 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds QUANT SIG MASS CONCENTRATIONS ON-COLUMN FINAL RT BXP RT RBL RT RBSPONSB (ng/mL) (ng/mL) Page 94 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HOlO426,b\HILLOO37.D D:\chem\Hillary,i\HOlO4; 1.0- b\HILL0037,D 3M Environmental Laboratory Project # E01-0768 Page 0.9- 0.8- 0.7- 0+6- H 1 THPFOS N 1.8; 1 lo 1.6; 1.4; ~ i.2: gIs) 1.0; vxi 0 , 8 { V > 0.6l 7 PFBS 2,71 2.41 2.11 c; 1+81 6 1+51 2d 1,2{ 0,9{ 0.6: h d Page 95 of 171 Data File: D:\chem\Hillary.i\HOlO426,b\HILLOO37.D 2 PFOS I on 1.01 0.9, 0.8j 0,7: qn * 0.6;: b. 0.5: 24 0.4; 0.1: , 1\ , . . ., ... , . .. ,. .. , .. . 4.4 4.8 5.2 5.6 6.0 BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 Page Page 96 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426.b\HILL0041.D 3MEnvironPmroejenctta#'LEa0b1or-a0to7f#age Report Date: 01-May-2001 11:16 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOO4l.D Lab Smp Id: PFOS-1.2.1 Inj Date : 27-APR-200100:16 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD tlHillarytHl0,10426.s Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426t.m Meth Date : 01-May-200111:12 terrell Quant Type: ISTD Cal Date : 26-APR-200121:40 Cal File: HILL0027.D Als bottle: 30 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Page 97of 171 BACK TO MAIN Data File: D:\chem\Hillary,i\HO10426.b\HILL0041.D 3M EnvironmentalLaboratory Project # E01-0768 Page 2 D:\chem\Hillary,i\HO10426.b\HILL0041.D 9.3: 9.0; 1 8,7; 8,41 8.11 7.8: 7.51 7.2; 6.91 6.6; 6.3: 6.0: 5.7; 5+4{ , 5.11 ; 4.8: 4.5: 3 4.21 - 3.9: v) 3.6; 0 LI 3.3; 3.01 1 2.7; 1 2.41 2.1: 1.81 1,5{ 1.2; 0.91 0,s: 0.31 . . .I. .I. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1. . . . . . 1. . . . . . . . .1 . . . . . . . . . I. I. . . . . . . . . . . . . . . . . . . . . . . . . . . . . ,1 . . 1 . . Ion 1.6: LD 1.4: 1,2{ ?A 1.0: . 0.84 " X > 0.6{ 0.4; 0.2: ..I I ."I '.' 1 I ' "I ' . . I . 4.4 ' 4.8 5.2 5.6 6.0 Min Y 7 PFBS 2.7: 2,4{ 2.1: 1.81 h 1,5{ 0 $" 1.2: > 0+9: 0.61 : I 0.3: 4.00 4.20 4.40 4,60 4,80 Page 98 of 171 Data File: D:\chem\Hi 1 lary. i\H010426,b\HILL0041.D 2 PFOS 8.0; 7.0; 6.0; A 5.0; 0 2 . 4.0; " ~ 3.0; 2.0; BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 99 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426.b\HILL0049.D 3M EnviroPnrmojencta#'LEa0b1o-0ra7%t age 1 Report Date: 01-May-200111:16 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOO49.D Lab Smp Id: PFOS-1.3.1 In] Date : 27-APR-2001 01:47 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD I1Hillaryf1H,010426.s Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426t.m Meth Date : 01-May-200111:12 terrell Quant Type: ISTD Cal Date . : 26-APR-2001 21:40 Cal File: HILL0027.D Als bottle: 34 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS ___-_-_ - 427 299 499 CONCBNTRATIONS ON-COLUMN FINAL RT BXP RT RBL RT RBSPONSB (ng/mL) (ng/mL) --_-_- --_--------_--_-- -_--------__--_-___-_ _____ __________-_______-______ 5.129 5.131 (1.000) 1269547 254.000 4.394 4.410 (1.000) 2195975 254.000 5.269 5.334 (1.027) 8267658 320.624 321 Page 100 of 171 Data File: D:\chem\Hillary.i\HOlO426,b\HILLOO43.D D:\chem\Hillary,i\HOIO4~ 1.0- 0.9- 0.8- 0.7- 0.6- A a5 : 0.5- " x : I 0.20,l- BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 n Ln 0 rl " X > 7 PFBS r 6 I on 2.71 2.4; 2.11 0*3:L 1.81 n <Ln 1 + 5 { 0 r"xl 1.2- 0,6- 0,3: . , . . . , . . . , . . . , . . . I . ' 4.0 4.2 4+4 4.6 4.8 Min Page 101 of 171 Data File: D:\chem\Hillary.i\HO10426.b\HILLOO49.D 2 PFOS IiIon 1.0: 0.9: 0.8; 0.7; 0.6; 0.5; 0.4; 0.3; 0.2; 0.1; , . . . , I. ./ . , . .\ . , .I . . , . . . 4.4 4.8 5,2 5.6 6.0 BACK TO MAIN 3M Environmental Laboratory Project ## E01-0768 Page Page 102 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HOlO5Ol.b\HILLOO37.D Report Date: 03-May-200109:17 3M Environmental Laborato Project # E01-076Tage 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO37.D Lab Smp Id: PFOS-2.1.1 In] Date : 01-MAY-200123:11 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD "Hillary1I,H010501.s Met hod : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Olt.m Meth Date : 03-May-200108:17 terrell Quant Type: ISTD Cal Date : 01-MAY-200121:19 Cal File: HILL0027.D Als bottle: 26 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS ---- 427 299 499 RT EXP RT REL RT RESPONSE ---- -------- -------- -------- 5.115 5.109 (1.000) 1215865 4.407 4.402 (1.000) 1912250 5.248 5.312 (1.191) 7420988 CONCENTRATIONS ON-COLUMN FINAL (ng/mL) (ng/mL) -----_--- -_-_-___-_ 254.000 254.000 311.873 312 Page 103of 171 Data File: D:\chem\Hillary.i\HOlO5O~.b\HILLOO37.D 9.31 9.0; 8,7{ 8.41 8.1; 7,81 7.5; 7.2; 6.91 6.6; 6.31 6.0; 5.7; 5.4f , 5.14 4.8: 4.51 ' - 4.2{ 3.91 D:\chem\Hillary.i\HOlO5Ol.b\HILLOO37.D BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 H 1 THPFOS 1.6; 1.4: 1+2{ c. 1.0: L$, o. 0.81 "X 0.61 >- 0.4: 0.21 p+ 1 Lo 2 PFBS 2.4: 2,1: 1.8: v 0.9; 2.. 0.6- h1m 0 4 e 4.0 4.2 4,4 4.6 4,8 Min Page 104 of 171 Data File: D:\chern\Hillary,i\HOlO5Ol.b\HILLOO37~D 3 PFOS BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 105 of 171 BACK TO MAIN DReaptoartFiDlaet:eD::\0c3h-eMma\yH-i2l0l0a1r0y9.i:\1H7010501.b\HILL0045.D 3MEnvironmentalLaborato~age 1 Project## E01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO45.D Lab Smp Id: PFOS-2.2.1 In] Date : 02-MAY-200100:40 Operator : KLT Inst ID: hi1lary.i Smp Info : 1 , O O O X dil 042301-PFOS-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD lfHillarylHf0,10501.s Method : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Olt.m Meth Date : 03-May-2001 08:17 terrell Quant Type: ISTD Cal Date . : 01-MAY-200121:19 Cal File: HILL0027.D Als bottle: 30 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT S I G MASS __ _- -- 427 299 499 CONCBNTTRATIONS ON-COLUMN FINAL RT BXP RT RBL RT RBSPONSB (ng/mL) E=== ---___-_-_---_-__-____-__-------_____ ------- (ng/mL) ------- 5.122 5.109 (1.000) 1256724 254.000 4.408 4.402 (1.000) 1900616 254.000 5.262 5.312 (1.194) 7021826 294.754 295 Page 106 of 171 Data File: D:\chem\Hillary.i\HOlO5Ol.b\HILLOO45.D 8.74 8.41 8.1: 731 7,51 7.21 6,94 6.61 6.31 6.04 5.71 5.4; 5.14 , ; 4.81 4.51 4.21 ' 3.9< 3.61 3.31 3.01 2.71 2.41 2.11 1.81 1.51 1,2{ 0.9: 0.6: D:\chem\Hillary,i\HOlO5Ol.b\HILLOO45.D n BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 W 1 THPFOS I M 2 PFBS Min Ion r 9 :/i > 0.8; 0,6{ 0,4; 0,2; .,...,...,...,. . 4.0 4.2 4.4 4.6 4.8 Min Page 107 of 171 Data File: D:\chern\Hillary,i\HOlO5Ol~b\HILLOO45+~ 3 PFOS BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 ' Page 108 of 171 BACK TO MAIN Data File,:D:\chem\Hillary.i\H010501.b\HILL0050.D 3MEnvironmenta'Laborato age 1 Report Date: 03-May-200109:17 Project # E01-0767 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO5O.D Lab Smp Id: PFOS-2.3.1 Inj Date : 02-MAY-200101:36 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD "HillaryllH,010501.s Method : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Olt.m Meth Date : 03-May-200108:17 terrell Quant Type: ISTD Cal Date : 01-MAY-200121:19 Cal File: HILL0027.D Als bottle: 34 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT S I G MASS ---- 427 299 499 RT EXP RT REL RT RESPONSE _ _ _ _ __________-___----------------- -------- 5.116 5.109 (1.000) 1211070 4.409 4.402 (1.000) 1958520 5.257 5.312 (1.192) 8177762 CONCENTRATIONS ON-COLUMN FINAL (ng/mL) ------- (ng/mL) ---------_-___ 254.000 254.000 339.416 339 Page 109 of 171 Data File: D:\chem\Hillary,i\HOlO5Ol.b\HILLOO5O~D D:\chem\Hillary.i\HOlO5Ol+b\HILLOO5O.D 1.1: 1.0: 0.9: 0.8: 0.7: BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 0.3: vm ) LL v) L LL I 0.2: 0.1: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I. . . . . . .I . . I . . .I . .1 . . . . . . . .I . .I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . M 1 THPFOS 1.6: 1.4; 1.2; ph 1.0: 20 0.8; V --! Lo 2 PFBS I on 2.7: c. Lo " 0 rl > Page 110of 171 Data File: D:\chem\Hillary,i\HOlO5Ol.b~HILLOO5O.D 3 PFOS BACK TO MAIN 3M Environmental Laboratory Project # E014768 Page ' Page 111 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010430 .b\HILL0064.D 3MEnvironmentalLaboratoQage 1 Report Date: 01-May-200113:29 Project # E01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO43O.b\HILLOO64.D Lab Smp Id: PFOS-3.1.1 In] Date : 30-APR-2001 20:15 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-3 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD I1HillaryffH,010430.s Method : D:\chem\Hillary.i\HOlO43O.b\HOlOO43Ot.m Meth Date : 01-May-2001 12:59 terrell Quant Type: ISTD Cal Date . : 30-APR-2001 13:21 Cal File: HILL0027.D Als bottle: 26 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds * 1 THPFOS * 7 PFBS 2 PFOS QUANT S I G MASS 427 299 499 CONCENTRATIONS ON-COLUMN FINAL RT BXP RT REL RT RESPONSB (ng/mL) (ng/mL) 5.097 5.108 (1.000) 1310521 254.000 4.375 4.380 (1.000) 2034585 254.000 5.237 5.318 (1.197) 6815087 267.605 268 Page 112 of 171 BACK TO MAIN Data File: D:\chem\Hi 1 lary, i\H010430,b\HILL0064.D 3M Environmental Laboratory Project # E01-0768 Page 2 D:\chem\Hillary.i\HOlO43 .b\HILL0064.D 8.11 7.81 7.5; 7.2{ 6.91 6.61 6.31 6,OL 5.7; 5,4; 5.11 4.81 ~ 4.51 d<Lfl 4.21 0 " 3.91 >- 3.61 2 3.31 v) 3.01 a 1 2.7i 2.41 2.1: 1.81 1.51 1.2; 0,s; 0.61 O,3: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1. . . . . . . . . . . . . I....,.... 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 Min x 1 THPFOS 1.61 1.41 1.2; G; l,O{ < t ?QI Ln W 7 PFBS Page113of171 Data File: D:\chem\Hillary,i\HOlO43O.b\HILLOO64.D 2 PFOS 8.01 7.01 6.0; 5.0; Lo 6 4.0; FP-J N, Lo ~~~ - BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 114 of 171 BACK TO MAIN DReaptoartFiDlaet:eD::0\c1h-eMma\yH-i2l0l0a1r1y3.:i3\0H010430.b\HILL0072 .D 3MEnvironPrrnoejenctat#'LaEbOorlat-oodage 1 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO43O.b\HILLOO72.D Lab Smp Id: PFOS-3.2.1 Inj Date : 30-APR-200121:45 Operator : KLT Inst ID: hil1ary.i Smp Info : 1,OOOX dil 042301-PFOS-3 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylHl0,10430.s Met hod : D:\chem\Hillary.i\HOlO43O.b\HOlOO43Ot.m Meth Date : 01-May-200112:59 terrell Quant Type: ISTD Cal Date : 30-APR-200113:21 Cal File: HILL0027.D Als bottle: 30 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds * 1 THPFOS * 7 PFBS 2 PFOS Q U A " SIG MASS ____ 427 299 499 RT EXP RT REL RT RESPONSE _ ___ ______-_________________ ________ 5.104 5.108 (1.000) 1305210 4.383 4.380 (1.000) 2128725 5.245 5.318 (1.197) 8117378 CONCENTRATIONS ON-COLUMN FINAL __(_n__g_/__m__L_)__-_-(__n_g__/_m__L_)_ _ _ 254.000 254.000 309.586 310 -- Page 115 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HOlO43O+b\HILLOO72.D D:\chem\Hillary,i\HOlO43O+b\HILLOO72~D 1.1- 3M Environmental Laboratory Project # E01-0768 Page 1.0- 0.9: 0.8: 0.7: 0+4- P 0.3- 0,2- 0.1- H 1 THPFOS 7 PFBS 3.0: 2.71 2,4{ 2.1: 1.81 1.5: d bCQ d Page116of171 Data File: D:\chem\Hillary.i\HOlO430.b\HILLOO72.D 2 PFOS 1.0: 0.9: 0.8; 0.7: ph 0.6; $; 0.5; 2 0.4; d r ?Lo BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 Page Page 117 of 171 Data File: D:\chem\Hillary.i\H010430.b\HILL0077 .D 3M Report Date: 01-May-200113:30 BACK TO MAIN Laborato!?1age Project#EI01-0768 3M Environmental Laboratory PFOS, SD-018, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO43O.b\HILLOO77.D Lab Smp Id: PFOS-3.3.1 Inj Date : 30-APR-200122:41 Operator : KLT Inst ID: hi1lary.i Smp Info : 1,OOOX dil 042301-PFOS-3 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD "Hillary", H010430.s Method : D:\chem\Hillary.i\HOlO43O.b\HOlOO43Ot.m Meth Date.: 01-May-200112:59 terrell Quant Type: ISTD Cal Date : 30-APR-200113:21 Cal File: HILL0027.D A l s bottle: 34 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS ---- 427 299 499 RT BXP RT REL RT RESPONSB -------- -------- -------- --_------------ 5.104 5.108 (1.000) 1325618 4.397 4.380 (1.000) 2084860 5.237 5.318 (1.191) 7558687 CONCENTRATIONS ON-COLUMN FINAL (ng/mL) -------------- (ng/mL) _-_-_-___ __ __ _ 254.000 254.000 292.440 292 Page118of171 BACK TO MAIN Data File: D:\chem\Hillary.i\HO1043O.b\HILL0077.D D:\chem\Hillary,i\HO104~ b\HILL0077.D 1.0- 3M Environmental Laboratory Project # E01-0768 Page 0+9- 0.8- 0.7- 0.6- 0 > 0.5- 5. 014- 0.3- 1 M 1 THPFOS 2,o: 1.8; 1.61 1.41 1,2{ l*O{ 0+8; 1 - 0 Is) M 7 PFBS I on 2.71 2.41 2.1: 1.81 <Is) 1.51 0 d "x 1.2; > 0.9: 0.61 1 ' 4.0 4,2 4.4 4.6 4.8 Hin Page 119 of 171 Data File: D:\chem\Hillary,i\HO1043O+b\HILL0077.~ 2 PFOS 1.0- 0.9; 0.8: 0.7; A 0.6, 6u) 0,s; 4 "X 0,4; > 0.3; 0.2: 0.1, *m N. Ln 1 II I BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 120 of 171 BACK TO MAIN Data File : D:\chem\Hillary.i\H010426.b\HILL0119 .D 3M EnvironmentalLaboratmge 1 Report Date: 30-Apr-200114:07 Project# E01-0768 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOll9.D Lab Smp Id: TN-A 4802 MeOH Inj Date : 27-APR-2001 14:51 Operator : KLT Inst ID: hil1ary.i Smp Info : TN-A 4802 MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD IIHillaryll04-26-01 Met hod : D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14:05 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 91 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS _____ _-_ 427 299 _R_T_ _ BXP RT ________ RBL RT RBSPONSB _-___-_-_______-___ _ _ _ _ Compound Not Detected. Compound Not Detected. CONCBNTRATIONS ON-COLUMN FINAL (ng/mL) (ng/mL) ___-_-_-_-_-_- --_-__--_---_ Page 121 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HOlO426.b\HILLOll9.D 3M Environmental Laboratory Project # E01-0768 Page D:\chem\Hillary,i\H0lO426,b\HILLOll9.D 7.0: 6.81: 6.6; 6.4: 6.2; 6.0; 5.8; 5.6: 5-4; 5.2; 5.01 4.8; 4,6: 4,4; 4.2; ? 4,O; > 3,s: 5 3.64 - 3.4: 3.2: 3.0: 2,s: 2.6: 2.4: 2,2; 1 2.0; 1.8: 1+6: 1.41 1.2; 1.0.; . ............................. I . . . . , ........................................................... 0.5 1.0 1.5 2+0 2.5 3+0 3.5 4.0 4+5 5.0 5+5 6.0 6.5 7.0 7+5 8+0 8.5 9,O Hin M 1 THPFOS (Undetected) I I on 7201 680; 640; 600; 560: 520j 2. 4 8 0 j 440; 400 1 360; 320< ":+' , . .. ,..., .. ., . 4.4 4.8 5.2 5,6 6.0 Min Y 7 PFBS (Undetected) ,.... 9.5 Page 122 of 171 Data File: D:\chem\Hillary,i\HOlO426.b\HILLOll9.D 2 FOSA Y 5.5, Lo 5.0; 4.5, h 4.0; 2 y 3.5, 3.0; BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 123 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426 .b\HILL0148.D 3MEnvironPmroejenctta#'LEab01o-r0a7to6?age 1 Report Date: 30-Apr-200114:08 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl48.D Lab Smp Id: TN-A 4802 MeOH Inj Date : 27-APR-200120:15 Operator : KLT Inst ID: hil1ary.i Smp Info : TN-A 4802 MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryl0l4-26-01 Met hod D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14:08 terrell Quant Type: ISTD Cal Date : 27-APR-200114:17 Cal File: HILL0116.D Als bottle: 92 Dil Factor: 1.00000 Intesrator: Falcon Compound Sublist: all.sub Targgt Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Page 124 of 171 Data File: D:\chem\Hillary,i\HOlO426,b\HILLOl48.D 3+1; 3.0; 2.9; 2.8; 2.7: 2.6: 2.5: 2,41 2.3; 2.2: 2.1; ; 2.0; 1> 1.9; 1.8; 1.7; 1,6; 1.5; 1+4; 1.3; 1.2; 1.1; 1.0; 0.9; 0.8; 1 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 H 1THPFOS I 1.00; 0.90: e 0.80; 0,70; 0 $"0.60; 1. 0.50{ 0,401 (Undetected) I on 1 Y 7 PFBS Ion 1.8; 1.4: 1.2: l.O{ 0.81 0.61 0.41 1 / .................... 4.0 4.2 4.4 4.6 4.8 Page 125 of 171 Data File: D:\chern\Hillary,i\HOlO426~b\H~LLOl48,D 2 FOSA (Undetected) I on 1.81 1.6: 1.41 ;; 1.2: 2< 1.0: " X 0.81 > . 0+6{ 0.4: ,.,...,...,...,..., . 5.2 5.6 6.0 6,4 6.8 Min BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 Page 3 Page 126 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010501.b\HILL0092.D 3MEnvironmenta1Laborato?age 1 Report Date: 03-May-200113:Ol Project # E01-076 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO92.D Lab Smp Id: TN-A 4802 MeOH Inj Date : 02-MAY-200109:26 Operator : KLT Inst ID: hi1lary.i Smp Info : TN-A 4802 MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryHl0l10501.s Met hod D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Ola.m Meth Date : 03-May-200111:02 terrell Quant Type: ISTD Cal Date : 02-MAY-200108:53 Cal File: HILL0089.D Als bottle: 93 Dil Factor: 1.00000 Intesrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS _-_---- 427 299 Page 127 of 171 Data File: D:\chem\Hillary,i\HOlO501.b\HILL0092,D D:\chem\Hillary.i\HO1O5Ol+b\HILLOO92.D 1 6.4: 6.1; 7.81 7.51 7.21 6.91 6,6{ 6,31 6.01 5.71 5,4; 5.1: 4.81 W Inw BACK TO MAIN Page 2 ...n.. ,.. , . .. . .~ ~ .- , M 1 THPFOS (Undetected) I on 5701 5404 510 { 480 4501 420{ ~ 390: 360: 3301 300 { 2701 240- . . , . . . , . . . , . . . , . . . , . 4.4 4,8 5,2 5.6 6.0 Y 2 PFBS (Undetected) 750 ; 700; 650; 600; 550 500 450; 400; 350; Ion \ Page 12sj@of 1%71o, Data File: D:\chem\Hillary.i\HOlO5O~.b\HILLOO92.D 3 FOSA n m 0 d " > BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 129 of 171 BACK TO MAIN Data File : D:\chem\Hillary.i\H010426.b\HILL0121 .D 3M EnvironmentalLaborat-ge 1 Report Date: 30-Apr-2001 14:07 Project# E01-0768 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl2l.D Lab Smp Id: FOSA-1.4.lblk In] Date : 27-APR-200115:13 Operator : KLT Inst ID: hil1ary.i Smp Info : 1OX dil 042301-FOSA-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD "Hillary1I04-26-01 Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14:05 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 55 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS __-_-- 427 299 498 RT EXP RT REL RT RBSPONSB ____ __ _ _ _ _ _ _ ___-_-__________________ 5.108 5.122 (1.000) 1379938 4.379 4.387 (1.000) 1893699 5.892 5.977 (1.345) 106755 CONCENTRATIONS ON-COLUMN FINAL (ng/mL) (ng/mL) __-____--_-_-_--_-_-_- -_ 254.000 254.000 2.50445 2.50 Page 130 of 171 Data File: D:\chem\Hillary.i\HOlO426.b\HILLOl2l.D 34 3,2{ 3.01 2.81 D:\chem\Hillar : : : i\H010426.b\HILL0121.D LLL L L i I- BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2.4: 2,2{ 2.0- 1.2: 1.01 0.81 0.61 0.4: 0.21 W 1 THPFOS 3.0: Ion -!z! 2.71 2,4{ 2.1: .-. 1.8; L&o 1,5{ 534 1.2; 0,9{ 0.6: 'm 0.3; 4,4 4.8 Hin M 7 PFBS m. 3,3{ m 3.0: 2+71 2,4; 2.11 ? 1.81 0 g 1,5j " 1,21 > 0.9; Page 131 of 171 Data File: D:\ohen\Hillary.i\HOlO426,b\HILLOl2l.D 2 FOSA . 1.1: co, Lo 1.0, 0.9: 0.8; 5wCI 0.7: V 0.6: ,X 0,s: BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page c Page 132 of 171 BACK TO MAIN DRaeptoartFiDlaet: eD::3\c0h-eAmp\rH-i2l0l0a1r1y4. i:\0H8010426.b\HILL0144 .D 3M EnvironmentalLabora1toT!age Project # E01-0768 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data fiie : D:\chem\Hillary.i\HOlO426.b\HILLOl44.D Lab Smp Id: FOSA-2.4.lblk Inj Date : 27-APR-200119:30 Operator : KLT Inst ID: hi1lary.i Smp Info : 1OX dil 042301-FOSA-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryl0l4-26-01 Method D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date.: 30-Apr-200114:08 terrell Quant Type: ISTD Cal Date : 27-APR-20011 4 ~ 1 7 Cal File: HILLO116.D Als bottle: 71 Dil Factor: 1.00000 Inteqrator: Falcon Com-pound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS --_- 427 299 498 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT REL RT RESPONSE (ng/mL) _ _ _ _ ______-_--___---_-----_-_--_-----_---- ------- (ng/mL) ------- 5.111 5.122 (1.000) 1308838 254.000 4.383 4.387 (1.000) 1875879 254.000 Compound Not Detected. Page 133 of 171 BACK TO MAIN Data File: D:\chem\Hillary,i\HOlO426.b\HILLOl44.D 4+0: D:\chem\Hillar .i\H010426,b\HILL0144,D 3M Environmental Laboratory Project # E01 -0768 Page . . I. .f. .iI. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0.5 1.0 1.5 2.0 2,5 3,O 3.5 4.0 4.5 5. 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 Nin H 1 THPFOS Ion 2.71 2.41 2*1{ EcI 1,s: 1.5{ v 4x 1*2{ > 0.91 0,6{ 0.3: . . . . . . .A . . . . . . . . .1 . . . . 4+4 4.8 5.2 5.6 6+0 Min W 7 PFBS 3,6; 3.2: 2.84 2,4{ 2,o: 1,6{ 1.2: cm M, d Page 134 of 171 Data File: D:\chem\Hillary.i\HO10426,b\HILLOl44.D 2 FOSA (Undetected) Ion 5,2 5,6 6,O 6,4 6.8 Hin BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 135 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010501.b\HILL0093.D 3MEnvironmentalLaboratorjBage 1 Report Date: 03-May-2001 13:Ol Project # E01-0768 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOO93.D Lab Smp Id: FOSA-3.4.lblk In] Date : 02-MAY-200109:38 Operator : KLT Inst ID: hil1ary.i Smp Info : FOSA Urine Solub Day 3 Re-diluted samples 5-01-01 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD lrHillaryHl0l10501.s Method D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Ola.m Meth Date : 03-May-2001 11:02 terrell Quant Type: ISTD Cal Date : 02-MAY-200108:53 Cal File: HILL0089.D Als bottle: 71 Dil Factor: 1.00000 Intesrator: Falcon Compound Sublist: all.sub TargGt Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG ' MASS ==== 427 299 498 RT BXP RT RBL RT RESPONSE _ _ _ _ ____________-_-_____ ________ 5.117 5.118 (1.000) 1236810 4.396 4.410 (1.000) 1629186 6.007 5.993 (1.366) 553440 CONCENTRATIONS ON-COLUMN FINAL (ng/rnL) (ng/mL) _-----_________ _ _ _ _ _ _ 254.000 254.000 19.8742 19.9 Page 136 of 171 Data File: D:\chem\Hillary,i\HOl05Ol.b\HILLOO93+D BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 3.41 3.21 3.0: 2.81 2.61 2,41 2.22,or $ 1.81 5 1.61 c 1.41 1.2- 1.0: 0.6: 0.4: 0.2: ....................................... . . . . . .L. . I. . I. . . .c1. . . . .I./ . . . .I. .I .I. . . . . . . . . . . . . . . . . . , . . . . , . . . . ,h . . . ., M 1 THPFOS H 2 PFBS Page 137 of 171 ~ ___~~ Data File: D:\chern\Hillary,i\HOlO5Ol~b\HILLOO93,D 3 FOSA - BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 138 of 171 BACK TO MAIN Data File : D:\chem\Hillary.i\H010426.b\HILL0126 .D 3M EnviPrroojnecmt#eEn01t-a07l6L8abora1toPage Report Date: 30-Apr-200114:07 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl26.D Lab Smp Id: FOSA-1.1.1 In] Date : 27-APR-200116:09 Operator : KLT Inst ID: hil1ary.i Smp Info : 1OX dil 042301-FOSA-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryl0l4-26-01 Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-200114:05 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 59 D i l Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Q U M SIG MASS _-_---- 427 299 498 CONCBNTRATIONS ON-COLUMN FINAL RT BXP RT REL RT RBSPONSB (ng/mL) (ng/mL) _ _ _ _ _---________--______-__-----______---- -__--_-_---- ------- 5.112 5.122 (1.000) 1223896 254.000 4.390 4.387 (1.000) 1863247 254.000 5.889 5.977 (1.341) 9090670 342.874 343 Page 139 of 171 Data File: D:\chern\Hillary.i\HO10426,b\HILLO126.D D:\chem\Hillary,i\HO10426,b\HI 1.1- 1.0: 0.9- 0,8- 0,7: , 0.6! ' 0.5- BACK TO MAIN 126. D 3M Environmental Laboratory Project # E01-0768 Page nin n 1 THPFOS 2.7: 2.41 2.1; 1,8{ r. Ln 1.5; b 4 X 12: Y > 0,9{ L4 -l Lo ,'."I".. Page 140 of 171 Data File: D:\chem\Hillary,i\HOlO426,b\HILLOl26+D 2 FOSA 1.1; LO4 0.9; 0.8; 7 0,7; 0.6; 0 $ 0.5; " 0,4: > 0.3; 0,2: 0.1: 0 (r\ r ?LD 1 I1 I BACK TO MAIN 3M Environmental Laboratoiy Project # E01-0768 Page Page 141 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426.b\HILL0130.D 3MEnvironPmroejnectat'#LaEbOolra-toO76Fage Report Date: 30-Apr-200114:07 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl3O.D Lab Smp Id: FOSA-1.2.1 Inj Date : 27-APR-2001 16:54 Operator : KLT Inst ID: hil1ary.i Smp Info : 1OX dil 042301-FOSA-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD "Hillary" 04-26-01 Method D:\chem\Hillarv.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14A:05 'terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 63 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS ___-_-- 427 299 498 RT EXP RT RBL RT RBSPONSB __--__-_-_-_-___________-_-_--__-_-_ --_-------_----- 5.118 5.122 (1.000) 1223960 4.382 4.387 (1.000) 1887871 5.888 5.977 (1.344) 8385467 CONCBNTRATIONS ON-COLUMN (ng/mL) ------- FINAL (ng/mL) ------- 254.000 254.000 309.497 309 Page 142of 171 Data File: D:\chem\Hillary,i\HO10426,b\HILLOI3O~D D:\chem\Hillary,i\HO10426.b\HILL013O.D 1.00.90.80.7- BACK TO MAIN 3M EnvironmentalLaboratory Project # E01-0768 Page 2 0,41 x 1 THPFOS 2.7: 2.4; 2.11 n d " X > t - ld Lo ft 7 PFBS 4.0: 3.6: 3.2; 2.8{ n 2.4: &Lo 2.0; 5d 1.6; >- 1.2; ta, 1 d . . . . .I .1. . . . . . . . . . . . . . . . . . . . . . . ,O 6.5 7.0 7.5 8.0 8.5 'I"'.,"" 9.0 9,5 Page 143 of 171 Data File: D:\chem\Hillary,i\HOlO426,b\HILLOl3O.D 2 FOSA Lo; 0.9: 0.8; - 0.7; 9 0.6; 5 0.5; !! 0,4; * 0.3; 0.2; 0.1; Lrn ro, Lo BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 144 of 171 BACK TO MAIN Data File: D :\chem\Hillary.i\H010426.b\HILL0138 .D 3M EnvironmentalLaboratoqage 1 Report Date: 30-Apr-200114:08 Project # E01-0768 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl38.D Lab Smp Id: FOSA-1.3.1 Inj Date : 27-APR-200118:23 Operator : KLT Inst ID: hil1ary.i Smp Info : 1OX dil 042301-FOSA-1 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryl0l4-26-01 Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14:05 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 6 7 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS ____ 427 299 498 CONCBNTRATIONS ON-COLUMN FINAL _R_T_ _ EXP RT REL RT RESPONSE ______-______ ____-______ ___-___----_--_------- (ng/rnL) ------- 5.111 5.122 (1.000) 1177514 254.000 (ng/mL) ------- 4.390 4.387 (1.000) 1900215 254.000 5.881 5.977 (1.340) 6883587 248.307 248 Page 145 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HOlO426,b\HILLOl38+D 3M Environmental Laboratory Project # E01-0768 Page 2 8.11 7.81 7.51 7.21 6.91 6.6; 6.3: 6.01 5.71 5.41 5.1: 4.81 , 4.51 ' 4.21 ; 3.91 1 3.6; 3.3: 3.01 2.71 2.4: 2.1; 1.8: 1.51 1.21 0.91 0.61 0.31 Ih I D:\chem\Hillary,i\HOlO426,b\HILLOl38.D 1 u) 0 InL. 1 I[I I M 1 THPFOS I on :::; I)\ - . .... ... .. .. .. 4;4 4;8 5;2 5.6 6.0 Hin M 7 PFBS Page 146 of 171 Data File: D:\chem\Hillary.i\HOiO426.b\HILLOl38.D 2 FOSA I on 8.0{ 7.0; I II 3,Oi II I . , . . . , ! . !.- 5.2 5,6 6.0 Min 6:4 . 6.8 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page Page 147 of 171 Data File : D:\chem\Hillary.i\H010426.b\HILL0152 .D 3M Report Date: 30-Apr-2001 14:08 BACK TO MAIN Project # E01-07gage 1 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl52.D Lab Smp Id: FOSA-2.1.1 In] Date : 27-APR-2001 21:OO Operator : KLT Inst ID: hi1lary.i Smp Info : 1OX dil 042301-FOSA-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD ttHillaryl0t4-26-01 Method D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14:08 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 7 5 Dil Factor: 1.00000 Inteqrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS __ ___ _-- 427 299 498 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT REL RT RESPONSE (ng/mL) (ng/mL) _ _ _ _ _ _ _ _ _ _ _ _ ________ _____________ ____-_-___ ___ ____------- 5.115 5.122 (1.000) 1110585 254.000 4.387 4.387 (1.000) 1783250 254.000 5.879 5.977 (1.340) 8103858 317.291 317 Page 148 of 171 Data File: D:\chem\Hillary,i\HOlO426,b\HILLO152,D D:\chem\Hillary.i\HOlO426,b\HILLOl52,D 1.0- 0.9- 0.8- 0.7- 0.6- A 0.5vai ) BACK TO MAIN 3M Environmental Laboratory Project # E01 -0768 Page H 1 THPFOS 2.42;; 2.0; 1.8< 1.6; c; i.4; 6 1.2r T"xI i+Qi 0.8; 0.6; 0.4: 0.22 M 7 PFBS t--!r( LD I. I Ion L-k . \. . .I.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .O 6.5 7.0 7.5 8.0 8.5 9.0 9.5 Page 149 of 171 Data File: D:\chem\Hillary,i\HOlO426,b\HILLOl52+D 2 FOSA . 0.9; 0.8; 0.7; 5 0.6; a 0.5; "X 0,4+ > 0,3; 0.2; 0.1; reoLo, \ I1 I- BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 150 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426.b\HILL0157.D 3MEnvironrnenta'Laboratogage 1 Report Date: 30-Apr-200114:08 Project# E01-07 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl57.D Lab Smp Id: FOSA-2.2.1 In] Date : 27-APR-2001 2 1 ~ 5 6 Operator : KLT Inst ID: hil1ary.i Smp Info : 1OX dil 042301-FOSA-2 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD ffHillaryf0l4-26-01 Method : D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-200114:08 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 79 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS _____ __- 427 299 498 RT ____ __B__X_P___R_-T___ ___R___E_L____R__T_ _-_R_B__S__P_O_N_S_B_ 5.116 5.122 (1.000) 1202436 4.387 4.387 (1.000) 1863768 5.886 5.977 (1.342) 7993752 CONCENTRATIONS ON-COLUMN FINAL (ng/mL) _______ (ng/mL) _______ 254.000 254.000 297.956 298 Page 151 of 171 Data File: D:\chem\Hillary.i\HOlO426.b\HILLOl57.D 9+6: 9.3: 9.0: 8.7f 8,4( 8.1{ 7.8{ 7*5: 7.21 6.9; 6.61 6.3; 6.0: 5.7: BACK TO MAIN 3M Environmental Laboratory Project ## E01-0768 Page 2 n 1 THPFOS D -.. II JI H 7 PFBS *1 Ion co 4,O: Q) 3.61 3.2; 2.8; c; 2.4; b 2.0; 5d 1.6; o*8Ll!L- 1.2: 0.4: 4.00 4.20 4,40 4.60 4.80 Page 152 of 171 Data File: D:\ohem\Hillary.i\HOlO426.bWILLOl57,D 2 FOSA 9.0; m, rn 8,O; 7.0; h 6.0; m& 5.0; rl r 3.0; 2,o; 1+0: BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 153 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426.b\HILL0165 .D 3M EnviroPnmroejenctta#'LaEboorlat-oodage 1 Report Date: 30-Apr-200114:08 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOl65.D Lab Smp Id: FOSA-2.3.1 In] Date : 27-APR-2001 23:25 Operator : KLT Inst ID: hil1ary.i Smr, Info : 1OX dil 042301-FOSA-2 Mist Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryl0l4-26-01 Method . : D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-2001 14:08 terrell Quant Type: ISTD Cal Date : 27-APR-2001 14:17 Cal File: HILL0116.D Als bottle: 83 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable QUANT SIG MASS ---- 427 299 498 RT EXP RT RBL RT RESPONSE ---__- -_______---------_-------- -------- 5.116 5.122 (1.000) 1158902 4.394 4.387 (1.000) 1883593 5.879 5.977 (1.338) 7850050 CONCENTRATIONS ON-COLUMN (ng/mL) -------------- FINAL (ng/mL) ______- 254.000 254.000 288.823 289 Page 154 of 171 BACK TO MAIN Data File: D:\chem\Hillary,i\HOlO426.b\HILLOl65.D 3M Environmental Laboratory Project # E01-0768 Page 2 9,6: 9,3: 9.0: 8,7: 8.4: 8.1: 7.81 7,5{ 7.2: 6.9: 6.6: 6.3: 6.0; 5.7: 5.4: ; 5.1: > 4.8: -$ 4.5: 4.2{ 3.9; 3.61 3.31 3,01 2,7{ 2,41 2.11 1.81 1.51 1.2{ 0.91 0.61 0,31 - . 0.5 n 1 THPFOS D:\chem\Hillary,i\HOlO426.b\HILLOl65.D r U B II 1 . . 1 . . . . . , . . . . , . . . . 6..0, . . . .6.5, . . . .7.,0. . . - 4:0 1,0 115 ' 2.0 2.5 3.0 3.5 4.5 5.0 5:5 Min 7:5 ' ' . . . . . , . . .- 8:5. 9.0 9:5 ' M 7 PFBS P. Is) 0 d " >- Page 155 of 171 Data File: D:\chem\Hillary,i\HOlO426,b\HILLOl65.D 2 FOSA ~ ~~ ~~ BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 156 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HOlO5Ol.b\HILLOO97.D Report Date: 03-May-200113:Ol 3M Environmental Laborato Project # E01-07aage 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file,: D:\chem\Hillary.i\HOlO5Ol.b\HILLOO97.D Lab Smp Id: FOSA-3.1.1 Inj Date : 02-MAY-2001 10:22 Operator : KLT Inst ID: hil1ary.i Smp Info : FOSA Urine Solub Day 3 Re-diluted samples 5-01-01 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillarylH0l10501.s Met hod D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Ola.m Meth Date : 03-May-200111:02 terrell Quant Type: ISTD Cal Date : 02-MAY-2001 08:53 . Cal File: HILL0089 .D Als bottle: 7 5 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds __________________________ * 1 THPFOS * 2 PFBS 3 FOSA QUANT SIG MASS ____ 427 299 498 RT EXP RT REL RT RESPONSE ____ ________ ________ ________ 5.117 5.118 (1.000) 1186398 4.403 4.410 (1.000) 1735419 5.909 5.993 (1.342) 7201703 CONCENTRATIONS ON-COLUMN FINAL (ng/mL) (ng/mL) _______ _______ 254.000 254.000 268.781 269 Page 157 of 171 Data File: D:\chem\Hillary,i\HOlO5Ol.b\HILLOO97.D 8,7{ 8.4: 8.11 7.8: 73: 7,2{ 6.9: 6.61 6.3: 6,Of 5.7: 5.41 5.1: , 4.8{ 5 4.51 , 4.2; D:\chem\Hillary,i\HOlO5Ol.b\HILLOO97.D BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 I Min H 1 THPFOS 2.41 1 Lo 2*11 1.81 5C; l,5{ 1.2: " X > 0.9: 0.6: 0.31 H 2 PFBS Page 158 of 171 Data File: D:\chem\Hillary,i\HOlO5Ol.b\HILLOO97.D 3 FOSA BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 159 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010501.b\HILLO101.D 3MEnvironPrrnoejenctat#'LEaboolra-tooi2age1 Report Date: 03-May-200113:02 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file,: D:\chem\Hillary.i\HOlO5Ol.b\HILLOlOl.D Lab Smp Id: FOSA-3.2.1 Inj Date : 02-MAY-2001 11:07 Operator : KLT Inst ID: hil1ary.i Smp Info : FOSA Urine Solub Day 3 Re-diluted samples 5-01-01 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD l1Hi11aryl1H010501.s Method : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Ola.m Meth Date : 03-May-2001 11:02 terrell Quant Type: ISTD Cal Date : 02-MAY-200108:53 Cal File: HILL0089.D A l s bottle: 78 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds QUANT SIG MASS CONCENTRATIONS ON-COLUMN FINAL RT EXP RT REL RT RESPONSE (ng/mL) (ng/mL) Page 160of 171 Data File: D:\chem\Hillary,i\H010501,b~HILL0101.D BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 8.4: 8.1: 7.81 7.51 7.21 6.9: 6.61 6.31 6.01 5.71 541 5.11 4,81 -? 4.5; 4.21 ?j 3.91 r 3.61 3.31 3.01 2.7: 2.41 2.11 1.81 1.51 1.21 0.9: 0,61 0.31 . . 1. 1.,. .I. . . . . . . . . . . . . . . . . . B m 0 LL k 1 i \ \ ) . . . . , . . . . , . .I . .1, . .I . . , . I . . . I . . . . ,.. . . . . . .I .1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . (.... H 1 THPFOS I I on ft 2 PFBS 0 * 4 j A 4.0 4.2 4.4 4.6 4.8 Page 161 of 171 Data File: D:\chem\Hillary,i\HO10501,b\HILLOlOl+D 3 FOSA BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 162 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\HO10501.b\HILL0108.D 3MEnvironmenta'Laborato Project # Eol-oi2age 1 Report Date: 03-May-2001 13:02 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO5Ol.b\HILLOlO8.D Lab Smp Id: FOSA-3.3.1 Inj Date : 02-MAY-200112:25 Operator : KLT Inst ID: hil1ary.i Smp Info : FOSA Urine Solub Day 3 Re-diluted samples 5-01-01 Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillary"H010501.s Method : D:\chem\Hillary.i\HOlO5Ol.b\HOlO5Ola.m Meth Date : 03-May-2001 11:02 terrell Quant Type: ISTD Cal Date : 02-MAY-200108:53 Cal File: HILL0089.D Als bottle: 82 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds QUA" SIG MASS CONCENTRATIONS ON-COLUMN FINAL RT EXP RT REL RT RESPONSE (ng/mL) (ng/mL) Page 163 of 171 Data File: D:\chem\Hillary,i\HOlO5Ol+b\HILLOlO8+D 8.41 8.1: 7.81 7.51 7.21 6.91 6.61 6.31 6.0; 5.71 5.41 5+11 4.8; ; 4.51 5 4.21 5 3,91 c 3.61 3.3; 3.0: 2.71 2.4; 2.11 1.8: 1.5: 1.2: (D LFL4 & (0 P LaL 1 I1 BACK TO MAIN .08,D 3M Environmental Laboratory Project # E01-0768 Page 2 t 2.41 I-cu 2.2; ? Lo 2.01 1.8: 1.6; 1.44 3 1.2; "x L O { ~ 0.8; 0,6{ 0,4: 0.2; - 11 1 W 2 PFBS 4+0: 3.61 3.2; 2.84 A 2*4; bIs) 2,0{ d5 1,6{ * 1.21 0.8i 0.4; eo 4 d Page 164 of 171 Data File: D:\chem\Hillary.i\HO10501,b\HILLOlO8.D 3 FOSA 8.01 7.0: 6.0{ A 5,Oi '50 : 4,Oj 3.04 >- 2.0; l,Oj Q: Lo 1 I1 I ~ - BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 165 of 171 BACK TO MAIN Data File: D:\chem\Hillary.i\H010426.b\HILL0114.D 3MEnvironmenta'Laborato age 1 Report Date: 30-Apr-2001 14:07 Project # E o A - o A 3M Environmental Laboratory FOSA, SE-027, Solubility in Urine Data file : D:\chem\Hillary.i\HOlO426.b\HILLOll4.D Lab Smp Id: 01003-25-10 Inj Date : 27-APR-2001 13:55 Operator : KLT Inst ID: hi1lary.i Smp Info : 249.6 ng/mL FOSA in MeOH Misc Info : Comment : Samples analyzed on HPLCllOO LC/MSD llHillaryl0l4-26-01 Method D:\chem\Hillary.i\HOlO426.b\HOlOO426a.m Meth Date : 30-Apr-200114:05 terrell Quant Type: ISTD Cal Date : 27-APR-2001 13:55 Cal File: HILL0114.D Als bottle: 10 'Calibration Sample, Level: 8 Dil Factor: 1.00000 Inteqrator: Falcon Compound Sublist: all.sub Target Version: 4.10 Processing Host: W19401 Concentration Formula: Amt * DF * CpndVariable Cpnd Variable Local Compound Variable Compounds * 1 THPPOS * 7 PFBS 2 POSA QUANT S I G MASS ==== 427 299 498 RT EXP RT REL RT RESPONSE ==== ======== =x====== === ==== 5.114 5.122 (1.000) 1459549 4.393 4.387 (1.000) 1999512 5.976 5.977 (1.360) 7439185 AMOUNTS CAL-AMT ON-COL (ng/mL) ======= (ng/mL) =------ 254.000 254.000 249.600 256 Page 166 of 171 Data File: D:\chern\Hillary,i\HOl0426+b\HILLO114.D D:\chern\Hillary.i\HOlO426.b\HILLOl14.D 1.1: 1.0: 0.9: 0.8: 0+7- ; 0.6: > !: 0.5: 0.4: BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 2 M 1 THPFOS 1.8; 1.61 1,4{ 1.21 P 6 . 1.0; 0 d ' : rl ui M 7 PFBS 24 1.81 " . , " " . . . . . . . . . . . . . . . . . . . . . . . . . . . . I. .I 6,5 7.0 7.5 8.0 8.5 9.0 9,5 Page 167 of 171 Data File: D:\chem\Hillary,i\HOlO426.b\HILLOll4.D 2 FOSA 1.1; 1.0; 0.9; 0.8; I on c 5.2 5.6 6.0 6.4 6.8 Min BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Page 3 Page 168 of 171 BACK TO MAIN 3M Environmental Laboratory Project# E01-0768 ATTACHMENDT: SAMPLECALCULATIONS Page 169 of 171 BACK TO MAIN 3M EnvironmentalLaboratory Project# E01-0768 Purity and dilution correction factors. 0 PFOS, SD-018 10 pL sample was diluted with 990 pL methanolfor a 1 : l O O dilution, followed by a second dilution to 1:1,000. 100 pL of 1:lOO diluted sample was diluted with 9OOpL methanol. Therefore the on-column concentrationwas multiplied by a factor of 1,000 for the final reported value. Then, the samples were corrected for the purity of the standards used to analyze the samples. The standard purity was 97.9%. The solubility reported is the amount of PFOS in standard material id# SD-018 that dissolves in water at 23-24 "C. 0 FOSA, SE-027: 10 pL sample was diluted with 990 pL methanol for a 1:lOO dilution. Therefore the on-column concentration was multiplied by a factor of 100 for the final reported value. The standard purity was unknown, therefore no purity correction was done. The solubility reported is the amount of FOSA in standard material id# SE-027 that dissolves in water at 23-24 "C. Mean and standard deviation were calculated using functions provided in Microsoft Excel@software. Standard deviation was used to measure the scatter about the mean of a data set; thus, it can be used to estimate the precision of a method. Relative standard deviation presents a measure of the magnitude of the standard deviation and is calculated by dividing the standard deviation by the mean. Means are calculated by adding individual entities and dividing the resultant sum by the number of individual entities. Standard deviation was calculated using the following equation: -Where n is the number of observations, and x is sample concentration. AveraaelStandard Deviation /%RSD examde calculations: Sample ID Repl Rep 2 Rep 3 Rep 3 Concentration vg/mL 302 325 287 146.7 Calculations Average (302+325+287)/3=305 Std Dev (see above equation)= 19 %RSD (19/305)= 6% %RSD (19.0/143)= 13.3% Page 170 of 171 BACK TO MAIN 3M Environmental Laboratory Project # E01-0768 Q-Test Example (Shaded number represents the suspected outlier): Q-Test example conclusion: Since Qobserve0d.,923, was greater than Qtabulated, 0.886, rep 1 may be excluded. Page 171 of 171