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BACK TO MAIN Study Title - Characterization Study of PFOS,Primary Standard Test Control Reference #TCR-00017-046 PHASE: SOLUBILITY OF PFOS IN OCTANOL Data Requirement 40 CFR 160.105(b) Author Mark E. Ellefson Phase Completion Date Date of signing Performing Laboratory 3M EnvironmentalLaboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Erivironmental Laboratory Study # FACT-TCR002 (LIMS #E00-1716) Centre Analytical Laboratories Study # 023-021 Total Number of Pages 109 BACK TO MAIN GLP COMPLIANCSETATEMENT Study Title: Characterization Study of PFOS, Primary Standard - Test Control Reference # TCR-00017-046, PHASE: Solubility of PFOS in Octanol. Study ldentificaition Number: FACT-TCR002, Centre Analytical Laboratories Study # 023-021 This phase of the study was conducted in compliance with Environmental Protection Agency (EPA) Good Laboratory Practice (GLP) Standards 40 CFR 160 with the exceptions in the bulleted list below. Exceptions to GLP compliance: 0 The electronic data systems in use have not been validated and there is not an electronic audit trail of corrections currently available (40 CFR 160.130 (e)).. Authenticated hardcopies of chromatograms and associated documents will be considered as the original raw data. William K. Reagen, Testing Facility Management 05?/JCA/ Date FACT -TCROO2 (LIMS #E00-1716), Page 2 of 109 BACK TO MAIN QUALITYASSURANCSETATEMENT Study Title: Characterization Study of PFOS, Primary - Standard Test Control Reference # TCR-00017,-046,PHASE: Solubility of PFOS in Octanol. Study Identification Number: FACT-TCR002, Centre Analytical Laboratories Study # 023-021 This phase of the study has been inspected by the 3M EnvironmentalLaboratory Quality Assurance Unit (QAU) as indicated in the following table. The findings were reported to the study director and lahoratory management. I 04/13/01 I DraR Phase Report QAU Reprebentative I 04/18/01 I 04/18/01 I L//23/ o/ Date ' FACT -TCROO2 (LIMS #E00-1716), Page 3 of 109 BACK TO MAIN TABLEOF (:ONTENTS GLP Complianice Statement............................................................................................................... 2 Quality Assurance Statement............................................................................................................. 3 Study Information ............................................................................................................................... 6 Summary ............................................................................................................................................ 7 Purpose .............................................................................................................................................. 7 Test Substance .................................................................................................................................. 7 Test System ....................................................................................................................................... 8 Method Summaries ............................................................................................................................ 8 Preparatory Methods.......................................................................................................................... 9 Sample Collection and Analysis ......................................................................................................... 11 Analytical Method............................................................................................................................... 11 Analytical Results........................................................................................ I...................................... 12 Data Summary ................................................................................................................................... 13 Statistical Methods ............................................................................................................................. 15 Statement of Conclusion .................................................................................................................... 15 References ......................................................................................................................................... 15 List of Attachments ............................................................................................................................. 16 Signature Page................................................................................................................................... 17 Attachment A: Extraction and Analytical Methods ............................................................................. 18 Attachment 8:Data Summary Tables ................................................................................................ 48 Attachment C: Sample Chromatograms ............................................................................................ 67 Attachment 0:Deviations from the Protocol ...................................................................................... ?03 Attachment E: Sample Calculations ................................................................................................... 108 FACT -TCROO2 (LIMS #E00-1716). Page 4 of 109 BACK TO MAIN LISTOF TABLES Table 1. Summary Table of Solubility of PFOS TCR-00017-046 in Octanol..................................... 7 Table 2. Characterizationof the Test Substance and Analytical Reference Substances ................. 7 Table 3. Description of the test system used in this study ................................................................ 8 Table 4. Solubility Screen Test Sample Preparation ........................................................................ 10 Table 5. Shake Flask Method Preparation for Study (FACT-TCR002)............................................. 10 Table 6. Sample Collection and Preparationfor Study FACT-TCR002............................................ 11 Table 7. Calibration Standard Reprocessing Summary ..................................................................... 13 Table 8. PFOS; TCR-00017-046 Solubility in Octanol Data Summary .............................................. 14 Table 9. Summary Table of Solubility of PFOS TCR-00017-046 in Octanol..................................... 15 FACT -TCROO2 (LIMS #E00.1716). Page 5 of 109 BACK TO MAIN ~~ ~~~ ~ STUDYINFCIRMATION Study Director John Flaherty Centre Ana1ytic:al Laboratory 3048 Research1Drive State College, PA 16801 Analytical Chemistry Laboratories Solubility Testing Certificate of Analysis 3M EnvironmentalTechnology and Safety Services (ET&SS) 3M Environmental Laboratory Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Centre Analytical Laboratory 3048 Research Drive State College, PA 16801 John Flaherty, Study Director Dr. William K. Heagen, Laboratory Manager Mark E. Ellefson, Principal Analytical lnvestigator Kristin L. Terrell, Analytical Chemist Mark L. Anderson, Analytical Chemist Cindy M. Carlson, Analytical Chemist Sponsor 3M EnvironmentalLaboratory 935 Bush Avenue Building 2-3E-09 St. Paul, MN 55106 Dr. William K. Reagen Experimental Dates Experimental Start Date: 29 September 2000 Experimental Termination Date: 30 March 2001 Archives Reserve samples of the reference standards will be maintained by the 3M Environmental Laboratory. all^ study data for this phase and a copy of the phase report are archived by the 3M Environmental Laboratory. FACT -TCROOP (LIMS #E00-1716), Page 6 of 109 BACK TO MAIN SUMMARY The approximate solubility of PFOS in octanol was determined by performing a solubility screen test, followed b y a quantitative solubility determination via Shake Flask Method. Results are presented in table 1: Table 1. Summary Table of Solubility of PFOS TCR-00017-046 in Octanol. 56.9 w/mL 0.9 Day .2 55.7 pg/mL 1.4 Day 3 55.4 pg/mL 2.5 Average 'Value 56.0 pglmL 0.8 1.6% 2.5% 4.5% 1.4 PURPOSE The purpose of:this phase of the study was to determine the solubility of PFOS in octanol as part of the characterization of this test, control, and reference substance. TESTSUBSTANCE PFOS, Test Control Reference #TCR-00017-046 Table 2. Chaiacterizationof the Test Substance II Expiration Date Re-uystallizedby George Moore,3M Specialty Materials, I Blda. 236-1R10 I I 01-01-2010 I Storage Conditions I Frozen I I 3M Laboratory Identification Number I TCR-00017-046 I white Powder 97.9% FACT -TCROO2 (LIMS #E00-1716), Page 7 of 109 REFERENCSEUBSTANCE Table 3. Characterization of the Analytical Reference Substances BACK TO MAIN * The purity of THPFOS and PFBS (currently being determined by Centre Analytical Laboratory) will not affect any analytical findings since it is used as a reference substance only. TESTSYSTEM Table 4. Descriptionof the test system used in this study I Source m1- Date Number I 2005 Flammable Cabinet TN-A 3353 99+% II II 11-01-05 Flammable Cabinet TN-A 4703 99.9+% HPLC Grade METHODSUMMARIES The solubility determination of PFOS TCR-00017-046 in octanol was performed according to 3M Environmental Laboratory methods ETS-8-170.0 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents;" and ETS-8-172.0 "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. Methods ETS-8-170.0 and ETS-8-172.0 were subsequently revised to include additional information/clarification. The new FACT -TCR002 (LIMS #E00-1716), Page 8 of 109 BACK TO MAIN revisions, ETS-8-170.1 and ETS-8-172.1, are procedurally the same as the predecessors and shall be included in Attachment A of this report. All analyses were performed according to the 3M EnvironmentalLaboratory method ETS-8-155.0 "Analysis of Perfluorooctanesulfonateor Other Fluorochemicallsin Waste Stream or Water Extracts Using HPLC-ElectrospraylMass Spectrometiy". The Solubility Screen Test Method (ETS-8-170.0) was used to determine the solubility range of PFOS TCR-00017-046 in octanol. Incremental amounts of octanol were added to PFOS neat material and a (qualitativedeterminationof the solubility point was made. The screen test indicated a solubility of PFOS in octanol of less than 10 pglmL at 23C. It is recommendedthat the solubility of substances having a solubility of less than 10 pg/mL be determined via the Column Elution Method. However, the Shake Flask Method was used for the quantitative determination of PFOS solubility because of the difficulty in finding compatible tubing (preliminary results demons'tratedthat the tubing swelled and cracked when in contact with octanol for extended periolds of time) and the possible explosion hazardof containing a non-sealedcontainer of a flammable solvent in an incubator equipped with electrical outlets on the inside. Detailed descriptions of the Solubility Screen Test Method and Shake Flask methods used in this study are located in Attac:hment A. PREPARATOMREYTHODS ETS-8-170.0 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents." This method is a prerequisiteto the shake flask and column elution methods and gives an estimate of the solubility point of the test substance in the solvent of choice. Approximately ten milligrams of PFOS was weighed into a glass vial, and varying amounts of octanol were added in a step-wise fashion. After each octanol addition, the vial was shaken, sonicated, and observed for particulate. When particulatewas observed in a final volume of 2 mL of octanol (approximately 5000 pg/mL), the final step of the screen test was initiated. Fourteen point one mg PFOS was placed into a 100 mL volumetric flask and octanol was added in a step-wise fashion up to 100 mL (with shaking, vortex-mixing, sonicating, and observations for particulatein between). Particulatewas still observed at 4 4 0 pg/mL, therefore 2.48 mg was weighed into a 250 mL flask and filled to the mark with octanol for a final concentration of approximately 10 pg/mL. The particulate did not dissolve after more than 24 hours of contact at 23 "C.The screen test estimated the solubility of PFOS in octanol to be less than 10 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-8155.0 (2.5 .- 1002 ng PFOS/mL). FACT -TCROOP (LIMS #EOO-1716), Page 9 of 109 BACK TO MAIN Table 4. Solubility Screen Test Sample Preparation 1 2 3 4 5 6 7 8 I I 10 mg test substance weighed intotared 4.0 mL glass screw-cappedvial 0.1 mL of octanol added to glass screw-capped vial Followingthe addition of octanol, vial was capped, shaken vigorously and vortexed, and sonicated. Mixingand sonicationtimes recordedon the worksheet. Sample checked visually for undissolvedparticlesof the test substance. (If all of the test substanceis dissolved,the solubilityis determinedto be greater than 10% (massholume) and no additionalsolubilitytesting is required. If a portionof the test substanceremains . undissolved,additionaldilutions are performed using the appropriatesolvent.) Observation made: undissolved particles present.. continue to step 5. Step2 repeatedwith total volume of octanolof 0.5, I,and 2 mL, respectively. Following octanol addition,vial was shaken, sonicated, and observedfor particulate. Followingthe final octanoladditionto bringthe volume of octanolto 2 mL, there was still undissolved PFOS. The shake flask method would therefore be continued to step 7. 14 mg were weighed into a 100 mL volumetric flask and octanolwas added stepwke up to 100 mL. Following each addition of octanol the flask was capped, vortex-mixed, sonicated, and allowedto settle. Particulatestill remained. Therefore2.5 mg PFOSwere diluted in 250 mL octanol. The flask was vortex-mixed, sonicated, and allowed to settle for >24 hours. The concentrationof octanolwas approximately 10 pg/mL and there was still particulate. A solubility I10@mL would requirethe column elution methodto quantitativelydeterminethe solubility. However due to difficulties with tubing selection and the potentially dangerous solvent conditionsof havinga non-sealedcontainerof flammable solvent in an incubator equippedwith electricaloutlets on the inside,it was decidedthat the shake flask method shall be used for the solubility determination. 0 ETS-8-172.0 "Shake Flask Method: Solubility Determinationof a Test Substance in Various Solvents." Approximately 0.0109 PFOS was weighed into a tared 15 mL plastic centrifuge tube. Octanol was added gravimetrically to approximately 8.5 g (approximately 10 mL). Nine centrifuge tubes were prepared, along with three method blanks (tubes with 8.5 g f 0.5 g octanol only). The centrifuge tubes were placed in a temperature-controlled orbital shaker set at approximately 3OOC. See the following table documenting the shake flask method preparation: Table 5. Shake Flask Method Preparationfor Study (FACT-TCROOZ) 12- Octanoladdedto centrifugetube and octanol is approximately 8.5 such g. that total weight of PFOS 9 Method Blank Preparation-Approximately8.5 g odanol addedto a 3 12 temperature-controlledorbital shaker set at approximately 30.0 OC, 01-22-01 01-22-01 01-22-01 FACT-TCR002 (LIMS #E00-1716), Page 10 of 109 BACK TO MAIN SAMPLE COLLECTION AND ANALYSIS On each of Pays 1, 2, and 3 (approximately 24, 48 and 72 hours after initial sample preparation) three centrifuge tubes (containing PFOS) and a method blank were removed for analysis. The centrifuge tubes with PFOS and method blanks were allowed to equilibrate at 22-23 OC for 24 hours, centrifuged, and aliquots of supernatant were collected. The aliquots were diluted 1OOX (990 pL methanol: 10 pL sample) and spiked with internal standard (17 pL of 15 pg THPFOS and PFBS/mL) and ,analyzedvia HPLCIESIMSaccording to analytical method ETS-8-155.0. Four injections of each final diluted sample were performed. The following table describes the sample collection and preparation regimen: Table 6. Sample Collectionand Preparation for Study FACT-TCROOZ I I 1 At 24,43, and 72 hours, 3 PFOS+ octanol, and 1 4 centri- Day1 01-23-01 octanol (method blank) centrifuge tubes are fuge tubes removed from the incubator and equilibrated at per day 4-25 OC for approximately 24 hours. ~ 2 Aliquots of the supernatant (octanol containing 4 per 01-24-01 dissolved PFOS or octanol only from the method blanks) were collected and dispensed into autovials. centrifuge 01-2501 tube I Day3 I 01-26-01 3 The aliquots were diluted 1OOX with methanol (using a dual syringe diluter). Day 1 Day 2 01-24-01 01-2501 Dav 3 01-26-01 InternalStandardwas added to the IOOX sample dilutions(17 pL THPFOS and PFBS solution at via HPLCESIMS on HP Day 1 Day 2 Day 3 Day 1 Day 2 Dav 3 01-24-01 01-26-01 01-26-01 through 01-27-01 01-27-01 01-30-01 ANALYTICALMETHOD ETS-8-155.O"Analysis of Potassium Pertluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-Electrospray/MassSpectrometry". Diluted samples (Le. diluted supernatants) were analyzed using HPLC/ES/MS in the negative ion mode. PFOS levels were evaluated versus calibration standards ranging in concentration from 2.5-1002 ng PFOS/mL. Internal Standard quantification was used to normalize the data. Target ions were 499 m/z (PFOS anion), 427 m/z (deprotonated THPFOS), and 299 mlz (PFBS anion). Analytical Equipment FACT-TCROO2 (LIMS #E00-1716), Page 11 of 109 BACK TO MAIN Liquid Chromatograph: Hewlett-Packard@Series 1100 Liquid Chromatographsystem Analytical column: KeystoneQBetasil" CIS 2x50mm, 5ym particle size Column temperature: Ambient Cycle Time:, 10 minutes Flow rate: 300pUmin Injection volume: 1OyL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in water Solvent C: Methanol Solvent Gradient: Time 0.00 0.50 0.80 3.50 6.50 7.00 9.75 10.00 %C 15 % 15 % 60 % 100 % 100 % 15 % 15 % stop Mass Spectrometer: Hewlett-Packard" Series 1100APVMass Spectrometer Detector Software: HP ChemStation" 6.0 FragmentorVoltage: m/z 299=100 V; m/z 427=100 V; m/z 499= 140V Capillary Voltage: 3500 V Gain = 2.0 EMV Mode: Electrospray Negative Gas Temperature: 350 OC Drying Gas: 10.0 L /min. Nebulizer Pressure: 25 psig Analysis Type: Single Ion Monitoring (SIM) ANALYTICALRESULTS Data quality objectives outlined in the 3M Environmental Laboratory method were met (see Appendix A). Regressions. Quadratic curve fit was applied to calibration standards and sample data to improve qiiantitation over the concentration range appropriate to the data. All calibration curves hac1least-squarefits of 0.980 or greater (R2values ranged from 0.9994-0.9997). Calibration Standards. Twelve standards ranging in concentration from approximately 2.5 to 1002 ng PFOSlmL 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 15% difference from the expected value. Sample Rleplicates. Samples were within the 15% precision requirement. FACT-TCROO2 (LIMS #E00-1716),Page 12 of 109 BACK TO MAIN 0 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 k15% deviation of the target concentrations. (Note: the method states a &30% deviation of the target concentrations, but to be consistent with the more stringent sample requirements of &5%I precision, the accuracy requirementof the CCVs was reprocessedas 15%). 0 Limit of Quantitation (LOQ).The LOQ is equal to the lowest standard in the calibration curve (25 ng/mL). Table 7. Calibration Standard Reprocessing Summary IIDay 1 data: 01-26-01 II 25- 'Oo2 ng/mL Day 2 and 3 data: 01-26-00,01-29-00 2% 1002 ng/mL I I Std 1(2.5 ng/mL), Std 2 (5 ng/mL), and Std 3 (10 I ng/mL)were excludedto better fit the data at the low I end of the curve and becausethe higMow standards did not meet the 4 5 % deviation criteria. Std 1 (2.5 ng/mL), Std 2 (5 ng/mL), and Std 3 (10 ng/mL) were excluded to be consistent with Day 1 data work UD. 0 Blanks. Method blanks (octanol taken through the entire sample preparation, dilution, and analysis process) provided a measure of laboratory contamination. Acceptable values for the blainks were less than 50% of the limit of quantitation (LOQ). 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 quarititation (LOQ). Acceptable solvent blanks were analyzed before each calibration curve. Specificity (according to OPPTS 830.7840): The solubility determination as stated in the EPA Guidelines follows. This method should only be applied to: 0 Pure substance. This study shows a purity of 97.9%. 0 Substances that are stable in water. Hydrolytic studies have been conducted at the 3M Environmental Laboratory showing stability (Report # Wl878). DATASUMMARY Table 8 summarizes individual sample data. Representativechromatograms are presented in Attachment C. The table displays PFOS concentrations for individual injections. Also included are the average concentrationsfor each CentrifugeTube replicate, standard deviation, and % coefficient of variation. FACT-TCR002 (LIMS #E00-1716), Page 13 of 109 BACK TO MAIN iaoie u. r r c m IGKUUUI 1u4tiSOIUDIII~i~n uctanoi Data summary D~ 1 centrifuge rube #I Rep 1: O122O10PFOSl.l.l RW2: 012201-0PFOS-1.1.2 Rep3: 0122010PFOS-1.1.3 Rep4: 012201-0PFOS -1.1.4 lay 1 centrifuge rube w Rep 1: 012201-0PFOS-1.2.1 Rep 2: 012201-0PFOS -1.2.2 Rep 3: 0122010PFOS -1.2.3 RW4: 0122010PFOS-1.2.4 lay Cenmge rube m Rep 1: 0122010PFOS -1.3.1 Rep 2: 0122010PFOS -1.3.2 Rep 3: 012201-0PFOS -1.3.3 Rep 4: 0122010PFOS -1.3.4 )ay 2 centrifuge rube #I Rep 1: 0122010PFOS -2.1.1 R e p 2: 0122010PFOS -2.1.2 Rep 3: 012201OPFOS-2.1.3 RV4: 012201-0PFOS-2.1.4 Rep 1: 0122010PFOS -2.2.1 D~ 2 enmeRep2: 012201-0PFOS -2.2.2 rube Rep 3: 0122010PFOS -2.2.3 Rep 4: 0122010PFOS -2.2.4 D~ 2 rube m Rep 1: 0122010PFOS -2.3.1 Rep 2: 0122010PFOS -2.3.2 Rep 3:012201-oPFOS-2.3.3 Rep 4: 0122010PFOS -2.3.4 D~ 3 centrifuge Tube# I Rep 1: 0122010PFOS 3.1.1 Rep 2: OI22OlOPFoS 3.1.2 Rep3: 0122O10PF0S 3.1.3 Rep4: 0122010PF0S3.1.4 D~ 3 centrifuge Tube #2 Rep 1: 0122010PFOS -3.2.1 . Rep 2: 012201OPFOS -3.2.2 Rep 3: 012201~PFOS3.2.3 Rep 4: 0122010PFOS 3.2.4 Rep I:012201oPFOS3.3.1 D~~ 3 c e n ~Rep2: 0122010PFOS 3.3.2 Tube #3 Rep3: 012201oPFOS3.3.3 Rep4: 0122OIOPFOS3.3.4 59.1 57.4 57.6 57.1 59.1 55.7 57.7 55.0 55.7 55.7 56.9 55.5 55.2 56.5 54.6 56.8 54.9 53.8 54.2 54.1 57.3 57.0 55.5 58.3 54.7 55.4 57.1 54.2 53.4 52.9 52.7 52.7 59.3 57.2 57.7 57.7 57.8 0.9 1.5% 56.9 1.9 3.3% 56.0 0.7 1.2% ~~ ~~~~~ 55.0 1 .o 1.9% 54.2 0.5 0.9% 57. I 1.2 2.1% 55.4 1.2 2.2% 53.0 0.3 0.6% 58.0 0.9 1.6% FACT-TCROOS (LIMS #E00-1716), Page 14 of 109 BACK TO MAIN The average PFOS concentration and standard deviation for each day, as well as the final overall average for ,all non-excluded replicates and time points were as follows: Table 9. Summary Table of Solubility of PFOSTCR-00017-046 in Octanol. Day 1 Day 2 56.9 pghL 0.9 55.7 p g h L 1.4 Day 3 55.4 p@mL 2.5 Average Value 56.0 pg/mL 0.8 1.6% 2.5% 4.5% 1.4% The solubility of PFOS TCR-00017-046 in octanol is 56.0 pg/mL at 22-23 "C. Attachment B contains data summary tables. STATISTICAML ETHODS Statistical methods were limited to calculating means and standard deviations. Refer to Attachment E for formulas and example calculations. STATEMENT OF CONCLUSION Under the conditions of the present study, the solubility of PFOS TCR-00017-046 in octanol is 56.0 pg/mL at 22-23O C . 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. FACT-TCROOP (LIMS #E00-1716), Page 15 of 109 ~~ ~ ~~ LISTOF ATTACHMENTS 0 Attachment A: Extraction and Analytical Methods 0 Attachment B: Data Summary Tables 0 Attachment C: Sample Chromatograms 0 Attachment D: Deviations from the Protocol 0 Attachment E: Sample Calculations BACK TO MAIN FACT-TCROOZ (LIMS#E00-1716), Page 16 of 109 BACK TO MAIN SIGNATURPEAGE We certify that this report is a true and complete representation of the data for this phase of the study: fl / Mark E. Ellrtfson Principal Analytical Investigator Wfiliam K. Reagen Laboratory Manager Date FACT -TCROO2 (LIMS #E00-1716), Page 17 of 109 BACK TO MAIN ATTACHMENAT: EXTRACTIOANND ANALYTICAML ETHODS (ETS-8-170.0 and ETS-8-172.0, the methods used in this study, were not included as an attachment. ETS-8-170.1 and ETS-8-172.1, which are procedurally the same, will be included for confidentiality reasons and further clarification of the procedures followed.) FACT-TCROOZ (LIMS #E00-1716), Page 18 of 109 BACK TO MAIN 3M ENVIRONMENTLAALBORATORY SOLUBILITSYCREENTEST:APPROXIMASTOELUBILITYDETERMINATIOFNA TESTSUBSTANCIEN VARIOUSOLVENTS Method Number: ETS-8-170.1 Adoption Date: 09/08/00 Effective Date: 03/14/01 Approved By: William K. Reag,&, Laboratory Manager Date 1.o SCOPE AND .APPLICATION 1.1 Purpose. .Accordingto 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 performhg solubility testing via the Column Elution Method and Shake Flask Methods. The EPA and OECD solubility testing guidelines state that if the preliminary solubility determination test 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 determination will be necessary for compounds having solubility greater than 10% (massholume). 1.2 Compatible Analytes. Test substance and degradation products for solubility testing. ETS-8-170.1 Solubility Determination: Screen Test Method FACT -TCROO2 (LIMS #EOO-1716), Page 19 of 109 Page 1 o f 9 BACK TO MAIN 1.3 Acceptable Matrices. Aqueous (e.g. Milli-Q water, 0.01M CaCl,), Acetone, Methanol, or other solve:nt(s) of interest. 2.0 SUMMARYOF 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 fidly 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 100mL glass stoppered volumetric flask or graduated cylinder. Add test solvent until visual confirmation of the substance dissolution has been achieved. Record all observations on a standardized preparation sheet or logbook. Once 100 mL solvent has been reached, the approximate concentration 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 cylinder/volumetric flask (concentration is approximately 10ppm). 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, fhe column elution method will be utilized (ETS-8-171.O). 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.4ND CAUTIONS 4.1 Health anid 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 FACT-TCROO2 (LIMS #E00-1716), Page 20 of 109 Page 2 of 9 BACK TO MAIN 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 EOUIPMENT 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 IxlL glass screw-top vial. 7.5 Glass volumetric flasks, 10mL to 1000mL. 7.6 10 &-1000 pL Pipettemanm manual pipettor and plastic pipette tips, or equivalent. 8.0 REAGENTS AND STANDARDS 8.1 Methanol (MeOH), HPLC/SPEC/GC grade fiom EM Science, or equivalent. 8.2 Acetone, IPLC/SPEC/GCgrade firom E M 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 Ca.Cl,, A 0.01 M CaCl, stock solution is prepared by weighing 1.5 g CaCl, Dihydrate in a weigh boat and transfemng to a 1 L volumetric flask and diluting to the mark with Milli-QTMwater. 8.6 Test substance of known purity. 9.0 SAMPLHEANDLING 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 FACT -TCROO2 (LIMS #E00-1716), Page 21 of 109 Page 3 of 9 BACK TO MAIN 10.0 QUALITCYONTROL 10.1 Not applicable. 11.0 CALIBRATIOANND STANDARDIZATION 11.1 The compounds of interest must be of known characterization according to laboratory specifications. 11.2 All equipment used, such as the analytical balance, should be calibrated daily prior to use. 12.0 PROCEDURES 12.1 Solubility screen. 12.1.1 Weigh 10 mg k 1 mg of test substance into a 4 mL glass screw-top vial or equivalent. Record the weight (refer to attachment A for an example of a standardized prep sheet). 12.1.2 Add solvent (e.g. water, methanol, acetone) according to the table below to the test substance. 4 mL vial or eauivalent Total volume solvent added (I&). .. Approximate Solubility Solvent addition 1 0.1 100000 Solvent addition 2 0.5 20000 I I Solvent Solvent 1121 addition 3 addition 4 (Pg/I-nLJ)*-* 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 adlow 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 pglmL, the shake flask method (ETS-8-172.0) will be used to determine the accurate solubility point. However, if the test substance dissolves after thefirst 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 Ihe test substance did not dissolve in the 2 mL of solvent, weigh 10 mg k 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 FACT -TCROO2 (LIMS #E00-1716), Page 22 of 109 Page 4 of 9 BACK TO MAIN -1'00mL volumetric1 Solventaddition I1 Solvent addition 2 Solvent addition 3 I $ask or equivalent Total volume solvent 10 50 100 added (mL).,. Approximate Solubility 1000 200 100 (Pg/mL). - * 12.1.7 If ithe solubility point is reached at or prior to the complete addition of 100 mL, the concentration estimation 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 100 mL solvent level has been reached without complete dissolution of test sulbstance, 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 &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 DATAANALYSIS AND CALCULATIONS 13.1 The solubility determination in this method is qualitative/semi-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-quantitativein 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 bl= the approximate amount of solvent added (mt). 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 Soliibility Determination: Screen Test hletliod FACT -TCROO2 (LIMS #EOO-1716), Page 23 of 109 Page 5 of 9 BACK TO MAIN 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. 1-1 7, 1996 18.3 United States Environmental Protection Agency. OPPTS 830.7840 Water Solubility: Column Elution Method; Shake Flask Method. Prevention, Pesticides and Toxic 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 .O, "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 Deterntinntion: Screen Test Method FACT -TCROO2 (LIMS#E00-1716),Page 24 of 109 Page 6 of9 BACK TO MAIN 20.0 RJ3VISIONS 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 Soltibility Determination: Screen Test Method FACT -TCROO2 (LIMS #E00-1716), Page 25 of 109 Page 7 of 9 I Attachment A: Example Preparation Worksheet,page 1 of 2 ' I BACK TO MAIN volume added (mLy approx conc. after addition (uglmL) Sample Prep OBSERVATIONSINOTES Analyst Datemm- SteD 2-1 ___ Total volume solvent added (mL) (1.1 3 r s Analyst ApproximateSolubility (uglmL) 100,000 DatdTim- SteD 2-2 - Total volume solvent added (mL) (1.5 Apomximate Solubilitv ludmL120.000 Flaskvortex-mixed? Y ~ ~ R/~~N~ ~ ~ ~ 1s there solutd e stlll presentles-wntinue to step 2-2 Flassoknicated? Yes'No Flasakllowetdo settle? Yes/No NO-The solutionIs at IO%, and Is considered"infinitely soluble " P further solubility testing Is required. Flask vortex-mixed?Y e a 0 RoomT e d Is there solute still present? Y e d o Flask sonicated?YesMo -Ti Flask allowed to settle? Yes/No T I M Analyst - DatelTim- SteD 2-3 Total volume solvent added (mL) 1 Approximate Solubilitv (ua/mL) iO.000 Flaskvortex-mixed?Yes/No RoomT e d Is there solute still present? Yes/No Flask sonicated?Yes/No T-i Flask allowed to settle? YesMo T-i Analyst -__ Datmm- SteD 2 4 Total volume solvent added (mL) 2 ADprOXimate Solubilitv (udmLI 5.000 Flask vortex-mixed?YesMo RoomT e u Isthere solute still present? Yes/No Flask sonicated?Yes/No T-I Flask allowed to settle? YesMo T-i ~~ h m l Didthe substance dissolve in 2 mL or less of solvent? r? Yes -Approximate concentrationat which the test substance is soluble in solvent: u g f d -. 0 The solubilityof the test substanceis to be determinedvia the shakeflask method. u) or No Continue to page two for further testing. Notes/ additional comments: I ETS-8-170.1 Solubility Deterniination: Screen Test Method Page 8 of9 ~~ ~ Attachment A Cont.: Example Preparation Worksheet,page 2 of 2 GLP Study Number: Test Substance: Solvent: 3M Environmental Laboratow SOLUBILITY SCREEN Sample preparation worksheet Sourcw Date: Iw: Source: BACK TO MAIN volume added (A)/ appmx conc. after Solubllity data addition (UdW Sample Prep OBSERVATIONSINOTES Analyst DateIlime I Step 4-1 Flaskvortex-mixed?YesMo FlounTemp 'G Isthere a t estilt present? Y&o - Totalvolumesdventadded(mL)... 10 Approximate Solubility(uglmL)... 1,000 23 ~naiyst DaleIlime I Step 4-2 4 - Tdalvolumesdvent addedtmL)... 50 0 Approximate Solubility(ugimL)...200 Bs I h) I A Flask sonicated? YeslNo Time Flask allowedIo settle? YesMo Time Flask vortex-mixed?YesMo RoomTemp FW sonicated?Y ~ N O Time Flask allowedIo settle? Y W oTime Flask vortex-mixed?YesMo RoanTemp Flask I miQ min "C 1s there solute still present? YeslNo min min 'C Is theresdutesUll present? YeslNo -. 2 Analfit Datame I Step5 4 Flask vorlex-mixed? YesMo Room Temp 'C 1s there solute &ll present? Y&o m - Y Totalvolume solventadded(mL)... 1OW f i s k swcated?Y&No Time min 2 Approumate Solubility(ughnL)... 10 Flaskallowed to settle? YeslNoTime mkj la (IDu Conclusions: -I Did the substance dlssolve In 1000 mL or less of solvent? 8r! Yes -Approximate concentration at which the test substance is soluble in solvent: udml A The solubility of the test substance is to be determined via the shake flask method. or No -The solubility of the test substance is to be determined via the column elution method. ETS-8- 170.1 Solubility Determination: Screen Test Method Page 9 of9 BACK TO MAIN 3M ENVIRONMENTALALBORATORY SHAKE FLASKMETHODS:OLUBILrrY DETERMINATIOOFNA TESTSUBSTANCE IN VARIOUS 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/p/, Date 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 p g / d 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 SoIubiIity Test: Shake Flask Method FACT-TCROO2 (LIMS #E00-1716),Page 28 of 109 Page 1 of' 11 BACK TO MAIN 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 qualitativelsemi-quantitativedata obtained from the preliminan! 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 incubator/shaker set to 30 "C k 2 'Cy 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 centrifuged 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. 1H,1Hy2H,2Htetrahydroperfluorooctarie 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 contamination resulting fiom 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-free medium (e.g. methanol/water/ac:etone 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 Solubility Test: Shake Flask Method FACT-TCROO2 (LIMS #EOO-l716), Page 29 of 109 Page 2 of 1 1 BACK TO MAIN 3.6 Calibration Standard: A dilution of various amounts of a stock, intermediate or purchased standard to achieve standard solutions in a concentration range 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. centrifuge tubes) are to be sufficiently rinsed 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 demonstrate that there is no interference under the conditions of the analysis. 6.0 EQUIPMENT - 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Centrifbge capable of holding 15 mL. centrihge tubes or equivalent. 6.3 Incubator with heating/cooling 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 Gastight0 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 FACT -TCROOP (LIMS #E00-1716), Page 30 of 109 Page 3 of 11 BACK TO MAIN 8.0 SUPPLIES AND MATERIALS - 8.1 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science, or equivalent. 8.2 Acetone, HPLC/SPEC/GCgrade 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 1.4g CaC12in a weigh boat and transferring to a 1L volumetric flask and diluting to the mark with MilIi-QTMwater. 9.0 SAMPLHEANDLING 9.1 Record times of initial preparation, set-up, and sample aliquotingkmalysis 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.o 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 oper.atingprocedure. ETS-8-172.1 Solubility Test: Shake Flask Method FACT-TCROOZ (LIMS #E00-1716), Page 31 of 109 Page 4 of 11 BACK TO MAIN 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 PROCEDURES 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 gravimetrically until the desired volume has been obtained (note: solvents have diflering densities, therefore the weight needed to achieve a spec@ 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 af 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 k2 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", "Day1,Rep3", and "Day1,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 2026 OC (record the actual temperature) for about a 24 hour period. 12.2.5 After about 24 hours of equilibration at 20 OC-26 OC, 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: Shake Flask Method Page 5 of 11 FACT-TCR002 (LIMS #E00-1716), Page 32 of 109 BACK TO MAIN 12.2.6 At approximately 48 hours of incubation time, the second set of test vessels may be pulled (labeled "Day2,Rep1", "Day2,Rep2", "Day2,Rep3", and "Day2,Rep4") arid 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. aakorption 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 1:10 or higher dilution of samp1e:extraction solvent (Le. 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 h4icrosoft Access0 to calculate standard deviation. The built in function contains the following equation which is based on the individual entities (n) being less than 30: ETS-8-172.1 Solubility Test: Shake Flask Method FACT-TCROOZ (LIMS #E00-1716), Page 33 of 109 Page 6 of 11 BACK TO MAIN 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= concentration of 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 1 below for Qtabulated values). QobserveF gap/rmge where: Gap= the difference between the questionable point and the nearest value. Range= total spread of the data. Table 1 Criteria for Reiection of Outlier Values: observations 3 4 5 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% E D , 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 Method FACT-TCROO2 (LIMS#E00-1716), Page 34 of 109 Page 7 of 11 BACK TO MAIN via the Q-Test, then the non-compliant shake flask data is not to be used for the timepoint. Include justification for dropping the shake flask (e.g. q-test results). 14.1.3 And finally, the %RSD between the three days should be 4 5 % 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, butt 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 iristrument 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. 1-17,1996 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 FACT -TCROO2 (LIMS #E00-1716), Page 35 of 109 Page 8 of 11 BACK TO MAIN 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`hed.; 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 Solubility Determination 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 Solubility Test: Shake Flask Method FACT-TCROO2 (LIMS #E00-1716), Page 36 of 109 Page 9 of 11 BACK TO MAIN ATTACHMENT A: Example Sample Preparation Sheet (page 1 of 2). Temperatureof Study: 0 Test Substance: Test Solvent: plml IW sourca: IW Source: st: Sample preparatlon worksheet Date: Study # Analyst(s): Test Vessel: l5mlcentribe hrbe/Other(speclfl): samplelD-3.4.1 thru -3.4.4 Day3. Smp4 blank Datflime: Initials: none added Test Vessels sealed w/ tape? YM Test Vessels shakenbortex-mixed? Y/N Test vessels placed on orbital incubator?Y/N Incubator ID: Speed: 1: 'Internal Standard: Int. Std. Conc.: CQLU Initial Temperature: aID#: 0 Four 'Day 1' 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 Visual observations: I DalOlme/lnllials: ETS-8-172.1 Solubility Test:Shake Flask Method Page 10 of 11 BACK TO MAIN Visual observations: lay 3 Four "Dav 3" samples are pulled and allowed to be placed in a temperature controlled environment for approx. 24 hours with no agitation. DalwTimdlnitals: I I Location of samples: Temperature of environment: 0 r ID: Visual observations: I'Day 2" Test Vessels Centrifuged: Cent. ID: Visual observations: WM or RCF readout: duration: oh Dale/T!mdlnitlalS: I I each of the four replicatesare to be made into glasdplasticautovials(also, check autovials for correct labeling). YedNo the solution 1-: with methanol. MethanolTN-A: Additional dilutions: Dilution factor (test so1ution:methanol): 1: X's how many dilutions: =final dilutionfactor: I: Dale/Timdlnitiab: Diluter ID used: Amounts used: uL methanol, uI samples -Store samples and extracts in a cooler at until time of analysis. Cooler ID: Cooler Temp: 0 Datfllmdlnltials: 9 0 I! va , . n-0l 0 (D s w 03 -L 8 Day 4: At approx. 24 hours of equilibration. the "Day 3" samples are to be allquoled/extracted. Test Vessels Centrifuged: Cent. ID: RPM or RCF readout: Visual observations: EnvironmentTemp.: duration: "C Therm. ID: min. DatflimdlnlUals: -Four aliquots from each of the four samples are to be made into glasdplastic autovials (also, check autovials for correct labeling). Yes/No -Immediately dilute the solution -1: with methanol. MethanolTN-A: Additional dilutions: Dilution factor (test so1ution:methanol): 1: X's how many dilutions: =final dilution factor: 1: Diluter ID used: Amounts used: uL methanol, %tore samples and extracts in a cooler at until time of analysis. Cooler ID: Cooler Temp: 0 Date/Time/lnllals: ul samples DatfllmdlniUals: ETS-8-172.1 Solubility Test:Shake Flask Method Page 11 of 11 BACK TO MAIN 3M ENVIRONMENTLAALBORATORY METHOD ANALYSIS OF POTASSIUM ~RFLUOROOCTANESULFONATEOR OTHER FLUOROCHEMCIANLWSASTE 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/OD Revision Date: Laboratory Manager Date: Word 97 ETS-8-155.0 Page 1 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayMass .Spectrometry FACT -TCROO2 (LIMS #E00-1716),Page 39 of 109 BACK TO MAIN 1.0 SCOPE AND APPLICATION - 1.1 Scope: This method describes the analysis of waste stream or water extracts using HPLC- electrospraylmass spectrometry. 1.2 Applicable Compounds: Fluorochemicals or other electrospray ionizable 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 fiom water, using HPLC-electrospray mass spectrometry (HPLC- ES/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 h = 499 or perfluorooctanoate (PFOA), m/z = 4 13. 3.0 DEFINITIONS 3.1 AtmosphericPressure Ionization (MI):The MicromassPlatform LCZ single quadrupole system and other commercially available LC/MS 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 (i.e., 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 electricalfield. 3.3 Mass Spectrometer (MS): The Platform LCZ and other commerciallymanufactured ESMS systems are equipped with a single quadrupole mass selective detector. Ions are selectively discriminated by mass to charge ratio ( d z ) and subsequentlydetected. 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 conventional components are not compatible with one another, but only with similar systems (i.e., 2-spray components are compatible with some orher Z-spray systems, etc.). Other commercially manufactured ES/MS 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 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayMassSpectrometry FACT -TCR002 (LIMS #E00-1716), Page 40 of '109 BACK TO MAIN 4.0 WARNINGSAND 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 appropriate protective clothing, gloves, and eyewear. 4.2 Cautions: 4.2.1 Operate solvent pumps below a backpressure of 400 bar (5800 psi). Ifthe 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 anypart of instrumentation that comes in contact with the samplecr 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 SUPPLIESAND 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 (50x2mmy5 pin particle size) or equivalent. 7.1.3 Capped autovials or capped 15 mL centrifbge tubes. ETS-8- 155.0 Page 3 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrosprayhlass Spectrometry FACT -TCROO2 (LIMS #E00-1716),Page 41 of 1109 BACK TO MAIN 8.0 REAGENTS AND STANDARDS 8.1 Reagents - 8.1.1 Methanol, I-IPLC 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 mM ammonium acetate solution: Weigh approximately0.300 g ammoniumacetate. 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 Typically two method blanks (Mdli-Q water), two matrix blanks, and solvent standards are prepared during the sample extraction procedure. 9.0 SAMPLHEANDLING - 9.1 Standardsand sampleextracts are stored in capped autovials or capped 15m;l centrifbge 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 OUALITYCONTROL 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 Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayMass Spectrometry FACT -TCROOP (LIMS #E00-1716),Page 42 of 109 BACK TO MAIN concentration of the analyte. Spike concentrations should fall in the llow-range of the initial calibration curve if extremely low levels are expected. 10.3 Continuing Calibration Verifications 10.3.1 Continuing calibration verifications (CCV) are analyzed to veriG 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 Standardhrrogate Standard 10.4.1 An internal standard (IS) may be used to quantify the target analytes hy 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. ,TheIS should be added after the extraction process and before analysis. 10.4.2 A surrogate standard may be used for quality control. The surrogate is used to quantitatively evaluate the entire analytical procedure including sample preparation and analysis. The surrogate should be spiked to hll within the low to mid-range of the calibration curve. 11.0 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 lk, or quadratic fit (y = ax2+ 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 consistingof the standards fiom 5 ppb to 100ppb 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 (04for April), and the day (TO01012 for October 12,2000). If more than cine 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 Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayMass Spectrometry FACT -TCROOP (LIMS #E00-1716),Page 43 of 109 BACK TO MAIN 12.1.1.2 Assign a method (MS) file. 12.1.1.3 Assign an HPLC program (Inlet file). . 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 Micramass MassLynx GUIDE TO DATA ACQUISITION for additional information. 12.1.3 Typically the analytical batch run sequence begins and ends with a set of solvent standa;ds. 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 Autosampler/Column Heater 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 pc~rts. 12.3 Using the Inlet Editor 12.1.1 Set-up the H P l l O O 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 Samplesize = 10p.L injection 12.1.1.2 Flow rate = 300 pLJmin. 12.1.1.3 Cycle time = 10.0 minutes 12.1.1.4 Mobile phase components: Solvent A: 2.0 mM Ammonium Acetate Solvent B: Methanol (MeOH) Solvent Gradient: Time (min.1 0.00 1.oo 4.50 8.00 8.50 10.0 %B 5.00 Yo 5.00 % 95.0 % 95.0 % 5.00 % stop ETS-8-155.0 Page 6 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Wa.terExtracts Using HPLC-Electrospray/Mass Spectrometry FACT -TCROO2 (LIMS #E00-1716), Page 44 of 109 BACK TO MAIN 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 Electrosprayhlass 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 initiatethe 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 flowrate. 12.4.5.3 Observe droplets coming out of the tip of the probe. A fine rnist 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 Drylng 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 pUmin 12.4.6.4 Pressure <400 bar (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-Electrosprayhdass Spectrometry FACT -TCROO2 (LIMS #E00-1716), Page 45 of 109 BACK TO MAIN % Recovery = Observed Result - Background Result x 100 Expected Result 13.1.2 Calculatepercent differenceusing the following equation: % Difference = Expected Conc. - Calculated Conc. x 100 Expected Conc. 13.1.3 Calculate actual concentration of analyte in matrix (pg/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 acceptable non-zero standard in the calibration curve. 14.2 Solvent and method blank values must be ?4that 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 zk 30% of the standard concentration. 14.5 Internal Standard recoveries should be within & 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 POLLUTIOPNREVENTIOANND WASTEMANAGEMENT 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 f?om 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 of9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-Electrosprayhfass Spectrometry FACT-TCROO2 (LIMS #E00-1716),Page 46 of 109 BACK TO MAIN 16.4 Print data integration summary, integration method, and chromatograms, fkom MassLynx, and store in the study folder. 16.5 Summarizedata 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, WythenshaweM23 9LZ; United Kingdom. 18.2 MassLynxNT User's Guide, Micromass UK Limited, Tudor Road, Altrincham, WA14 5RZ; or Floats Road, WythenshaweM23 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 Electrosprayhlass 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 Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrosprayhlass Spectrometry FACT -TCROOP (LIMS #E00-1716), Page 47 of 109 BACK TO MAIN ATTACHMENBT: DATASUMMARYTABLES FACT -TCROO2 (LIMS #E00-1716), Page 48 Of 109 La !I n!tial -~ Date PFOS in Octanol Day 1 Hillary, H010126 BACK TO MAIN OpFos-l ' TCR-002, 012201-0PFOS-1.1.1,100X Dil. 45 H010126A.M HILL0045.D 18 SFM 01-24-01 MEE(MLA) TCR-002, 012201-oPFOS-1.1.2, l O O X Dit. 46 H010126A.M HtLL0046.D 19 0pFos-1'1.2 SFM 01-24-01 MEE(MLA) TCR-002. 012201-oPFOS-1.1.3, lOOX Dil. 47 H010,26A.M HtLL0047.D 20 0pF0s-1.i3 SFM 01-24-01 MEE(MLA) TCR-002. 012201-oPFOS-1.1.4. 100X Dil. 48 H010126A.M HtLL0048.D' 21 OPFOS-1.1.4 SFM 01-24-01 MEE(MLA) TCR-002. 012201-oPFOS1.2.1, lOOX Dit. 52 H010126A.M HtLL0052.D 22 0pFos-1'2.1 SFM 01-24-01 MEE(MIA) TCR-002. 012201-oPFOS-1.2.2. lOOX Dil. 53 H010126A.M HILL0053.D 23 0pFos-1'2.2 SFM 01-24-01 MEE(MLA) TCR-002, 012201-oPFOS-1.2.3, 100X Dit. 54 H010126A.M HILL0054.D 24 0pF0s-1'23 SFM 01-24-01 MEE(MLA) TCR-002, 012201-oPFOS-1.2.4, lOOX Dit. 55 H010126A.M HILL0055,D 25 0pF0s-1'2'4 SFM 01-24-01 MEE(MLA) TCR-002, 012201-oPFOS-1.3.1, 1OOX Dil. OpFos-l 3" SFM 01-24-01 MEE(MLA) 57 HO10126A.M HtLL0057.D 26 TCR-002. 012201-oPFOS-1.3.2,100X Dit. 58 HO10126A,M HILL0058.D 27 0pF0s-1'3.2 SFM 01-24-01 MEE(MLA) TCR-002.012201-oPFOS-1.3.3, l O O X Dil. 59 H010126A,M HILL0059,D 28 0pF0s-13.3 SFM 01-24-01 MEE(MLA) , TCR-002, 012201-oPFOS-1.3.4, lOOX Dit. 6o H010126A.M HILL0060,D 29 0pF0s-1'3.4 SFM 01-24-01 MEE(MLA) 5.14 , 15147632 5.14 14930350 5.14 14776497 5.14 14962900 5.14 14992721 5.14 15098625 5.14 14869642 5.14 14727877 5.14 14778751 5.14 14632795 5.14 14831350 5.14 14666605 603.6 586.0 588.4 583.7 603.3 569.0 589.4 561.3 568.5 568.9 581.5 567.3 Day 1 average: std dev: %C.V.: 59.1 57.8 57.4 0.88 57.6 1.5% 57.1 59.1 56.9 55.7 1.87 57.7 3.3% 55.0 55.7 56.0 55.7 0.65 56.9 1.2% 55.5 56.9 p g h L 0.9 1.6% PFOS in Octanol Day 2 Hillary, H010126 BACK TO MAIN TCR-002, 012201-oPFOS-2.1.1, lOOX Dit. 25 H010126B.M HILLO103.D 34 0pFos-2.1'1 SFM 01-25-01 MEE(MLA) TCR-OO2,012201-0PFOS-2.1.2lO,OX Dil. 26 H010126B.M HlLLO104.D 35 0pFos-2.1.2 SFM 01-25-01 MEE(MLA) TCR-OO2,012201-0PFOS-2.1.3, l O O X Dil. 27 H010126B.M HILL0105.D 36 0pF0s-2'1.3 SFM 01-25-01 MEE(MLA) TCR-002. 012201-oPFOS-2.1.4, 100X Dil. 28 H010,26B.M HILL0106.D2 37 0pFos-2'1.4 SFM 01-25-01 MEE(MLA) TCR-002, 012201-oPFOS-2.2.1. lOOX Dil. 32 H010126B.M HILLOl 38 0pFos-2'2.1 SFM 01-25-01 MEE(MLA) TCR-002, 012201-oPFOS-2.2.2. 1OOX Dil. 33 H010126B.M HILLOl .D 39 0pFos-2'2.2 SFM 01-25-01 MEE(MLA) 0pF0s-2.2.3 TCR-OO2.012201-oPFOS-2.2.3,100X Dil. SFM 01-25-01 MEE(MLA) H010126B.M HILL0112.D 4o TCR-002. 012201-oPFOS-2.2.4, 100X Dil. 35 H010126B.M HILLOl 13.D 41 0pF0s-2'2*4 SFM 01-25-01 MEE(MLA) TCR-002, 012201-oPFOS-2.3.1, l O O X Dil. 37 H010126B.M HILLOl 15.D 42 0pF0s-2.3.1 SFM 01-25-01 MEE(MLA) TCR.OO2.012201-oPFOS-2.3.2. l O O X Dil. 38 H010126B.M HILLOl 16.D 43 0pF0s-2'3.2 SFM 01-25-01 MEE(MLA) TCR-002, 012201-oPFOS-2.3.3, lOOX Dil. 39 H010126B.M H,LL0117.D 44 0pF0s-2'3.3 SFM 01-25-01 MEE(MLA) TCR-O02,012201-oPFOS-2.3.4, lOOX Dil. 4o H010126B.M HILL0118.D 45 0pF0s-2'3'4 SFM 01-25-01 MEE(.MLA.) 5.14 14,706,785.0 564.2 5.14 14,920,516.0 577.2 5.14 14,680,691.0 557.4 5.14 14,975,314.0 579.8 5.14 14,469,565.0 560.9 5.14 14,290.466.0 549.1 5.14 14,221,117.0 553.7 5.14 14,265,295.0 552.5 5.14 14,756,722.0 585.6 5.14 14.880.485.0 582.6 5.14 14,413,925.0 567.0 5.14 14,903,941.0 595.9 I Day 2 average: std dev: %C.V.: SohblUy. Shako Fhak Molhod 4llM11 55.2 55.8 56.5 1.04 54.6 1.9% 56.8 54.9 54.2 53.8 0.48 54.2 0.9% 54.1 57.3 57.1 57.0 1.17 55.5 2.1% 58.3 1 55.7 pglmL 1.4 2.5% Pop. 2 Of 3 PFOS in Octanol Day 3 Hillary. H010129 BACK TO MAIN TCR-002,012201-0PFOS-3.1.1lO,OX Dil. 45 H010129A.M HILL0045,D 31 0pF0s-3'1.1 SFM 01-26-01 MEE(MLA) TCR-002,012201-0PFOS-3.1.2, lOOX Dil. 46 H010129A.M HILL0046,D 32 0pF0s-3'1*2 SFM 01-26-01 MEE(MLA) TCR-002, 012201-oPFOS-3.1.3, lOOX Dil. 47 H010129A.M HILL0047.D 33 0pF0s-3'13 SFM 01-26-01 MEE(MLA) TCR-002,012201-oPFOS-3.1.4,100XDil. 48 HO10129A,M H1LL0048.d 34 0pF0s-3'1.4 SFM 01-26-01 MEEIMLA) TCR-002,012201-oPF0S-3.2.1,100X Dil. 52 H010129A.M HILL0052.D 35 0pF0s-3'2-1 SFM 01-26-01 MEE(MLA) TCR-002,012201-oPFOS-3.2.2,100DXil. 53 H010129A.M HtLL0053,D 36 0pF0s-3'2.2 SFM 01-26-01 MEE(MLA) TCR-002,012201-0PFOS-3.2.3, lOOX Dil. 54 H010129A,M HILL0054.D 37 0pF0s-3'23 SFM 01-26-01 MEE(MLA) TCR-002,012201-0PFOS-3.2.4, lOOX Dil. 55 H010129A,M HILL0055.D 38 0pF0s-3`2.4 SFM 01-26-01 MEE(MLA) TCR-002, 012201-oPFOS-3.3.1, lOOX Dit. 57 H010129A.M HtLL0057,D 39 0pF0s-33-1 SFM 01-26-01 MEE(MLA) 0pF0s-3'3.2 TCR7002; 012201-0PFOS-3.3.2, lOOX Dil. SFM 01-26-01 MEE(MLA) HO10129A.M HILL0058.D 4i TCR-002, 012201-oPFOS-3.3.3. lOOX Dil. 59 H010129A.M HILL0059.D 41 0pF0s-33.3 SFM 01-26-01 MEE(MLA) TCR-OO2,012201-oPFOS-3.3.4,100X Dil. 6o H010129A.M HILL0060.D 42 0pF0s-33'4 SFM 01-26-01 MEE(MLA) 5.14 12,608.096.0 5.14 12,676,281.0 5.14 12,836,214.0 5.14 12,513,073.0 5.14 12,316,146.0 5.14 12,252,723.0 5.14 12,205,875.0 5.14 12,170,612.0 5.14 12,979,064.0 5.14 12,939,967.0 5.14 12,913,836.0 5.14 12,899,195.0 559.0 565.6 582.9 554.1 545.9 540.8 538.7 538.4 605.8 584.4 589.6 589.7 Day 3 average: std dev: %C.V.: SolubMy, Shake Fbsk Ydwd Ulaml 54.7 55.4 55.4 1.23 57.1 2.2% 54.2 53.4 53.0 52.9 0.34 52.7 0.6% 52.7 59.3 58.0 57.2 0.91 57.7 1.6% 57.7 55.4 pglmL 2.5 4.5% 2F40 A 0 0 N 6 1 9 i -J A 0) Y UI N 3, -. 8 BACK TO MAIN Yy 11 11 1 a a 4 * 5 1 S 10 11 *a 11 1 1 a 4 D e 1 e 0 10 11 (a 91 e 1 2 1 4 5 I 7 e S 10 11 12 11 CE4.U11m 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 411 4.11 411 4.11 4.11 411 411 411 411 4.11 411 4.11 4.11 4.11 4.11 4.11 4.1t 411 4.11 4.11 411 411 411 4.11 4.11 7 m.4 m. m4 an4 a544 m4 mi m4 a544 . m4 a541 m4 a544 as44 m.4 m.4 m.4 m.4 m.4 254.4 254.4 w4.4 m.4 m.4 m.4 m.4 254.4 m4 m.4 254.4 m.4 m.4 an4 m.4 m.4 m.4 254.4 254.1 154.4 254.4 m.4 254.4 Itm?QUn 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M .M 4M 4.M 4M 4n *y 4.M .I 4M 4- 4M 4.88 4.M 4M 4.a 41 4- 4.M 4.M 4M .Cd 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M BACK TO MAIN 4 4 0 A 0 I0u h f A 0 (D S W # FACT-TCRW2 Cwrpwnd PFOS S W s odlnol Sciubility M Y 1 .I*o.-1*1 18 m o 1m 254.0 a0 m n 11 mo n 254.0 n m.0 a4 a I m.0 n m.0 n wu am -0 n m.0 14 mn 15 m.0 1m 2540 17 X4.0 4.11 4.11 4.1% 4.11 4.1% 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 0.0 00 00 00 0.0 00 00 00 0.0 00 0.0 00 0.0 00 0.0 00 00 00 0.0 00 0.0 00 00 00 0.0 00 ao 00 00 00 0.0 00 0.0 LO 0.0 00 0.0 00 0.0 00 m4 4m my1 4.Y 25&4 4.a 2544 4Y m4 4.88 m4 4Y IyI( 461 my1 4.88 ls44 1.w 254.4 u n4.4 4m m4 4m m4 4.) m4 4.m 2544 488 m4 4.88 BACK TO MAIN 4 0 A 0 0 r u 2 Q) Y ACT-TCRWZ Insliumnt: t wnd: PFOS I CI1 Studv: Odnnd SolubililY I3AY 1 ia 01nm1I bawl HOlIZSlj!.b mu I1 I1 I 1 1 4 $ I ? I I IIOI I1 I1 I 1 1 4 5 B ? B D IO I1 11 I1 B S I 2 1 4 5 e 7 I t 10 11 11 11 -5.14 5,5 000 114 &14 8.14 5.14 5.14 5.14 5.14 5.14 114 5.14 5.14 5n om 5.14 5.14 6.14 5.14 5.14 6.14 115 5.14 5.11 5.14 5.14 5.14 5.14 5.14 0.00 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 ccIs.Dsulw.5 & IPUJ 0.0 0.0 1.1 LO a1.4 me 51,5 Io.( IWl 110.1 5U.n 50.0 1p1u 00 0.0 a7 5.B Pd *.I aa n.r 1m0 15.1 Q.? m.5 ms nn.4 1043 09 I ao 0.1 1.4 9.1 l?.S u1 Iu 1011 m.8 y1.4 UJ 1.011.1 NA E+10.4% 1.7% 1.a 01.3% 2.4% 1.1% 1.1% 0.m BACK TO MAIN M km E R m I a 1 I L14 48 1. 5.14 a 10 s.14 I 11 L14 a n 5.14 53 a LU Y 14 5.14 5 I s.14 n 1 6.14 3 17 s.14 A P 11 1.14 f m m 6.14 m 0 0 0 u) BACK TO MAIN 3 A 0 73 0 0 N * m 8 a 2 0, Y LfpL.u 00 ddas OD 0.0 M 00 0.0 0.0 010 * n 0.0 0.0 2 (1 0.0 0.0 5 N ao 0.0 I #I LO 0.0 n I W 0.0 OD 21 P 0.0 OJ P OJ oa n 82 0.0 OJ 2? P OJ 0.0 an P 00 0.0 s P 00 ao 44 P 0.0 0.0 a P OJ 0.0 a11 P 0.0 0.0 I u 0.0 0.0 P u 0.0 00 u 0.0 0.0 u u 0.0 00 on n m 0.0 m m OD OB BACK TO MAIN PFOS SchbBty FACT-TCROO2 Study#FACT-TCRoo1 Corn@: PFOS Study: OdandSOkrbility AnaMical Instrument: Hillaryon 01-2601 RepocessingWokWkn: D i m 01/30/01batch H01017.6b.b .mpkD..ak(Dn liwll5&Ew!aMkm!e 3 !z&ukta 2540 mo ernesr 4.11 4.11 - Itrereear DE%?&wa 2514 4m 1.1ffi.llO.I 2544 4m 1.187.274.1 n40 4.11 a44 4m l.W.710.3 I 2540 4.11 2544 4m 1.azOQo 7 zuo 4.11 2544 4m l.lIl,~.O 8 zuo 4.11 2544 4m i.iei.imo 0 2uo 4.11 2544 4m 1,1~.SU3.0 10 2540 4.11 2544 4m 1,im.as.i 11 n40 4.11 2544 4m 1.1m.m.e 12 2540 4.11 2544 4m i.im.mm 13 2510 4.11 2544 4m 1,181.rn.l 14 2uo 4.11 2544 4m l.lPpo8 1s 2uO 4.11 2544 4m 1,124,WI.O 29 n40 4.11 2544 4m 1.21o.op.s mo 4.11 2544 4m 1.1E3.543.0 2540 4.11 n44 4m 1.100.~.4 4 WO 4.11 2544 4m ODTS1.4 47 2540 4.11 2544 488 1m.mi.i 4 n40 4.11 2544 400 1W.ZZ.e 48 2540 4.11 n44 4m 1.1M.p9.0 3 0 n40 4.11 2544 4m 1.1m.oL11.9 61 2540 4.11 2544 4m 1.1u2.m.1 ;XI 52 2540 4.11 2544 4m i.im,isi.4 0 N 0 53 2540 4.11 2544 488 1.lBLgB.O Y 2540 4.11 m4 4m l.178.?W3 56 2540 4.11 2544 41y) 1.1ffi.JgeB 58 2540 4.11 1544 4m 1,152.965.0 s7 2540 4.lt 2544 4m 1,l17,018.0 T C R W 012Xl14FOS-2.1.1. 1mX W.0 OPFOS-21 1 SFM 01- MEE(Ml.4 25 TCRmZ OlpOl4FOS-21.2 1ooXMO opFOS212 SFM 01-ZSUI MEE(MW W T C R ~ Zm m i - e F o s a 1 . a imxw.o Q oPFO621.3 SFM W-ZS41 MEE(MW 27 nl TCR002.OlPO14F05214,lmXW D a oPFOS-2 1 4 6FMOl-ZS.W MEE(MIA) W (D TCR002.OlPD14FO&221. 1WW.0 SFM 01-=1 M E E O T C R m Z O l 9 D 1 4 F O & 2 2 2 1COXWO SFM Ol-ZSUI YEEIMU) a3 34 ss 2540 4.11 2540 4.11 2540 4.11 2yO 4.11 2540 4.11 2540 4.11 mo 4.11 2540 4.11 2544 M4 2544 2544 2544 2544 2544 2544 4.m I.Is(I.5wI 4.m 1.151.CO45 4.M l.le.8.3098 4.m 1.150.5904 4.m 1.14.rn4 4 4.M l.lY.OM0 4.- 1.19).148 3 4.m 1.145.588 9 oPFOS23.1 I 2540 4.11 2544 4.00 1,123,414.5 oPFOW.3.2 Fobl.3 3 fiob23.4 I n40 4.11 2544 4.00 1.150.2m8 ae 2540 4.11 2544 4.05 1,1303444 Q 2540 4.11 n44 4.m 1.114?2&5 PAGE 1 d l PFOS so*astlFKT-TCRm2 111 BACK TO MAIN m m.0 4.11 223821S.3 11 m.0 4.M zID(,w.o m.4 m.4 4.M 4.M . 1.2023200 1.W8.ooLE 52 ZXO 411 M.3m3.8 m.4 4.00 l.lW.771 3 3Y m.0 4.11 2mcMS m.4 4.00 l.lM.lXL1 A-u xcv.: 4.11 Om1 owl% 21W.818 Q.J.1)sl 24% 4M om12 om* 1n.17.4.a1%s8 233% i 0.0 om 0.0 00 om 00 0.0 am 0.0 an om 00 16 0.0 om 0.0 00 om 00 17 00 om an 00 om OD 18 00 om 0.0 00 om 00 19 00 om 00 00 om 00 24 an am an 00 om 00 m 00 om 00 00 om 00 51 00 om 00 00 om 00 XI 00 om OD 00 om 00 u 00 om 00 00 om 00 43 00 om 00 00 om 00 44 00 om 00 00 om 00 s 00 om 00 00 om 00 5Q 00 om 00 00 om 00 PFOS ScAhHy FACT-TCROPZ BACK TO MAIN 4 5 e 7 a 0 10 11 12 11 14 15 29 41 4 be 47 be 8B W 51 52 53 54 40 PFOS M.W P F B ~ 55 WfL W 1 , W R O S . 254.- THPFOS. SM.W PFOS 254.- PfBS 58 SM 33,lOmoPpb WOS. 2y.WTHPFOS. 1mIppbPFOS z4.wPFBS 57 LLPULI o.m 5.15 5.14 5.14 6.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 o.m 5.14 5.14 5.14 5.14 L14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 ELgav 0.0 87.7123 1saasa.e slsrr.6 msiaa 124O.ZSUS lS4777.4 z.?4S,3ms 30.717.Qs. 7.m.em.o ll.llE,ISID 143M.Ml.O 24ZZ.WO 3.W7.616.0 awmu 0.0 87,17111 lY.Ol3.5 28o.SU.1 7es.m4 1.217.m.l 1,497.*.3 zzs3.e34.5 l.rn4.363.6 7.m2416.0 1o.QB.1oB.o 1zBzmo a.i9WIZo 00 00 ai 5.2 215 3B5 404 me 1043 ag2 4m.4 e83 l.mz.3 lLU3 KQI 00 00 01 4B 21 6 575 bea 75s 101 7 251.7 3%5 4eae $845 ---HA 14.40% 1.S 4.m 4.1~ 5.1~ 1.92 0.5m 20% -zim 241 tu EE*d.o Euw4 13.7~ e.4~ 4.2% 1.0% 1.5% 0.5~ 1.51 urn 1.7% PME3d5 PFOS F*CT-TUIO(II BACK TO MAIN Study X FACT-TCROOZ Compand: PFOS Study: odarolSohrbility Ar!aiyticallnshmmt Hiilmycn01-2M)l Repc8sshg Wuicatakm Doana01/3M)1 batch H010126b.b A 0 8 0 N h f 6 53 A 2 va, I - I- --- TCROOT M P D I 4 F O S Z Z l . l& Ci 0 I . TCROOZ OlpOl4FOb23.3,1& M d IoPFOS-23.3 SFU OI-Z501 MEE(MU1 0 TCRODTOlpO18F05-L34.lOOXMO k w eEplltl 34 5.14 I4.7W.7SSO S4.2 58.4 m 5.14 14.OXI.SlkO 5n2 n.7 HOlOlZ8.M HlLLMC6.D m 5.14 14.m.Wl.O W.4 56.7 HOlO12W.M HIUO1O.D 37 5.14 l4,675,314.0 m.8 58.0 HOlOl2W.M MUD1IO.D 3u 5.14 14,469.Sed.O CdO.0 58.1 HOl0llss.M HIUO1ll.D 30 5.14 1 4 3 9 0 , a . O W.1 54.0 HOlO1lss.M HILUllZD 4 5.14 14,22l,117.0 6Q.l 56.4 MlOlZ8.M W 1 3 . D 41 5.14 1 4 , ~ . ~ 0 S25 55.2 Q 6.14 14,?3#.mO W.6 5.8 43 5.14 1 4 , 0 M . ~ . O 562.8 5.3 K1101Z8.M HILLCH17.D 44 5.14 14,413,925.0 W.0 58.7 45 5.14 14,8133.Ml.O 57.0 1 .m tm 56.4 0.48 0.- 58.3 1.m 2.1% a, 0 0, A 0 (0 BACK TO MAIN ~PdlBlylllTQKisILLPLpl~ moim,~wumss.o XI 0.m ao 0.0 n m m m . ~HO-. n 0.m 0.0 0.0 P m1oim.u nnunm.o sa 0.m 0.0 0.0 4 m m a u . ~wunm.o 5) 0.m a0 0.0 -n 1 0.m 0.0 00 2 om ao ao 10 om 0.0 0.0 17 om 0.0 ao 18 0.m 0.0 0.0 (a 0.m 0.0 0.0 24 0.m 0.0 0.0 30 am 0.0 0.0 31 0.m 0.0 0.0 31) 0.m 0.0 0.0 42 o.m 00 0.0 43 om 0.0 0.0 44 am 0.0 0.0 $4 om 00 0.0 59 o.m 0.0 0.0 2 4-. 0 W PFOS ocbnolWbFA.CT-TCRW BACK TO MAIN 3 0 i f h 0) h) %-. 0 u) Study # FACT-TCROOZ Compound: PFOS Study: Octanol Solubility AnalvticalInstrument Hilarv on 01-29-01 ReDmcessingWorkstation: Dioana 01/31/01 batch H010129a.b EwIfEeRblt4~Ybll e l 3 3 HOlOl2SA.M HlUWO3.D 13 01WM7-13 4 HOlO120A.M HllLC0M.D 13 01WM7-01 8 H0101ZSAM HILLWM.0 1 010034742 0 HOIOt29AM H-lUW-.0 2 0100347-03 OlWM7.M 01WM7-05 10 Ho10129AM HlUWlOO 3 11 HolOlBAM HlLLMll D 4 12 HOlOlZPAM HlUWlZ 0 5 0100507-06 13 HOlOlZ9AM HILLW13D 6 01003-01-07 0100347-08 OlooM7-09 0100347-10 01W347-11 01WM7-12 01WM7-13 01W347-01 01W M 7 4 2 OlWM743 14 75.lppb PFOS svlcnn 15 * 1 w mPFOS SIdmw- 18 uO.sppb PFOS SdhtavE- 17 4004p4 PFOS Sphrn -QmmeW 18 501.oppb PFOS Sy8temWm 19 1W.oppb PFOS svlcnn 20 - oppbPFos 24 2 . m PFOS Y a w - W P F O S 20 HO~OIZSAM m u ~ i 4 . 0 7 HOlO129AM HlllDOlSD 8 H01012SA.M HKLW16.D 0 HO10129A.M HUW17.D 10 HO10129AM HIUWl8.0 11 HOlO129AM HILLW1O.D 12 HO101WM HlUW2O.D 13 H01012hM W 4 . D 1 mioizsu WUOOW.~ 2 HO1012SA.M HlUWZE.D 3 010034744 OlW347M OlWM7~ 01Ws-07-07 OlW3-0748 01w347-09 01003-07-10 01WM7-11 01OOM7-12 OlW347-13 0100107-09 0100307-09 0100M7-01 01003-0742 01003-07-03 01WM7-M 01WM7-05 0100M7-06 01w3-07-07 OlWM7-08 01003-0709 01W M 7 - 1 0 Oloc107-11 OlOOM7-12 01W M 7 - 1 3 1OppbPFOI Std s,n.MppbPFOS.2 5 4 . w 2Sppb PFOS THPFOS.254.W PFBS LO4 SM 6,4 0 0 . 0 ~ P F O S25.44ppb .27 HO1012SAM HlUW27.D 4 28 H O l O l W M HlLLW28.D 5 .(oppbPFOS THPFOS, 2 5 4 . W PFBS 29 H O l O l W M HiLLW23.D 6 SM 7,M.loppb PFOS. 254.4ppb m.ippb PFOS WFOS, 2 5 4 .P~FBS 24 Ho101WM HIl.LOB0.D 7 SM 8.75.1- PFOS, 254Appb 75.1ppb PFOS THPFOS, 254.W PFBS 31 HO101WM HlUW3l.D 8 Side. 1002OppbPFOS.254.4ppb 1WSppb PFOS THPFOS. 7.54.W PFRS SM io. m.5oppbPFOS. 254.4ppb I HOl012SA.M HUW32.D 0 2S.WPFOS W F O S . 254.WPFBS 33 HOlOlZ9AM HlLLW33.D 10 Sld 11,4W.~PFOS.254.4ppb 4W.Eppb PFOS W F O S , 254.W PFBS 34 HOlOl2SA.M HlLLW34.D 11 SM 12.501.- PFOS. 254.4ppb - 501.Oppb PFOS THPFOS,254.WPFBS SM13.1Om.oppbPFOS.254.~ 1W.Oppb PFOS THPFOS,254,oPpbPFBS Sld 9. ~OO~CQF+PFOS2.54.4ppb 100.2PpbPFOS. CCV THPFOS,2 5 4 . W PFBS Sld 9, IMPoppbPFOS, 254.1W.zppb PFOS, CCV THPFOS, 2 5 4 . W PFBS Owb PFOS W 35 HO10128A.M HIUW35.0 12 38 HOl0120AM HILLW.36.D 13 49 HOlOlBAM HlLLW49.0 9 61 HO10129A.M HILLW31.0 s 65 H010129AM HlLLW65.0 1 V P 2 . m PFOS 66 HOlO120AM HILLW60.0 2 P Sppb PFOS 67 H010129AM HlLL0067.D 3 P 1OppbPFOS 6a H010129AM HILLWB8.D 4 Sld 25,25.osppb PFOS. 254.4ppb 25wb PFOS MPFOS, 254.0W PFBS LOP 89 H01012SA.M HlLLW69.0 5 Std 20.40 Osppb PFOS, 254Appb 4oppb PFOS THPFOS, 2W.oppb PFBS 70 HOlO129A.M HlLL0070.0 6 SLd 27.50.1oppb PFOS. 254Appb M.lppb PFOS THPFOS. 2 5 4 . W PFBS 71 HOlOl29A.M HlLLW71.0 7 Sld 28.75.15ppb PFOS. 254.4ppb 75.1ppbPFOS MPFOS, 2 5 4 . W PFBS 72 H01012OA.Y HILLW72.0 8 Sld29.1002qrpb PFOS. 2 5 4 . W 1WZppb PFOS THPFOS. 2 5 4 . W PFBS 73 H010129A.M HlLLW73.D 0 Sld 30. '254.- PFOS. 254.4ppb 250.5Ppb PFOS THPFDS. 254.oppb PFBS 74 H010129AM HlLLW74.D 10 Sld 3 1 . 4 W . W PFOS. 254Appb 4W m . PFOS THPFOS. 254.oppb PFBS 75 HO101WM HlU0075.D 11 SMU. % l . ~ P F O S , 2 5 4 . 4 & 5Ol.owbPFOS THPFOS.254.WPFBS m H0101ZSAM HILLW76.D 12 Std 33,lOmappbPFOS, 254.1W.Oppb PFOS W F O S , 254.oppb PFBS n H010129AM nnuxl77.D 13 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 254.0 ?? 4.11 4.1 1 4.11 4.11 4.11 4.11 4.11 4.1 1 4.11 4.1 1 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.1 1 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.1 1 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 4.11 l?EMam 2.185.510.8 2;174;015.0 2.082po.e 2,118,167.8 2.122.751.3 2,093.721.4 2,109.798.8 2.~8.m.4 2,nKo,409.8 2.073.1923 2.CUl,078.4 2.024.400.1 2,CUl,840.5 2.055,938.8 2.MOZO4.1 1.11.51.8 2,014,828.0 2,024,E48.6 2,WS,sD(.4 l.ffl.0~4 2.0lO.mO.5 1.083,18.1 1~61.870.5 2,004.324.4 1W,Jb.8 1,991.97e.8 1,964,675.8 1,860.1 13.1 1,9162M.5 1,840.462.3 1.842.181.5 1,858.774.6 1,853,1544 I,e41,~.1 1,836225.1 1,842,147.3 1,842.W.9 1.860.163.4 1.8IJ82.6 1.848.SOO.3 1,856,547.8 1,851,9274 i,uo.sm.3 254.4 254.4 254.4 254.4 254.4 254.4 254.4 2541 254.4 254.4 254.4 254.4 2544 254.4 254.4 254.4 254.4 254.4 254.4 . 254.4 254.4 254.4 2544 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.4 254.1 254.4 254.4 4.92 4.92 4.92 4.93 4.92 4.93 . 4.92 4.92 4.93 4.92 4.92 4.93 4.93 4.92 4:92 492 4.93 4.03 4.92 4.93 4.92 4.92 4.92 4.93 4.92 4.93 4.92 4.93 4.92 4.92 4.92 4.92 '4.93 4.92 4.92 4.92 4.93 4.93 4.93 4.93 4.92 4.93 Iwms3Qa 1.060.177.5 1,064.676.6 1.0(13.910.8 1.080.905.1 1.1 02.536.4 1,097,531 3 1.18.412.5 1,07733.8 1.0(18,394.4 1.081p77.8 1,073.770.8 1,061,0525 1,047256.6 1,039.712.0 1.0llbl8.3 l.Cd0.712.1 1.Cd3.416.9 1,094,411.O 1,087.423.1 1,005.605.8 1,079JS4.4 1.ow.75e.e 1,079.623.0 1.079205.6 1.073.789.1 1 ,OsJ,604.4 1.M2.784.3 998220.0 1.M6.334.4 1.042.889.3 1.032201.5 1,036,027.6 1,043.668.0 1,043.395.3 1,035.1 59.4 1,032.746.5 1.030.456.4 1,034.624.0 1,036.166.8 1,014,587.7 1,006.745.6 1,010,418.6 962.453.6 PFOS Omml SOU.FACT-TCRW2 BACK TO MAIN Study # FACT-TCR002 Compound: PFOS Study: Octanol Solublllty Analytical Instrument Hillaryon 01-29-01 ReprocessingWorkstation: Dloana 01/31/01 batch H010129a.b oPFOS3.1.1 oPFosJ.12 oPFOSJ.1.3 oPFOS3.1.4 TCR-002. 012201-oPFOS3.1.1. l00X W.0 SFM 01-2601 MEE(Mu) Day 3. &mpb 1. Rep1 TCR~0122014F055.12.1WXDL0 SFM 01-264 MEEF(U) Day3.0.- 1,R.pz TCR-002.OlpOl-oPFOsJ.1.3,1WXW.0 SFM0 1 2 M 1 MEE(UU) Day3 , S m p * 1. RW3 TCR-WZ. 0 1 2 2 0 14 SF F0SJ.1.4.100X M 01-2501 M E DI. E 0 O Day 3, &eI, R.P) 4S H010129AM HlUW4S.O 31 48 H01012BAM HUW46.D 2'3 47 H01012SA.M HIUOM7.D 33 48 HOIO~XAM HIW.D Y oPFosS2.1 52 H010129AM ldLWW52.0 3S oPFOSJ.22 63 HOlOl2SA.M HlllooS3.D 30 oPF08-32.3 OPFosS2.4 TCR-002.0laOl-oPFOS32.4, H.0 54 HO1012SA.M H I W . 0 37 SS HOlO12SA.M HIUWSS.D 38 oPFOS3.3.1 57 HOlOl2SAM HlUWS7.0 39 oPF~.32 5U HOlOlTOhM HlLLW.0 40 oPF055.3.3 59 HO1012SAM HIIlWS9.D 41 oPFosS.3.4 (10 WIO129AM Hlll0060.D 42 254.0 254.0 254.0 254.0 zY.0 254.0 254.0 254.0 254.0 W.0 2U.O 254.0 4.11 4.11 4.11 4.11 4.11 4.tl 4.11 4.11 4.11 . 4.11 4.11 4.11 1,864,087.8 1.872270.8 1,8SS,lS1.8 l,M3,739.0 1,834.71S.8 l,MO,M4,0 1.7W.M3.3 1.827.SO8.S 1,798,130.1 1,783.8M.O 1.812,W3.8 1,824,825.8 254.4 254.4 254.4 254.4 254.4 2n.4 254.4 254.4 254.4 254.4 254.4 254.4 4.93 . 4.92 4.92 4.02 4.03 4.93 4.92 4.83 4.92 4.03 4.92 4.92 908.381.8 981,228.1 ow.812.S 966~834.1 961,672.3 088.M8.8 087,871.7 085.S092 Y3.755.3 972,202.3 m.445.7 961252.4 oPFOsJ.4.1 40 H01012SA.M WUW4O.D 21 254.0 4.11 1.87S281.S 254.4 4.92 1.039101.3 OPF055.42 41 WlOl2SA.M HlLLW(l.0 22 254.0 4.11 IPBO,~~.O 254.4 4.92 1.047255.1 oPFOS3.4.1 42 H01012SA.M HlUW42.D 23 254.0 4.11 l,M3,163.3 254.4 4.93 1.032.528.6 oPFOSJ.4.4 43 H01012SA.M MLLOM3.D 24 254.0 4.11 1,853,592.0 W.4 4.92 1.031.346.7 Am- SWh xc.v.: 4.11 0.0012 0.024% 1,890,788.5 70.M2.S 3.75% 4.92 0.0011 0.022% i.0272n.o 42.009.1 4.09% McOH Blank TN-A-4740. 04 Des 00 SOLVENT BUNK 1 H010129A.M HILLOW1.D 91 0.0 0.00 0.0 0.0 0.00 0.0 MeOH 0llnk TN44740. 04 Doc00 8OLVENT B U N K 2 H010129A.M HIU0002.D 91 0.0 0.00 0.0 0.0 0.00 0.0 MeOH Blank TN-A-4740. M Da 00 SOLVENT BUNK 5 H010129A.M HIUOW5.D 91 0.0 0.00 0.0 0.0 0.00 0.0 * MeOH Blank WA4740. M Dec 00 SOLVENT BUNK 6 H010129A.M HILLO0W.D 91 0.0 0.00 0.0 0.0 0.00 0.0 rn MeOHB h k WA-4740. 04 Du00 80LVENlB U N K 7 H010129A.M HIUOW7.D 91 0.0 0.00 0.0 0.0 0.w 0.0 0 MeOH Bhnk W - 4 7 4 0 . 04 Du00 SOLVENT B U N K 21 H010129A.M HlUW21.0 92 0.0 0.00 0.0 0.0 0.00 0.0 9L MMeeOOHHBBlaInnLL TTNK-AM47744O0..0044 D Dauc 0000 SSOOLLVVEENNTT BB UU NN KK 2232 HH001100112299AA.MM HHILl LU02022300 9922 00..00 00..0w0 00..00 00..00 00..0000 00..00 2 MeOH @tank TKM740. 04 Da00 SOLENT B U N K 37 H010129AM HlLL0037D 92 0.0 0.w 0.0 0.0 0.00 0.0 rn m MsOn Blnk TNMI~O. 04 0c. SOLVENT BUNK 38 H010129AM HlLL0030.D e2 0.0 0.00 0.0 0.0 0.00 0.0 Y MsOn BM: T K M ~M~D..S00 WLVENTBUNK 39 H010129AM HlLLW39.D 03 0.0 0.00 0.0 0.0 0.00 0.0 MoOnB*nlr TKM740. 04 Du00 SOLVENT B U N K 44 H01012SAM MLOM4.D 93 0.0 0.w 0.0 0.0 0.00 0.0 2fmn MsOn Blank McOH Blnk TNM74O. 04 Du00 WLVENT B U N K TKM740. 04D u 00 SOLVENT B U N K M HOlOl29A.M HILLW5O.D 93 0.0 0.00 0.0 51 HOlOlZ9AM HlLLW51.0 93 0.0 0.W 0.0 0.0 0.00 0.0 0.0 0.00 00 MeOH 0- WA-4740. M O u 00 SOLVENT B U N K 56 HO10129AM HILLWS6.D 94 0.0 0.w 0.0 0.0 0.00 0.0 MeOH M TKM740, 04 Dac 00 SOL- B U N K 62 HO10129AM HILLCM2.D 64 0.0 0.00 0.0 0.0 0.00 0.0 MeOH BInL TKM740.04 Da00 S O L W B U N K 61 HOlOl29AM rml0Ml.D 64 0.0 0.00 0.0 0.0 0.00 0.0 MeOH Blank -740. 04 h e 00 SOLVENT B W K 64 H010129AM W W . D M 0.0 0.w 0.0 0.0 0.00 0.0 MeOH Blank TKM740. 04 Da00 SOLVENT BUM 78 HOlOl29AM MLW78.D 9s 0.0 0.00 OD 0.0 0.00 0.0 MaoHBbnL TKM740.04 Du00 SOLVENT BLANK 79 H010129AM tilUW79.D 85 0.0 0.00 0.0 0.0 0.00 0.0 MoOnEhk W - 4 7 4 0 . 0 4 Du00 SOLVENT BUNK 80 H01012QAM MUOOO0.D 95 0.0 0.00 0.0 0.0 0.00 0.0 PFOS odM(sdub.FACT-TCRW2 BACK TO MAIN Study # FACT-TCR002 Compound PFOS Study: Octand Solubility AnalyticalInstrument: Hillaryon 01-29-01 ReprocessingWorkstatlon: Dloana 01/31/01 batch H010129a.b s%QlIwQ 0100347-13 0100347-13 ? ? H O * M H X D O =a NA . 4 H010129A.M HlUWO4.D 13 5.14 22.420.566.0 083.5 NA ~ ~ 010034741 010034742 OppbPF0SswIam-E-M 2 Sppb PFOS S.mMbl*tbon 8 H010129A.M HlUWJ8.D 1 0.00 0.0 0.0 NA 9 H01012SA.M HKLOOO9.D 2 5.14 85275.0 0.0 NA 010034743 01003470) 010034741 )ppb PFOS S&bNyiE-ala, 10 H010129A.M HIUWlO.0 3 5.14 155,562.0 2.3 NA IOppbPFOS~sSb*y~*~ 14 H010129A.M Hl_L_LMll D- 4 5.14 283,8314 7.1 NA 25ppb PFOS Sydun SJ(.bylEplbralm 12 H010129AM HlU0012D 5 5.14 741.962.1 241 NA 010030TX% 4- PFOS Wrn SdUtiWE@brasOn IS H010129AM HlLUIOl3.0 6 5.14 l.lrnW.5 41.4 NA 010034747 010034748 010034749 0100347-10 0100347-11 0100347-12 0100347-13 01OD34741 - 5O.lppbPFOS Syslem S&aKt#E#bnla, 14 H010129AM HIUW14.0 7 5.14 1.470.558.1 52.1 NA 75.lppb PFOS S- SdbbWfEquBnla, 15 HOlO129AN HIUWISD 8 5.14 2211.%7.5 30.8 NA I002ppbPFOS Syalan m d b r d c n 16 HOlO129AM HIUW160 0 5.14 2.854.703.0 1W.4 NA 250 )ppbPFOS S w r n s u l M E I / E m 17 HOlOl2SAU HILL0017 D 10 5.14 6,799.18.0 285.3 NA 400.- PFOS S w m WhWE#bmM 18 H010129A.M HIUWl8D 11 5.14 10210.912.0 417.0 NA MlOppbPFOS-- 10 HOlOl29AM HlUWlOD 12 5.14 12.352.513.0 518.1 NA lW2.OppbPFOS Syslm -gBnc&l 20 HOlOl29A.MHlU0020D 13 5.14 22.M9.0)2.0 1,024.9 NA Oppb PFOS 24 H010129A.M HlU0024D 1 0.00 0.0 0.0 NA 010034742 P 2.- PFOS V 26 H01012SA.M HIU0025D 2 5.14 W,837.8 0.0 Ex&jad OIOOU)~~~ P WPFOS HOlO129A.M H l l l 0 0 1 D 3 5.14 145.%9.8 2.0 vAdsd 010034744 P I 1OppbPFos- 21 H010129AM HILLW27.0 4 5.14 83,650.3 8.4 sm 5.zs.05ppb PFOS. ZW.~PPC, Ex&jad C~IO3474s 2 5 5 PFOS THPFOS. 254- PFBS L W 26 H01012SA.M H!UW28D 5 5.14 7Q.Mn.3 Z.8 0.1% Sld 8,40.08@ PFOS. 254.4ppb' 0100347010034741 010034143 mw 4Oppb #OS THPFOS.Z%@pb PFBS HO10129A.M WJAGZRD 8 6.14 1.1t2,scea 33.8 su 7, m.icppa PFOS. a4.w W.lppbPFOS THPFOS. 1Y.WPFB3 30 H01012SA.M HIUW3OD I 5.14 1,387,411 .e 403 m~8, PFOS. in.4ppb 75.lppb PFOI THPFOS. 2Y.Oppb PFBS 31 HOlO12M.M HlUW31 D 8 5.14 2.101.189.8 764 3.3% 1.E% 2.0% sM0,lm~PFos.w.4ppb OlW34749 1002ppb PFOS THPFOS. 254.WPFBS 32 HOlOl29AM HIUW32 D 0 5.14 2.785P0.5 103.1 2.w Sld 10.250.5Wb PFOS.254.4ppb 0100347-10 2SO.)ppbPFOS THPFOS. 2% oppb PFBS 33 H01012OAM HMW31D 10 5.14 8.W7P9.0 250.0 2.2% Sld11,4W.~PFOS.254.~ 0100341-11 400.8ppbPFOS THPFOS,254+ffBS 34 HOlOl29AM HlUW34 D 11 5.14 9.909.537.0 4012 0.1% SM 12. 5Ol.Wppb PFOS, 254.- 2 0100347-12 Ml.OppbPFOS THPFOS. 2 Y . W PFBS 35 HO1012SA.M MU0035 0 12 5.14 12.095,Mo.o 502.9 0.4% sm 13. too?- PFOS. 2n.rppb 0100347-0 1002.Oppb PFOS THPFOS. 2 W . W PFOS M H010120AM HlUWM D 13 5.14 21.541272.0 1.012.4 1 .OK 0 hl Sld O.lOOI(Ppb PFOS. 254.4ppb 010034749 100 2ppb PFOS. CCV THPFOS. 254 Cwb PFBS 49 H010129A M HILL0049 D 0 5.14 2.565.C64.8 101.7 1.5% 010034749 Sld 9.1002Wb PFOS. 254 4ppb i w 2ppb PFOS. ccv THPFOS. 254 oppb PFBS 61 H010129AM HILL0061 0 9 5.14 2.647.ffi5.8 101.4 1.2% - 010034741 Oppb PFOS 65 H010129AM H I U W S D 1 0.00 0.0 0.0 NA 01003-0742 010034743 P P 2.Sppb PFOS ffi H010129AM HlLLMX6D 2 5.14 73.876.5 0.0 Sppb PFOS 67 H010129AM H l U W 7 D 3 5.14 135.444.5 1 .8 Exchded Exc*ldcd P Q) Otw3-0744 1- PFOS 68 H010129AM H I U W 8 D 4 5.14 254.129.7 6.5 Exduded Y sm 25,25.~ppbPFOS. 254 4ppt. 0100347QS ' 25ppb PFOS W F O S . 2% Qpb PFBS LOP 69 HOlOl29AM HILU10690 5 5.14 657.037.1 22,s 10.2% Sld2%4O~PFOS.~.4ppb 0100347X% 4Oppb PFOS THPFOS. 254 lbpb PFBS 70 H010129AM HMM7OD 6 5.15 1.050.159.5 38.2 4.7% Sld 27,M lW PFOS. 254.4ppb 2 010034747 4 010034748 A 50 lppb PFOS THPFOS. 254 $pb PFBS SM28.75.15WbPFOS.254.4ppb 75.lppb PFOS THPFOS. 254 Dppb PFBS Sld 29,lW 2 W PFOS. 254.4ppb 71 H010129AM HlU0071D 7 72 H010129AM HlUW72 0 8 5.14 1,316268.3 48.9 5.14 l.99l,ms.9 75.7 2.3% 0.8% 0 010034749 100- PFOS THPFOS. 254@pb PFBS 73 H010128AM HMO073 D 9 5.14 2837,5243 101.7 1.5% u) SM 30,250.5QPbPFOS. 254.4ppb 0100347-10 2 M m P F O S W F O S . 254.oppbPFBS 74 HOlOl29A.M HILL0074 D 10 5.14 6243,7443 256.0 22% SM 31.4W.W~bPFOS. 254.4ppb 0100347-11 01wM7-12 m . w 400.8ppb PFOS THPFOS. 254lbpbPFBS sld 32.mi.ooppbPFOS. 5Ol.Oppb PFOS THPFOS. a4.WPFBS 75 HOlOl2QA.M W 7 5 D 11 5.15 0,534,313.0 4042 78 HOlOl29AM H1110078D 12 5.14 11.346.7M.O 4w.5 0.8% 3.1% SM 33,1002.W PFOS. 254.4ppb 0100347-13 lW2.OppbPFOS WFOS.ZYQpbPFBS 77 HO1012opIM HUW77D 13 5.14 20,449.734.0 994.0 0.8% Paw3015 PFOSOQml soclb.FACT-TCROO2 BACK TO MAIN 3 sA31 0 0 N 8It 0 i 2 Q) - Study # FACT-TCROO2 Compound PFOS Study: Octand Solubility Analytical Instrument: HiLry on 01-29-01 ReprocessingWorkstation: Dioana 01/31/01batch H010129a.b WmhJD TCROO?.Oltzol-oPFOS-3.1.1,lWX M.0 BMll BumdlKMkfIl? oPFo53.1.1 SFMO!-W! M E E ~ D r / 3 . S m n p h l . R e p l 45 H010tZSAM WLLW5.D 31 oPFOS3.12 T C R ~ Z O l P O 1 -oPF SFM OS-3.111WX M.0 01-28-01 MEE(MLA)Day 3. w 1. Rep2 46 HOlOlZOAM HIUW48.D 32 TCROO?.O122Ol4PFOS-3.1.3,1WXDI.0 oPFOS3.1.3 SFM 01-28-01 MEE(MU) D y 3, &mp* 1, Rep3 47 HOl0lrJA.M HILLW7.D 33 oPFOSJ.1.4 &I+ TCROO?. 0122O1-aPFosJ.1.4,1WXDL0 SFM 01-2601 MEE(MU) Day 3. 1. Rap4 48 M10120AM HlLLW48.D 34 TCROO?,Ol22Ol-oPFOS3Z1.1WX M.0 oPFOSJ2.1 SFMOl-ZWl MEE(MU)Day3,SMp*2.R.p1 52 H0101ZOAM HllL0052.D 35 TCR-002.0122O1-oPF0S322.1WX DI.0 oPFOS-322 SFM 01-2601 M E W Day 3, sunp*2. Rep2 53 HO10129AM t I l U M 3 . D % TCR-w2. DlPOl.oPFOS-321.1WX M.0 opF0532.3 SFM 01-28-01 MEE(MU) Day 3, &mp*2. Rep3 54 H010129AM Hlll0051.D 37 TCROO?.011201.aPFOSJ2.4,l W X D L O oPFOS-32.4 SFM 01-28-01MEE(MLAJDay 3,88mp*Z.Rep( 55 M10129AL1 HILLW55.D 38 TCROO?. 0l22O1-oPF0SJ.3.1.1WXDI.0 oPF053.3.1 SFM 01-2601 M E Y M W Day 3.Samp*l.Rep1 57 H010129AM HIUO057.D 35 TCROO?, 0122O1-0PFOS3.32.1ODX M.0 oPF063.3.2 SFM 01-28.01 biEE(Ml-4) Day 3. Smnp4e1.Rep2 l H010129AM HILLOO58.D 40 TCR-GQ2.012201-oPF063.3.3.1WX M.0 oPF093.3.3 SFM 01-26-01MEE(MLA) Day 3.Sam* I, Rep3 TCR-OO2.Ol22Ol-oPFOS3.3A,lWX DI.0 59 H010129AM HlLLOO59.D 41 oPF053.3.4 SFMOl-26-01MEYMWDay3,SampleI. Rep4 60 H010129AM HILL00gO.D 42 l?EQ&El 5.14 5.14 5.14 0.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 5.14 eEpecm !lmmxa 1 2 , W , ~ . O 559.0 !mufnkm sQnYakQ u9h1 55.0 Aumss B4.DH nm!. w1.5 12676281.0 565.6 50.6 1.26 12,838214.0 Mz.0 1.3 2.2% 12.513,073.0 W.1 55.4 12,316,146.0 545.9 54.8 54.1 12,2u.m.o 540.8 54.1 0.35 12,205,875.0 538.7 53.9 0.8% 12.1m,612.0 m.4 53.8 12,979,~.0 W.8 60.e 592 12.939.887.0 554.4 58.4 0.93 12813,838.0 5119,s 59.0 1.6% 12,899.195.0 589.7 59.0 1 0.v 1 i mnge: skl due xc.v.: 66.8 WmL ar 4.5% PFOSocbmlS o b . FACT-TCR002 BACK TO MAIN Study # FACT-TCR002 Compound: PFOS Study: Octanol Solubility Analytical Instrument Hillat-von 01-29-01 ReDrocessinaWorkstation: Dioana01/31/01 batch H010129a.b SmMbJQ ' .mo*d M bR.la.dpILLLillYbls e.QuI l?E.QmmlnlhuEB oPFOsJ.4.1 40 HO10129A.M H i U W . 0 21 0.00 0.0 0.0 QPFOS-3.42 :DI.0 D q 3,8mmp*4,Rep3 41 HOIOIZWCM HIUOM1.D 22 0.00 0.0 0.0 oPFOsJ.4.3 MOWBlu*.S N O l - ~ Ml E E W ) YCTIK)D BUNK 42 M1012W.M HlUW42.D 23 0.00 0.0 0.0 TCR4D2,Olp01.0PF0~.4.4.100XDI.O Dq3,Smmp*4,Rop4 oPFOSJ.4.4 MeOH Blank M e W E h k , SFM 01-2S.01MEE(MU) METHODBUNK ~ ( ~ 7 4D.C0 w. a~oLvENIBUNK 43 HolOl29AM HlUW43D 24 0.00 0.0 0.0 1 HOlOl29AM H1- 0 01 0.00 0.0 0.0 M.oHBlu* W 7 4 0 . 04 ha00 SOLVENT BUNK 2 H010129AU HIIlW02D 91 0.00 0.0 0.0 MeDH BhnL MeDH Bht MeOH Ebnk T K M 7 4 0 . M D.000 SOLVENT BUNK -710. M DOC w aoLVEwT BUNK W 7 4 0 , M Des 00 SOLVENT BUNK 5 HOlOl29AM HIUW(H0 91 0.00 0.0 0.0 8 ~ 0 1 0 1 n2 u~w~oo~s~ 01 0.w 0.0 0.0 7 HOlOl29AM HlllWO7D 91 0.w 0.0 0.0 MeOH Blnt TKA4740,M D.000 SOLVENT BUNK 21 HO10120A.U HIUW21.D 02 5.14 41,422.1 0.0 A MeOH Blnt W 7 4 0 , MD.000 MLVEWTBUNK 22 H010129AM HILUIOP.D 92 0.00 0.0 0.0 MeDH Blmk ~ 7 4 0 M. haw a o L mBUNK 23 H01012EAM HlllW23D 92 0.00 0.0 0.0 M.oH Bbnlr ~ 7 4 0 . 00 ~aoLvtw BUNK 37 H01012W.M HlU0037D 02 0.00 0.0 0.0 ;o MeOHM -740, MD.C w aowmBUNK 38 H010129AM HlUW38D 02 0.00 0.0 0.0 0 MeOHBlnt ~ 4 . 4 7 1 0 .M DOG00 a o L m BUNK 3s H010129AM HlLLWlSD 93 0.00 0.0 0.0 0 MeOH Bbnt T K M 7 4 0 . M D.s00 SOLVENT BUNK U H010129AM HlUW44 D 93 0.00 0.0 0.0 N MUOH Blu* TN-740. M DOG00 SOLVENT BUNK sa H010129AM HIUWMO 93 0.00 0.0 0.0 MeOHE h k ~ ~ - 4 7 4 M 0 .D.C 00 aoLvErnBUNK 51 H010129AU HlUWSl D B3 0.00 0.0 0.0 MeOH Wank TKA-4740. 04 Dee00 SOLVENT BUNK 56 H010129AM HILL00560 94 0.00 0.0 0.0 MeOH Ehk W 7 1 0 , MDec 00 SOLVENT BUNK 62 H010129AM HIUW6tD M 0.00 0.0 0.0 McOH 6h-A TN44740. MDes 00 SOLVENT BUNK 63 H010129AM HILLW63.D M 0.00 0.0 0.0 MeOH Ehk TKM740, MDo0 00 SOLMNT BUNK e4 H01012OAU HIUW64D M 0.00 0.0 0.0 %0 MMeeOOHHBBIlUuI*(I TTNN-4A--44774400.. 0M4 DDeeo0 0W0 SSODLLVMENNTT BBUUNNKK 7789 HH0O1l0O1l22O9AAMM HHlUUW007789DD 9O5S 00..0000 00..00 00..00 53& WHBlnt TKA4740. M Doc 00 SOLVENT BUNK 80 ~ 0 1 0 1 2 9n ~i u~w 8 o ~ as 0.00 0.0 0.0 BACK TO MAIN ATTACHMENCT: SAMPLECHROMATOGRAMS FACT-TCROOZ (LIMS #E00-1716), Page 67 of 109 Batch Run # 37 of 79 Data File C:\HPCHEM\1\DATA\HolOl26\HILLOO37.D BACK TO MAIN Sample Name: MeOH Blank T-_-N_---A_--_-4-7---4--0--,-----0--4---D-_e--c--0_0--_-___________________--_--------------------------- --- Injection Date : 1/26/01 8:44:34 PM Seq. Line : 37 Sample Name : MeOH Blank Vial : 92 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj Volume : 10 p1 Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed : 1/25/01 9:13:04 AM by M I A Analysis Method : C:\HPCHEM\l\METHODS\HOl0126A.M Last changed ': 1/30/01 8:17:48 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on "DioanaI* (LT MSDl 299, ElC=298.7:299.7(H010126WlLL0037.D) API-ES, Neg, SIM, Frag:Var 2500 1 : 2000 1 1; I I 14000 : 12000 : 8000 y 6000 - 30 012000 1000 : I - ---,.A-/ Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name Signal 1/30/01 8:17:46 AM 1.0000 1.0000 - t u 4-49I P. Initial _yIL_ Date Instrument 1 1/30/01 8:21:09 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 68 of 109 Page 1 of 3 Batch Run # 37 of 79 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOO37.D - - -#- I - - - -[-n-g-/- -m-I- - - I - - - - - - - - - - - - - - - - - - - - - - - - - 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1: MSDl 299, EIC=298.7:299.7 BACK TO MAIN Sample Name: MeOH Blank Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type Area , Amt/Area --[-m-i-nl-- 1------1----------1----------1----------1--1------r--a---t--i---o-- 4.878 - - Totals without ISTD(s) : Amount [ng/-l - G r p Name THPFOS 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 1 Warnings or Errors : Warning : ISTD compound(s1 not found Sorted By Calib. Data Modified : Multiplier Dilution Signal 1/30/01 8:17:46 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298 7 :299.7 Totals : 0.00000 Instrument 1 1/30/01 8:21:09 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716),Page 69 of 109 Page 2 of 3 Batch Run # 37 of 79 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOO37.D Signal 2: MSDl 427, EIC=426.7:427.7. BACK TO MAIN Sample Name: MeOH Blank Totals : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 Peak RetTime Type Width Area Area Name - - - - I#- - - - - - - [minl I - - - - - - I --[-m-i-ni-- I - - - - - - - - - - I- - - -%- - - - I -. . . . . . . . . . . . . . . . . . . . 1 5.144 0.0000 0.00000 0.0000 PFOS Totals : 0.00000 1 Warnings or Errors : Warning : ISTD compound(s) not found Instrument 1 1/30/01 8:21:09 AM KLT/MLA FACT-TCR002 (LIMS#E00-1716), Page 70 of 109 Page 3 of 3 Batch Run # 36 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOll4.D BACK TO MAIN Sample Name: MeOH Blank T-. .-N.--.-A.--.-4.-7.-4.-.0-.,-.-.-0.-4._.D._e.c_. ._0. .0_. ._._. ._. ._._. ._. ._._. ._. ._. _. ._. ._. ._._. ._. ._._. ._. ._._. .-.-.-.-.-.-.-.-.-. -. ------------------- Injection Date : 1/27/01 11:01:20 AM Seq. Line : 114 Sample Name : MeOH Blank Vial : 96 Acq. Operator : MLA Acq. Instrument : Hillary Inj : 1 I n j Volume : 10 pl Acq. Method : C:\HPCHEM\l\METHODS\HO10125.M Last changed : 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126B.M Last changed ': 1/30/01 9:16:52 AM by JCLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on I1Dioana1* LT MSDl 299, ElC=298.7:299.7(H010126\HILL0114.D) API-ES, Neg, SIM. Frag:Var 22550000 : 2000 - 1500 I 1000 500 4 ij I 4 I I I I 2 4 6 8 MSD1427,ElC426.73427.7(HOlOl26\HILLO114.D) API-ES. Neg, SIM, Frag: Var 14000 12000 L 10000 1 8000 { 6000 4000 { ,. 2000 { 0- - -__---I__-- mil A 2 4 6 8 min Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD 'Amount Name 1.. 0000 1.0000 Instrument 1 1/30/01 9:20:18 AM KLT/MLA FACT-TCROO2 (LIMS #EOO-l716),Page 71 of 109 Page 1 of 3 Batch Run # 36 of 59 Data File C:\HPCHEM\1\DATA\HolOl26\HILLOll4.D _ _# - - ~ -[-n-g-/m-L-l- - - - -I-_-_ _ _ _ _ _ _ _ _ - _ _ _ _ _ _ _ _ - - - - ~ - 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1: MSDl 299, EIC=298.7:299.7 Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 Totals without ISTD(s) : 0.00000 Totals without ISTD(s) : 0.00000 BACK TO MAIN Sample Name: MeOH Blank Sorted By Calib. Data Modified : Multiplier Dilution Signal 1/30/01 9:16:50 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298.7:299.7, Totals : 0.00000 Instrument 1 1/30/01 9:20:18 AM KLT/MLA FACT-TCROO2 (LIMS #E00-1716), Page 72 of 109 Page 2 of 3 Batch Run # 36 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOll4.D Signal 2: MSDl 427, EIC=426.7:427.7 BACK TO MAIN Sample Name: MeOH Blank Signal 3: MSDl 499, EIC=498.7:499.7 Peak RetTime Type ----1#-------1--[---m-1i--n--l---1---------- 1 5.144 Width [minl 0.0000 Area 0.00000 Totals : 0.00000 1 Warnings or Errors : Warning : ISTD cornpound(s1 not found Name Batch Run # 56 of EO Data File C:\HPCHEM\1\DATA\HOlOl29\HILLOO56 .D BACK TO MAIN Sample Name: MeOH Blank \ 2000 I 1750 1500 2 4 6 ,8 min MSDl 427. ElC=426.7:427.7 (HOlOl29WILLOO56.D) API-ES, Neg, SIM, Frag: Var 1 1 12000 10000 8000 6000 4000 7 - 2000 : 0- - -__--_I_- 1600 1200 1000 800 : 600 { sorted By signal Calib. Data Modified : 1/31/01 10:09:45 AM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: --- ISTD ISTD Amount Name Instrument 1 1/31/01 10:30:50 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 74 of 109 Page 1 of 3 BACK TO MAIN Sample Name: MeOH Blank Signal 1: MSDl 299, EIC=298.7:299.7 Totals without ISTD(s1 : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 Totals without ISTD(s) : 1 Warnings or Errors : Warning : ISTD compound(s) not found 0.00000 Sorted By Calib. Data Modified : Multiplier Dilution Signal 1/31/01 10:09:45 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298 7 :299.7 Peak RetTime Type Width - - -#- I - -[-m-i-n-l- I - - _ _ _I -- -[-mi_n_l - - 1 4.110 I 0.0000 Totals : 0.00000 Instrument 1 1/31/01 10:30:50 AM KLT/MLA FACT-TCROO2 (LIMS #E00-1716), Page 75 of 109 Page 2 of 3 Batch Run # 56 of 80 Data File C:\HPCHEM\1\DATA\HolOl29\HILLOOS6.D Signal 2: M S D l 427, EIC=426.7:427.7 Totals : 0.00000 Signal 3: M S D l 499, EIC=498.7:499.7 Totals : 1 Warnings or Errors : 0.00000 BACK TO MAIN Sample Name: MeOH Blank Instrument 1 1/31/01 10:30:50 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 76 of 109 Page 3 of 3 Batch Run # 40 of 79 Data File C :\HPCHEM\l\DATA\HO10126\HILLO040.D BACK TO MAIN Sample Name: oPFOS-1.4.1 TCR-002, 012201-oPFOS-1.4.1,lOOX Dil. M_. e._t. h._o. d._. _.B.l-.a_.n.k_.,._.S._F.M-._.0._1.-._2. .4_.-_.0.1-.-M._E.E._(.M._L. A_. )._. ._. ._. _. ._. ._. ._. _. ._. ._. ._._. ._. ._. ._._. ._. ._. .-.- - - - - - - - - - - - - - - - - - - - - - - - - - Injection Date : 1/26/01 9:18:01 PM Seq. Line : 40 Sample Name : OPFOS-1.4.1 Vial : 14 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary I n j Volume : 10 p1 Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed ,: 1/25/01 9:13:04 AM by M L A Analysis Method : C:\HPCHEM\l\METHODS\HOl0126A.M Last changed : 1/30/01 8:17:48 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil Sum 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on IIDioanal' . KLIT MSDl 299, ElC=298.7:299.7 (HOl0126V11LL0040.D) API-ES, Neg, SIM, Frag: Var 300000 250000 200000 150000 100000 50000 0 175000 150000 125000 A I 2 MSDI 427, ElC=426.7:427.7 (H010126WILL0040.D) k" 1 4 6 API-ES, Neg, SIM, Frag: Var I I 8 min I Instrument 1 1/30/01 8:21:25 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 77 of 109 Page 1 of 2 Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----I#------------[-I-n---g--/---m---~--l----------- 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1/30/01 8:17:46 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298.7:299.7 RetTime Type Area Amt/Area Amount - -[-m-in-l- - I - - - - - - i - - - - - - - - - - l - -r-a-t-i-o- - - - - _[n-g- -/-a-l- - - 4.108 VBA+ 12.16967e6 1.00000 254.00000 Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, ~1~=426.7:427.7 RetTime Type [minl Area Amt/Area ratio Totals without ISTD(s) : 0.00000 1 Warnings or Errors : Warning : Calibrated compound(s) not found BACK TO MAIN Instrument 1 1/30/01 8:21:25 AM KLT/MLA FACT-TCROOP (LIMS #E00-1716),Page 78 of 109 Page 2 of 2 ~~ Batch Run # 20 of 59 Data File C:\HPCHEM\1\DATA\HOlol26\HILLOO98.D BACK TO MAIN Sample Name: oPFOS-2.4.1 TCR-002, 012201-oPFOS-2.4.1, 1OOX Dil. M. e. .t.h.o.d. .B.l..a..n..k..,....S..F..M.....0..1..-...2..5..-..0.1.M. E. E. (. .M.L.A.). . . . . . ................................ . . . . . . . . . . . Injection Date : 1/27/01 8:03:06 AM Seq. Line : 98 Sample Name : OPFOS-2.4.1 Vial : 30 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj Volume : 10 p1 Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed ,: 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126B.M Last changed : 1/30/01 9:16:52 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on I1Dioana1l LT MSDI 299, ElC=298.7:299.7 (H010126WlLL0098.D) API-ES, Neg, SIM. Frag: Var 1 300000 I + 250000 200000 I 150000 4 100000 I - I I I I 225000 200000 175000 150000 125000 100000 75000 50000 25000 I k05 '1$' 2 4 6 a mir Instrument 1 1/30/01 9:18:47 AM KLT/MLA FACT -TCR002 (LIMS #E00-1716), Page 79 of 109 Page 1 of 2 Batch Run # 20 of 59 Data File C:\HPCHEM\l\DATA\HOlOl26\HILL0098.D Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----I#------------[-In--g--/--m--L--l--------------- 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1/30/01 9:16:50 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298.7:299.7 RetTime Type - - -[- - -m- Ii- - -d- - - 4.110 VBA+ I Totals without ISTD(8) : 0.00000 Signal 2: M S D l 421, EIC=426.7:427.7 Area &nt/Area - - _ _ - - - _I_---_--r--a---tio 1.20233e6 1.00000 . Totals without ISTD(s) : Signal 3: MSDl 499, EIC=498.7:499.7 RetTime Type [minl Area &nt/Area ratio Amount G r p Name [ng/fil BACK TO MAIN Sample Name: oPFOS-2.4.1 1 Warnings or Errors : Warning : Calibrated cornpound(s) not found Instrument 1 1/30/01 9:18:47 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 80 of 109 Page 2 of 2 Batch Run # 40 of 80 Data File C:\HPCHEM\1\DATA\HolOl29\HILLOO4O.D BACK TO MAIN Sample Name: oPFOS-3.4.1 250000 200000 L 150000 { 100000 1 50000 4 05 * I $f?*+ a%' Q> , - 175000 &% 150000 125000 100000 75000 50000 25000 0, I .. 2 4 6 MSDI 499. EIC=498.7:499.7 (H010129WILL0040.D) API-ES. Nea. SIM. Frag: Var 1800 1200 1000 8 mil Instrument 1 1/31/01 10:29:28 AM KLT/MLA FACT-TCR002 (LIMS #E00-1716), Page 81 of 109 Page 1 of 2 Batch Run # 40 of 80 Data File C:\HPCHEM\1\DATA\HOlOl29\HILLOO4O.D Sorted By Calib. Data Modified : Multiplier Dilution Sample I s m Information: ISTD ISTD Amount Name ----I#-------------I[--n---g---/---m---~--l--------- 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1/31/01 10:09:45 AM 1.0000 1.0000 BACK TO MAIN Sample Name: oPFOS-3.4.1 Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type Area Amt/Area ---[--m--iI-n---l--I---------- I------r--a--It--i--o------I 4.924 VBA+ 11.03940e6 1.00000 Amount Grp Name [ng/al -- I --------------____ 254.40000 THPFOS Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 1 Warnings or Errors : Warning : Calibrated compound(s) not found Instrument 1 1/31/01 10:29:28 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 82 of 109 Page 2 of 2 Batch Run # 48 of 79 Data File C:\HPCHEM\1\DATA\Holol26\HILLOO48.D BACK TO MAIN Sample Name: oPFOS-1.1.4 TCR-002, 012201-oPFOS-1.1.4, lOOX Dil. SFM 01-24-01 MEE (MLA) ==I=P=======IP======I=P===X=IIEPI===================~=====~==================== Injection Date : 1/26/01 10:47:12 PM Seq. Line : 4 8 Sample Name : OPFOS-1.1.4 Vial : 21 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj Volume : 10 pl Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed ,: 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126A.M Last changed : 1/30/01.8:17:48 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on "Dioana1I KLT MSDl 299, ElC=298.7:299.7(H010126\HILL0048.D) API-ES, Neg, SIM, Frag: Var , 300000 L e' ,,> < 250000 200000 I 150000 L 100000 1 50000 ! 0; I i I I I 175000 150000 125000 100000 75000 50000 25000 I 4% .ci.Q I -. 2000000 1750000 1500000 1250000 1000000 750000 500000 250000 Instrument 1 1/30/01 8:22:08 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 83 of 109 Page 1 of 2 Batch Run # 48 of 79 Data File C:\HPCHEM\1\DATA\HolOl26\HILLOO48.D Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTI) Information: ISTD ISTD Amount Name ----I#------------[- ng/m~l 1 254.00000 2 254.40000 Signal . 1/30/01 8:17:46 AM 1.0000 1.0000 Signal 1: MSDl 299 EIC=298.7 :299.7 RetTime Type - ; - [- -m- -iI -n- -l- - - 4.110 VBA+ I Totals without ISTD(s) : ' G r p Name 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type [minl Area &nt/Area ratio Amount G r p Name BACK TO MAIN Sample Name: oPFOS-1.1.4 Instrument 1 1/30/01 8:22:08 AM KLT/MLA FACT-TCROOZ (LIMS #E00-1716), Page 84 of 109 . Page 2 of 2 Batch Run # 55 of 79 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOO55.D BACK TO MAIN Sample Name: oPFOS-1.2.4 TCR-002, 012201-oPFOS-1.2.4, lOOX Dil. S. .F.M. .0.1.-.2.4.-.0.1. .M.E.E.(.M.L.A.). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Injection Date : 1/27/01'12:05:06 AM Seq. Line : 55 Sample Name : OPFOS-1.2.4 Vial : 25 Acq. Operator : MLA Acq. Instrument : Hillary Inj : 1 Inj Volume : 10 p1 Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed ,: 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126A.M Last changed : 1/30/01 8:17:48 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on "Dioanafl LT MSDI 299, ElC=298.7:299.7 (H010126WlLL0055.D) API-ES, Neg, SIM, Frag: Var 1 300000 250000 200000 L I50000 L 200000 175000 150000 125000 100000 75000 50000 25000 -I 2000000 : 15Ooooo 1 1000000 1 500000 - 0, I 2 I 49 .c;.Q 69' I' I L - I I 4 6 , I I I I 8 mir Instrument 1 1/30/01 8:22:45 AM ICLT/MLA FACT -TCR002 (LIMS #E00-1716), Page 85 of I09 Page 1 of 2 Batch Run # 55 of 79 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOO55.D Sorted By Signal Calib. Data Modified : 1/30/01 8:17:46 AM Multiplier 1.0000 Dilution Sample ISTD Information: . 1.0000 ISTD ISTD Amount Name - - #- - I - - -[-n-g-/-f-i-l- - - - I - - - - - - - - - - - - - - - - - - - - - - - - - 1 254.00000 PFBS 2 254.40000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 Signal 3: MSDl 499, EIC=498.7:499.7 Totals without ISTD(s) : 561.33095 BACK TO MAIN Sample Name: oPFOS-1.2.4 Instrument 1 1/30/01 8:22:45 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 86 of 109 Page 2 of 2 Batch Run # 60 of 79 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOO6O.D BACK TO MAIN Sample Name : oPFOS-1.3.4 TCR-002, 012201-oPFOS-1.3.4, lOOX Dil. SFM 01-24-01 MEE (MLA) .... ............................ .. .... ......................... .. .... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Injection Date : 1/27/01 1:00:45 AM Seq. Line : 60 Sample Name : OPFOS-1.3.4 Vial : 29 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj Volume : 10 pl Acq. Method : C:\HPCHEM\l\METHODS\H010125.M Last changed e 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126A.M Last changed : 1/30/01 8:17:48 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on DioanaIt MSDl299, ElC=298.7:299.7 (H010126WlLL0060.D) API-ES, Neg, SIM, FEU: Var , ' i" 300000 250000 200000 150000 + 100000i 50000 I 0; - 200000 175000 I50000 125000 100000 75000 50000 25000 I (p" 4 c$0' I 2000000 1 1500OOO : 1000000 1 500000 1 I ;" !, Instrument 1 1/30/01 8:23:11 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 87 of 109 Page 1 of 2 Batch Run # 60 of 79 Data File C:\HPCHEM\1\DATA\HOlOlZ6\HILLOO6O.D Sorted By signal' Calib. Data Modified : 1/30/01 8:17:46 AM Multiplier 1.0000 Dilution : ' 1.0000 Sample ISTD Information: ISTD ISTD Amount Name - -#- - I - - -[-n-g-/-f-i-1- - - - I - - - - - - - - - - - - - - - - - - - - - - - - - 1 254.00000 PFBS 2 254.40000 THPFOS Totals without ISTD(S) : 0.00000 Signal 2: MSDl RetTime Type - -[-m-in-l- - I - - - - - 4.878 VPA+ I Totals without Signal 3: MSDl 499, EIC=498.7:499.7 RetTime Type [minl Area Amt /Area ratio Amount [ng/mtI BACK TO MAIN Sample Name: OPFOS-1.3.4 Instrument 1 1/30/01 8:23:11 AM KLT/MLA FACT-TCROO2 (LIMS #E00-1716), Page 88 of I09 Page 2 of 2 Batch Run # 28 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOlO6.D BACK TO MAIN Sample Name: oPFOS-2.1.4 TCR-002, 012201-oPFOS-2.1.4, lOOX Dil. S.-F.-.M-.-.0-.1-.--.2-.5-.--.0_.1._.M.-E.-E.-(._M.L._A.)_. ._. ._. _. ._. ._. ._._. ._. ._. ._._. ._. ._._. ._. ._. ._._. ._. ._. _. ._. .-.-.-.-.-.-.-.-. --------------------- Injection Date : 1/27/01 9:32:10 AM Seq. Line : 106 Sample Name : OPFOS-2.1.4 Vial : 37 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj Volume : 10 pl Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed ,: 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126B.M Last changed : 1/30/01 9:16:52 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil Sum 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on ttDioanatl 1" LT MSDl 299, ElC=298.7:299.7 (H010126WlLL0106.D) API-ES, Neg, SIM, Frag:Var 1 4 300000 250000 200000 ; ! I 15OOOO I I 100000 I ~ I 50000 I I 0; \, -. 200000 175000 150000 125000 100000 75000 50000 25000 6%. )* I ~ - \ -- 2000000 - 1500000 7 1000000 7 500000 - 01 1 2 ilI \ , I I 4 6 I 8 mir Instrument 1 1/30/01 9:19:35 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 89 of 109 Page 1 of 2 Batch Run # 28 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOlO6.D Sorted By Signal Calib. Data Modified : . 1/30/01 9:16:50 AM Multiplier 1.0000~ Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name - - -#- I - - - -[-n-g-/-r-n-L-l- - I - - - - - - - - - - _ _ _ _ _ - _ - _ _ _ _ _ _ _ 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1: MSDl 299, EIC=298.7:299.7 BACK TO MAIN Sample Name: oPFOS-2.1.4 Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type Area AmtjArea Amount G r p Name ----[--m-Ii--n--l--I----------l----------1----------1--1---------.-------r--atio [ng/dI 4.878 VBA+ 11.15060e6 1.00000 254.40000 THPFOS Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 RetTime Type Area Amt/Area ---[--m--iI-n---l--I----------l----------l----------l--l------------r--a---t- io 5.144 PVA+ 1.49753e7 1.75101e-1 Amount G r p [ng/rn~l Name 579.77372 PFOS Totals without ISTD(s) : 579.77372 Instrument 1 1/30/01 9:19:35 AM KLT/MLA FACT -TCROOP (LIMS #E00-1716), Page 90 of 109 Page 2 of 2 Batch Run # 34 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOll2.D BACK TO MAIN Sample Name: oPFOS-2.2.3 300000 I 250000 I 100000 2 50000 I 0; 200000 175000 150000 125000 100000 75000 50000 25000 2000000 1750000 1500000 1250000 1000000 750000 500000 250000 ' qQ@ I $0' I " 1 I ( 1 I .cl.Q49 69' L' Q 8 I Instrument 1 1/30/01 9:20:08 +M KLT/MLA FACT-TCROO2(LIMS #E00-1716),Page 91 of 109 Page 1 of 2 Batch Run # 34 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOll2.D BACK TO MAIN Sample Name: oPFOS-2.2.3 Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount ----I#------------[- ng/fil 1 254.00000 2 254.40000 Signal 1/30/01 9:16:50 AM 1.0000 1.0000 Signal 1: MSDl 299 EIC=298.7:299.7 Totals without Amount G r p Name _[-n-g- -/-a-I- - - - - I - - - - - - - - - - - - - - - - - - 254.00000 PFBS 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type [minl Area Amt/Area ratio Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 Totals without ISTD(s) : 553.69686 Instrument 1 1/30/01 9:20:08 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 92 of 109 Page 2 of 2 Batch Run # 35 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOll3.D BACK TO MAIN Sample Name: oPFOS-2.2.4 TCR-002, 012201-oPFOS-2.2.4, lOOX Dil. SFM 01-25-01 MEE (MLA) ==========P=PP====pI='=====z=========xI============================== Injection Date : 1/27/01 10:50:10 AM Sample Name : oPFOS-2.2.4 Acq. Operator : MLA Acq. Instrument : Hillary Seq. Line : 113 Vial : 41 Inj : 1 Inj Volume : 10 p1 Acq. Method : C:\HPCHEM\l\METHODS\HOl012S.M Last changed : 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0126B.M Last changed : 1/30/01 9:16:52 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on "Dioana" LT MSDl299, EIC=298.7:299.7(HOl0126WILL0113.D) API-ES, Neg, SIM, Frag: Var 300000 250000 200000 4 150000 II 100000 50000 '-L---------- I I I I 200000 175000 150000 125000 100000 75000 50000 25000 I 4% I 1 4 I fl I 1 I. 2000000 1 1750000 1500000 1250000 1000000 750000 500000 250000 Instrument 1 1/30/01 9:20:13 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 93 of 109 Page 1 of 2 Batch Run # 35 of 59 Data File C:\HPCHEM\1\DATA\HolOl26\HILLOll3.D Sorted By Signal Calib. Data Modified : , 1/30/01 9:16:50 AM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name ~ _# ~ - ~[n- g-/-m-~-l- - - -I .-.-. .-.-. . . . . . . . . . . . . . . . . . . . 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1: MSDl 299, EIC=298.7:299.7 Totals without ISTD(s1 : 0.00'000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime - -[-m-i-n-l- BACK TO MAIN Sample Name: oPFOS-2.2.4 Totals without ISTD(s) : 552.49733 Instrument 1 1/30/01 9:20:13 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 94 of 109 Page 2 of 2 Batch Run # 40 of 59 Data File C:\HPCHEM\1\DATA\HOlOl26\HILLOll8.D BACK TO MAIN Sample Name: oPFOS-2.3.4 TCR-002, 012201-oPFOS-2.3.4, lOOX Dil. S..F.M..0.1.-.2.5.-.0.1..M.E.E..(.M.L.A.)............................................................. Injection Date : 1/27/01 11:45:54 AM Seq. Line : 118 Sample Name : OPFOS-2.3.4 Vial : 45 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj Volume : 10 pl Acq. Method : C:\HPCHEM\l\METHODS\HOl0125.M Last changed ,: 1/25/01 9:13:04 AM by MLA Analysis Method : C:\HPCHEM\1\METHoDs\HOlOl26B.M Last changed : 1/30/01 9:16:52 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium' Acetate Reprocessed on "Dioana" LT MSDl 299, ElC=298.7:299.7(H010126WlLLOll8.D) API-ES, NW, SIM, Frag:Var - 250000 { ,6"' 200000 : < 150000 100000 4 < 50000 0; L I I I I < 175000L 1 150000 125000 2000000 1750000 1500000 1250000 1000000 750000 500000 250000 Instrument 1 1/30/01 9:20:38 &M KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 95 of 109 Page 1 of 2 ~ ~ ~~~~ Batch Run # 40 of 59 Data File C:\HPCHEM\1\DATA\Holol26\HILLOll8.D sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name - - -#- I - - - -[-n-g- -/-m-~- l- - I - - - - - - - 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1/30/01 9:16:50 AM 1.0000 1.0000 Signal 1: MSDl 299I EIC=298.7:299.7 RetTime Type Area Amt/Area Amount G r p - -[-m-i-nl- - I- - - - - _ - - - - - - - - - - I - -r-a-t-i-o- - - - i -t-n-g-/-m-~-l- - - 4.109 VBA+ I 2.11655e6 1.00000 254.00000 Totals without ISTD(r3) : 0.00000 Name Signal 2: MSDl 427, EIC=426.7:427.7' RetTime Type Cminl Area Amt/Area Amount G r p Name Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 BACK TO MAIN Sample Name: oPFOS-2.3.4 Totals without ISTD(s) : 595.91123 Instrument 1 1/30/01 9:20:38 AM KLT/MLA FACT -TCROOP (LIMS #E00-1716), Page 96 of 109 Page 2 of 2 Batch Run # 4 8 of 8 0 Data File C:\HPCHEM\1\DATA\HOlOl29\HILLOO48.D BACK TO MAIN Sample Name: oPFOS-3.1.4 TCR-002, 012201-OPFOS-3.1.4, lOOX Dil. SFM 01-26-01 MEE (MLA) ..................................................................... Injection Date : 1/29/01 10:26:40 PM Seq. Line : 4 8 Sample Name : oPFOS-3.1.4 Vial : 34 Acq. Operator : MLA Inj : 1 Acq. Instrument : HiLlary Inj Volume : 10 pl Acq. Method Last changed : C:\HPCHEM\l\METHODS\HOl0129.M ,: 1/29/01 1:35:12 PM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0129A.M Last changed : 1/31/01 10:09:47 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on 18Dioana1' JT MSDl 299, ElC=298.7:299.7 CH010129WlLL0048.D) API-ES, Neg, SIM, Fmg: Var I I 250000 1 I +@ Q 200000 1 150000 1 100000 7 1 [bJ - 50000 : 0- I 1 I I 175000 < 150000 125000 100000 75000 50000 25000 I 455 +'4 I L 1750000 1500000 1250000 1000000 750000 500000 250000 Instrument 1 1/31/01 10:30:09 AM KLT/MLA FACT-TCROO2 (LIMS #E00-1716), Page 97 of 109 Page 1 of 2 Batch Run # 48 of 80 Data File C:\HPCHEM\1\DATA\HolOl29\HILLOO48.D Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----I#-------------[I--n---g---/---m--~---l--------- 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1/31/01 10:09:45 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298.7:299.7 BACK TO MAIN Sample Name: oPFOS-3.1.4 Totals without ISTD(s) : 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type Area ' Amt/Area Amount G r p Name . ---[--m--iI-n--l---I----------~----------~----------~--~--------r---a--t--i--o- [ng/m~l 4.925 VBA+ I 9.86834e5 1.00000 254.40000 THPFOS Totals without ISTD(s) : 0.00000 Totals without ISTD(s) : * 554.13421 Instrument 1 1/31/01 10:30:09 AM KLT/MLA FACT -TCROO2 (LIMS #EOO-i716), Page 98 of 109 Page 2 of 2 ~ ~ ~ ~~~~~~ Batch Run # 55 of 80 Data File C:\HPCHEM\1\DATA\HolOl29\HILLOO55.D BACK TO MAIN Sample Name: oPFOS-3.2.4 TCR-002, 012201-oPFOS-3.2.4, lOOX Dil. S.F..M..0.1.-.2.6.-.0.1..M..E..E...(...M..L..A..)................................................................. . Injection Date : 1/29/01 11:44:43 PM Seq. Line : 55 Sample Name : oPFOS-3.2.4 Vial : 38 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj volume : 10 pl Acq. Method : C:\HPCHEM\l\METHODS\HOl0129.M Last changed ; 1/29/01 1:35:12 PM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0129A.M Last changed : 1/31/01 10:09:47 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um 50x2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on "Dioana*I LT MSDl 299, ElC=298.7:299.7 (H010129WlLL0055.D) API-ES, Neg, SIM, Frag: Var 1 200000 150000 < 100000 50000 I 0- - I I I I 100000 < 80000 < 60000 L 40000 I /-I 1750000 1500000 1250000 1000000 750000 500000 250000 Instrument 1 1/31/01 10:30:45 AM KLT/MLA FACT -TCROO2 (LIMS #E00-1716), Page 99 of 109 Page 1 of 2 Batch Run # 55 of 80 Data File C:\HPCHEM\1\DATA\Holol29\HILLOO55.D Sorted By Signal Calib. Data Modified : 1/31/01 10:09:45 AM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name _ -#_ - I - _ -t-n-g-/-m-L-I- - - - I . . . . . . . . . . . . . . . . . . . . . . . . . 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1: MSDl 299, EIC=298.7:299.7 BACK TO MAIN Sample Name: oPFOS-3.2.4 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type Area Amt/Area ----[--m-Ii--n--l--I----------l----------l----------l--l----------------r--atio 4.927 VBA+ 19.85509e5 1.00000 Amount G r p Name [ng/aI 254.40000 THPFOS Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 Totals without ISTD(s1 : 538.37559 Instrument 1 1/31/01 10:30:45 AM KLT/MLA FACT -TCROOP (LIMS #EOO-1716),Page 100 of 109 Page 2 of 2 Batch Run # 60 of 80 Data File C:\HPCHEM\1\DATA\HOlOl29\HILLOO6O.D BACK TO MAIN Sample Name: oPFOS-3.3.4 TCR-002, 012201-oPFOS-3.3.4, lOOX Dil. S---F---M-----0---1-------2---6------0---1-----M--E---E-_-(--_M--L-_A--)_---_-=_---_---_---_---_---_--_---_---_---_-.--_.._.._.._.._.._.._.._.._............ Injection Date : 1/30/01 12:40:27 AM Seq. Line : 60 Sample Name : OPFOS-3.3.4 Vial : 42 Acq. Operator : MLA Inj : 1 Acq. Instrument : Hillary Inj volume : 10 p1 Acq. Method : C:\HPCHEM\l\METHODS\HOl0129.M Last changed ,: 1/29/01 1:35:12 PM by MLA Analysis Method : C:\HPCHEM\l\METHODS\HOl0129A.M Last changed : 1/31/01 10:09:47 AM by KLT/MLA (modified after loading) PFOS Analysis Column: C18 Betasil 5um SOX2 mm Eluents: Methanol and 2 mm Ammonium Acetate Reprocessed on l1Dioanat1 . 250000 : 200000 7 150000 1 qe95 20%' 1 100000 : 50000 7 0-- A \ I I I I - < 140000 120000 L 100000 2 80000 I 60000 L 40000 I 20000 I I G5 4 I 92"' b. I J 1600000 1 1400000 I I 800000 4 600000 I 400000 I 200000 - Instrument 1 1/31/01 10:31:10 AM KLT/MLA FACT -TCROOP (LIMS #EOO-1716), Page 101 of 109 Page 1 of 2 Batch Run # 60 of 80 Data File C:\HPCHEM\1\DATA\HOlOl29\HILLOO6O.D Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----#I-----------[--nI-g---/--m---~--l-------------- 1 254.00000 PFBS 2 254.40000 THPFOS Signal 1/31/01 10:09:45 AM 1.0000 1.0000 Signal 1: MSDl 299, EIC=298.7:299.7 RetTime Type - -[-m-in-l- - I - - - - - - 4.108 VBA+ I Totals without ISTD(s) : G r p Name 0.00000 Signal 2: MSDl 427, EIC=426.7:427.7 RetTime Type [minl Area &nt/Area ratio Amount [ng/m~l 4.924 VBAc 19.61252e5 1.00000 254.40000 Totals without ISTD(s) : 0.00000 Signal 3: MSDl 499, EIC=498.7:499.7 G r p Name Totals without ISTD(s) : 589.66683 BACK TO MAIN Sample Name: oPFOS-3.3.4 Instrument 1 1/31/01 10:31:10 AM KLT/MLA FACT-TCROO2 (LIMS #E00-1716), Page 102 of 109 Page 2 of 2 BACK TO MAIN ATTACHMENDT: DEVIATIONFSROM THE METHOADND PROTOCOL FACT -TCROO2 (LIMS #E00-1716), Page 103 of 109 BACK TO MAIN Record of Deviation I. Identification - Study / Project No. PFOS Octanol Solubility/ FACT-TCR002 ---____l__ Deviation Type: 0 SOP RMethod 0 Equipment Procedure (Check one) 0 Protocol 0 Other: ~ ----_-_.I -~~ Document Number: ETS-8-155.0 - ----------__- i Date(s) of occurrence: 26 Jan, 01 to 29 Jan, 01 i ! II. Description: . -R--_e_quired Procedure/*process: ETS-8-155.0 section 11.3 states "For m. ma ose-s--o_f_a_c_c_ur_a-cy when quantitating low levels of analyte, it may be necessary to use the`lowend of the - ca_ liI b_ ra_ tio_ n c_ urve rather than the full range." The method does not discuss fu_r_th_er_m__et-hods of - ex- c_ lud_ in_ g s_ tanI d_ ards from the calibration-curve. -- Actual Procedure/process: Since the exact range of the sample concentration was unknown, a large range of standards were analyzed fiom 2.50-1002 ng/mL. And as section 11.3 eludes to, it was necessary to use the part of the curve that was more appropriate to the data range, rather than the full range for purposes of accuracy when quantitating samples in that da-ta-range. --_I--___ -Theref__o- re the higher end-of the curve was used to quantitate the hiiher levels~of-a-na-ly-te- (-550610 ng/mL on column) to provide better accuracy at the respective data range. It was documented in the raw data that the 2.5,5, and 10 ng/mL standards were not included in the ~ l__l__ -I___----_..- .c...a..l..i..b...r.a...t..i.o...n....c...u...r..v...e...s...i.n...c...e....t.he sample range was at _th_e_u_p_p-e_r_r_a_nge of th- e curve. Also, _.I--__-.-.----.-__..___..___ not all standards at the low end of the curve satisfied the +/- 15% accuracy requirement, and therefore . ----.-----___..... --.--.-I___- were not included. Subsequent batcheqi.e. Days 2 and 3) were ;precessed the same way to allow ----I__.--~__I_-- for consistency in the way the data wa-s reprocessed and reported. ___ ~ Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) -A_ method deviation was written. Recorded - - - - - A _ - 7-___-- __ Date: 1 6@/4/ - IV. ImDact on Studv/ Proiect - - - - - I - - - - -~ The deviation improves the quality of ;he data becausi the czkbratio; standard range used to quantify the data is more appropriate to the sample range, and is thus more accurate. A u t h o r 6 T B D u d y Director / p j e c t Lead) -7- 3M Environmental Laboratorq! Deviation No. (assigned by Study Director or Pkject Lead at the end of study or project) FACT-TCROOZ (LIMS #E00-1716), Page 104 of 109 BACK TO MAIN Record of Deviation 1. Identification ~ __ Study / Project No. -- ................. ~ -__ Deviation Type: PFOS Octanol Solubility/ FACT-TCR002 ______^---------l___l__~l--__ -I___--.._-___I_____-_._ 0 SOP Fa Method 0 Equipment Procedure (Check one) 0 Protocol 0 Other: . ......... ....... ~ -.-_____._._I_ Document Number: ETS-8-170.0 - i Date(s) of occurrence: -----____ 22 Jan 01 to 26 Jan 01 I 11. Description: -R-- equired Procedure/process: ETS-8-170.0, section 12.1.8 states " If the undissolved_p_a_rt_ic-l-es are still observed, the column elution method s l l be utilized (E'jTS-8-171.O). If no visible particulates are observed, the shake flask me-thod (ETS-8-172.0) shall be used to determine --_ I---___ the solubility." - Actual Procedure/process: Results of the solubility screening test indicated that the column elution method shall be used, however the shake flask method was used due to issues -w- ith octanol. The column elution method requires the test solvent to be compatible -with the _ __I_"__ P tubingI u_ sed,_ and - pre. lim. inary method_ de- v_ eiI o_ pm_ e_ nt_ de_ m_ on_ str_ at- ed-_ tha_ t L tub_ in_ g _ would not be ab--le_- -_t_o withstand exposure to octanol for the anticipated duration of the experiment. _ I _ _ _ _ - - _ ____-__ Another factor to consider is volatility of the solvent. The test solvent may not be volatile, since it is e_-nclosed in an incubator f_o_ r an extended am___o-u-n-t-o-f-tim--e and is not in a sealed - - - - " ~ I _ ___. container. The incubator has electrical outlets on the inside, and would Dose a safety threat in the event of octanol leakage. --- ______- - Ill. Actions Taken: --_--_ (such as ame--n-_d_m__e-n- t issued, SOP revision, etc.) - A method deviation was written. A_no_te_to_f_ile_was al_so_w_rit-t-en and included in the study - ___I-- binder to document that the shake flask was used as an alternative to the column elution method. -- Recorded By: - , -I__--_________ &I@! /I Date: - ---_ IV. Impact on Study/ Project --_- - -- This deviation does not adversely &ect the quality of the data. -- The solubility determination of PFOS in octanol via shake flask method met all data quality objectives. Since the quality control passed, the results are valid and may be -re-po_rte-_d _w_ith confidence. __ _ _ _ _ _ _ _ _ - I_ - _ - I _ --I_----___I_-________________ Authorized By: ($tu& Director / P r o j d Lead) l Date: 3M Environmental Laboratoly / Deviation No. 3 (assigned by Study Director or Project Lead at the end of study or project) FACT-TCROO2 (LIMS #E00-1716),Page 105 of 109 BACK TO MAIN Record of Deviation 1. Identification No. Study Project / .............................................. h i a t i o n Type: (Check one) ~ ~ TF&&x P- FOS Octanol Solubility/ FAC-_T__-_T- CR002 0 SOP 0 Equipment Procedure 0 Protocol 0 Other: ., -.^__l_l_l__ _---- Document Number: ETS-8-155.0 Date(s) ___7.--- j of occurrence: 2 X O T & 2 9 - J 5 3 ? - 8 , 11. Description: R-e- quired Procedure/process: ETS-8-155.0 section10.2.1 states, " Matrix spikes are prepared - I _ - - - for each sample set and analyzed to dete&ne the matrix effect on the recovery efficiency." Section 10.2.3 states, "analyze the-m--a-tr-ix spike and the matrix spike duplicate -~-.-_--_.-.___ ( if prepared) in &e same run as the original sample." Actual Procedure/process: The matrix spikes were not prepared, therefore, they were not Ill. Actions Taken: (such as amendment issued, SOP revision, e- tc.)--- -- - --___-__I_ n e ETS-8Li55.0 method has b e ~ n ~ ~ ~ ~ s o - ~ e s may --_.-- be prepared at the I - - _ _ c I - _ _ Discretion of the project lead. Recorded By: --- 1 Date: - - - ~ [ oy-l'b-#/ IK Impact on Study/ Project msdeviationdoes not adversely affect the quality of the data. --__----_I_-_ _ - _ _ _ _ - - - _ _ _ - - - - I _ - - - - __ %e matrix is well defined and characterized. The creation of matrix spikes are n_o_t a--ctually possible in this study since the_a_n_a_ly-te_of inter_e_s_t_i_s__a_lr_e-_a_d-y_-at saturation concentrat_io_n_s_.-_____ ~ 3M Environmental Laboratory Deviation No. 4 (assigned by Study Director or Project Lead at the end of study or project) FACT -TCROOP (LIMS #E00-1716), Page 106 of 109 BACK TO MAIN Record of Deviation itudy / Project No. _____-______.__-____._.____._._l-l-_-l_-.I_ -_---leviation Type: (Check one) 1. Identification ~ I ~ - ~ -__..-_. ~ L- -.I__ PFOS Octanol Solubility/ FACT-TCR002 0 SOP hp Method 0 Equipment Procedure 0 Protocol 0 Other: --.__ - --Iocument I ____VI__- Number: ETS-8-155.0 -Date(s) of occurrence: 22 Jan 01 to 26 Jan 01 - I/. Description: iequired Procedure/process: ETS-8-155.0 ,section 14.4states "Continuing Calibration-__ derification (CCV)p-e--rc-e--n-t_re_c_ov-eries must be f 30% of the s&ndard concentration. ~~~ __I___._____ _ c - - - - p - - - l 4ctual Procedurehocess: The criteria used for reprocessin-g data was that all CCVs and xilibration standards had to have a % accuracy o f f 15%. This was done to be consistent with he requirement of the s h a k e G k samples to b-e_accurate to wi-thin 15% (according to ETS-8- Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) A method devi_-_a_-_-tion was written. -- Recorded By: 1 Date: I i! &&/ IV. Impact on Study/ Project - .___.I__-________- __._______ I______ The deviation provides better quality, a~ nd more controlled data. The m- ore stringent precision - - - - - - - I -re-quirement of the calibration standards provided a higher level of confidence for the reported samde data. -___-- Authorized By: (Study Director /Project p a d ) 7Date: ~- 3M Environmental Laboratory 5 Deviation No. (assigned by Study Director or Project h a d at the end of study or project) FACT -TCR002 (LIMS#E00-1716), Page 1.07of 109 ATTACHMENET: SAMPLECALCULATIONS BACK TO MAIN FACT -TCROO2 (LIMS #EOO-1716), Page 108 of 109 BACK TO MAIN 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 individualentities. Standard deviation was calculated using the following equation: - f -Where n is the number of observations, and x is sample concentration. Sample ID Rep1 Rep 2 Rep 3 Rep 4 Concentration C19/mL 59.1 57.4 57.6 57.1 Calculations Average (59.1+57.4+57.6+57.1)/4=57.8 Std Dev (see above equation)= 0.88 %RSD (0.88/57.8)= 1.5% FACT -TCR002 (LIMS #E00-1716), Page 109 of 109