Document 4Qp8DVeg1j7G18ar89bD6o5Va

~~ ~ 27) OECD 105-OPPTS 830.7840, solubility studies (water methanol, acetone), #FACT-TCR009 SANITlZED - Study Title DEC 0 9 2003 CharacterizationStudy PFBS: a.) Primary Standard Test Control Reference#TCR- 99030-023 (Lot I O I ) , b.) Test Control Reference#TCR-00017-071 (Lot 2) PHASE: SOLUBILITY DETERMINATION Data Requirement 40 CFR 160.105(b) ~ -~ Author Mark E. Ellefson Phase Completion Date Date of Signing Performing Laboratory 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St.Paul, MN 55106 Project Identification 3M EnvironmentalLaboratory Study ## FACT-TCROO9 (LIMS'#EOO-I877) Centre Analytical Laboratories Study # 023-032 Total Number of Pages 98 3M EnvironmentalLaboratory . Study NO. FACT-TCROOS GLP COMPLIANCSETATEMENT - Study Title: a.) Primary Standard - Test Control Reference #TCR-99030-023 (Lot 1O l ) , b.) Test Control Reference #TCR-00017-071 (Lot 2) PHASE: SOLUBILITY DETERMINATION - Study Identification Number: FACT-TCROOS, Centre Analytical Laboratories Study # 023-032 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. 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. rn Some corrections were not made in the raw data per GLP requirements. The stability for all test and reference substances has yet to be determined (40 CFR 160.185 (a)@)). rn The purity of THPFOS has yet to be determined (40 CFR 160.185(a)(4)). Testing Facility Management 0 3 /J&/ Date ' LIMS# 00-1877 Page 2 of 98 3M Environmental Laboratory Study N O . FACT-TCROOS ~ ~~ QUALITYASSURANCSETATEMENT Study Title: a.) Primary Standard - Test Control Reference#TCR-99030-023 (Lot 1O I ) , b.) Test Control Reference #TCR-00017-071 (Lot 2) PHASE: SOLUBILITY DETERMINATION Study Identification Number: FACT-TCROOS, Centre Analytical Laboratories Study # 023-032 This phase of the study has been inspected by the 3M Environmental Laboratory Quality Assurance Unit (QAU) as indicated in the following table. The findings were reported to the study director and laboratory management. I 07/17/00 I 09/13/00 02/06-08/01 and 02/12-14/01 Sample Prep I Solubility Screen Data Audit I Draft Report #2 I 07/26/00 I 07/26/00 I 0211m1 0315/01 I O~ROIO~ 1 03/20/0i I LIMS# EOO-1877 Page 3 of 98 . 3M Environmental Laboratory Study N O. FACT-TCROOS TABLEOF CONTENTS . GLP Compliance Statement............................................................................................................... 2 Quality Assurance Statement................................................................. ..................................... 3 Study information .... ...................................................................................................................... 6 Summary ............................................................................................. :.............................................. 7 Purpose.............................................................................................................................................. 8 Test Substance .................................................................................................................................. 8 Reference Substance......................................................................................................................... 8 Test System ....................................................................................................................................... 9 Method Summaries ............................................................................. ............................................... 9 Preparatory Methods.......................................................................................................................... 9 Sample Collection and Analysis ......................................................................................................... 11 Analytical Method............................................................................................................................... 12 Analytical Results............................................................................................................................... 13 Data Summary and Discussion.......................................................................................................... 14 Statistical Methods ............................................................................................................................. 17 Statement of Conclusion .................................................................................................................... 17 References......................................................................................................................................... 17 List of Attachments............................................................................................................................. 18 Signature Page................................................................................................................................... 19 LIMS# E00-1877 Page 4 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS LISTOF TABLES Table l a. Summary Table of Solubility of PFBS TCR-00017-071 in Water...................................... 7 Table 1b. Summary Table of Solubility of PFBS TCR-99030-023 in Water...................................... 7 Table IC . Summary Table of Solubility of PFBS TCR-0017-071 and TCR-99030-023 in Methanol and Acetone........................................................................................................................ 7 Table 2. Characterization of the Test Substance and Analytical Reference Substance ...................8 Table 3. Characterization of the Analytical Reference Substance.................................................... 8 Table 4. Description of test systems used in this study .................................................................... 9 Table 5. Summary of Screen Test method Sample Preparation...................................................... 10 Table 6a. Shake Flask Method Preparation for Study (FACT-TCROOS), PFBS TCR-00017-071 ....10 Table 6b. Shake Flask Method Preparation for Study (FACT-TCROOS), PFBS TCR-99030-023 .... 11 Table 7. Sample Collection and Preparation for Study FACT-TCR-009 for both PFBS lots studied...................................................................................... ................................ Table Ea. PFBS TCR-00017-071 (PFBS "a") Data Summary............................................................ 14 Table 8b. PFBS TCR-99030-023 (PFBS "b")Data Summary............................................................ 15 Table 9a. Summary Table of Solubility of PFBS TCR-00017-071 in Water...................................... 16 Table 9b. Summary Table of Solubility of PFBS TCR-99030-023 in Water...................................... 16 Table 9c. Summary Table of Solubility of PFBS TCR-0017-071 and JCR-99030-023 in Methanol and Acetone........................................................................................................................ 17 LlMSt EOO-187 Page 5 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS STUDYINFORMATION Study Director John Flaherty Centre Analytical Laboratory 3048 Research Drive State College, PA 16801 Analytical Chemistry Laboratories Solubility Testing 3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory 935 Bush Avenue Bldg. 2-3E-09 St. Paul, MN 55106 Laboratory Manager Principal Analytical Investigator Analytical Chemist Analytical Chemist Analytical Chemist Sponsor 3M Environmental Laboratory 935 Bush Avenue Building 2-3509 St. Paul, MN 55106 Certificate of Analysis Centre Analytical Laboratory 3048 Research Drive State College, PA 16801 John Flaherty, Study Director Experimental Dates Experimental Start Date: 13 September 2000 Experimental Termination Date: 24 September 2000 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. LlMSt 00-1877 Page 6 of 98 3M Environmental Laboratory Study No. FACT-TCROOS Solubility of PFBS in water is a significant parameter because the spatial and temporal movement (mobility) of a substance is largely determined by its solubility in water. Water soluble substances readily gain access to humans and other living organisms; furthermore the knowledge of the solubility in water is a prerequisite for testing biological degradation, bioaccurnulation, and other tests. Solubility of PFBS in methanol and acetone is a significant parameter for general laboratory knowledge in the preparation and analysis of PFBS. The approximate solubility of PFBS in water was determined by performing a solubility screen test (ETS-8-170.1), followed by a quantitative solubility determination via the Shake Flask Method (ETS-8-172.1) and HPLClMS analysis (ETS8-155.0). The approximate solubility of PFBS in acetone and methanol was determined by petforming a solubility screen test. Results are presented in table 1: Table la. Summary Table of Solubility of PFBSTCR-00017-071inWater. Day 1 Day 2 Day 3 Final Value ~ 57300 pg/rnL 55300 pg/mL 57300 pg/rnL 56600 pglrnL 1 160 pg/rnL 32100 W L 3820 ps/mL 1150 pglmL 2.0% 5.8% 6.7% 2.0% Table lb. Summary Table of Solubility of PFBS TCR99030-023inWater. I 53300 pg/mL I 651 pghL I 1.2% I Day 2 I 52100 pg/rnL I 346 WrnL I 0.7% I Day 3 52500 pe/rnL 702 pg/rnL 1.3% Final Value 52600 VglmL 611 pglrnL I .2% Table IC.Summary Table of Solubility of PFBS TCR-0017-071 andTCR-99030423 in Methanol and Acetone. Methanol Acetone >lo% > 10% The solubility of PFBS TCR-00017-071 in water was determined to be 56,600 pglmL, and PFBS TCR-99030-023 in water was 52,600 p g h L at 22.5-24 'C. The solubility of PFBS TCR-00017-071 and TCR99030-023 in both methanol and acetone was LIMW EOO-1877 Page 7 of 98 3M EnvironmentalLaboratory Study NO. FACT-TCROOS determined to be '10% mass/volume at 22.5 "C. PURPOSE The purpose of this phase of the study was to determine the solubility of PFBS in water, methanol, and acetone as part of the characterization of this test, control, and reference substance. TESTSUBSTANCE PFBS, Test Control Reference # TCR-00017-071, and #TCR-99030-023 Table 2. Characterizationof the Test Substance and Analytical ReferenceSubstance source 1 Wildlife. Lot 2 Cottage Grove, Lot 101 I I Expiration Date I 1 I c Storage Conditions 3M Laboratory Identification Number I 1 1 Physical Description I Not Available Frozen TCR-00017-071 I Fluffywhite crystals I Not Available Frozen TCR-99030-023 F-844211-7038)"D~~ITIZIIVstandard" I IT I white powder I 1 1 Purity 97.3% 96.7% I 'PFBS TCR-99030-23 was designated "primarystandard"at the time of analyses and was therefore utilized as the reference substance to quantify the samples. ~~ REFERENCSEUBSTANCE Table 3. Characterizationof the Analytical ReferenceSubstance I Source I Expiration Date Storage Conditions I SvnQuest Labs I Not Available Frozen -1 I 3M Laboratory Identification Number TCR-00017-047 Lot M75-30 I 1 Physical Description White Powder I I Purity * The punty of THPFOS (currently being determined by CentreAnalytical Laboratory) will not affect any analytical findings. The same lotslsource of both compounds were used throughout the entire study. UMSX EOO-1877 Page 8 of 98 3M Environmental Laboratoty Study NO. FACT-TCROOS TESTSYSTEM Table 4. Descriptionof test systems used inthis study I source I 1 I I I ExpirationDate 1 I 1 I I Storage Conditions , I I Not applicable 2005 2005 Ambient Ambient, Flammable Ambient, Flammable Cabinet Cabinet I 1 1 1 3NMumLbaebroratory ldenmcationNot applicable TN-A: 4604 TN-A: 4616 I Purity/Grade ' I ASTM Type I I HPLC Grade I HPLCGrade I ~~ METHODSUMMARIES The solubility determination of PFBS TCR-00017-071 and TCR-99030-023 in water, methanol, and acetone was performed according to United States Environmental Protection Agency OPPTS 830.7840, "Water Solubility: Column Elution Method; Shake Flask Method" and OECD 105, "Water Solubility" Guidelines, as implemented by 3M Laboratory in methods ETS-8-170.1 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents;" and ETS-8-172.1 "Shake Flask Method; Solubility Determination of a Test Substance in Various Solvents". All analyses for the water solubility portion were performed according to the method ETS-8-155.0 "Analysis of Perfluorooctane Sulfonate or Other Fluorochernicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry". The Solubility Screen Test Method was used to determine the solubility range of PFBS TCR00017-071 and 99030-023 in water, methanol, and acetone. The screen test indicated a solubility of PFBS in methanol and acetone to be greater than 10% at 22.5 "C, and no further testing was required. The screen test indicated a solubility of PFBS in water of greater than 20,000 pg/mL at 22.5 "C. Therefore the Shake Flask Method was used for the quantitative determination of PFBS solubility in water. Detailed descriptions of the Solubility Screen Test Method and Shake Flask methods used in this study are located in Attachment A. PREPARATOMREYTHODS ETS-8-170.1 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents." This method is a pre-requisite to 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 10 mg of PFBS was weighed into a glass vial, and varying amounts of solvent were added in a step-wise fashion. After each solvent addition, the vial was shaken, sonicated, and observed for particulates. The screen test estimated the solubility of PFBS TCR-0017-71 and TCR-99030-023 in methanol and acetone to be greater LlMSlY E00-1877 Page 9 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS than 10% at 22.5 "C.The screen test also estimated the solubility of PFBS in water to be greater than 20,000 pg/mL at 22.5 "Cfor both PFBS lots. This concentration estimationwas 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 extract concentration into the analytical range of 1.24 - 1235 ng PFBS/mL. Table 5. Summa of Screen Test method Sample Pmpamtion. 10 mg test substancewasweighed into tared glass screw-capped vial. 0.1 mL of test solvent (water, methanolor acetone)was addedto glass 9/13/00 9/13/00 Following the addition of test solvent. the vial was capped, shaken vortexed and then sonicated. 4 Following sonication, the sample was checked visually for undissolved 9/13/00 particlesof the test substance. Inacetoneand methanol, all of thetest substanceappearedto be dissolved, andthe solubilitywas then greater than 10%(massholume) and noadditional solubilitytestingwas required. A portion of the test substance remained undissolved in water for both lots,therefore further dilutionswere necessary. 5 0.4 mL additional water was added and the vial was vortexed and 9113/00 sonicated. No undissolvedparticleswere observedat this dilution level. The solubilitywas estimatedto begreater than 20,000@nL andthe shakeflask methodwould then be usedto determinethe exact solubility in water for PFBS TCR-00017-71 and TCR-9903G23. ETS-8-172.1 "Shake Flask Method: Solubility Determination of a Test Substance in Various Solvents." 0 PFBS TCR-00017-071: Approximately 400 mg PFBS was weighed into a tared 15 mL plastic centrifuge tube. Water was added gravimetrically to a total weight of 4 g L- 0.3 g. Nine centrifuge tubes (with PFBS and water) were prepared, along with three method blanks (with 4.0 g k 0.1 g water only). The centrifuge tubes with test substance and the method blanks were placed in a temperature-controlled orbital shaker set at 30 "C. See the following table documenting the shake flask method preparation: Table 6a. Shake Flask Method Prepaationfor Study (FACT-TCROOS). PFSS TCR-00017-071 I temperature-mntrolled o LIMW E00-1877 Page 10 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS PFBS TCR-99030-023:When it was noticed that the amount of PFBS TCR-99030-023 was limited, the total volume for the samples was scaled down so as to not use up the remaining PFBS stock. The volume difference has no affect on the integrity of the results, since only a small aliquot from the supernatant is actually analyzed and the concentration of PFBS is identical. Approximately 140-180mg PFBS was weighed into a tared 15 mL plastic centrifuge tube. Water was added gravimetrically to approximately I .5 g +_ 0.5 g. Nine centrifuge tubes with PFBS and water were prepared, along with three method blanks (tubes with 4.0g 5 0.1g water only). The method blanks were prepared immediately prior to the samples. The centrifuge tubes were all placed in a temperature-controlled orbital shaker set at 30 OC. See the following table documenting the shake flask method preparation: Table 6b. Shake Flask Method Preparationfor Study (FACT-TCROOS), PFBS TCR-99030.023 1 >I45 mg PFBS weighed into tared 15mL centrifugetube 9 2 Water added to centrifugetube such that total weight of PFBS and 9 wateris>1.4g 3 MethodBlank Preparation-Approximately4 rnLwater added to a 3 tared centrifugetube gravimetrically. 4 Samples sealed with tape, vortex-mixed, and placed in 12 temperature-controlledorbiial shaker controlledat 29.530.0 OC, rotationspeed set at approximately250 rpm. 09118/00 0911 8/00 09/18/00 091.18/00 SAMPLE COLLECTION AND ANALYSIS On each of Days 1 , 2.and 3 (approximately 24. 48 and 72 hours after initial sample preparation) three centrifuge tubes containing test substance and one method blank tube were removed for analysis, The four centrifuge tubes were allowed to equilibrate at 22.5-24"Cfor 24 hours, centrifuged, and approximately 200 pL aliquots of supernatant were collected into plastic autovials. The aliquots were diluted serially using two IOOX dilutions (990 pL methano1:lO pL sample) to a dilution factor of 10,OOOX. Then one 1OX dilution (900 pL methano1:lOO pL sample) was performed from the 10,OOOX samples for a final dilution factor of 100,OOOX. The final diluted samples were spiked with internal standard (11pL of 22 pg THPFOSlmL) and analyzed via HPLC/ES/MS according to analytical method ETS-8-155.0. The following table describes the sample collection and preparation regimen: LlMSlE00-1877 Page 11 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS Table 7. Sample Collectionand Preparationfor Study FACT-TCR-009for both PFBS lots studied. 1 At 24,48, and 72 hours,3 PFBS+water, and 4 centrifuge 9/19/00,9R0/00,9/21/00 1 water (method blank) centrifuge tubes are tubeddayAot removedfrom the incubator and equilibrated at 22.5-24 OC for approximately 24 hours per PFBS lot. 2 Aliquots of the supernatant (water containing 4lcentrifuge 9/20/00.9121/00,9/22/00 dissolved PFBS) were collected and tube dispensed into autovials. 3 The aliquots were diluted two times l00X Day 1 AwitIhOmX edtilhuatinoonli(nusminegthaadnuoalwl saysrinpgeerfodrimluetedr)to. . Day produce the final dilution factor of 100,000X. Day 3 9/2wm 9/21/00 9/22/00 4 InternalStandardwas addedto the sample Day 1 dilutions (11 pL.22.2 pgTHPFOSImL methanol, soh. oooO1-24-03) Day 2 Day 3 9L?WOO(100,OOOX dilution) 9/21/00 ( l ~ , o o O Xdilution) 9LWOO (100,ooOX dilution) 5 Samples were analyzed via HPLCIESIMS on Day 1 HP 1100 LChlSD 'Hillary." Day 2 9i201oo (SequenceH092000.s) 9122/00(SequenceH092200.s) Day 3 9R2/00 (SequenceH092200.s) ANALYTICAML ETHOD ETS-8-155.0"Analysis of Pemuorooctane Sulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrospraylMassSpectrometry". Diluted samples (i.e. diluted supernatants) were analyzed using HPLCIESIMS in the negative ion mode. PFBS levels were evaluated versus standards ranging in concentration from 2.5-1000 ng PFBSlmL. Internal Standard quantification was used to best fit the data. Target ions were 499 m/z (deprotonated PFBS), and 427 m/z (deprotonated THPFOS). Analytical Equipment Liquid Chromatograph: Hewlett-Packard@S' eries 1100 Liquid Chromatograph system Analytical column: Keystone@B' etasil" Cte2x50mm, 5pm particle size Column temperature: Ambient Cycle Time: 10.5 minutes Flow rate: 300pUmin Injection volume: 5pL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in water Solvent 8: Methanol LIMS# E00-1877 Page 12 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS Solvent Gradient: Time 0.00 1.00 6.00 9.00 9.50 14.50 %B 10 % 10 % 95 % 95 % 10 % stop Mass Spectrometer: Hewlett-Packard@Series Software: HP ChemStation" 6.0 Fragmentor Voltage: m/z 427=100 V; m/z 299= 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) 1100 APVMass 100 V Spectrometer Detector 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 quantitation over the concentration range appropriate to the data. All calibration curves had least-square fits of 0.990 or greater (Rz values ranged from 0.9946 - 0.9999). Calibration Standards. Standards ranging in concentration from 12.4 to 1235 ng PFBSlmL methanol were used for the calibration curves. Calibration curves were run before and after every analytical sequence. Sample Replicates. Samples 091800-PFBSbl.1.4 and 091800-PFBSb-3.2.4 were excluded from the reported data using Dixon's Q-Test. All reported values had a % Coefficient of Variation of <15% as required by OECD guidelines. Continuing CalibrationVerification: For quantitative determinations, a mid-level matrix calibration check was analyzed every five samples to monitor instrumental drift, with a limit of rt: 30% of the target concentrations. Limit of Quantitation (LOQ). The LOQ is equal to the lowest standard in the calibration curve (12.4 ng PFBSImL). Standard 1 (1.24 ng PFBS/mL) and standard 2 (618 ng PFBSlmL) were excluded as they did not meet the 30% deviation requirements, and by dropping the lower standards from the curve, the data range was better fit. Blanks. Method blanks (Milli-Q water taken through the entire sample preparation, dilution, and analysis process) provided a measure of laboratory contamination. Acceptable values for the blanks were less than 50% of the limit of quantitation (LOQ). Two out of four PFBS TCR-00017-071 Day 1 samples had a level of PFBS >50%. One out of four PFBS TCR99030-023 Day 1 samples had a level of PFBS >50%. The elevated level of PFBS in these samples is inconsequential since there was no instrument contamination (demonstrated in the solvent blanks) and no contamination of the methanol used to dilute L I M S 00-1877 Page 13 of 98 3M EnvironmentalLaboratory Study NO.FACT-TCR009 the samples (demonstrated in the remaining method blanks that did not have significant levels of PFBS. The source of PFBS was therefore a random error and the precision of the samples demonstrates that any significantly elevated levels of PFBS in the actual samples (091800-PFBSb-1.1.4,and 091899-PFBSb-3.2.4) was identified and controlled accordingly. The initial solvent blank in the sample run sequence for PFBS TCROOOl7-071Day 2 data had background levels of PFBS >50% of the LOQ. The background PFBS detected was insignificant because any significant amount PFBS was subsequently flushed from the column and no samples/standardswere affected by it. Specificity (accordingto 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 97.3% for PFBS TCR-00017-71, and 96.7% for PFBS TCR-99030-23. Substances that are stable in water. Hydrolytic stability has yet to be determined. DATASUMMARY AND DISCUSSION Tables 8a and 8b summarize individual sample data. Because both lots of PFBS were tested at the same time, PFBS TCR-00017-071 was designated as 'a", and PFBS TCR-99030-023 was designated " b to help clear confusion. Representativechromatograms are presented in Attachment C. The table displays PFBS "a", TCR-00017-071 and PFBS "b",TCR-99030-023 concentrations for individual injections. Also included are the average concentrations for each shake flask vessel replicate, standard deviation, and % coefficient of variation. Table 8a. PFBS TCROOOl7-071 (PFBS "a")Data Summary LlaylShokeFkask Vesselw1 DaylShakeFW Vesselx2 Day 1 Shake Flask V&#3 Day 2 Shake Flask vessel #l Rep 1 Rep2 Rep 3 Rep 4 Rep 1 Rep2 Rep 3 Rep 4 Rep 1 Rep Rep 3 Rep 4 Rep 1 Rep R ~ 3Q Rep 4 63,700 54.300 56,200 57,400 54,700 55,700 57.500 56,100 56,600 58,900 58,100 56,400 ~~~ 53,100 53,700 54,100 53,000 4070 7.0% 56000 1160 2.1% 57900 1200 2.1 % 53600 51 9 0.97% LlMSl E00-1877 Page 14 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS Day 2 shake Fbsk V&#2 DayZShakeFbsk V&#3 Day3ShakeFbsk vessel 61 oay3shakeFbsk VeSselX? Day3shakEFlask Vessel= ~ Rep 1 Rep2 Rep 3 Rep 4 Rep 1 Rep 3 Rep 4 Rep 1 Rep 3 Rep 4 ~ Rep 1 R+2 Rep 3 R ~ 4D Rep 1 Rep2 Rep 3 -4 51,900 53,800 54,100 54,200 54,100 67,200 55,800 55,400 54,100 55,400 54,900 54,800 54,500 61,200 69,300 56.700 54,700 56,100 55,400 55.500 Table 8b. PFBSTCR-99030-023(PFBS "b") DataSummary 53300 1080 - 2.03% 59000 6090 - 10.3% ___ 54800 535 0.98% 61700 6540 10.60% -~ 55400 574 1.04% DayIShaksnask vessel #1 WlShakeFlaSk Vessei#2 IOa,1 Shake Flask VesJel#3 Day 2 Shake Flask vessel 61 Rep 1 Rep 3 Rep 4 Rep 1 Rep 3 Reo 4 Rep 1 Rep Rep 3 Rep 4 Rep 1 Rep Rep 3 Rep 4 53,900 51,400 56,600 69,600 53,300 52,400 53,000 52.000 52,000 53,300 52.600 55,300 54,000 52,200 52,000 50,800 54000 2600 4.8% 52700 458 0.87% 53300 651 1.22% 52300 1100 2.10% LIMS# E00-1877 Page 15 of 98 3M Environmental Laboratory Study NO. FACT-TCROO9 Day 2 Shake Flask Vessel #2 Day 2 Shake Flask V&#3 Day 3 Shake Fbsk vessel #l llay3ShakeFlask V&#2 Day 3 Shake Flask V&#3 Rep 1 Rep 2 Rep 3 Rep 4 Rep 1 *Rep Rep 3 Rep 4 Rep 1 Rep Rep 3 Rep 4 Rep 1 Rep2 Rep 3 Rep 4 Rep I * Rep 3 Rep 4 51,300 51,200 51,900 52,200 52,700 52,700 52,600 51,300 53,000 52,000 52,400 52.000 52,100 51,300 52.100 147.000 57,600 51,900 51,500 51,900 51700 379 0.73% 52300 57.7 0.11% 52400 503 0.96% 51800 462 0.89% 53200 3410 6.41 % The average PFBS 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 9a. Summary Table of Solubility of PFBS TCR-00017-071 in Water. Day1 ' Day 2 nay 3 Final Value 57300 pg/mL 55300 pg/mL 57300 w/mL 56600 pglmL 1160pg/mL 3210 pg/mL 3820 pg/mL 1150 udmL 2.0% 5.8% 6.7% 2.0% Table 9b. Summary Table of Solubility of PFBS TCR-99030023 in Water. I Day 1 I 53300 p@mL I 651 udmL I 1.2% I Day 2 52100 pg/mL 346 P!WL 0.7% Day 3 52500 j.@mL 702 pg/mL 1.3% Final Value 52600 p g h L 611 &mL 1.2% 3M EnvironmentalLaboratory Study NO. FACT-TCROOS Table 9c. Summary Table of Solubility of PFBS TCR4017-071 and TCR-99030-023 in Methanol and Acetone. I I Methanol I Acetone ~ > 10% The solubility of PFBS TCR-00017-071 in water is 56,600 pg PFBShL at 22.5-24 "C. The solubility of PFBS TCR-99030-023 is 52,600 pg PFBSlmL at 22.5-24 "C. And the solubility of PFBS TCR-00017-071 and TCR-99030-023 in methanol and acetone is >lo% at 22.5 "C. Attachment B contains data summary tables for the water solubility test, and sample preparation worksheets/resultsfor'the methanol and acetone solubility tests. ~ ~~~ - ~ STATISTICAMLETHODS Statistical methods were limited to calculating means and standard deviations. Data that did not meet acceptance criteria as described in the OPPTS and OECD guidelines was excluded using statistical justification provided by Dixon's Q-test. Refer to Attachment E for formulas and example calculations. STATEMENT OF CONCLUSION Under the conditions of the present study, the solubility of PFBS TCR-00017-071 in ASTM Type I water is 56,600 pg PFBSlmL, and the solubility of PFBS TCR-99030-023 in ASTM Type I water is 52,600 pg PFBSlmL at 22.5-24 "C. Under the conditions of the present study, the solubility of PFBS TCR-00017-071 and TCR- 99030-023 in methanol and acetone is >lo% at 22.5 "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. 3. Harris, Daniel C. Statistics. Quantitative Chemical Analysis. 4' ed.; W.H. Freeman and Company: New York, 1982; p 70-71. LIMSX 00-1877 Page 17of 98 3M EnvironmentalLaboratory Study NO. FACT-TCROOS 4. 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. LISTOF ATTACHMENTS Attachment A: Extraction and Analytical Methods 0 Attachment B:. Results Attachment C: Sample Chromatograms 0 Attachment D: Deviations Attachment E: Sample Calculations LlMSt 00-1877 Page i 8 of 98 SIGNATURPEAGE 3M EnvironmentalLaboratory Study NO. FACT-TCROOS - We certify that this report is a true and complete representation of the data for this phase of t h e study: n Principal Analytical Investigator Laboratory Manager Date LlMSI E00-1877 Page 19 of 98 3M Environmental Laboratoly Study NO.FACT-TCROOQ ATTACHMENAT: EXTRACTIOANND ANALYTICALMETHODS LlMSl E00-1877 Page 20 of 98 3M ENVIRONMENTLAALBORATORY 3M EnvironmentalLaboratoly Study NO.FACT-TCROOS SOLUBILITY SCREEN TEST: APPROXafATE SOLUBILITYDETERMINATIOONF A TESTSUBSTANCEIN VARIOUS SOLVENTS Method Number: ETS-8-170.1 Adoption Date: 09/08/00 Effective Date: 03/14/01 Approved By: - Laboratory Manager Date ProjZ t L e 9 - Exact CoFy of Q~iytila! fl&+ lsitial iLzZL4L Date 1.0 SCOPE AND APPLICATION 1.1 Purpose. According to methods set forth by the United States Environmental Protection Agency (US EPA) and the Organization for the Economic Cooperation and Development (OECD) a preliminary study, which will be outlined in this method, serves as a prerequisite to performing solubility testing via the Column Elution Method and Shake Flask Methods. The EPA and OECD solubility testing guidelines state that if the preliminary solubility -= determination test indicates a solubility of > 1O'* gfL (10 ppm), the Shake Flask Method (ETS-8-172.0) is to be used. If preliminary testing indicates a solubility of l o 2 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 particuIar solvents, no further solubility determination will be necessary for compounds having solubility greater than 10% (mass/volume). 1.2 Compatible Analytes. Test substance and degradationproducts for solubility testing. ETS-8-170.1 Solubility Determination: Screen Test Metliod Page 1 of9 LIMSX00-1877 Page 21 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS 1.3 Acceptable Matrices. Aqueous (e.g. Milli-Q water, 0.01M CaCl,), Acetone, Methanol, or other solvent(s) of interest. - 2.0 SUMMARY OF METHOD 2.1 Preliminary Solubility Determination in solvent. Weigh approximately 10 mg of test substance into a 2.5 mL-4 mL glass screw-capped vial. Add test solvent in varying increments to the vial. After each solvent addition, vortex mix for approximately 15 sec., and sonicate 2-5 minutes. Make observations of the solution and visually confirm whether or not particles are present; it may be necessary to allow the solution to sit for up to 5 minutes. Add test solvent in a stepwise fashion up to 2 mL, If the substance hlly dissolves after the first solvent addition of 100 pL,the test substance shall be considered highly soluble, and no further testing is required. If there are still undissolved particles in the vial, weigh 10mg of test substanceinto 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 10 ppm). The container should be sonicated and allowed to sit overnight to allow for maximal dissolution. If the undissolved particles are still observed after sitting > 12 hours, the column elution method will be utilized (ETS-8-171.0). If the test substance dissolves in 1 L or less of solvent, the shake flask method (ETS-8-172.0) will be used to determine the solubility. 3.0 DEFINITIONS 3.1 Test substance. A liquid or solid material for which the relative solubility in a specified solvent will be determined. 3.2 Test solvent. The matrix to which the test substance of interest is introduced. The test solvent may include but is not limited to aqueous matrices, including ASTM Type I Water and vsUious salt matrices (e.g. 0.01M CaCI,(,J; and organic solvent matrices, including methanol, and acetone. 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Wear the proper lab attire for all parts of these procedures. Wear gloves and eye protection at all times. 4.1.2 Handle all solvents in a hood for all parts of the described sample preparation procedure. 4.1.3 For potential hazards of each chemical used, refer to material safety data sheets, packing materials, and 3M Environmental Laboratory's Chemical Hazard Review. 4.1.4 No mouth pipetting is allowed. ETS-8-170.1 Solubility Determination: Screen Test Method Page 2 of 9 LIMS# 00-1877 Page 22 of 98 3M EnvironmentalLaboratory Study NO. FACT-TCROOS 4.2 Cautions: 4.2.1 Glassware in which standards are prepared should be triple rinsed with acetone and methanol to reduce the possibility of accidental contamination. 5.0 INTERFERENCES 5.1 Impurities may significantly affect the solubility of the test substance. The purity of the test substance should be known and documented prior to starting the screening procedure. 6.0 EQUIPMENT 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Vortex mixer. 6.3 Sonicating device. 7.0 SUPPLIES AND MATERIALS 7.1 Disposable glass graduated pipettes, 1 mL to 100 mL. 7.2 Disposable glass Pasteur pipettes and rubber bulbs. 7.3 Glass beakers, various sizes. 7.4 2.5 mL-4 mL glass screw-top vial. 7.5 Glass volumetric flasks, 10 mL to 1000&I 7.6 10 pL-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, HPLC/SPEC/GC grade from EM Science or equivalent. I 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 CaCl,, A 0.01 M CaCI, stock solution is prepared by weighing 1.5 g CaCI, 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 Page 3 of 9 LIMS# E00-1877 Page 23 of Q8 3M Environmental Laboratory Study NO.FACT-TCROOS 10.0 QUALITYCONTROL 10.1 Not applicable. 11.0 CALIBRATION AND 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. addition 1 addition 2 addition 3 addition 4 Total volume solvent 0.1 added (d) .. . . I 0.5 1 2 Approximate Solubility 5000 O0O0 /LTbf'\ 12.1.3 Following addition of solvent, vortex mi pproximately 15 seconds, sonicate 2-5 minutes. Visually check for undissolved particles. If undissolved test substance remains, continue adding solvent according to the above chart. It may be necessary to allow the solution to settle for up to 5 minutes before making observations. Record observations on the standardized prep sheet. 12.1.4 If all particles dissolve, then estimate the approximate solubility and document the concentration. Because the concentration is >10 pg/rnL, 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 >IO% (mass/volume). The test substance shall be regarded as "highly soluble," and no further testing is required. 12.1.5 If the test substance did not dissolve in the 2 mL of solvent, weigh 10 mg +_ 1 mg of test substance into a 100 mL glass volumetric flask or equivalent. Record the weight (refer to attachment A for an example of a standardized prep sheet). 12.1.6 Add solvent according to the table below to the test substance to deliver the appropriate amount of solvent. As described in 12.1.3, vortex mix, sonicate, and allow the test solution to settle. Record all observations and procedures on the preparation sheet. ETS-8-170.1 Solubility Determination: Screen Test Method Page 4 of 9 LIMW 00-1877 Page 24 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS 100 mL volumetric Solvent addition 1 Solvent addition 2 Solvent addition 3 * flask or equivalent I Total volume solvent 10 added (mL). .. I 50 100 Approximate Solubility 1000 200 100 (Pg/mc>.- * 12.1.7 If the 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 100mL solvent level has been reached without complete dissolution of test substance, weigh out 10 mg k 1 mg test substance and transfer to a 1 L volumetric flask, graduated cylinder, or equivalent. Dilute to 1 L with test solvent. The approximate concentration of test substance is at 10 pglmL. 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-8- 171.O). If no visible particulates are observed, the shake flask method (ETS-8- 172.0) shall be used to determine the solubility. 13.0 DATAANALYSIASND CALCULATIONS 13.1 The sohbility determination in this method is qualitative/semi-quantitative. 13.2 The point to which the substance dissolves insolvent is confirmed visually. The solubility point is therefore a qualitative determination. 13.3 The concentration estimation is semi-quantitative in that the approximate concentration is calculated by the following equation: c=a/b Where: c= the semi-quantitative concentration pg/mL, a= amount of substance weighed out (pg), and b= the approximate amount of solvent added (mL). 14.0 METHODPERFORMANCE 14.1 Limitation of data. The accuracy to which the solubility is determined is subject to a large margin of error due to the way in which the solvent is added to the solute. Since large and varying increments of solvent are being added to the solute, this error margin must be considered when reporting the concentration estimation. 14.2 The data obtained through this study is an estimation only and should be treated as a qualitative estimation of the solubility of test substance in a given solvent. ETS-8-170.1 Solubility Detertitination: Screen Test Method Page 5 of 9 LlMSX 00-1877 Page 25 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS 14.3 For an accurate measurement of the substance solubility concentration, the shake flask method, or column elution method will be used. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 16.0 RECORDS 16.1 Sign and date all observations and calculations. 17.0 ATTACHMENTS 17.1 Attachment A-Example of a Standardized Sample Preparation Sheet: Solubility Screen Prep Sheet. 18.0 REFERENCES 18.1 Organization for Economic Cooperation and Development. OECD Guideline for Testing of Chemicals. Water Solubility -0ECD Guideline 105: pp. 1-7, Adopted 1995 18.2 United States Environmental Protection Agency. OPPTS 830.7860 Water Solubility (Generator Column Method). Prevention, Pesticides and Toxic Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-042: pp. 1-17, 1996 18.3 United States Environmental Protection Agency. OPPTS 830.7840 Water Solubility: Column Elution Method; Shake Flask Method. Prevention, Pesticides and Toxic Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-041: pp. 1-12,1996 18.4 3M Environmental Laboratory Method ETS-8-171.O,"Column Elution Method: Solubility Determination of Test Substance in Various Solvents." 18.5 3M Environmental Laboratory Method ETS-8-172.0, "Shake Flask Method: Solubility Determination of Test Substance in Various Solvents." 19.0 AFFECTEDOCUMENTS 19.1 ETS-8-171.0, "Column Elution Method: Solubility Determination of Test Substance in Various Solvents." 19.2 ETS-8-172.0, "Shake Flask Method: Solubility Determination of Test Substance in Various Solvents." ETS-8-170.1 Solubility Determination: Screen Test Method Page G of 9 LlMSl EOO-1877 Page 26 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS 20.0 REVISIONS Revision Number Reason For Revision 1 It was desirable to make the method more universally applicable by removing references to specific test substances. Revision - Date 03/13/01 ETS-8-170.1 Solubility Determinotion: Screen Test Metliod Page 7 of 9 LIMS# E00-1877 Page 27 of 98 I AttncJinientA: Example Preparation Worksheet,page I of 2 GLP Study Number: Sample prepamtion worksheet Test Substance: lm Date: Source: Solvent: ID# Source: Step I Welgh approximately 10 mg f 1 mg of lest subsfance into a 2.5 mL-4 mL glass screw-top vial. Weight of test substance: . m-.g Balance ID: 2nd replicate optional: mg Thermometer ID: Datellnltials: 2 Add test solvent according to the fable below. Following each addition of solvent, shake vigorouslyhortex, and sonicate volume added (mL1 appror conc. after Analyst __ Daldime---L-Tolal volume solvent added (mL) 's considered 'infinilely soluble.' Dld the substance dissolve In 2 mL or less of solvent? Yes -Approximate concentration at which the test substance Is soluble in solvent: -qhl The solubilib of the test subslance is to be determined via the shake flask method. or No Continue to pagetwo for furher testing. Notedadditional comments: ETSd- 170.1 Solubility Determination: Screen Test Method Page 8 of 9 ~~ - Attachment A Conk Examps Preparation Worksheet,page 2 of 2 SO^. Step (continued from page I) -Did the test substance dissolve In 2 mL7 YedNo If YES, tho test 1% ComDlete. and t h e n Is no need to continue with the screen f w test. If NO,continue to step 3 to determlnethe approxlmate salubillty of the substance. 3 Weigh approximately 10 mg f I mg of lest SubStaf1~i8nla a mL gfadUated stoppered cylinderholumefric flask. Weight of test substance7 mq B a l a n c e ID: Date/lnltials: 2nd repriels oplhdr m0 Thermometer I D 4 Add lest solvenf accordinglothe tabla below. Fdlowingeech eddition d sdvent, Shake vigorouslyhdex, andsonicab ____ - Voluma added (mLy approx COM. after SolublllhFdata u H o n ( u g l m L ) Sample F- OBSERVATIONS/NOTES 1 =5 8B Om OX o- mf. 83 Analysl - Datfima I TOWvolme sdvenl a d d d i L ) Apprciumale Sdublity (uglmL) step 4 4 100 1W Flask mriurmxed?YerMo RoomTemp W ~ tme msk allowed to x ~ e ? YCSNO Tim *C Is there solute sOll pml?Y&No ~ rn O min fwalyst - Weight of test subslance: rnq Balance ID: Datdlnitials: Dalm I Step 5.1 T W volume Mlvenl addeU(u!l) ... loo0 Appaumaie Sdublily ( u r n ) . 10 Flaskmriex-n-d=ad?YerMo RwmTemp ' FbrkronuZesW T i Flask allowed lo S0tUe'/ Y&No lima 'C Is lhera sol& rmq rrrm s1iU m o l 7 Y&No Concluriw Dld the substance dissolve in 1000 mL or less of solvent? Yes -Approximatecmcentrabon at whlch the test substanceis soluble In solvent: - The solubllltyof the test substancdslo be determinedvia the shake flask method. udml or No -The solubilityOf Lhe teslsubstance IS to be determlnedvla the column elution method. - ~ Notes/ addltlonal comments: ~ ETS-8-17 0 . 1 Solubility Determinaiion: S-creen TestMethod Page 9 of 9 3M ENVIRONMENTALLABORATORY 3M Environmental Laboratory Study NO. FACT-TCROOS SHAKE FLASKMETHOD: SOLUSILITYDETEFWNATIOONF A TESTSUBSTANCEIN VARIOUS SOLVENTS Method Number: ETS-8-172.1 Adoption Date: 09/08/00 Effective Date: 03/27/0 1 Authors: Approved By: - / Laboratory Manager d+&/, Date 1.0 ScoDe and ADplication 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/methanoI) of greater than 100 pg/mL must be analyzed via the shake flask method (OECD Guideline 105, and OPPTS Guideline 830.7840). The prerequisite to this . method is a preliminary screen of the test compound for its approximate solubility level. Refer to ETS-8-170.0 for the preliminary solubility screen test procedures. ETS-8-172.1 Solubility Test: Shake Flask Method Page I of 11 LIMS# 00-1 877 Page 30 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS 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 tiom the preliminary solubility screen test, weigh out more than five times the estimated soluble concentration oftest substance into each of twelve screw-cappedvessels (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 OC-26 "C) for approximately 24 hours. The equilibrated samples are centrihged 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:methanoVacetoneserial dilutions), and an appropriate internal standard (e.g. lH,1Hy2H,2Htetrahydroperfluorooctane sulphonic acid (THPFOS)) will be added to the final dilution prior to analysis. Samples are to be analyzed via LCMS against a standard curve containing the test substance and an appropriate internal standard. 3.0 DEFINITIONS 3.1 Method blank: An analyte-free matrix (e.g. methanol, water, or acetone) to which all reagents agree are added in the same volumes or proportions as used in the sample processing. The method blank is used to document contamination resulting from the entire sample treatment and analytical process. The method blank is canied through the complete sample preparation, treatment, and analytical procedure. 3.2 Solvent blank: A sample of analyte-free medium (e.g. methanol/water/acetone solution) that is not taken through the sample treatment process. This blank is used to evaluate instrument and reagent contamination. 3.3 Shake Flask Sample Triplicates: Three test vessels taken from and representative of the same sample source carried through all steps of the treatment, extraction, and analytical procedures in an identical manner. 3.4 Sample replicates: Replicate samples (two or more) taken from and representative of the same sample source (e.g. test vessel containing test analyte and solvent) and separately carried through analytical procedures in an identical manner. 3.5 Internal Standard (IS): A known amount of a compound similar in analytical behavior to the compound(s) of interest, added to all samples and standards, and camed through the entire measurement process. ETS-8-172.1 Solubility Test: Shake Flmk Metliod Page 2 of 11 LIMS# EOO-I 877 Page 31 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS 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. . - 4.12 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 stMing the preliminary solubility screening procedure. 5.2 Contaminants in solvents, reagents, glassware and other sample processing OT 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 Centrifuge capable of holding 15 mL centrihge tubes or equivalent. 6.3 Incubator with heatinglcooling 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 O C - 4 0 'C. 7.2 5-250 mL polypropylene centrifuge tubes, or equivalent. 7.3 Disposable glass graduated pipettes, 1 mL 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 Gastight@syringes (precisionk 1% of the total volume), 5 pL to 1000 pL. 7.8 Pipette-man manual pipettor. ETS-8- 172.I Solubility Test: Slmke Flask Method Page 3 of 11 L I M S E00-1877 Page 32 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS 8.0 SUPPLIES AND MATERIALS 8.1 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science, or equivalent. 8.2 Acetone, HPLC/SPEC/GC grade fiom EM Science or equivalent. 8.3 Water, ASTM Type 1. 8.4 Test substance of known purity. . 8.5 Calcium Chloride Dihydrate, Approximately 99% or better, from Sigma. 8.6 0.01 M CaC12 solution, Example: A 0.01 M CaC12 stock solution is prepared by weighing 1.4 g CaCl2 in a weigh boat and transferring to a 1 L volumetric flask and diluting to the mark with Milli-QTMwater. 9.0 SAMPLHEANDLING 9.1 Record times of initial preparation, set-up, and sample aliquotinghalysis on a sample preparation sheet or logbook. 9.2 Once the samples have been diluted, they may be analyzed via LCMS. AItematively, 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 QUALITCYONTROL 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 (fromeach 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 contaminationaccrued 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 (i.e. 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 operating procedure, ETS-8-172.1 Solubility Test: Sliake Flask Method Page 4 of 11 LIMS# EOO-1877 Page 33 of 98 3M EnvironmentalLaboratory Study NO. FACT-TCROOS 11.3 All samples analyzed will be run against a standard curve containing varying amounts of test substance, and a fixed amount of internal standard. 12.0 CALIBRATION AND STANDARIZATION 12.1 Preparation of Test Vessel. Record all data and observations on the example sample preparation worksheet (Attachment A) or equivalent. 12.1.1 Prepare a solution at least fivetimes-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 cenbifbge 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 specijc volume will be dependent on the density.). Record all weights on a standardized preparation sheet or logbook. 12.1.4 Visually confirm that there are undissolved particles. If no particles are present, repeat 12.1.2 with more test substance or add more test substance and record the additional weight. 12.1.5 Prepare the solution described in 12.1.1-12.1.4 nine times and label (at minimum) the test vessels as Day 1, -Rep 1, -Rep 2, and -Rep 3; Day 2, -Rep 1, -Rep 2, and - Rep 3; and Day 3, -Rep 1, -Rep 2, and -Rep 3. Also include the day of initial sample prep, the person(s) responsible for the sample, the test compound, nature of the study (e.g. water solubility), the solvent utilized, and the study number. 12.1.6 Prepare three test vessels as described in 12.1.3 without the test substance and label (at minimum) as Dayl,Rep4; Day2,Rep4; and Day3,Rep4 along with the information described under 12.1.5. These three test vessels are the "method blank" samples. 12.1.7 Seal the cap to the test vessel. Midshake test vessel to ensure contact between the test substance and the solvent, And wrap the cap with tape. 12.2 Equilibration of samples. 12.2.1 Place all of the test vessels on their side (horizontal) in an orbital incubator set to approximately 30 "C k 2 "C and rotating at a considerable speed to ensure sufficient contactlmixing between the test substance and solvent. 12.2.2 At approximately 24 hours, four test vessels are removed: three containing test substance labeled "Day l,Repl", "Dayl,Rep2", "Dayl,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 20- 26 OC (record the actual temperature) for about a 24 hour period. 12.2.5 After about 24 hours of equilibrationat 20 "C-26 "C,centnfbge 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 fbrther sample preparation. ETS-8-172.1 Solirbiliiy Test: Shake Flusk Method Page 5 of 11 LIMM E00-1877 Page 34 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS 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") a d 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,Repl", "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 emulsiondor a concentrated layer of solute at the surface of the solution, it may be necessary to pipette offthe top layer of solution (or an aliquot of sample may be centrifbged 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. 1.5mL polypropylene or glass crimp-cap vials). 12.3.2.1 Sample aliquots may be taken by submergingthe 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. adrorption 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 LabeI the four autovials and record the sample id. 12.3.4 Using a diluter, make a 1:lO or higher dilution of samp1e:extraction solvent (i.e. methanol, acetone) into a labeled autovial. 12.3.5 Using the solubility information obtained in the preliminary screen test, estimate the appropriate dilutions, if any, required to bring the concentration of sample into the appropriate analytical range (e.g. approximately 3 ng/mL to 3000 ng/mL test substance). Utilize the diluter to make serial dilutions of the sample using a suitable analytical solvent. 12.3.6 To the final analytical sample, add internal standard at a concentration suitable to .the analytical method. 12.3.7 Samples will be analyzed via L C N S 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 c w e . 13.2 Means will be calculated by adding the individual entities and dividing the resultant sum by the number of individual entities. 13.3 Standard deviations will be calculated using either Microsoft Excel@ or Microsoft Access@ to calculate standard deviation. The built in fimction 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 Page 6 of 11 LIMW E00-1877 Page 35 of 98 3M Environmental Laboratory Sttidy NO. FACT-TCROOS 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 Sample RPD will be calculatedusing the following equation: A-B I /((A+B)/2)= Sample RPD where: A= concentration of first replicate B= concentration of second replicate Number of observations 3 4 i 5 Qt.bulntcd, 90% Confidence Range 0.886 0.679 0.557 ETS-8-172.1 Soltibility Test: Shake Flask Metliod Page 7 of 11 LlMSd E00-1877 Page 36 of 98 _. 3M Environmental Laboratory Study NO. FACT-TCROOS 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-diluteire-shoot samples to check for error. 14.2 Limit of Quantification. For this study, the LOQ will be equal to the lowest calibration standard used in the calibration curve containing more than twice the area counts of the highest Quality Control Blank. 14.3 Quality Control Blanks: Method and Solvent Blank samples. The level of analyte (test analyte OR internal standard analyte) shall be less than 50% of the area counts of the LOQ. If background levels of analyte exist in the initial method blanks, but subsequent . blanks prior to the calibration standard curve are clean, the data may be accepted (provided that there are no other indicators that either the samples or the instrument contain significant background levels of analyte). 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 16.0 RECORDS 16.1 Sign and date all observations and calculations. 16.2 Fill out all appropriate sample preparation worksheets. 17.0 ATTACHMENTS 17.1 Attachment A-Example Sample Preparation Worksheet. 18.0 REFERENCES 18.1 Organization for Economic Cooperation and Development. OECD Guideline for Testing of Chemicals. Water Solubility -0ECD Guideline 105: pp. 1-7, Adopted 1995 18.2 United States Environmental Protection Agency. OPPTS 830.7860 Water Solubility (Generator Column Method). Prevention, Pesticides and Toxic Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-042: pp. 1-17, 1996 18.3 United States Environmental Protection Agency. OPPTS 830.7840Water Solubility: Column Elution Method; Shake Flask Method. Prevention, Pesticides and Toxic ETS-8-172.1 Soliibility Tesf:Sltnke Flask Method Page 8 of 11 LIMS# EOC-1877 Page 37 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS Substances: Fate, Transport and TransformationTest 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: Slinke Flask Method Page 9 of 11 LIMS# 00-1 077 Page 38 of 90 A TTACHMENTA: Example Sample Preparation Sheet (page 1 of 2). Temperature o f Study: C .. &F%nI v i m v Test: Sample preparallon worksheet Test Substance: la: Sovs.: Date: Test Solvent: IW. Test Vessd: dni SaU-: Was the sdubifity s c m n lest p e r f m d p r i o r to start7 Ym /f so, whafIs the approximate COrICeflbaUon? U Ba&e ID: Thermometer ID: . RoarnTemp: SFM vials ID& . welght test subthnc. Total mass (solute+ solvent) 'Internal Standard addition uU Study X Analyst(s): 151& cenlnfupe lub&iIWrpedfy): Tesl Vessels sealed w/ tape? YM Test Vessels shakenlvortex-mked? Y M Test vessels pbced on orbital Incubator?Y/N Incubator ID: Speed: - lint ~ t dC. OW.: rpm InitialTemperature: ~~ uplml c J .. I Location of samples: visual observalions: Temperatureof e n v i r o r m e n t " C Thwmome(erlD: I J ETS-8-172.1 Solubilily Test:Shake Flask Method Page 10 of 1 I ATTACHMENTA CONTINUED (page2 of 2). I J""F'2r "Day T samples are pulled and allowed to be placed in a t e m p h e conlroned environment forappmx. 24 hourswith no agitation. I"Day 1" Test Vessels Centrifuged: Cant I D Visual obswaticm% RPMor RCF readout dualion: -Four aliquots from each of the four samples are to be made into glasslplasUc autovials (also, check autovials for correct labeling). -Immediately dilute the solution I:-with methanol. Methand TN-A' Additional dilutions: Dllutim factor (lest soIution:methand): 1: X's how many dilutbns: Diluter ID used: Amounts used: uL methanol. -Store samples and extracts in a cooler at until time of analysis. Cooler I D Ccoler Temp: mia YesMo =final dilutionfa*. 0 DaMimdlritirk. Dalenkldnllbb: 1: ul samples DaWhvlniLh: Day 2" Tesl Vessels Centrifuged: C f i D [ Visual observations: RPM or RCF readout duration: mh. I I Four aliquots from each of the four replicatesare to be made into glasslplastic autovials (also. check autovials for correct labeling). YesMo Immediately dilule the solution 1-: with methanol. MelhanolTN-A' I Additional dilukns: Dilution factor (test soluti0n:methand): 1: X's how many dilutions: =final dilution factor. 1: Diluter ID used: Amounts usad: UL methanol, h-Store samples and extracts In a coder at untiltimed enalysls. I__.-__ cader ID: Cwler Temp: 0 DaLsCT!mc/lniUlls: DMinilik. ul samples DaleJTlndWUslr: `Day 4: AI approx. 24 hours of equilibration. the 'Day 3' samples are to he aliquotdextraded. Test Vessels Centrifuged: Cent. ID: I1I II visual observatiions: RPM or RCF readout EnvironmenlTemp.: duration: O C ThermID: @. -Four aliquots from each of the four samples are to be made into glasslplasticautovials (also, check autovials for correct labeling). YesMo -Immediately dilute the solution 1:-with methand. MelhanolTN-A` Additional dilutions: Dilution factor (Lest so1ution:methand): 1: X`s how many dilutions: =final dilution factor: 1: Diluter ID used: Amounts used: uL methanol. `Store samples and extracts in a m l e r at untiltime of analysis. Coder ID: Cooler Temp: 0 @dlmdNb!S Dal.=frmdl~WI~: ul samples Dnlcfrlmf#lniUPb ETS-8- 172.1 Solubility Test: Shake Flask Method Page 11 of 11 1 1 1 .a 1 w K I 3s g 5. -z g Pi 3h4 ENVIRONMENTAL LABORATORY 3M Environmental Laboratory Study NO.FACT-TCROOS METHOD ANALYSXOSF POTASSIUPMERFLUOROOCTANESULFOONRAOTTEIER FLUOROCHEMICIAN WLSASTE STREAM OR WATER EXTRACTS USING HPLC-ELEC~RO~PRAY/MSPAESCSTROMETRY Method Number: ETS-8-155.0 Author: Approved By: -__ Laboratory Manager - Gr'oup Le'ader - / Adoption Date: l(/p/Oo Revision Date: ,. Date bate ' Word 97 ETS-8- 155.0 Page 1 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Exbacts Using HPLC-Electrosprayfif ass Spectrometry LIMS# 00-1877 Page 41 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS 1.0 SCOPE A N D 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 fluorochemicalsor other electrosprayionizable compounds extracted from water, using HPLC-electrospraymass spectrometry(HPLCES/MS). The analysis is performed by the mass selection of a single ion characteristic of a particular compound, such as the perfluorooctanesulfonate(PFOS) anion, m/z = 499 or perfluorooctanuate (PFOA), m/z = 413. 3.0 DEFMITIONS 3.1 Atmospheric Pressure Ionization (API): The Micromass Platform LCZ single quadrupole system and other commerciallyavailable LCMS systemsallow for various methods of ionization by utilizing a variety of sources, probes, and interfaces. These include but are not limited to: ElectrosprayIonization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization in these processes occurs at atmospheric pressure (Le., not under a vacuum). y' 3.2 Electrospray Ionization @S, ESI): A method of ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These droplets are produced by the application of a strong electrical field. 3.3 Mass Spectrometer(MS): The Platform LCZ and other commerciallymanufactured ES/MS systems are equipped with a single quadrupole mass selective detector. Ions are selectivelydiscriminatedby mass to charge ratio ( d z ) and subsequentlydetected. 3.4 Conventional vs. Zspray probe interface: The Micromass Platform LCZ system utilizes a "Z-spray" conformation. The spray emitted fiom 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, Z- spray components and conventional components are not compatible with one another, but only with similar systems (i.e., 2-spray components are compatible with some other Z-spray systems, etc.). Other commerciallymanufacturedESMS systems may have similar features. 3.5 Mass Lynx Software: System sofhvare designed for the specific operation of Micromass LCZ Mass spectrometer. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details see the manual specific to the instrument (MassLynx NT User's Guide or Micromass Platform LCZ User's Guide). ETS-8-155.0 Page 2 of9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry LlMSAr 00-1877 Page 42 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage of approximately5000 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). If the backpressure exceeds 400 bar, the HP1100 will initiate automatic shutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 Interferences 5.1 To minimize interferences when analyzing samples,Teflon should not be used for sample storage or any part of instrumentation that comes in contact with the sample or extract. , 6.0 EOUIPMENT 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 HPllOO 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 100psi (or house air system.). 7.1.2 HPLC analytical column, such as a Betasil C18 column (5Ox2mm, 5 pm particle size) or equivalent. 7.1.3 Capped autovials or capped 15 mL centrifuge tubes. ETS-8-155.0 Page 3 of9 Analysis of PotassiumPerfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-ElectrosprayhfassSpectrometry LlMSll E00-1877 Page 43 of 98 3M Environmental Laboratory Study NO. FACT-TCROOS 8.0 REAGENTS AND STANDARDS 8.1 Reagents 8.1.1 Methanol, HPLC grade or equivalent. 8.1.2 Milli-QTMwater (ASTM type I), all water used in this method should be Milli-QTM water or equivalent, and may be provided by a Milli-Q TOC Plus system or other vendor. 8.1.3 Ammonium acetate, reagent grade or equivalent. 8.1.3.1 When preparing different amounts than those listed, adjust accordingly. 8.1.3.2 2.0 mh4 ammonium acetate solution: Weigh approximately0.300 g ammonium acetate. Pour into a 2000 L volumetric flask, add the appropriate volume of Milli-Q water, mix until all solids are dissolved. .Store at room temperature. 8.2 Calibration Standards 8.2.1 Typicallytwo method blanks (Milli-Q water), two matrix blanks, and solvent standards are prepared during the sample extraction procedure. 9.0 SAMPLEHANDLING 9.1 Standards and sample extracts are stored in capped autovials or capped 15 mL centrifkge tubesuntil analysis. 9.2 If analysis will be delayed, standards and sample extracts may be refiigerated at approximately4"C until analyses canbe performed. 10.0 QUALITY CONTROL 10.1 Solvent Blanks, Method Blanks and Matrix Blanks 10.1.1 Solvent blanks, method blanks, and matrix blanks are prepared and analyzed with each sample set to detemrhe 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 duplicateconcentrationsshould fall in the mid-range of the initial calibration curve or should be prepared at 1.5-5 times the endogenous ETS-I- 155.0 Page 4 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-EIectrospray/Mass Spectrometry LIMS# E00-1877 Page 44 of 98 3M EnvironmentalLaboratory Study NO. FACT-TCROOS concentration of the analyte. Spike concentrations should fall in the low-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 verify the continued accuracy of the calibration curve. 10.3.2 Analyze a mid-range calibration standard after every tenth sample, with a minimum of one per sample set. 10.4 Internal StandardlSurrogateStandard 10.4.1 An internal standard (IS) may be used to quanti@the target analytes by establishing a relationship between the ratio of analyte response to IS response and a known concentration of the analyte of interest. The IS should be spiked at an amount that will fall within the mid-range of the calibration curve. The IS should be added after the extraction process and before analysis. 10.4.2 A surrogate standard may be used for quality control. The surrogate is used to quantitativelyevaluate the entire analyticalprocedure including samplepreparation and analysis. The surrogate should be spiked to fall 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 l/x, 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 acceptancecriteria perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. 11.3 For purposes of accuracy when quantitating low levels of analyte, it may be necessary to use the low end of the calibration curve rather than the full range. Example: when attempting to quantitate approximately 10 ppb of analyte, generate a calibration curve consisting ofthe standards kom 5 ppb to 100 ppb rather than the full range ofthe curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear regression weighting of high concentration standards. 12.0 PROCEDURES 12.1 Acquisition Set up 12.1.1 Set up the sample list. 12.1.1.1 Assign a sample list filename using the first letter of the name of the instrument (T for Tucker), the year (00 for 2000), the month (04 for April), and the day (TO01012for October 12,2000). If more than one list is made on the same day, use increasing letters of the alphabet starting with A at the end of the list. ETS-8- 155.0 Page 5 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochernicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayIMass Spectrometry LlMSX 00-1877 Page 45 of 98 3M Environmental Laboratory Study No. FACT-TCROOS 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 descriptionsand vial position numbers. 12.1.2 To create a method,click on method in the Acquisition control panel then mass spectrometer headings and select SIR. Set ionization mode as appropriate and mass to 499 or other appropriate masses. A full scan is usually collected in addition to the SIRs. Save acquisition method. See the Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information. 12.1.3 Typically the analytical batch run sequence begins and ends with a set of solvent standards. 12.1.4 Samples are analyzedwith a continuing calibration verification (0 injected after every tenth sample. Solvent blanks should be analyzed periodically to monitor for possible analyte carryover. 12.2 Using the AutosampIer/ColumnHeater 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 appropriateports. 12.3 Using the Inlet Editor 12.1.1 Set-up the HPI100using the following conditionsor at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook 12.1.1.1 Sample size = 10 pL injection 12.1.1.2 Flow rate = 300 pUmin. 12.1.1.3 Cycle t h e = 10.0minutes 12.1.1.4 Mobile phase components: Solvent A: 2.0 mM Ammonium Acetate Solvent B: Methanol (MeOH) Solvent Gradient: Time (rnin.) 0.00 1 .oo 4.50 8.00 8.50 10.0 - Yo B 5.00 % 5.00 % 95.0 "io 95.0 % 5.00 % stop ETS-8-155.0 Page 6 of 9 Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry LIMS# E00-1077 Page 46 of 98 3M EnvironmentalLaboratory Study NO. FACT-TCROOS 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 EIectrospraylMass 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, disassemblethe probe and replace the stainless steel capillary. 12.4.4 Turn on the nebulizing gas. 12.4.5 Open the tune page. Click on `Operate'to initiate the desolvationheaters. 12.4.6 Open the Inlet Editor. 12.4.5.1 Set HPLC pump to "On". 12.4.5.2 Set the solvent flow to the desired flow rate. 12.4.5.3 Observedroplets coming out of the tip of the probe. A fine mist should be expelled with no nebulizing gas leaking around the tip of the probe. Readjust the tip of the probe if no mist is observed. 12.4.5.4 Allow to equilibrate for at least 10 minutes. 12.4.6 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response: 12.4.6.1 Drying gas 250-425 literdhow 12.4.6.2 ESI nebulizing gas 10-15 literdhow 12.4.6.3 HPLC constant flow mode, flow rate 10 - 500 pLumin 12.4.6.4 Pressure 4 0 0 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,'samplelist, 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 followingequation: - ETS-8- 1.LSL Page 7 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochernicals in Waste Stream or Water Extracts Using HPLC-ElectrosprayRvlassSpectrometry LlMSt E00-1877 Page 47 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS % Recovery = Observed Result - Background Result x 100 Expected Result 13.1.2 Calculate percent difference using the foIlowing equation: % Difference = Expected Conc. - Calculated Conc. x 100 Expected Conc. 13.1.3 Calculate actual concentration of analyte in matrix @g/mL): On-Column Concentration (pg/mL,) x Dilution Factors = Calculated Concentration 14.0 METHOD PERFORMANCE 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 (r') value for the calibration curve must be greater than or equal to 0.980. 14.4 Continuing CalibrationVerification (CCV) percent recoveries must be k 30% of the standard concentration. 14.5 Internal Standard recoveries should be within k 50% of the spiked concentration. 14.6 If criteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the raw data. 14.7 If data is to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 RECORDS 16.1 Each page generated for a study must have the following information included either in the header or hand written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. 16.2 Print the tune page, sample list, and acquisition method tiom 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 .O Page 8 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using I-IF'LC-Electrospray/Mass Spectrometry LIMW 00-1877 Page 48 of 98 3M Environmental Laboratory Study No. FACT-TCROOS 16.4 Print data integration summary, integration method, and chromatograms, fiom MassLynx, and store in the study folder. 16.5 Summarize data using suitable software and store in the study folder. 16.6 Back up electronic data to appropriatemedium. Record in study notebook the file name and location of backup electronic data. 17.0 ATTACHMENTS 17.1 None 18.0 REFERENCES 18.1 Platform LCZ User's Guide, Micromass UK Limited, Tudor Road, Altrincham, WA14 5RZ;or Floats Road, Wythenshawe M23 9LZ; United Kingdom. 18.2 MassLynx NT User's Guide, Micromass UK Limited, Tudor Road, AItrincham, WA14 5Rz;or Floats Road, Wythenshawe M23 9LZ,;United Kingdom. 18.3 MassLynx NT Guide To Data Acquisition, Micromass UK Limited, Tudor Road, Altrincham, WA14 5RZ;or Floats Road, WythenshaweM23 9LZ; United Kingdom. 18.4 ETS-9-36.0,"Operation and Maintenance ofthe Micromass Platform LCZ EIectrospray/Mass Sprectrometer". 19.0 AFFECTEDDOCUMENTS 19.1 None 20.0 REVISIONS Revision Number. Reason For Revision - Revision Date ETS-8-155.0 Page 9 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrospraylMass Spectrometry LIMSt E00-1877 Page 49 of 98 3M Environmental Laboratory Study NO.FACT-TCROOS ATTACHMENBT: RESULTS -WATERSOLUBILITY:DATASUMMARSYHEETS -METHANOANLD ACETONESOLUBILITY: SAMPLE PREPARATION WORKSHEETS LIMS# E00-1877 Page 50 of 98 __ PFBS TCR-W017-71 ,Iy ,I Ii I n 9 m.1 I*L :jl a/$?/( arrnm e m a1 STUDY FACT-TCROOS PFBS Solubility Study X FACT-TCROOS PFBS Solubilitv Study, Shake Flask Method inWATER DAY 1. PFBS TCR-99010-23BATCH H92WO C DATA EXPORT aam 1 " m LBULL NEW4 L!kW.h W W.WB.#. P!E!2UI IJIF'FOO.AW E f L u l z m Ems!e?. 1 244 2 7s 11m 2 244 1 7.w IP I 1u 1 im 6W 4 2442 7s 60 6 am 1 7- 6U I 144 1 7m LW 7 144 1 7.m aU 6 244 1 7.w sw S 2u 2 7.92 CW *10 144 1 7.m &U 1u 1 7.92 LP S 244 1 7.m CU 9 144 1 7.m em 2 1ua 7m CP 3 144 1 7.- SU 4 a u1 7m am * 6 lU1 1u 1 1.- 7m su Mil 7 au 2 70 CW I 244 1 7m 6W 8 2u1 7.m sm 10 244 2 7.91 IO n W2 7.m MU m 1441 7 e2 am 2s 144 1 7.m 6U y1 244 1 7n 6W 31 1443 7.- SW sa 144 1 7.a *m 33 244 1 7w CU 34 lU1 7.m (112 31 1441 7.01 4w Y 144 1 1Y 6U 37 au I 7.m 6U s wz 7m 6P m 244 1 IS3 6P (0 IU I 7 e3 I I2 a41 244 2 7pI 112 144 2 7u IIl om om om OW M 00 Dm w 00 om M 00 om SS DO om 05 00 om SS 00 om %OEVmT@tj 4.5% Id% 2.4% %I% O.% 0.4% 1.1% 4.4% 1.7% 10.0% 11.7% 0.5% 1.1% 1.8% 03% 0.w ACTUAL PPM 55747 5zwl m m M Y I 0.b. PFBS 1CR-a w I -- E$ Study #FACT-TCR-OOS PFBS Solublllty . ~ ~~ SludV 1 F A C T - T m 9 PFBS Solubilllv Study-Shake .SsmoIaDce Fksk Method In WATER. DAY 1. PFBS TCR-99030-23BATCH H922DD b DATA EXPORT Run # BUrhMI6$ W IHPFOSDPb JnPfOSRT ~pFBsI\RI~ an PO 00 IOI na US SWV 0142 -9 11117 "1 64 00 00 LO 101 27 I MI 11.1 1113 Val ,2111 173% 11.1% 4.2% 1.2% 2441 2441 w2 2u1 77P 7M --I_ 244 2 7- I 00 om 00 2 00 om 00 I, 00 om 00 I. L n m LO IS 01 om OD om X 00 0- 00 m 00 om 00 3I 0- 00 17 00 om 00 Y 00 om 00 Q 00 om 00 Y) M om Od M 51 YSQ u 7m 514 #O PIS SUY -6 L19 74 m in ylm sun u104 OB 00 OD OB 00 on 00 OD 00 00 OD no 00 00 00 OQ Study IFACT-TCR009 PFBS Solubility Study*FACT-TCROOSPFBASolobilih, Study. Shake Flask MethodIn WATER. DAY 3. P F B S I C H 1 7 - 7 1 BATCH H92200 C DATA EXPORT THPFOSmb mpFo81R1 , auz 7w .7. 2 W1 71 an I 2.u TY en 4 244 2 748 6 75 I 24u IY 676 I 2441 71 5 71 7 2441 11 Lm I 2441 TP 6 70 I 2441 71 a 76 ID 1441 7w 6lI 8 2U 2 71 I7s e 144 2 7Y an 1 auz 7m In 2 w 2 7Y en a 2441 1N en 4 2U2 7- an I 2442 1P an 6 2Ul 724 sn 1 2U I 7- an I 2441 in em I 1U2 7" an IO W1 7Y 6n -1 en 0 1441 71 sn 44 2441 IY am 43 2442 1N I ad4 1M an 47 W1 7Y an u 2441 71 an a 2441 7P 4 76 0 ani 1- cn II 2441 la 6n II W1 7m an s aru 7w 6.n u 2u3 7m 015 s 2441 TY rn m a442 an 6la an om on an om On an om 00 en om DO En om 00 a 7I om 00 an om 00 sn om 00 en om 00 an Em ao an om 00 an om 00 om o* on 06 I1 I ld W2 113 a174 WJ 1m5 UI u4 00 09 00 10 5 no 671 1,lJ OOJ SI, I Ollb ms% 1.5% 1.1% 4.1% 0.0% 23% zm 43% 43% (%I% 11.1% 1.m 0.8% Ll% l+% 13% urn 5m ,I %n %Is2 G9Y 7llY ma SI ma M 371 1 00 ao 00 DO 00 00 DO 00 00 00 00 OD 00 00 00 00 !r!i a iaI StudylFACt-TCROOOPFBS Solubility Study I FACT-TCROW PFBS Solubiliw Study. Shake Flask Helhod In WATER DAY 3.pFBs TCR-99030-23 BATCH HS21M) D DATA EXPOET I 7M 1 7Y 1 7" 4 7Y I 7M m 7M 7 7s I 7M S 7M 10 711 I 71 I 7Y I 7m 2 71 a 7M 4 7M I 7e I 7M ? 71 I 7M s 7Y IO 7" 5s 7M .I 7M I! 7M rl 7M I 7M u 7m I 7Y Y Lu n 71 M 71 1 - 1 m 7M m 7M n71 7Y n 00 OW PO OW 00 om 00 om on Om 00 DOD 00 om 00 Pm OD om no OW no om 00 OW 02-21 - %s!Ewdw 12.6% 10.6% s.lu rrx O.y* 02% 02% 6.354 6.2% izau 10.4% 4.1% 2.4% 11% 02% 0 1% rcruAcPP~ 547Q 3M Environmental LaboratorySOLUBILITY SCREEN GLp Study Number: TCR-009 Test Substance: PFBS ID# Lot 101,TCR-00030-23 Souno: 3M Internal ? - / m Sample preparation worksheet Date: Solvent: Methanol Step 3 , 8/$?8/ I 1 Weigh approximately 10 mg f I mg of tesf substance Into a 2.5 mL-4 m l glass screw-to vial. Weight of test substance: mq Balance ID: 1 . - . zndmpkate OptiMl/: mg Thermometer 1D:Wmc.U . Datehitials: 2 Add test solvent according to the table below. Following each addition of solvent. shake vigomuslyhortex, and sonicate the mixture and visually check for undissolvedparticles. 9-/363 haiyrt Total volume S O ~ V M ~added (mL)... ApproximateSdubility (ughL)... Datfltme Flask allowed to Analyst w d Dalflime ... v e n i added (mL) 2 ApproMmale Solubility(ug/mL)... 5 . W Flask allowadto retUe? Y e m o Tlma Conclusions: (2 Did the substance dissolve in 2 mL or less of solvent? Yes Approximate concentrationat which the test substance is soluble in soIvent?\Qm he solubility of the test substance is to be determined via the shake flask method. or No Continuetopage two for further testing. Notes1 additional comments: Exact my uslrnl -sOm .\9~1ou/3-/Ib &Y#Qf Qf Qti<.;+,:gj Solubility Screen Test Page 1 of 2 3M ETS Laboratory 3M Environmental La- %?torV SOLUBILITY SCREEN GLP Study Number: TCR-009 Sample preparation worksheet Test Substance: PFBS ID* Lot 101.1CRt.9~030 23 source: 1Y Illlrnunrl Date: Solvent Methanol - ID# Source: 5, Step (continued from page 1) '*Did the test substance dissolve in 2 mL? YesfNo IYES, the test Is complete, andthere b no need to continue with the screen '\\.\ test. If NO,continue to step 3 to detemlne the approxlrnate solubility of the substance. - Analyst - Dalflune Totalvdume rolvenl added [mL). la thae aoMe still weacnl?'.Yes/No - Weight of test substancej BalanceID: \ \ Datellnitials: \ olve In 1000 mL or less of solvent? Yes-approximate concentration at which the test substance is sol le in solvent: The solubilityoflhe lest substance Is to be determinedvia the No -The solubilityof the test substance is to be determined via the SolubilityScreen Tesl Page 2 of 2 3M ETS Laboratory 3M Environmental Laboratow SOLUBILITY SCREEN GLP Study Number: TCR-009 Sample preparation worksheet Test Substance: PFBS IW Lot 101.TCR-99010-15 Date: l?&f#) Saurca: 1M Intarnal Solvent: Acetone Step IDX: Ill source: f.n~s.xNQ? 4-.4 b )~kf%W) 1 Weighapproximately 10mg f 1 mg of @stsubstmcs Into 8 2.5 mL-4 mL glass screw-top vlat. Weight of test substance: mq Balance ID: 2nd repiicab op#ona/: mg Thermometer ID: '7hfacU . 2 Add test solvent according to the table below. Following each addition of solvent, shake vigornuslyhodex, and sonicate the mixture and visuallycheck for undissolvedparticles. I 1 I I volume added (mLV appiux COUL Iftar Solubllitv datz addltlon (uphnL) Sample Prep OBSERVATlONSlNOTES 3 Daemme%fiUI)/ \\)'a& s t e p 2-1 Total volume solvent added (mL).. 0.1 Approximate Solubility (uglmL).. 1oo.oO0 Analyst / - Analysl Delmme I Total vdume solvent added (mL).. Approximate Solubillty (ugjlmL).. Step 2-2 Flask vo~iex-mixed'Yl erMo RoomTemp 'S q +/"q" 0.5 2o.m Flask ronlcated? YerMo T i m Flask aUo~6dto .CKk? YerMo lln" Flask vortex-mixed? YerMo Room T m p Flask micated? ~ e n~me o Flask allowed to settle? YeaMo T i min min 'E &I; soIuI0rlil present? Yemo DalelFie I Step 1-4 \ Tokt volume solventadded (mL).. 2 ApproximaleSolubiliiy (ughnL).. 5.000 Is l h sol~ute a~611present? Ye& 1 I \ Did the substance disso 2 mL or less of solvent? d/'\ @pprcrinute concentration at which the test substance is soluble in solvent:)f&L'w) The solubility of the test substance is to be determinedvia the shake flask method. * or No Continuelo page two for further testing. us/ml >\c3'jc .. \i')f' . %kih Solubility Screen Test Page 1 of 2 3M ETS Laboratory 3M Env;ronmental Laboratow SOLUBlLlN SCREEN GLP Study Number: TCR-009 Sample'preparation worksheet Test Substance: PFBS IM Lot 101. TCR-gSOU)-Zt SOUK.: 3M InWaJ Date: Solvent: Acetone ID#: ered cylinderholurnefric flask. Is Ihnm solUte still prerent7 Yes/Na I Step 5-1 ' Flask vatexmhd? Y d N o Is Vlsn soMe still present? Y e f l o Flask Sonicated7Yer/No Flask slbwed lo astlh? Y e s M TmS m Conclusions: Did the substance d SSOlVe In I000 mL or less of solvent? \ Yes -Approximateconcentrationat which the test substanceIs solublfh The solubility of the test substanceIs to or No -The solubility of the test substance is to uqlml Solubility Screen Test Page 2 of 2 Cnltiaf 3M ETS Laboratory L;>;p 3M EnvironmentalLaboratow SOLUBlLlN SCREEN GLP Study Number, TCR-009 Test Substance: PFBS ID# Lot Z.TCR00017-71 Sourno: 111lnbrnrl Sample preparation worksheet Date: 7 - / 3 a Solvent: Methanol 4mu m'zc IDY: source: Step 1 Weigh apptuximately 10 mg f I % 3 l/f?/-lS) Weight of test substance: - m m 2ndmpUca1eO N r n L of fedsubstance into a 2.5 mL-4mL glass screw-top vial. mq Balance ID: mg Thermometer ID: ~ ~~~ ~ 2 Add test solvent accordingto the fable Mow. Followingeach addition of solvent, shake vigomuslyhortex, and sonicate the mixture and visually check for undissolved parficles. Fhrk vortex-mixed? YesNo Flask ronicsled? Y e 4 0 Flask alJowedto ael.7 Y w N o It hem solule dill presenl? Y e M o halyrt DalelTime Approximale Solobility (uglmL)... 5.000 Flask vorlex-mixed? Y w N o Raun Temp Flask sonicated? Yes'No T i Flask allowedto settle? Y e M o Time 'c Is then sdule rcill present? YesNo mh min Conclusions: Did the substance dissolve In 2 rnL or less of solvent? B d p p r o x i r n a t e concentration at which the test substance is soluble in solvent:7J@J$ us/rnl $CIQbb0 @ ,cC/o l The solubility of the test substance is to be determined via the shake flask method. r or No Continue lo page two for further testing. Exaet copy af 0 ; .,I Notes1additional comments: Initial L,,,I, Solubility Screen Test Page 1 of 2 3M ETS Laboratory 3M E n v i r o n m e n t a l LaboratowSOLUBILITY SCREEN GLP Study Number: TCR-009 Sample preparation worksheet Test Substance: PFBS ID# Lot 2. TCRPM)I?-?l Source: 3yI lntmrl Date: Solvent Methanol IDf Source: Step (continuedfrom page 1) "'Did the test substance dlssolve In 2 mL? YeslNo If YES, the test Is completepnd there Is no need to continue with the screen test. IfNO,contlnue to step 3 to determine the approximate solubility of t t ~ esubstance. / 3 Weigh approximately 10mg f 1mg of test substance into a IJlOn5L graduatedstopperedcylinder/lolumetnc flask. Weight of test substance: mq Balance ID: /e . 2nd mpacata opbond r?W/ Thermometer ID: ,/- 4 Add test solvent accordingto the teble b e ? F o l l o w i n g each eddition of sdvenf. shake vigomuslyhrfex. and sonicate the mixture and visually check for undissolvedparticles. /' .' I /' volume added (nLy /'appmr cono. d(n . ," SoMhlty data addwon (WW Sample Prep ,' OESERVATlONSlNOTES Flask va-texmhed7 YerINo Fhtk MnMed7 Y e a l o Flask allowed lo settle'? J%' Is there solute SUI prssenl7 Y e f l o ,/' Total volume solved a d a d (mL)... e soUe slill present? YaslNo / Old the substance dlsso\v#ln 1000 ml o r lms of solvent? es -Approximateconcentration at which uslml 7 The solubilityof the test substance is the shake flask method. N o -The solubilityof the to be determined via the column elution method. Solubility Screen Test Page 2 of 2 3M ETS Laboratory 3M Environmental Laboratory SOLUBILITY SCREEN GLP Study Number: TCR-009 Sample prepaatlon worksheet Test Substance: PFBS IDY Lot Z.TCR40017-71 Sourcn: 3Y Internal Solvent: Acetone Iwf: Tl;l?'t Step f Weigh approximately I O mg f 1 Weight of test substance: 2ndll@CEtb O@OtlEt - ,r/7-s/ /? g ottest substance into 8 2.5 mL-4 mL glass screw-top vial. mq Balance ID: I mg Thermomet~D-.*IIDL( Date/lnitials: 9 - / 3 - y ! f I E Analyst - - Analyst vobmn added (mLY approx cone. aRor Solubilitv data addlUon(uphnL) oalalrima+i3Ml io:20 SlBp2-1 Total v d u m sokenl added (mL)... 0.1 Appmxlmate Sdubility (ughnL) ... 1CQ.wO Datfime I step 2-2 Total volume solvent added (mL)... 0.5 - Approldmate Solubility (ugtmL)... 20.m TOWvolume solvent added (rnL)... Approximate Solubility (ughnL) .... Oatfime I Sample Prep OBSERVATIONSINOTES Flask vortu-mixd? &o Flask MnlcaIed? e y o Flask anowed lo L e ? b R- T~~ o Time A,lj .r3f;i~~\tdy - %% 1sthern sdute stin p m e n p Y e s a t i n e lastep 2-2 *c .>io./) CLT 4 1 ( e n # d u b is at 10%.and is can d e d infinilely duble. fu solubilityiesthg Isrequired. Flask vorlu-mixed? Y e M o RoomTemp Flask Sonicaled?YerMo l i m e Flarlr allowedto rnffln? YesNo Timn ------- i9'etlNo Room Temp Flask allowedlo retlle? YerMo Time 'C Is them solute still present? YerMo rnin dn *C Is there rolute ailpresent? Y e w mln rnin _ - _/--- i Flask vorkx-mlxed? Y e M o Room Temp Flaskronicstod?Yec/No Tim Flask allowed to setue? Y e m o Time *C Is there d u t e still present? Y e w min 4 Did the substance mL or less of solvent? proximate concentrationat which the test substance is soluble in solvent: >/Lva?S,q/rnl solubility of the test substance is to be determined via the shake flask method. or No Continueto page two for further testing. Ex>-,;.t i29 b 2 f MU Noted additional comments: [>lo$) .I Solubility Screen Test Page 1 of 2 3M ETS Laboratory 3M Environmental Laboatow SOLUBILI7Y SCREEN GLP Study Number: TCR-009 Samplepreparation workrheet Test Substance: PFBS ID# L o t t fCR00017-71 Source: 3W intimal Date: Solvent Acetone IM: Source: Step (continuedfrom page 1) "'Did the test substance di66OlVe In 2 mL? Is no need to contlnue with the screen test If NO, continue to step 3 to determlne 3 Weigh approximately 10 mg f 1 rng of test substance into a 1W mL graduatedstopperedcylinderholurnetric flask. Weight of test substance: mq Balance ID: Date/initials: 2nd rcplrmle optimal: mg Thermometer ID: 4 Add test salvent accOming to the table below. Fdlowing each addition of sdvenf, shakevlgomus/y&ortex. andsonicate I 1 I I 1 the mixture and visually check for undissolvedpadicicles. volume added (&)I appmx cone. allu t Solublilty data addluon l@W Sample Prep OBSERVATIONSINOTES Flask albwed (0 re*? YssrNo Time mk Total volume rdvenl added (mL)... Apptox'mate Solubiliy [ Total volume dwnl added (mL)... ed7YesNo Tms - M y s t Dsl~me I Total volume solvent add& (mL) Appmxmale Soiubiity( W L ) SlepI-1 loOa , 10 Flask vutar-musd? YerMo Roun Tamp F k r k sonrated7 YerMo Tme Fhrke(bwsdLO.stlle7 YsslNoTme Datallnitials: 'C Is here rdutl,~~ p rIaIIm ? YES,NMO mh rnQ Solubility Screen Test Page 2 of 2 3M ETS Laboratory I I I I I I I I I I I I P I 0 H H !ii H d P s 4 I I F I 0 3 P I f ID i b Batch Run # 5 of 61 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOOO5.D Sample Name: MeOH TN-A4604 MSDl 427, ElC=426.7:427.7(HO92XO\HILLMX35.D)API-ES Negative l " ' I " ' l " ' ~ ' ' ~ ~ " ' ~ " ' ~ 2 4 6 8 10 12 14mii Instrument 1 9/26/00 2:09:50 PM KLT Page 1 of 2 LIMS ~ 0 0 - 1 a n Page 68 of 98 Batch Run # 5 of 67 Data File C:\HPCHEM\1\DATA\HO9ZZOO\HILLOOO5.D ~~ ~ Sample Name: MeOH TN-A4604 Acq. Operator : KLT Inj : 1 Inj Volume : 5 p l Acq. Method : C:\HPCHEM\~\METHODS\PFBS~~~.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H9220O-A.M Last changed : 9/26/00 2:06:27 PM by KLT PFBS Analysis study# FACT-TCROO9 water solubility Reprocessing on Lefty 9-25-00 by KLT Sorted By Signal Calib. Data Modified : Tuesday, September 26, 2000 2:02:46 PM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name ~ #~ ~ ~ ~rPP-b-1- ~ - - - - - - - - _ ~ - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~ ~ ~ ~ 1 244.20000 THPFOS Totals without ISTD(s) : Signal 2:.MSD1 299, EIC=298.7:299.7 0.00000 Totals without I S T D ( s ) : 0.00000 2 Warnings or Errors : Warning : ISTD compound(s) not found Warning : Negative results set to zero (cal. curve intercept), (PFBS) Instrument 1 9/26/00 2:09:50 PM KLT Page 2 of 2 LIMS# ~ 0 0 - i a 7 7 Page 67 of 98 Batch Run # 52 of 67 Data File C:\HPCHEM\l\DATA\HO922OO\HILL0052.D Sample Name: 91800aPFBS2.4.4 mFm-TCRW 1:100,000 sample- DAY 2 in water diluted 3M EnvironmentalLaboratory ............................................................ Injection Date : 9/22/00 10:00:40 PM Sample Name : 91800aPFBS2.4.4 Seq. Line : Vial : 26 Acq. Operator : KLT Inj : 1 Inj Volume : 5 pl Acq. Method : C:\HPCHEM\l\METHoDs\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200 -A.M Last changed : 9/26/00 2:06:27 PM by KLT PFBS Analysis study# FACT-TCR009 water solubility __R_e__p_r__o_c__e_s__s__i_n_-g_----o_-n_-_-L_-e_-f_-t_-y_-_-_9_--_2-5_--_-0_-0_-_-_b-_y-_-_K-_L_-T_-_--------------_---_---------------_----------- ------------_-------- MSD1 427,ElC=426.7:427.7(H092MO\HILLco52.D) A PI-ES Negative 0 I ~ " I " ~ I ~ ~ ' I ' ' ~ I ~ ~ . I r 2 4 6 8 MSOl 299, ElC=298.7:294.7(H092200\HILLM)52.D) API-ES Negative 10 12 14m lXc0 Instrument 1 9/26/00 2:16:37 PM KLT c. Page 1 of 2 LlMSC E00-1877 Page 68 of 08 Sorted By Signal Calib. Data Modified : Tuesday, September 26, 2000 2:02:46 PM Multiplier 1,0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name ---#-I--------[-P--P-b-lI-------_----------------- 1 244.20000 THPFOS Signal 1: MSDl 427, EIC=426.7:427.7 Totals without ISTD(s) : 0.00000 Signal 2: MSDl 299, EIC=298.7:299.7 RetTime Type [minl -------I------I----------I----------l---------- 6.166 BB Area 9.31987e4 Amt/Area ratio 0.00000 Totals without ISTD(s) : Amount Grp Name [PPbl ( _ _ ) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 0.00000 PFBS 0.00000 1 Warnings or Errors : Warning : Negative results set to zero (cal. curve intercept), (PFBS) Instrument 1 9/26/00 2:16:37 PM KLT Page 2 of 2 LIMS# E00-1877 Page 69 of 98 MSDl 427. ElC=426.7:427.7(HW2COVllLL0087.D) API-ES Negative m- 6oowo5Mmo400000XKXxnXcCcD- layxx3- 0 I, 11Mw Instrument 1 9/26/00 4:11:16 PM KLT LlMSIl E001877 Page 70 of 98 Batch Run # 34 of 50 Data File C:\HPCHEM\l\DATA\HO922OO\HILL0087.D Sample Name: 91800bPFBS2.4.3 _1:_10_0,_0_00_d_i_lu-t--e--d---s_-a-m-p-l-e-_---D_-A-Y----2---i-n_-w--a--t--e--r-------------_-------3-M--En-v-iro--n-m--e-n-ta-l L-a-bo-ratory ~WFFReFTCROO9 Injection Date : 9/23/00 7 : 1 6 : 0 2 AM Seq. Line : 87 Sample Name : 91800bPFBS2,4.3 Vial : 41 Acq. Operator : KLT Inj : 1 Inj Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9 / 2 0 / 0 0 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200-B.M Last changed : 9/26/00 4:06:02 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-25-00 by EUT ...................................................................... Sorted By Signal Calib. Data Modified : 9/26/00 4:06:00 PM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name __# __~_[P_Pb_1 _--___-_--~-_____________________--_ 1 244.20000 THPFOS Signal 1: M S D l 427, EIC=426.7:427.7 RetTime Type [min] _______1_--__-1__________)__________ Area 7.896 BB I 2.80044e6 Amt/Area Amount Grp Name ratio _-[-P-P-b_l ----_ _ ) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1.00000 244.20000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2 : MSDl 299, EIC=298.7:299.7 RetTime Type [min] Area Amt/Area ratio - 6.777 BB 6.56607e4 0.00000 Totals without ISTD(s) : Amount Grp Name [ PPb 1 0.00000 PFBS 0.00000 1 Warnings or Errors : - Warning : Negative results set to zero (cal. curve intercept), (PFBS) Instrument 1 9/26/00 4:11:16 PM KLT I LIMSW E00-1877 Page 71 of 98 Batch Run # 1 4 of 67 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOOl4~D Sample Name: 00027-44-07 7oMoo8oMoosmax,4ooMoJrXx30- amax,- 10amQ- zcc#xxl- 175MM; 1Smm125000010M000- 750000- 5aTrx)- 25oMoQ 2to5 i 1 " ' 1 ' ~ ' I ~ ~ ' I " ~ I ~ ' ~ I ' I b I " ~ / " I " ~ 1 ~ ' ~ 1 " ' I a"l 1 Instrument 1 9/26/00 2:11:08 PM KLT Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----I__-#----------I------_--__--r--P_------_P-- I ~ 1 244.20000 THPFOS Signal Tuesday, September 26, 2000 2:02:46 PM 1.0000 I.0000 Signal 1: MSDl 427, EIC=426.7 :427.7 RetTime [min] --___--(-___-- Type Area Amt/Area Amount Grp Name ___-______)__r_a-_t_io____I___[P_P_b_I____ _ _ I _ _ _ _ - - - _ _ _ _ _ _ _ _ _ _ _ 7.896 BB I 2.95464e6 1.00000 244.20000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2: MSDl 299, EIC=298.7 :299.7 Totals without ISTD(s) : 614.19952 Instrument 1 9/26/00 2:11:08 PM KLT Page 2 of 2 LlMSl E00-1877 Page 73 of 98 Batch Run # 35 of 67 Data File C:\HPCHEM\l\DATA\HO922OO\HILL0035.D Sample Name: 91800aPFBS2.1.4 1 : 1 0 0 , 0 0 0 diluted sample- DAY 2 in water ----------------------------------------------------------~~~~~~laborato~ Injection Date : 9/22/00 5:31:05 PM Seq. Line :StugyS10.FACT-TCROOS Sample Name : 91800aPFBS2.1.4 Vial : 14 Acq. Operator : KLT Inj : 1 Inj Volume : 5 pl Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200-A.M Last changed : 9/26/00 2:06:27 PM by KLT PFBS Analysis study# FACT-TCR009 water solubility __Re__pr__oc__e__ss__in__g_-o-n_---L--e--f--t--y---9-----2--5-----0--0---b--y----K--L--T-------------------------------------------------------------------------------- MSDI 427, ElC=426.7:427.7(HCQZXIJHILL0035.D) API-ES Negative 7Wa30- aMM0- 5wooo- 4xMoo- 300000- m- 1UXR3- 04 I\ I ~ " I ' " l ' ' ~ l ' ~ ' l ~ " I ' ' ' I 2 4 6 8 10 12 1h i MSDl 299,EIC=ZB.7:299.7(H092200WILL0035.D) API-ES Negative 175(3ooo15umo12XiXQ- laxX300- 75ax)o- 500000- mooo- 7 Instrument 1 9/26/00 2:14:09 PM KLT Page 1 of 2 LIMS# 00-1877 Page 74 of 98 Batch Run # 35 of 67 Data File C:\HPCHEM\l\DATA\HO922OO\HILL0035.D 1:100,000 diluted sample- DAY 2 in water Sample Name: 91800aPFBS2.1.4 =========================================================-~i~m~~~borato~ Injection Date : 9/22/00 5:31:05 PM Seq. Line 0. FACT-TCROOQ Sample Name : 91800aPFBS2.1.4 Vial : 14 Acq. Operator : KLT Inj : 1 Inj Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200 -A.M Last changed : 9/26/00 2:06:27 PM by KLT PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-25-00 by KLT Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----I----#---------I--------------[---P----P----b1 1 244.20000 THPFOS Signal Tuesday, September 26, 2000 2:02:46 PM 1.0000 I.0000 Signal 1: MSDl 427, EIC=426.7:427.7 Totals without ISTD(s) : 0.00000 Signal 2: M S D l 299, EIC=298.7:299.7 Totals without ISTD(s) : 545.35188 Instrument 1 9/26/00 2:14:09 PM KLT Page 2 of 2 LIMS# E00-1877 Page 75 of 98 Batch Run # 4 0 of 67 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOO4O..D Sample Name: 91800aPFBS2.2.4 1:100,000 diluted sample- DAY 2 in water ===============================---------=================~~~~borato~ Injection Date : 9/22/00 6:50:20 PM Seq, Line :StUq#O. FACT-TCROOS Sample Name : 91800aPFBS2.2.4 Vial : 18 Acq. Operator : KLT Inj : 1 Acq. Method Inj Volume : 5 p l : C:\HPCHEM\l\METHoDs\FFBS92O.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHoDS\H92200-A.M Last changed : 9/26/00 2:06:27 PM by KLT PFBS Analysis study# FACT-TCR009 water solubility _Re_pr_oc_e_ss_in_g_o_n _Le-f-t-y---9-----2--5-----0--0---b--y----K--L--T---------------------------------------------------------------- 7cccm- 6Mwo- 500000- m- Tx)(xx3- 2ulum- 1oaxx)- 0 J 17500001sO: 125000010- 750000sO250M30- 0 3 4+ \ I ~ ~ . l ' ' ~ ( ' ~ r l ~ ~ ' l . ~ Instrument 1 9/26/00 2:14:52 PM KLT Page 1 of 2 LlMSIlE00-1877 Page 78 of 98 Batch Run # 40 of 67 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOO4O.D Sample Name: 91800aPFBS2.2.4 _-1-: 1-_0_0__, 0__0 _0_---d-_i_l_u-t-_e-d_---s--a--m--p--l--e------D--A--Y----2----i--n----w--a-t-e-r------------------3M--E-nv--ir--o-n-m--e-ntalLaboratory fiiiVWfi.mtf-~~~009 Injection Date : 9/22/00 6:50:20 PM Seq. Line : 40 Sample Name : 91800aPFBS2.2.4 Vial : 18 Acq. Operator : KLT Inj : 1 Inj Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200-A.M Last changed : 9/26100 2:06:27 PM by KLT PFBS Analysis study# FACT-TCR009 water solubility _Re_pr_oc_e_ss_in_g_o_n -_L_e_f-t-y--9---2-5---0-0--_b-y----K-L--T-------------------------------------------------------------------------------_--- Sorted By Signal Calib. Data Modified : Tuesday, September 26, 2000 2:02:46 PM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name _ _#_ _ ( _ _ _ _[_PP_b_l- _ - _ - - I - - - - - - - - - - - - - - - - - - - - - - - - - 1 244.20000 THPFOS Totals without ISTD(s) : 0.00000 Totals without ISTD(s) : 557.05679 Instrument 1 9/26/00 2:14:52 PM KLT &&-, Initial bS ? O / iy Dah? Page 2 of 2 LIMS# 00-1 877 Page 77 of 98 Batch Run # 4 4 of 67 Data File C:\HPCHEM\l\DATA\HO922OO\HILL0044.D Sample Name: 91800aPFBS2.3.1 _1:_10_0,_0-0-0--d--i--l--u--t--e--d----s--a--m--p--l--e------D--A--Y---_2---i--n----w-a--t--e--r-------------------------3-M-E-n-vi--ro--nm-e-ntal Laboratory SEW~~~~-TCROOS Injection Date : 9/22/00 7:53:47 PM Seq. Line : 44 Sample Name : 91800aPFBS2.3.1 Vial : 19 A c q . Operator Acq. Method : KLT Inj : 1 I n j Volume : 5 pl : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200 -A.M Last changed : 9/26/00 2:06:27 PM by KLT PFBS Analysis study# FACT-TCR009 water solubility _Re_pr_oc_e_ss_in_g--o-n--L--e--_f-t--y----9-----2--5-----0--0----b--y----K--L--T---------------------------------------------------- 7axm8owoo- Eman- 400000Jma,- 1 *$qo% *BB' 2oawx)- 1oaxM- 0 I, 1 , 1 ' ~ ' 1 ~ ' ' 1 ~ ' ~ -I 1 ' ' 1750a3015Mooo: 125oax31OMoM)75ooco5MMo250000- 0 i,..,9y." I ' " l ' ~ ' l ~ " I ~ " I " ~ l ~ ' ~ Instrument 1 9/26/00 2:15:27 PM KLT - ... Page 1 of 2 LIMS# E00-1877 Page 78 of 98 Sorted By Signal Calib. Data Modified : Tuesday, September 26, 2000 2:02:46 PM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name # r PPb1 ___-I___--________ I______-__________________ 1 244.20000 THPFOS Signal 1: MSDl 427, EIC=426.7 :427.7 RetTime Type Area Amt/Area Amount Grp Name [ min] ----_--I---_-_ _ _ _ _ _ _ _ _ _ _ I _r_a_t_i_o_ _ _ ___1[-P-P-b-l----- - I - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 7.888 BB I 2.87261e6 1.00000 244.20000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2: MSDl 299, EIC=298.7:299.7 RetTime T y p e [ min] Area Amt/Area ratio Totals without ISTD(s) : 556.36504 Instrument 1 9/26/00 2:15:27 PM KLT Page 2 of 2 LIMS# EOO-1877 Page 79 of 98 MSD1 427,ElC=426.7:427.(7H0922W\HILLOlO4.D) API- ES Negative I I " ' I " ' 1 ' ~ ' I " ' I ' ~ ~ I 8 ~- I 2 4 6 8 MSDl 299,EIC=2!?8.7:299.7(H092203VllLLOlW.D) API-ES Negative 10 12 1 hi l7XCUl- - 1- 125axx)10000CO- 750000- 5a3aT)- 25oaM- Y l " ~ l ' " I " ' l " ' I " ' I ~ ' ' I 2 4 6 8 10 12 14mii Instrument 1 9/27/00 10:03:33 AM KLT L hitiad J&L( oat6 Page 1 of 2 LlMSt 00-1877 Page 80 of 98 Sorted By Signal Calib. Data Modified : 9/27/00 9:51:42 AM Multiplier 1.0000 Dilution 1.0000 Sample I S T D Information: ISTD ISTD Amount Name # [PPbl ~ ~ ~ ~ ~ ~ _ ~ ~ ~ ^ - - - - - - - ~ - - ~ ~ ~ 1 244.20000 THPFOS Signal 1: MSDl 427, EIC=426.7:427.7 RetTime Type --[-m-i-n-]- I ------ 7.879 BB I Totals without ISTD(s) : 0.00000 Signal 2 : M S D l 299, E I C - 2 9 8 - 7 :299.7 RetTime T y p e [minl Totals without ISTD(s) : 555.67175 Instrument 1 9/27/00 10:03:33 AM KLT Page 2 of 2 L I M S E00-1877 Page 01 of Q8 Batch Run # 22 of 50 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOlll.D Sample Name: 91800aPFBS3.2.3 -_1-:__10_-0_,_0--0--0---d--i--l--u--t--e-_d---s--a--m--p--l--e-------D--A--Y----3----i--n---w--a--t--e--r------------------------------------3--M--E--n-v--ir-o--nmentalLaboratory =?FR?f-TCROO9 Injection Date : 9/23/00 1:36:47 PM Seq. Line : 111 Sample Name : 91800aPFBS3.2.3 Vial : 49 A c q . Operator Acq. Method : KLT Inj : 1 Inj Volume : 5 pl : C:\HPCHFX\1\METHODS\PFBS92O.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHoDs\H92200-C.M L a s t changed : 9/27/00 9:54:51 AM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-27-00 by KLT 700000- gX3a30- soaxx,- 400000- rx3a)(3- 2(30000- 1m- 0 I . * , 1 - I , , r 1 8 I , I I , , 2 4 6 8 MSDl 299,EIC=298 72997 (H0922M1\HILL0111 0)API-E5 Negative 10 12 I 14m MMOOO1 15OoQ30- I0M)O- 5oaxx)- 1 I! Instrument 1 9/27/00 10:04:35 AM KLT Page 1 of 2 LlMSt E00-1877 Page 82 of 98 Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----1__#___-____---1-_______[___P___P___b___l_---- 1 244.20000 THPFOS Signal 9/27/00 9:51:42 AM 1.0000 1.0000 Signal 1: M S D l 427, EIC=426.7:427.7 7.885 BB 12.91948e6 Totals without ISTD(s) : 1.00000 244.20000 0.00000 THPFOS S i g n a l 2 : M S D l 299, EIC=298.7:299.7 Totals without ISTD(s) : Amount Grp _ _r P_Pb_1 - - - - - - 711.83532 711.83532 Name Instrument 1 9/27/00 10:04:35 AM KLT Page 2 of 2 LIMS# E00-1877 Page 83 of 98 Batch Run # 17 of 50 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOO7O.D ~ Sample Name: 91800bPFBS2.1.3 MSDl 427, ElC=426.7:427.7 (H092200\HILL0070.D) API-ES Negative I , ,, I ~ ~ ~ ~ " ' I ' ~ " ' ' ' I ' 2 4 6 8 10 12 1 hi MSD1 299, EIC=298.7:293.7 (H092XO\HlLL0070.D) API-ES Negative Instrument 1 9/26/00 4:08:50 PM KLT Page 1 of 2 LlMSlY E00-1077 Page 04 of 90 Batch Run # 17 of 50 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOO7O.D Sample Name: 91800bPFBS2.1.3 1:100,000 diluted sample- DAY 2 in water _____-----_-_____________________c______----------------------------- ______--________________________________------------------- 'io Injection Date : 9/23/00 2:46:14 AM Seq. 3M Environmental Laboratory Sfu?d-RKFRCT-TCROOQ Line : Sample Name : 91800bPFBS2.1.3 Vial : 29 Acq. Operator : KLT Inj : 1. Inj Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200-B.M Last changed : 9/26/00 4:06:02 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility _____---------___ Reprocessing on Lefty 9-25-00 by KLT .................................................................... Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: r ISTD ISTD Amount # PPb 1 ____j_--------__--J_------------------------ Name 1 244.20000 THPFOS Signal 9/26/00 4:06:00 PM 1.0000 ' 1.0000 Signal 1: MSDl 427, EIC=426.7:427.7 Totals without ISTD(s) : 0.00000 S i g n a l 2: MSDl 299, EIC=298.7:299..7 Totals without ISTD(s) : 537.55839 Instrument 1 9/26/00 4:08:50 PM KLT Page 2 of 2 LIMS# E00-1877 Page 85 of 98 Batch Run # 20 of 50 Sample Name : 91800bPFBS2.2.1 Vial : 31 Acq. Operator : KLT Inj : 1 I n j Volume : 5 pl Acq.. Method : C:\HPCHEM\l\METHODS\PFBS920.M L a s t changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200 -B.M Last changed : 9/26/00 4:06:02 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility R_e_pr_oc_e_s_s__in__g__o_-n-_-L_ef-t-y---9-----2--5----0--0----b--y----K--L--T---------------------------------------------------------- MSDl 427, ElC=426.7:427.7 (H092XD\HILL0073.D) API-ES Negative - 6oaxx)- 500000- 400000- 300000- 2oooM)- 1Oax10- 0 I ~ ' ' l ' ~ ~ l ' r ' I ~ ' - 1l , . I *, , - 2 4 6 8 MSDI 299, ElC=298.7:299.7 (HC?32XO\HILL0073.D) API-ES Negative 10 12 1h 2 16Oa300- f-KxXC012oaTx)- 1000WO8oaMo600000400000MOOOO- 0 Instrument 1 9/26/00 4:09:16 PM KLT Page 1 of 2 LlMSW E00-1877 Page 88 of 98 Batch Run # 20 of 50 Data File C:\HPCHEM\l\DATA\HO922OO\HILLOO73.D Sample Name: 91800bPFBS2.2.1 _1:_10_0,_0_00_d-i-l-u-t-e-d--s-a-m-p-l-e----D-A-Y--2--i-n-w-a-t-e-r----------------------3--M--E--n--vi--ro--n-m-e-n-ta-l L-a-bo-ra-to-ry-- -SfUXf-No7FAFT-TCROO9 Injection Date : 9/23/00 3:33:53 Seq. Line : y13 Sample Name Acq. Operator : 91800bPFBS2.2.1 : KLT Vial : 31 Inj : 1 Inj Volume : 5 p l A c q . Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by U T Analysis Method : C:\HPCHEM\l\METHODS\H92200-B.M Last changed : 9/26/00 4:06:02 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility R_e_pr_oc_e_-s_s_i_n_g__o__n__L_e_f--t--y----9-----2--5---0-0-b-y--K-L-T----------------------------------------------------------------------- Sorted By Signal Calib. Data Modified : 9/26/00 4:06:00 PM Multiplier 1 * 0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name _ _# _ _ ~ [_P_Pb_l_ _ - - - - - - - - ~ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~ ~ ~ ~ 1 244.20000 THPFOS Signal 1: MSDl 427, EIC=426.7:427.7 7.889 BB I 2.80381e6 Totals without ISTD(s) : 1.00000 244.20000 0.00000 Signal 2: MSDl 299, EIC=298.7:299.7 RetTime Type Area Amt/Area Amount Grp [minl ratio _______.1-_----1_---______I__________ _'_[P-P-b-l----- 6.756 BB 7.41853e6 8.19801e-1 529.69166 ' Name Totals without ISTD(s) : 529.69166 Instrument 1 9/26/00 4:09:16 PM KLT Page 2 of 2 LIMSX E00-1877 Page 87 of 98 Batch Run # 28 of 50 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOO8l.D Sample Name: 91800bPFBS2.3.2 Sample Name : 91800bPFBS2.3.2 Vial : 36 Acq. Operator : KLT Inj : 1 Inj Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H922OO-B.M Last changed : 9/26/00 4:06:02 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility R__e_p_r_o_c_e_s_s_i_n__g___o__n___-L___e__f__t__y__9_-_2_5_-_0_0__b_y__K_L_T__-_-------_--------------------- --- MSDl 427, ElC=426.7:427.7(H092XO\HILL0081. O ) API-ES Negative 7wooo- 600000- 2 to5 5uXx30- m- 3ooax)- Mwoo- 1oowO0 I 1 ' ~ ' 1 ' ' ~ 1 ~ ~ ' 1 ~ " 1 " ' l ' ' ' J 18MMM- 1600000- 14oma3- 12QXXO1000000- 8oayx)8Oa300- 400000XoM30- 0 $." p ' Instrument 1 9/26/00 4 : 1 0 : 2 5 PM KLT Page 1 of 2 LIMS# 00-1877 Page 08 of 98 Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: I S T D ISTD Amount Name # [ PPb1 ____1_____--_____-1-____________________---- 1 244.20000 THPFOS Signal 9/26/00 4:06:00 PM 1.0000 1.0000 Signal 1: MSDl 421, EIC=426.1:427.7 Totals without ISTD(s) : 0.00000 Totals without ISTD(s) : 544.66468 Instrument 1 9/26/00 4:10:25 PM KLT Page 2 of 2 LIMSIT E00-1877 Page 89 of 98 Batch Run # 15 of 50 Data File C:\HPCHEM\1\DATA\HO922OO\HILLOl4O.D Sample Name: 91800bPFBS3.1.1 1:100,000 diluted sample- DAY 3 in water _____------------------------------------------------3M--En-v-ir-on-m-entalLaboratory Injection Date : 9/23/00 9:16:49 PM Sea. Line 140 SfGTNOTFm-TCRO09 : Sample Name : 91800bPFBS3.1.1 Acq. Operator : KLT Inj : 1 Inj Volume : 5 p l A c q . Method : C:\HPCHEM\l\METHODS\PFBS92O.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHoDS\H92200-D.M Last changed : 9/27/00 2:04:35 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-27-00 by KLT MSD1 427, ElC=426.7:427.7 (H092XO\HILL0140.D) API-ES Negative 4 700000- 600MXl- 5oaxx3- #xMoo- i 300000- 200000- ia3YXl-l 0- - - - __ - . - I1 . I 1 _, . - I - ____- I 0 'I - I ' 2 '-, I . . I - I ' ~ ' l ' ' ' 1 " ' 1 ' ' ' ~ 4 6 8 10 12 I 14mi Instrument 1 9/27/00 2:04:35 PM KLT . ' LlMSAt E00-1877 Page 90 of 98 ~~ Batch Run # 15 of 5 0 Sample Name : 91800bPFBS3.1.1 Vial : 59 Acq. Operator : KLT Inj : 1 I n j Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200-D.M Last changed : 9/27/00 2:04:35 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-27-00 b y KLT Sorted By Calib. Data Modified : Signal Wednesday, September 27, 2000 1:55:19 PM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name _ _#_ _ I _ _ _ _[P_P_b_l_ _ _' _ _ _ ) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - _ 1 244.20000 THPFOS S i g n a l 1: MSDl 427, EIC=426.7:427.7 RetTime Type Area ___[__m_-)i--n--]--I-_--------~__-_-_____~--_------- 7.883 BB I 2.96550e6 Amt/Area ratio 1.00000 Totals without I S T D ( s ) : Amount Grp Name [PPbl 1--1-----______-______ 244.20000 THPFOS 0.00000 S i g n a l 2: M S D l 299, EIC=298.7:299.7 RetTime Type Area Amt/Area Amount Grp Name _ _[_mi_n_]_ _ I _ _ _ _ _ _ ( _ _ _ _ _ _ _ _ _ _ l _ _ra_t_i_o_ _ _ _ _ l _[_P_P_b-1- _ _ - - ~ - - ~ - - - - _ _ _ - - - _ _ _ _ _ - - _ 6.761 BB 7.79224e6 8.53438e-1 547.62323 PFBS Totals without ISTD(s) : 547.62323 Instrument 1 9/27/00 2:04:35 PM KLT Page 2 of 2 LlMSt 00-1877 Page 91 of 98 Batch Run # 22 of 50 Data File C:\HPCHEM\l\DATA\HO922OO\HILLOl47.D Sample Name: 91800bPFBS3.2.3 __1_:__1_0__0_,-_0-_0-_0-__-d_-i_-l_-_u--t--_e--d-_-_s-_a_-m_-p_-_l-_e-_--_-_D-_A-_Y-_-_3_-_-i_-n_-_-w_-a---t---e--r----------------------- ----------3-M-E-n-vi-r_o-n_-m_entalLaboratory S~~VIWTFEVROTO^C Injection Date : 9/23/00 11:07:57 PM Seq. Line : 147 Sample Name : 91800bPFBS3.2.3 Vial ; 65 Acq. Operator : KLT Inj : 1 I n j Volume : 5 p l Acq. Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H92200-D.M Last changed : 9/27/00 2:05:30 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-27-00 by KLT MSD1 427, ElC=426.7:427.7 (H0922DJHILL0147.D) API-ES Negative Q - . . .. 3 1750000- fsxyxx)1250000 100M0075owoscafio- ....... ......... ...-..... ..__ ... .. . ~ . ....I .L - - - . . .... , . . .. .. Instrument 1 9/27/00 2:05:31 PM KLT Page 1 of 2 LlMSX E00-1877 Page 92 of 98 ~ Batch Run # 22 of 50 Data File C:\HPCHEM\l\DATA\HO922OO\HILLOl47.D Sample Name: 91800bPFBS3.2.3 _1_: 1_0_0_,-_0--0_--_0____--d_-_i-__l__u__t__e__d____s__a___m__p__l__e___-___D___A__Y____3____i__n__-_-w-_a-_-t---e---r----------------- 3M EnvironmentalLaboratory %W~WCFF~-TCROO~ Injection Date : 9/23/00 11:07:57 PM Seq. Line : 147 Sample Name A c q . Operator : 91800bPFBS3.2.3 : KLT A c q . Method : C:\HPCHEM\l\METHODS\PFBS920.M Last changed : 9/20/00 5:29:49 PM by KLT Analysis Method : C:\HPCHEM\l\METHODS\H922OO-D.M Vial : 65 Inj : 1 Inj Volume : 5 pl Last changed : 9/27/00 2:05:30 PM by KLT (modified after loading) PFBS Analysis study# FACT-TCR009 water solubility Reprocessing on Lefty 9-27-00 by KLT _____________ ______-___________________________L_____---------------------------- Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name -___1_-#_--___-----1-_______[___P___P___b___l_---- 1 244.20000 THPFOS Signal Wednesday, September 27, 2000 1:55:19 PM 1.0000 1.0000 Signal 1: MSDl 427, EIC=426.7:427.7 Totals without ISTD(s) : 0.00000 Signal 2 : MSDl 299, EIC=29e.7:299-7 Totals without I S T D ( s ) : 538.76369 Instrument 1 9/27/00 2:05:31 PM KLT Page 2 of 2 LIMW E00-1877 Page 93 of 90 - Attachment D Deviationsfrom the Protocol 3M EnvironmentalLaboratory Study NO.FACT-TCROOS L l M S l E00-1877 Page 94 of Q8 Record of Deviation 1. Identification 3M Environmental Laboratory Study NO. FACT-TCROOS FACT-TCR-009__ __-----__I___--.__-_- ---- --_-I_ .__ __________ Deviation Type: ~ ~ 0Method 0Equipment Procedure (Check one) x o c o l 0 Other: ___-- --.--- ._______ - ---- Document Number: ETS-8-155.0 i Date(s) of occurrence: 13 Sep 00 to 24 Sep 00 I I /I. Description: Required Procedu~rdp-ro.ce-ss:_ E-TS----8_-1_5_5_.0-.s_ec_t_ io-n10.2.1 states, " Ma-t-ri-x-spike-s_a_re-.p.r_e_ pa_re_d__ for each sa- mpIe- set an~ d analyzed to determine the matrix effect----o-----n the recovery efficiency." n Section 10.2.3 states, `:analyze the matrix sp~ike and t-he matri-x spike duplicate ( if prepared)~ ----____I.-___ in the same run as the original sample." ~ -__---.___ Actual Procedure/proces- s- :- - T. - h- e- I m_ atrix spikes were not prepared, theref-o-re, they were no__t- - - anal- yzed.-- --..._^__ ---- _- ---I----._ Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) The ETS-8-155.0 method has been revised so that matrix spikes may b- e prepared aL- t the iiscre_ti_o-n.-o-f_t_he project-.l-e_ad-.-_____ -- - -___ --__ Xecorded By: ---__-._-- -- ./!/OI Date: i I I _,___._.I /". Project Impact on Study/ __ ~ ______.________..L_IC___. ~~ -________. rhis deviation does not adversely affect the quality of the data. _ .__I-.__-.I ~ ____-.-I-- -___________ k e matrix is well defined and characterized. Matrix spikes are not actually ___._.-..._-__-----.--.-~_-_-_--__--~_..- I- .-.__-_- _______I__.I iossible in this s-tudy since the analyte of interest --____-.----- is already at-saturation concentrations. _. ........ .C c - - --- ~ _____._.I_______.-- - - - .- - ..Date:. ~ . . .. iutborized By: (Study Director I!P h c t Lead) ! 3M Environmental Laboratory Form ETS-4-8.0 / Deviation No. 1. (assigned by Study Director or Project Lead at the end of study or project) L I M S ~EOO-Ian Page 95 of 98 ATTACHMENET: SAMPLECALCULATIONS 3M EnvironmentalLaboratory Study NO. FACT-TCROOS LIMS# E00-1877 Page 96 of 98 3M Environmental Laboratory Study No. FACT-TCROOS 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 magnitudeof the standard deviation and is calculatedby dividing the standard deviation by the mean. Means are calculated by adding individualentities and dividing the resultant sum by the number of individual entities. Standard deviationwas calculated using the following equation: -Where n is the number of observations, and x is sample Average/StandardDeviation/%RSD example calculations: Sample ID Rep1 I Rep 2 I Rep 3 1 Rep 4 Concentration PglmL 53300 52400 53000 52000 Calculations Average (53300+52400+53000+52000)14=52700 Std Dev (see above equation)= 458 %RSD (2600/54000)=0.87% Final reported values were corrected for purity. For PFBS TCR-99030-23, the oncolumn concentration was multipliedby the purity (96.7%). Example calculation: 55700 pglrnL * 0.967= 53900 pglrnL For PFBS TGR-00017-71, the oncolumn concentration was multiplied by the purity of the standard used to quantify the sample (96.7%). Then the concentration was corrected for the purity of the test substance (97.3%) which was a 0.6% difference (a correction factor of 1.006). Example calculation: 65500 pg/mL * 0.967 *1.006= 63700 pg/rnL Data that did not meet acceptance criteria as described in the OPPTS and OECD guidelines was excluded using statisticaljustification provided by Dixon's Q-test. A data point may be excluded if "ClobSmedis. 'greaterthan "Qlabulalewd'ith 90% confidence. = QobWerved gaplrange where "gap" is the difference between the questionable data point and the closest value of the data set, and "range" is the difference between the highest and lowest value of the data set. Q-tabulated (90% confidence) for 4 observations (or data points) is 0.679 and for 3 observations is 0.886. LIMS# EOO, 1877 Page 97 of 98 3M Environmental Laboratoty Study No. FACT-TCROOS Q-Test Example (Shaded number represents the suspected outlier): h Description Sample Conc., Rep1 Sample Conc.. R e ~ 2 uo/mL 53896 53056 Retain? No Q-Test example conciusion: Since QobJewed, 0.992,was greater than Qlsbulatedl 0.679, excluded. 4 may be LIMS# 00,1877 Page 98 of 98