Document mXvYJdyEBex5KyDjrN13QD50

BACK TO MAIN Study Title Characterization Study of PFOS, Primary Standard - Test Control Reference #TCR-00017-046 PHASE:SOLUBILITY OF PFOS IN WATER 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 935BushAvenue St. Paul, MN55106 Project Identification 3M Environmental Laboratory Study # FACT-TCR002 (LIMS #E00-1716) Centre Analytical Laboratories Study # 023-021 Total Number of Pages 67 BACK TO MAIN GLP COMPLIANCE STATEMENT ~ ~~ ~ ~~ Study Title: Characterization Study of PFOS, Primary Standard- Test Control Reference # TCR-00017-046, PHASE: Solubility of PFOS in Water. Study Identification Number: FACT-TCR002, Centre Analytical Laboratories Study # 023-021 This phase of the study was conducted in compliancewith Environmental Protection Agency (EPA) Good Laboratory Practice (GLP) Standards 40 CFR 160 withthe exceptions in the bulleted list below. Exceptions to GLP compliance: 0 Some corrections were not made in the raw data per GLP requirements 0 The electronic data systems in use have not been validated and there is not an electronic audit trail of corrections currently available (40CFR 160.130 (e)). Authenticated hardcopies of chromatograms and associated documents will be considered as the original raw data. 0 Not all data was dated on the day of entry and signed or initialed by the person entering the data. 0 The SOPS (methods) used in the study were not approved by management prior to using them. .- v Mark E. Ell&fson, Prin */A , -CY William K. Reagen, Testing Facility ManagDeamteent bate ' &+& FACT -TCR002 (LIMS #E00-1716), Page 2 of 67 BACK TO MAIN QUALITY ASSURANCE STATEMENT Study Title: Characterization Study of PFOS, Primary Standard - Test Control Reference # TCR-00017-046, PHASE: Solubility of PFOS in Water. Study Identification Number: FACT-TCR002, Centre Analytical Laboratories Study # 023-021 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. Iwarn, I ~ ~ , O ~ I O OData 01l23l01,01l25l01, 01/26l01,01l29l0101l31l01 Draft Phase Report QAU Represedtative 12/11/00 0210 1 I0 1 12/11/00 02/01I01 <+3/30I /~ 'Date FACT -TCR002 (LIMS #E00-1716),Page 3 of 67 BACK TO MAIN TABLEOF CONTENTS GLP Compliance Statement............................................................................................................... 2 Quality Assurance Statement ............................................................................................................. 3 Study Information ............................................................................................................................... 6 Summary ............................................................................................................................................ 7 Purpose .............................................................................................................................................. 7 Test Substance .................................................................................................................................. 7 Test System ....................................................................................................................................... 8 Method Summaries ............................................................................................................................ 8 Preparatory Methods.......................................................................................................................... 8 Sample Collection and Analysis ......................................................................................................... 9 Analytical Method ............................................................................................................................... 10 Data Summary, Analyses, and Results.............................................................................................. 11 Analytical Results ............................................................................................................................... 11 Data Summary ................................................................................................................................... 12 Statistical Methods ............................................................................................................................. 14 Statement of Conclusion .................................................................................................................... 14 References ......................................................................................................................................... 14 List of Attachments ............................................................................................................................. 15 Signature Page................................................................................................................................... 16 FACT -TCR002 (LIMS #E00.1716). Page 4 of 67 BACK TO MAIN LISTOF TABLES Table 1. Summary Table of Solubility ofPFOS TCR-00017-046 in Water........................................ 7 Table 2. Characterizationof the Test Substance and Analytical ReferencSeubstances.................7 Table 3. Descriptionof test systems usedin this study .................................................................... 8 Table 4. Solubility Screen Test Sample Preparation........................................................................ 9 Table 5. Shake Flask Method Preparatiofnor Study (FACT-TCR002)............................................. 9 Table 6. Sample Collection and Preparation for Study FACT-TCR002..*......................................... 10 Table 7. CalibrationStandard Reprocessing Summary..................................................................... 12 Table 8. PFOS TCR-00017-046 (PFOS) Data Summary................................................................. 13 Table 9. Summary Table of Solubility ofPFOS TCR-00017-046 in Water........................................ 14 FACT -TCROOP (LIMS #E00.1716). Page 5 of 67 BACK TO MAIN STUDYINFORMATION Study Director John Flaherty Centre Analytical Laboratory 3048 Research Drive State College, PA 16801 Analytical Chemistry Laboratories Solubility Testing Certificate of Analysis 3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Centre Analytical Laboratory 3048 Research Drive State College, PA 16801 John Flaherty, Study Director Dr. William K. Reagen, Laboratory Manager Mark E. Ellefson, Principal Analytical Investigator Kristin L.Terrell, Analytical Chemist Mark L. Anderson, Analytical Chemist Cindy M. Carlson, Analytical Chemist Sponsor Dr. William K. Reagen 3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Experimental Dates Sample Analysis Initiation: 13 July 2000 Sample Analysis Completion: 29 August 2000 Archives Reserve samples of the reference standards will be maintained by the 3MEnvironmental Laboratory. All study data for this phase and a copy of the phase report are archived by the 3M Environmental Laboratory. FACT -TCR002 (LIMS #E00-1716),Page 6 of 67 BACK TO MAIN SUMMARY The approximate solubility of PFOS in water wasdetermined by performing a solubility screen test, followed by a quantitative solubility determination via Shake Flask Method. Results are presented in table 1 Table 1. Summary Tableof Solubility of PFOS TCR-00017-046 in Water. Day 1 621 lg/mL 23 Day 2 717 pg/mL 91 Day 3 702 u d m L 2 I Average Value 680 pglmL 43 4% 13% 0.3% 6% I The solubility of PFOS TCR-00017-046 in water wasdetermined to be 680 pg/mL at 24-25 "C. PURPOSE The purpose of this phase of the study was to determine the solubility of PFOS in water as part of the characterization of this test, control, and reference substance. TEST SUBSTANCE PFOS,Test Control Reference #TCR-00017-046 Table 2. Characterization of the Test Substance and Analytical Reference Substances Source SRpee-ccriyasllytalMlizeadterbiayls,GBeoldrgg.e2M36o-oB1r-e1,03M SynQuest Labs Expiration Date 01-01-2010 Not Available Storage Conditions Frozen Frozen 3M Laboratory Identification Number I DPeshcyrsipictaioln I Puritv TCR-00017-046 I PowdWerhite I 97.9% TCR-00017-04L7ot #Q75-30 IPowdWerhite I I* I 'The purity of THPFOS (currently being determined by Centre Analytical Laboratory) will not affect any analytical findings. The same lothource of THPFOS was used throughout the entire study. FACT-TCR002 (LIMS #E00-1716), Page 7 of 67 BACK TO MAIN TEST SYSTEM Table 3. Descriptionof test systems usedin this study Conditions Source Expiration Date I Storage I 3NMIdueLmnabtbifeiocrraattiorny I Purity/Grade Millipore System NA I Ambient I I NA I Type 1 ASTM I I METHOD SUMMARIES The solubility determination of PFOS TCR-00017-046 in water was performed according to United States Environmental Protection Agency OPPTS 830.7840, "WaterSolubility: Column Elution Method; Shake Flask Method" and OECD 105, "Water Solubility" Guidelines, using the3M Environmental Laboratory methods ETS-8-170.1 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents;" and ETS-8-172.1 "Shake Flask Method; Solubility Determination of a Test Substance in Various Solvents". All analyses were performed according to the 3M Environmental Laboratory method ETS-8-155.0"Analysis of Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or WaterExtracts Using HPLC-ElectrospraylMass Spectrometry". The Solubility Screen Test Method (ETS-8-170.1) was used to determine the solubility range of PFOS TCR-00017-046 in water. Incremental amounts of water were added to PFOS neat material and a qualitative determination of the solubility point was made. The screen test indicated a solubility of PFOS in water of greater than 5,000 pg/mL. The Shake Flask Method was used for the quantitative determinationof PFOS solubility. Detailed descriptions of the Solubility Screen Test Method and Shake Flask methodsused in this study are located in Attachment A. PREPARATOMRYETHODS 0 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 anestimate of the solubility point of the test substance in the solvent of choice. Approximately ten milligrams of PFOS was weighed into a glass vial, and varying amounts of water were added in a step-wise fashion. After each water addition, the vial was shaken, sonicated, and observed for particulates. The screen test estimated the solubility of PFOS in water to be greater than 5,000 pg/mL. This concentration estimation was used as a starting point for the shake flask method, and as a guide to determine the amount of dilution necessary to bring the final sample concentration into the analytical range of method ETS-8-155.0 (2.5- 1002 ng PFOS/mL). FACT -TCROOP (LIMS #E00-1716), Page 8 of 67 BACK TO MAIN Table 4. Solubility Screen Test Sample Preparation 1 10 mgtestsubstanceweighedintotared 4.0 mLglassscrew-cappedvial 2 0.1 mLofwateradded to glassscrew-cappedvial 3 Following the addition of water, vial was capped, shaken vigorously and vortexed, and sonicated. Mixing and sonication times recorded on the worksheet. 4 Sample checked visually for undissolved particles of the test substance. (If all of the test substance is dissolved, the solubility is determinteodbe greater than10% (mass/volume) and no additional solubility testing is required. If a poortfiothne test substance remains undissolved, additional dilutions are performed using the appropriate solvent.) Observation made: undissolved particles present..continue to step5. 5 Step 2 repeatedwith total volumeofwaterof 0.5, 1, and 2 mL,respectively.Followingwater addition, vial was shaken, sonicated, and observed for particulate. 6 Following the final water additionto bringthevolumeofwater to 2 mL, itappearedasthough the PFOS had dissolved. The concentration was approximate5,ly000 pg PFOS/mL and used as a starting point for the shake flask method. 0 ETS-8-172.1 "Shake Flask Method: Solubility Determination of a Test Substance in Various Solvents." Greater than 0.045 g PFOS was weighed into a tared 15 mL plastic * centrifuge tube. Water was added gravimetrically to approximately 10 g. Nine centrifuge tubes were prepared, along with three method blanks (tubes with 10 g 0.5 g water only). The centrifuge tubes were placed in a temperature-controlled orbital shaker set at approximately 3OOC. See the following table documenting the shake flask method preparation: Table 5. Shake Flask Method Preparation for Study (FACT-TCROOZ) 2 Water addedto centrifuge tube such that total weight of PFOS 9and water is > 9.9-10.4 g. 3 Method Blank Preparation- Approximately 10 mL water adtodead 3 tared centrifuge tube gravimetrically. 4 Samples sealed with tape, vortex-mixed, and placed in a 12 temperature-controlled orbital shaker set at approxima3tOe.lOy 0C, rotation speed set at approximat2e2ly5 rpm. 07-31-00 07-31-00 07-31-00 SAMPLE COLLECTIONAND ANALYSIS On each of Days 1, 2, and 3 (approximately 24, 48 and 72hours after initial sample preparation) three centrifuge tubes (containing PFOS) and a method blank were removed for analysis. The centrifuge tubes with PFOS and method blanks were allowed to equilibrate at 24-25OC for 24 hours, centrifuged, and aliquots of supernatant were collected. The aliquots were diluted serially using two 1OOX dilutions (990pL methanol:lO pL sample) to a final dilution factor of 10,OOOX. The final diluted samples were spiked with internal standard (1 1pLof 22.2 pg FACT -TCROO2 (LIMS #E00-1716), Page 9 of 67 BACK TO MAIN THPFOS/mL) and analyzed via HPLC/ES/MS according to analytical method ETS-8-155.0. Four injections of each final diluted sample were performed. The following table describes the samplecollection and preparation regimen: Table 6. Sample Collection and Preparatiofnor Study FACT-TCROO2* 1 At24,48,and72hours, 3 PFOS+water,and 1 4 centri- Dayl 8-1-00 water (method blank) centrifuge tubes are fuge tubes removed from the incubator and equilibrated D8a-dat2ay-p02ye0r 4-25 OC for approximately24 hours. 8D-3a-y030 2 Alsitqhuuepooeftrsna(tawnactotenrtaining 4 per dissolved PFOS or water only from the method centrifuge blankws)erceollecteadnddispenseindtuobe autovials. Dayl Day2 8-2-00 8-3-oo 84-00 3 aTlihqweudoeilutrseted IOOXmweitthhDaanyol (using a dual syringe diluter). A second IOOX dilution in methanol was performed to prodauce of facdtoilurtiofninal 10,OOOX. 4 lnteSmtaanwldadrsedd to the 10,OOOX sample dilutions (11 pLTHPFOS solution at 22.2 pg ImL in methanol). 5 SampwalensraelHyvzPieaLdClESloMnS HP Day LC/MSD 1100 1 Day 2 Day 3 Day 1 Day 2 . Day 3 Day 1 2 az7-00 8-27-00 8-4-00 827-00 8-27-00 (method blks only), and 8-2400 (samples only) ~ ~~ ~ 87-00 827-00 8-27-00, and a 2 4 0 0 Day 3 a2500 *Information pertainsto sample preparation and dates of final reported data only. ANALYTICAML ETHOD ~ ~ 0 ETS-8-155.0"Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-ElectrospraylMass Spectrometry". Diluted samples (i.e. diluted supernatants) were analyzed using HPLC/ES/MSin the negative ion mode. PFOS levels were evaluated versus calibration standards ranging in concentration from 2.5-1002 ng PFOS/mL. Internal Standard quantification was usedto normalize the data. Target ions were499 m/z (PFOS anion), and427 m/z (deprotonated THPFOS). Analytical Equipment Liquid Chromatograph: Hewlett-Packard@ Series Analytical column: Keystone@ Betasit" Cq82x50mm, Column temperature: Ambient Cycle Time: 10.5 minutes Flow rate: 300pL/min Injection volume: 5pL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in water 1 100 Liquid Chromatograph 5pm particle'size system FACT -TCR002 (LIMS #E00-1716), Page 10 of 67 BACK TO MAIN Solvent B: Methanol Solvent Gradient: - Time 0.00 1.oo 4.50 7.50 8.00 10.50 - % B 40 % 40 % 95 OO/ 95 % 40 % stop Mass Spectrometer: Hewlett-Packard@Series 1100 APVMass Spectrometer Detector Software: HP ChemStation" 6.0 Fragmentor Voltage: m/z 427400 V; m/z 499= 140 V 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) DATASUMMARY, ANALYSESA, ND RESULTS ANALYTICARLESULTS 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 orgreater (R2values ranged from 0.9960 - 0.9998). Calibration Standards. Eleven standards ranging in concentration from approximately 2.5 to 1000 ng PFOS/mL methanol wereused for the calibration curves. Calibration curves were run before and after every analytical sequence. Sample Replicates. Samples 073100-wPFOS-1.1.4, 073100-wPFOS-2.2.4, and 073100-wPFOS-2.3.4 were excluded from the reported data using Dixon's Q-Test. Day 2, sample 1 was barely above the warning limit (RSD of 15.9%). Continuing Calibration Verification. For quantitative determinations, a mid-level matrix calibration check was analyzed every five samples to monitor instrumental drift, with a limit of +5I% deviation of the targetconcentrations. Limit of Quantitation (LOQ). The LOQ is equal to the lowest standard in the calibration curve (either 2.5 or 5.0 ng/mL). FACT -TCR002 (LIMS #E00-1716),Page 1 1 of 67 BACK TO MAIN Table 7. Calibration Standard Reprocessing Summary Day 3 data: 08-25-00 Day 1 data: 08-27-00 Day 2 data: 08-29-00 5 - 500 ng/mL 2'5 - 500 ng/mL 5 - 500 ng/mL Std 1 (2.5 ng/mL), and Std 11 (1002 ng/mL) were excluded to better fitthe data at the low end of the mNe. Std 11 (1002 ng/mL) was excluded to better fit the data at the lowend of the cuwe. Std 1 (2.5 ng/mL),and Std 11 (1002 ng/mL) were excluded to better fit the data at tlhoew end of the 0 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). 0 Solvent blanks (methanol injections) provided a measure of instrument contamination. Acceptable solvent blanks must contain levels of target analyte less than 50% of the limit of quantitation (LOQ). The initial solvent blank in each of the sample run sequences for days 1 and 2 data had background levels of PFOS >50% of the LOQ. Day 1, run #9I exhibited a background level of PFOS>50% of the LOQ. Acceptable solvent blanks were analyzed before each calibration curve. Specificity (accordingto OPPTS 830.7840): The solubility determination as stated in the EPA Guidelines follows. This method should only beapplied to: Pure substance. This study shows a purity of 97.9%. Substances that are stable in water. Hydrolytic studies have been conducted at the 3M Environmental Laboratory showing stability (Report # W1878). DATASUMMARY Table 8 summarizes individual sample data. Representative chromatograms are presented in Attachment C. The table displays PFOS concentrations for individual injections. Also included are the average concentrations for each Centrifuge Tube replicate, standard deviation, and % coefficient of variation. FACT -TCR002 (LIMS #E00-1716), Page 12Of 67 BACK TO MAIN Table 8. PFOS TCR-00017-046 (PFOS) Data Summary Rep 1: 731 OO-wPFOS-1. 1.1 D~~1bntrifugeRep2:731oo-WPFOS-1 .I.2 731030:R-Wep#PT1FuObeS-1.1.3 Rep4: 731OO-wPFOS-1.1.4 Rep 1: 73100-wPFOS-1.2.1 Day 73100-W2P:FOSR-1e.2p.2 Tube #2 Rep 3: 73100-wPFOS-1.2.3 Rep 4: 73100-wPFOS-1.2.4 Day Centrifuge Tube #3 Rep 1: 73100-wPFOS-1.3.1 R7e23p:1OO-wPFOS-1.3.2 Rep 3: 73100-wPFOS-1.3.3 Rep4: 73100-wPFOS-1.3.4 Rep 1: 73100-wPFOS-2.1.1 Day2bntribge Rep2: 73100-~PFOS-2.1.2 Tube# I Rep 3: 731OC-wPFOS-2.1.3 Rep 4: 731 OO-wPFOS-2.1.4 Rep 1: 73100-wPFOS-2.2.1 D~~2 Centrifuge Re#pT2ube Rep27:31OO-wPFOS-2.2.2 3: 731OO-wPFOS-2.2.3 Rep 4: 73100-wPFOS-2.2.4 Rep 1: 73100-wPFOS-2.3.1 D~~2 bntrifugeRep2: ~~IOO-WPFOS-2.3.2 Tube #3 Rep 3: 73100-wPFOS-2.3.3 Rep4: 73100wPFOS-2.3.4 ~~~3 Centrifuge Tube # I D~~ Mtri73130R:0-ew#pTP2uFbOeS3.2.3 Rep 1: 73100-wPFOS3.1.1 Rep27:3100-wPFOS3.1.2 Rep3: 73100-wPFOS3.1.3 Rep 4: 731oo-WPFOS3.1.4 Rep 1: 731OO-wPFOS3.2.1 Re72p3:100-wPFOS3.2.2 Rep 4: 73100-wPFOS3.2.4 Rep 1: 731OO-wPFOS3.3.1 D~~3 Centrifuge Rep2: 73100-~PFOS3.3.2 Tube #3 73130R:0e-wpPFOS3.3.3 Rep 4: 73100-wPFOS3.3.4 594 579 613 2593' 659 653 599 556 667 659 595 680 646 71 8 617 91 1 580 572 656 1446' 051 932 696 3702' ~~ 648 723 723 717 705 702 712 702 683 685 71 8 71 1 596' 14 2% 'Rep 4 excluded via Q-Test 617 42 7% 650 33 5% 723 115 16% 603' 38 6% 'ReD 4 excluded via Q-Test 826' 98 12% 'Rep 4 excluded via Q-Test 703 32 5% 705 4 1% 699 15 2% FACT -TCR002 (LIMS #E00-1716), Page 13 of 67 BACK TO MAIN The average PFOS concentration and standard deviation for each day, as well as the final overall average for all non-excluded replicates and time points were as follows: Table 9. Summary Tableof Solubility of PFOS TCR-00017-046 in Water. I Day 1 I Day 2 Day 3 Average Value 621 pg/mL 717 ps/mL 702 ps/rnL 680 pglmL I 23 I 91 2 43 4% ~ 13% 0.3% 6% I ~~ ~~ The solubility of PFOS TCR-00017-046 in water is 680 pg/mL at24-25 OC. Attachment B contains data summary tables. STATISTICAMLETHODS Statistical methods were limited to calculating means and standard deviations. Data that did not meet acceptance criteria as described in the OPPTS and OECDguidelines was excluded using statistical justification provided by Dixon's Q-test. Refer to Attachment E for formulas and example calculations. STATEMENT OFCONCLUSION Under the conditions of the present study, the solubility of PFOS TCR-00017-046 in ASTM Type I water is 680 pg/mL at 24-25O 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. OECD105: Water Solubility. Adopted27July,1995. FACT -TCROO2 (LIMS #E00-1716), Page 14 of 67 LISTOF ATTACHMENTS 0 Attachment A: Extraction and Analytical Methods 0 Attachment B: Data SummaryTables 0 Attachment C:SampleChromatograms 0 Attachment D: Deviations fromthe Protocol 0 Attachment E: Sample Calculations BACK TO MAIN FACT -TCR002 (LIMS #E00-1716), Page 15 of 67 BACK TO MAIN SIGNATURE PAGE We certify that this report is a true and complete representation of the data for this phase of the study: Mark E. Ellefson Principal Investigator Date ` Wiiliam K. Reagen Laboratory Manager Date FACT -TCROO2 (LIMS #E00-1716), Page 16 of 67 BACK TO MAIN ATTACHMENAT: EXTRACTIOANND ANALYTICAL METHODS FACT -TCR002 (LIMS #E00-1716), Page 17 of 67 BACK TO MAIN 3M ENVIRONMENTLAALBORATORY SOLUBILITY SCREEN TEST: APPROXIMATE SOLUBILITY DOEFTAERMINATION TEST SUBSTANICNEV m o u s SOLVENTS 09/08/0D0EaAtTedN:Sou-p8mt-Mi1bo7en0trh.:1od Effective Date: 03/14/01 Approved By: Wilfiam K. Reaggn, Laboratory Manager Date 1.0 SCOPE AND APPLICATION 1.1 Purpose. According to methods set forth by the United States Environmental Protection Agency (US EPA) andthe Organizationfor the Economic Cooperation and Development (OECD) apreliminary study, which will be outlined inthis method, serves as a prerequisite to performing solubility testing via the Column Elution Method and Shake Flask Methods. The EPA and OECD solubility testing guidelines state that if the preliminary solubility determinationtest indicates a solubility of > g/L (10 ppm), the Shake Flask Method (ETS-8-172.0) is to be used. If preliminary testing indicates a solubility of < g/L (10 ppm) the Column Elution Method (ETS-8-171.O) will be used todetermine the solubility in the solvent of interest. Additionally, due to the tendencyof certain compounds to be highly soluble in particular solvents, no further solubility determinationwill be necessary for compounds having solubility greater than 10%(mass/volume). --Exact db2P- yc&f@ff@f&nalytes. Test substance and degradationproducts for solubility testing. fl& hitiall 3bYhLv Date ETS-8-170.1 Solubility Determination: Screen Test Method FACT -TCROOP (LIMS #E00-1716),Page 18 of 67 Page 1 of 9 BACK TO MAIN 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 substanceinto a 2.5 mL-4 mL glass screw-cappedvial. Add test solvent in varying increments to the vial. After each solventaddition, vortex mix for approximately 15 sec., and sonicate 2-5 minutes. Make observationsof thesolution and visuallyconfirmwhether or notparticles are present;it may be necessary to allowthe solution to sit for up to 5 minutes. Add test solvent in a stepwise fashion up to 2 mL. If the substancehlly dissolves after the first solvent addition of 100 pL, thetest 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 substance intoa 100 mL glass stoppered volumetric flask or graduated cylinder. Add test solvent untilvisual confirmationof the substance dissolution has been achieved. Record ali observations ona standardized preparation sheetor logbook. Once 100 mL solvent has been reached, the approximateconcentrationis at 100 pg/mL. If the solubility limit has not yet been achieved, weigh approximately10mg of test substance and transfer to a 1 L graduated cylinder/volumetricflask (concentration is approximately 10ppm). The container should be sonicated and allowed tosit overnight to allow for maximal dissolution. If the undissolvedparticles are still observed after sitting > 12 hours, the columnelution method willbe utilized (ETS-8-171.0). If the test substance dissolves in 1L or less ofsolvent, the shakeflask 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 therelative solubility in a specified solvent will bedetermined. 3.2 Test solvent. The matrix to which thetest substance of interest is introduced. The test solvent may include but is not.limitedto aqueous matrices,including ASTM Type I Water and various salt matrices (e.g. 0.01M Cacl,,,,,); and organicsolvent matrices, including methanol, andacetone. 4.0 WARNINGSAND CAUTIONS 4.1HealthandSafetyWarnings: 4.1.1 Wear the proper lab attire for all partsof these procedures. Wear gloves and eye protection at all times. 4.1.2 Handle all solvents in a hood for all partsof the describedsample preparation procedure. 4.1.3 For potential hazards of each chemical used, refer to materiaslafety data sheets, packing materials, and 3M Environmental Laboratory's Chemical HazaRrdeview. 4.1.4 No mouth pipetting is allowed. ETS-8-170.1 Solubility Deterntination: Screen Test Icriethod FACT-TCROOZ (LIMS #E00-1716), Page 19 of 67 Page 2 of 9 BACK TO MAIN C4.a2utions: 4.2.1 Glassware in which standards are prepared should be triple rinsed withacetone and methanol to reducethe 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 beknown and documentedprior to startingthe screening procedure. 6.0 EQUIPMENT 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Vortexmixer. 6.3 Sonicating device. 7.0 SUPPLIESAND MATERIALS 7.1 Disposable glass graduated pipettes, 1mL to 100mL. 7.2 Disposable glass Pasteur pipettes and rubber bulbs. 7.3 Glass beakers,various sizes. 7.4 2.5 mL-4 mL glass screw-top vial. 7.5 Glass volumetric flasks, 10 mL to 1000 mL. 7.6 10 pL-1000 pL Pipettemanm manual pipettor and plasticpipette tips, or equivalent. 8.0 REAGENTS AND STANDARDS 8.1 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science, or equivalent. 8.2 Acetone, HPLC/SPEC/GC grade from EM Science or equivalent. 8.3 Water, ASTMTypeI, 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 CaC1, stock solution is prepared by weighing 1.5 g CaCl, Dihydrate in a weigh boat andtransferringto a 1L volumetric flask andiluting to the mark with Milli-QTM water. 8.6Testsubstance of known purity. 9.0 SAMPLE HANDLING 9.1 Record times of initial preparation and dilution on a sample preparationsheet or logbook. 9.2 Once the preliminary testing has been completed and the appropriateinformation is extracted, the screen testsamples should be disposed into theproper waste stream. ETS-8-170.1 Solubility Determination: Screen Test Method FACT -TCROOP (LIMS #E00-1716),Page 20 of 67 Page 3 of 9 BACK TO MAIN 10.0 QUALITYCONTROL 10.1 Not applicable. 11.0 CALIBRATION AND STANDARDIZATION 11.1 The compounds of interest must beof known characterization according tolaboratory 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 a4mL glass screw-top vial or equivalent. Record the weight (refer to attachmeAntfor an example of a standardizedprep sheet). 12.1.2 Add solvent (e.g. water, substance. 4 mL vial or equivalent1 Total volume solvent added (.m~L)... Approximate Solubility methanol, Solvent addition 1 0.1 acetone) according 1 Solvent 1 addition 2 0.5 thoe table below to the test Solvent Solvent addition 3 addition 4 1 /Kbi'\ 12.1.3 Following addition of solvent, vortemxi pproximately 15 seconds, sonicate 2-5 minutes. Visually check for undissolved particlesI.f undissolved testsubstance remains, continue adding solventaccording to the above chart. It may be necessary to allow the solutionto settle for up to 5 minutes before making observations. Record observationson the standardized prep sheet. 12.1.4 If all particles dissolve, then estimatethe approximate solubility and document the concentration. Because the concentrationis >10 &mL, the shake flask method (ETS-8-172.0) will be usedto determine the accurate solubilitypoint. However, if the test substancedissolves after thefirst solvent addition of100 pL the solubility is determined to be>lo% (rnass/volume). The test substance shallbe regarded as "highly soluble," and no furthertesting is required. 12.1.5 If the test substancedid not dissolve in the 2 mL of solvent,weigh 10 mg k 1mg of test substance into a 100 mL glass volumetric flask or equivalent. Record the weight (refer to attachment A for an exampolfea standardized prepsheet). 12.1.6 Add solvent according to thteable below to the test substance todeliver the appropriate amount of solvent. As described in 12.1.3,vortex mix, sonicate, and allow the test solution to settle. Record all observations and procedures on the preparation sheet. ETS-8-170.1 Solubility Determination: Screen Test Method FACT -TCROO2 (LIMS #E00-1716), Page 21 of 67 Page 4 of 9 BACK TO MAIN 100 mL volumetric!Solvent addition1 Solvent addition 2 Solvent addition 3 flask or equivalent Total volume added .I solvent (mL) . . 10 50 100 I Approximate Solubility1 1000 200 100 12.1.7 If the solubilitypoint is reached at or prior to the complete addition of 100mL, the concentrationestimation at which thseubstanceis soluble should be documented and shall be usedas a startingpoint for the shake flask method (ETS-8-172.0). 12.1.8 If the 100mL solvent levelhas been reachedwithout complete dissolution of test substance,weigh out 10mg k 1 mg testsubstance and transfer toa 1 L volumetric flask, graduated cylinder, or equivalent. Dilute to I L with test solvent. The approximate concentrationof test substanceis at 10 pg/mL. The flask shouldbe allowed to sit 12-24 hours to allow for maximadlissolution. If the undissolved particles are still observed, the column elution methodwill be utilized (ETS-8171.O). If no visible particulates are observed,the shake flask method (ETS-8172.0)shall be used to determinethe solubility. 13.0 DATAANALYSIASND CALCULATIONS 13.1 The solubility determination in this methodis qualitativehemi-quantitative. 13.2 The point to which the substance dissolves in solvent is confirmed visually. The solubility point is therefore a qualitative determination. 13.3 The concentrationestimation is semi-quantitative in thathe approximateconcentration is calculated by the following equation: c=a/b Where: c= the semi-quantitativeconcentration pg/mL, a= amount of substanceweighed out(pg), and b= the approximate amounot f 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 addedthtoe solute. Since large and varying increments of solvent are being added ttohe solute, this error margin must be considered whenreporting the concentration estimation. 14.2 The data obtained throughthis study is an estimation only and shouldbe treated as a qualitative estimation of the solubility of test substance ina given solvent. ETS-8-170.1 Solubility Detemination: Screen Test hletltod FACT -TCROO2 (LIMS #E00-1716),Page 22 of 67 Page 5 of 9 BACK TO MAIN 14.3 For an accurate measurementof 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 placingin high or low BTU containers asappropriate. 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-Exampleof a Standardized Sample Preparation Sheet: Solubility Screen Prep Sheet. 18.0 REFERENCES 18.1 Organization for Economic Cooperation and Development. OECDGuideline for Testing of Chemicals. Water Solubility -0ECD Guideline 105: pp. 1-7, Adopted 1995 18.2 United States Environmental Protection Agency. OPPTS 830.7860 WaterSolubility (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 WateSr olubility: 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 ElutionMethod: Solubility Determination of Test Substance in Various Solvents." 18.5 3M Environmental Laboratory MethodETS-8-172.0, "Shake Flask Method: Solubility Determination of Test Substance in Various Solvents.'' 19.0 AFFECTEDDOCUMENTS 19.1 ETS-8-171.0, "Column Elution Method: Solubility Determination of Test Substance in Various Solvents." 19.2 ETS-8-172.0, "ShakeFlask Method: Solubility Determination of Test Substance in Various Solvents." ETS-8- 170.1 Solubility Dcterlllimtiort: Screen Test Method FACT -TCROOZ (LIMS #E00-1716), Page 23 of 67 Page G of 9 Reason 20.0 REVISIONS Revision Number Revision 1 It was desirable tomaktehme ethodmorueniversallyapplicablbey removing references to specific test substances. BACK TO MAIN Revision D- ate 03/13/01 ETS-8-170.1 Solubility Determination: Scseen Test Method FACT-TCR002 (LIMS #E00-1716), Page 24 of 67 Page 7 of 9 I Attachment A: Example Preparation Worksheet,page 1 of 2 I BACK TO MAIN GLP Study Number: Test Substance: 366 F n v 1olf. Source: F l IN 8C:RFFIV Sample preparation worksheet Date: Solvent: ID#: Step Source: 1 Weigh approximately10 m g f 1 m g of test substance into a 2.5mL-4 mL glass screw-top vial. subWsteaingchet:of test m g Balance ID: Datellnitials: replicate2nd opfional: mg Thermometer ID: 2 Add test solvent accordingfo the table below. Following each additionof solvent, shake vigorouslyhortex, and sonicate the mixture and visually chefocrkundissolvedpadicles. Analyst - volume added(mL)l .. approx conc. after addition (uglrnL) Sample prep OBSERVATIONSINOTES Datmm Totavlolumseolvenatdded P Step 2-1 (mL) (1.1 Approximate Solubility (ug/mL)100,000 Flask vortex-mixed?Y ~ ~ R/~~N~ ~ ~ ~ Is there solud testill presentlescontinue to step 2-2 Flassoknicated?Yes/No Flaskallowed to settle?YesMo T-i NO- The solutionis at IO%, and is considered "infinitely soluble.," further solubility testing is required. DatAenrimaly- st - - Analyst Analyst __ ,Step 2-2 Totavol lumseolvenatdde(dmL) 0.5 ADDroximate Solubilih, fudmL) 20.000 Datflim- Step 2-3 Totavol lumseolvenatdde(dmL) 1 ADDroximate Solubility (udmL) 1.0,OOO DateflimTotal volumseolvenatdde(dmL) Step 2-4 2 ApDroximate Solubility fuq/mL) 5,000 Flask vortex-mixed?Y e m o Room T e d Is there solute still present? YeslNo Flasksonicated?Yes/No -Ti Flaskallowed to settle?YeslNo T-i Flask vortex-mixed? YedNoRoomTernp-2C Flasksonicated?YesINo T - i Flaskallowed to settle?YedNo T-i Is there solutestill present? YedNo , Flask vortex-mixed? YeslNoRoom Temp% Flasksonicated?Yes/No -Ti Flaskallowed to settle?YeslNo T-i Is there solute still present? Y e a 0 D i d the substance dissolvein 2 mL or lessof solvent? Yes -Approximateconcentration at which the test substancise soluble in The solubilityof the test substance is to be determinedvia the shake or No Continue to page two for further testing. solvent: flask method. Notes/ additional comments: C ETS-8- 170.1 Solubility Determination: Screen Test Method Page 8 of 9 BACK TO MAIN 1 Attachment A Cont.: Example Preparation Worksheet,page 2 of 2 I I GLP Study Number: Test Substance: 3M Environmental Laboratow SOLUBILITY SCREEN Sample preparation worksheet ID#: Saurce: Date: Solvent: IDk Source: Step (continuefrdopmage I) -Did the test substance dlssolvein 2 mL? YedNo If YES, the test is complete, and thereIs no need to contlnue withe screen test. If NO, continue to step 3 to deterrnIne the approximate solubilityof the substance. 3 Weigh approximately10 mg f I mg of test substance intoa 100 mL graduated stopperedcylinderholumetricflask. substancWee: ight of test mg Balance ID: Datdlnitials: 2nd replicate optional: Thermommegter ID: 4 Add test solvent accordingto the table below.Followingeach additionof solvent, shake vigorouslyhortex,andsonicate the mixtureand visually checkfor undissolvedparticles. I I I I v o l u m added (mL)l approx conc. after Solubilitv data addition (UglmL) Sample Prep OBSERVATIONSINOTES Analyst __ Datame I Step 4-1 Total volume solvent added(mL)... 10 Approximate ... Solubility(ue/mL) 1,000 Flaskvoctex-mixed?YeslNo Fkmn T e m P ' C Flask sonicated? Y d N o lime rniq Flask allowetdsoettleY?esMToime . min Isstohleurte still present? YeJNo __ Approxirr Analyst - Datfllme I Step 4 3 (vsamoodTlLludvo)metadnelt ... 100 Approximale ... Solubility(ug/mL) 100 FlaskvortexmTYRieexomseoMdpm?o sofnlaicsakted? Y&No Flask allowed to settle? mTiqime Y d N o Time 5 Weigh approximatelyIO mg k 0.5 mg of test substance into a 1 L graduated stoppered "S Isothluetere still present? YesJNo min cylindedvolumetrifclasklorequivalent. substancWee: ight of test mq ID: Balance Date/initials: Analyst __ DaleTTtme / Step 5-1 Total vsaoodlludvmendet (mL)... 1000 Approximate Solubility (ug/mL.).. 10 Flask vortexmixed?YesJNo Room T e m p 2 Is there solute still present? Y&NO Flask sonicated? Yldi mN oe min Flask allowed to settleY?es/No lime min Conclusions: Did the substance dissolvein io00 mL o r less of solvent? Yes -Approximateconcentrationatwhichthetestsubstance is solubleinsolvent: ucVml The solubilityof the test substanceis to be determined via the shakfelask method. or No -The solubility of the test substanceis to be determinedvia the column elution method. Notes1additional comments: I ETS-8- 170.1 Solubility Determination: Screen Test Method Page 9 of 9 BACK TO MAIN 3M ENVIRONMENTAL LABORATORY SHAKE FLASKMETHODs:OLuSlLITY DETERMINATION OF A TESTSUBSTANCE IN VAIUOUS SOLVENTS NumMbetrh: od ETS-8-172.1 Adoption Date: 09/08/00 Effective Date: 03/27/01 Authors: KristinL. Terrell, Mark L. Anderson, andMark E. Ellefson Approved By: Date WMilalniLaamagbeKorr.aRtoeraygen, d&>/, bate ' 1.0 Scope anAd pplication 1.1 Purpose. AccordingtotheUnitedStatesEnvironmentalProtectionAgency(U.S.EPA) and Organizationfor the Economic Cooperation and Development (OECD) guidelines, solubility determinationof substances that have solubilities in a givtesnt matrix (e.g. water/acetone/methanol) of greater than100 pg/mL must be analyzedvia 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 solubilitylevel. ,Refer to ETS-8i170.0 for the preliminary solubility screen test procedures. E.i C' . - r >- 1 , .a;. ; _1 ETS-8- 172.1 Solubility Test: Shake Flask Method FACT -TCROO2 (LIMS #E00-1716), Page 27 of 67 Pag-e 1 of 1I BACK TO MAIN 1.2 Compatible Analytes. 1.3 Acceptable Matrices. Testsubstanceanddegradation products for solubility testing. Water, Acetone, and Methanol, or other solvents of interest. 2.0 SUMMARY OF METHOD 2.1 Using the qualitative/semi-quantitativedata obtained from the preliminary solubility screen test, weigh out more than five timethse estimated soluble concentration of test substance into eachof twelve screw-capped vessels(three time points with four test vessels each-sample, duplicate, triplicate, andmatrix blank). Add the appropriate amount of test solvent gravimetrically. Capthe tubes, seal the capswith tape, andplace horizontally on an incubatodshaker set to 30 "Cf2 "C, shaking for approximately 24 hours. At approximately 24 hours, the first time point will be pulled and equilibrated at roomtemperature(about 20 "C-26"C) for approximately 24 hours. The equilibrated , samples are centrifuged and aliquoted into autovials.Samples are then diluted 1:10 or higher with a suitable solvent for analysis via LC/MS. At approximately48 hours and 72 hours, the second and third sets, respectively,will be pulled, equilibrated, centrifuged, aliquoted and dilutedwith a suitable analytical solventfor LCMS analysis. Depending on the analytical range,dilutions may be made using adiluter or syringe to make successive serial dilutions using a suitable analytical solvent to reatchhe desired concentration (e.g. 1:10 or higher samp1e:methanoVacetone serial dilutions), and an appropriate internal standard (e.g. lH,lH,2H,2H tetrahydroperfluorooctane sulphonic acid (THPFOS)) willbe added to the final dilution prior to analysis.Samples are tobe analyzed via LC/MS against a standardcurve containing the testsubstance and an appropriate internal standard. 3.0 DEFINITIONS 3.1 Method blank: An analyte-fiee matrix (e.g. methanol, water, or acetone) to which all reagents agree are added in the same volumesor proportions as used in the sample processing. The method blankis used to document contaminationresulting from the entire sample treatment and analytical processT. he method blankis carried through the complete sample preparation, treatment, and analytical procedure. 3.2 Solvent blank:A sampleof analyte-free medium (e.gm. ethanol/water/acetonesolution) that is not taken throughthe sample treatment process. Thisblank is used to evaluate instrument and reagent contamination. 3.3 Shake Flask Sample TriplicatesT: hree test vessels takenfiom and representative of the same samplesource carried through all steps of the treatmenext,traction, and analytical procedures in an identical manner. 3.4 Sample replicates:Replicate samples (twoor more) taken from andrepresentative of the same sample source (e.g. test vessel containing test analyte ansdolvent) and separately carried throughanalytical procedures in an identical manner. 3.5 Internal Standard(IS): A known amount of a compounsdimilar in analytical behavior to the compound(s) of interest, added to all samples andstandards, and carried through the entire measurementprocess. ETS-8- 172.1 Solubility Test: Shake Flask Method FACT -TCROO2 (LIMS #E00-1716),Page 28 of 67 Page 2 of 11 BACK TO MAIN 3.6 Calibration Standard: A dilution of various amounts of a stock, intermediate or purchased standard to achieve standarsdolutions in a concentration rangeof interest. 4.0 WARNINGS AND CAUTIONS 4.1 Health andSafetyWarnings: 4.1.1 Wear the proper lab attire forall parts of these procedures. Wear gloves and eye protection at all times. 4.1.2 Handle all solvents in a hood for allparts of the described sample preparation procedure. 4.1.3 For potential hazards of each chemical used,refer to materialsafety data sheets, packing materials, and3M Environmental Laboratory's Chemical HazardReview. 4.1.4 No mouth pipetting is allowed. 4.2 Cautions: 4.2.1 Glassware in which standards are preparedare to be triple rinsed with acetone and methanol to reducethe possibility of accidental contamination. 4.2.2 All test vessels (e.g. centrifbgetubes) are to be sufficientlyrinsed with solvent prior to their usage. 5.0 INTERFERENCES 5.1 Impurities may significantly affect the solubility of the test substance. The purity of the test substance should be known and documentedprior to startingthe preliminary solubility screening procedure. 5.2 Contaminantsin solvents, reagents, glassware and other sample processing or analysis hardware may cause interference. The routine analysis of laboratory method blanks must be used to demonstratethat there is no interferenceunder the conditions of the analysis. 6.0 EQUIPMENT 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Centrifuge capable of holding 15 mL centrifuge tubes or equivalent. 6.3 Incubator with heatingkooling capabilities. 6.4 Diluter, Hamilton Microlab@500 Series, or equivalent. 6.5 Vortex-mixer. 7.0 SUPPLIES AND MATERIALS 7.1 Thermometer capable of reading at least 15OC-40 "C. 7.2 5-250 mL polypropylene centrifbge tubeso,r equivalent. 7.3 Disposable glass graduated pipettes, 1mL to 10mL. 7.4 Disposable glass Pasteur pipettes and rubber bulbs. 7.5 Glass beakers, varioussizes. 7.6 Crimp cap autovials-1.5 mL, caps, crimper, and decapper. 7.7 Hamilton Gastight@syringes (precision k 1% of the total volume),5 pL to 1000 pL. 7.8 Pipette-man manualpipettor. ETS-8-172.1 Solllbility Test: Sl~nkeFlask Method FACT -TCR002 (LIMS #E00-1716), Page 29 of 67 Page 3 of 11 BACK TO MAIN 8.0 SUPPLIES AND MATERIALS 8.1 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science, or equivalent. 8.2 Acetone, HPLC/SPEC/GC grade from EM Science or equivalent. 8.3 Water, ASTMType 1. 8.4 Test substance of knownpurity. 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 CaClz in a weigh boat andtransferring to a 1L volumetric flask and diluting to the mark with Milli-QTM water. 9.0 SAMPLHEANDLING 9.1 Record times of initial preparation, set-up, andsample aliquotinglanalysison a sample preparation sheet or logbook. 9.2 Once the samples have been diluted, theymay be analyzed via LCMS. Alternatively,the diluted samples may be kept in coldstorage (e.g. approximately1-5 "C) in crimp-capped autovials until the time of analysis. 10.0 QUALITYCONTROL 10.1 Shake Flask Sample Triplicates. Set up each test vessel in triplicate to provide a measure of the precision on sample preparation. 10.2 Sample Replicates. Prepare and analyze allsamples (from each testvessel 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 testvessel without test substance to measure any contamination accrued throughouthe sample preparation process. The method blank is carried throughthe same sample preparation procedures athse three test vessels with test substance, only ntoest substanceis added throughoutthe 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 anpyossible contamination of the solvent(s) used duringthe sample preparationprocess, and to detect any instrumental contaminatioonr background interferences. 11.0 CALIBRATIONANDSTANDARIZATION 11.1 The compounds of interest must be standardized according tloaboratory specifications. 11.2 All equipment used,such as the analytical balance and automateddiluter, should be calibrated prior to use(daily, weekly, etc.) as specifiedin its standardoperating procedure. ETS-8-172.1 Solubility Test: Shake Flask Metllod FACT -TCROOP (LIMS #E00-1716), Page 30 of 67 Page 4 of 11 BACK TO MAIN 11.3 All samples analyzed willbe run against a standardcurve containing varying amounts of test substance, and a fixed amouonft internal standard. 12.0 CALIBRATION AND STANDARIZATION 12.1 Preparation of Test Vessel. Record all data andobservationson the example sample preparation worksheet (Attachment A)or equivalent. 12.1.1 Prepare a solution at leasftive times more concentrated thanthe concentration at which the substancewas known to be dissolvedat in the preliminary solubility screen test (ETS-8-170.0). 12.1.2 Weigh outtest substance into a tared test vessel (e.g1.5 mL polypropylene centrifuge tube). 12.1.3 Add the test solvent to the test vessel gravimetricallyuntil the desired volume has been obtained (note: sohents have differing densities, therefore the weight needed to achieve a spec$c volume will be dependenton the density.). Record all weights on a standardizedpreparation sheetor logbook. 12.1.4 Visually confirmthat there are undissolved particles. If no particles are present, repeat 12.1.2 with more test substanceor add more testsubstance and recordthe 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 ;3Day 2, -Rep 1,-Rep 2, and - Rep 3; andDay 3, -Rep 1, -Rep2, and -Rep 3. Also include the day of initial sample prep, the person(s) responsible forthe sample, the test compound,nature of the study (e.g. water solubility), the solvent utilized, andthe study number. 12.1.6 Prepare three test vessels as describedin 12.1.3 without the test substance and label (at minimum) as Dayl.,Rep4; Day2,Rep4; and Day3,Repa4long with the information described under 12.1.5. These three tesvt essels are the "method blank" samples. 12.1.7 Seal the cap tothe test vessel. Midshake test vessel toensure 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 vesselson their side (horizontal) in an orbital incubator set to approximately 30OC 4 2 OC and rotating at a considerablsepeed to ensure sufficient contactlmixingbetween the test substance and solvent. 12.2.2 At approximately 24 hours, four test vessels are removed: threceontaining test substance labeled "Day1,Repl", "Dayl,Rep2", "Dayl,Rep3", and "Day1 ,Rep4". 12.2.3 The samples are shaken to ensure no test substance istuck to the side of the tube. 12.2.4 The tubes are then placedin a stable temperature environmentof approximately 20- 26 OC (record the actual temperature) for about 2a4 hour period. 12.2.5 After about24 hours of equilibration at20 "C-26 `Cy centrifuge the samples until the solution is visibly clear. If micelle formation is suspected, an additional highspeed centrifugation step maybe added (20,000 RCFfor approximately 1 hour). The test vessels are now ready for dilution. See section 12.3 for further sample preparation. ETS-8- 172.1 Solubility Test: Sltnke Flnsk Method FACT -TCROO2 (LIMS #E00-1716), Page 31 of 67 Page 5 of 11 BACK TO MAIN 12.2.6 At approximately 48 hours ofincubation time, the second set of test vessels may be pulled (labeled"Day2,ReplY'",Day2,Rep2", "Day2,Rep3",and "Day2,Repq') and equilibrated as described in 12.2.3-12.2.5. 12.2.7 At approximately 72 hours, the final set of samples is pulled (labeled"Day3,ReplY', "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 otfhe solution, it may be necessary topipette off the top layer of solution (or an aliquot ofsample may be centrifugedas stated in 12.2.5). The use of a Pasteur pipetteor equivalent is recommended. 12.3.2 Aliquot the test solution into eachof four autovials (e.g. 1.5 mL 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 typeof pipette to be used, take into consideration thetest substance's tendency to adsorb to certain materials(i.e. adsorption of test substance to glass or plastic). 12.3.2.2 Prior to aliquoting sampleto the autovial, the pipette tip must be equilibrated/saturated with the test solution by drawing andgently expelling the test solutionin and outof the pipette tip, taking care notto disturb the solid particulate. 12.3.3 Label the four autovials and recordthe sample id. 12.3.4 Using a diluter, make a 1:lO or higher dilution of samp1e:extractionsolvent (i.e. methanol, acetone) into a labeled autovial. 12.3.5 Using the solubility informationobtained in the preliminary screentest, estimate the appropriate dilutions, if any, required to bring tchoencentrationof sample into the appropriate analytical range (e.g. approximately 3 ng/mtLo 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 willbe analyzed viaLCMS against a standardcurve of varied, known concentrationsof test substance with a constant concentratioofninternal standard equal to the concentration in the samples. 13.0DATAANALYSIS AND CALCULATIONS 13.1 Samples will be analyzed via a calibration curve. The amount of test substance in the sample will be quantified against a standard curve. 13.2 Means will be calculated by adding the individual entities and dividing the resultant sum by the number of individual entities. 13.3 Standard deviations will be calculated using either Microsoft Excel@or Microsoft Access@ tocalculate standard deviation. The built in functioncontains the following equation which is based on the individual entities (n) being less than 30: d n Cx2 -( n(n-1) ~ x ) * ETS-8-172.1 Solubility Test: Shake Flask Method FACT -TCROO2 (LIMS#E00-1716),Page 32 of 67 Page 6 of 1 1 BACK TO MAIN 13.4 Sample precision will be reported as %RSD (or %CV). Sample precision will be calculated using the following equation: Ah3 X 100 = Sample %RSD where: A= standard deviation of averaged samples B= average of samples I I Sample RPD will be calculated usingthe following equation: A-B /((A+B)/2)= SampleRPD where: A= concentration of first replicate B= concentration of second replicate 13.5 Exclusion of an outlier data point may be performed by utilizing Dixon's Q-Test. The questionable data point may err to the higohr low endof the data set. Calculate the variable "Qobserved." If Qobserved > Qtabulated, thenthe data point may be rejected with 90% confidence (see table 1 below for Qtabulated values). Qobserved= gap/range where: Gap=the difference betweenthe questionablepoint and the nearest value. Range= total spreadof the data. Table 1 Criteria for Rejectionof Outlier Values: Numboerf ()tabulated, 90% Confidence 0.679 0.557 14.0DATAANALYSIS AND CALCULATIONS 14.1 Precision of data. Sample data must have a percent relative standard deviation (%RSD) (or relative percent difference) of < 15%. Non-compliant data must be evaluated for obvious outliers. The Q-Test may-be applied to exclude questionable data points. If an outlier value exists, sample average and precisionis re-calculated andreported without the questionable data point. Document the non-compliant dataon data summary sheets, and include results of the statistical analysis with the final results. 14.1.1 14.1.2 Sample replicate %RSD's should be4 5 % . If the average of the sample triplicate data is >15% RSD, evaluate thethreevalues for anoutlier value. To questionable data points, apply the Dixon's Q-Test.If the outlier value is rejected with 90% confidence, exclude the sample from furthedrata calculations andcalculate the average and RPD for the remaining two values. If no data points can be excluded and the data does not meet the criteria, thenthe data set may not be used in the final reported data. The remaining shake flasks for the timepoinsthall be used to report the data. The %RSD between shake flasks should agree within 15%I.f this criteria is not met, applythe Q-test tooutlier datum. If the questionable datapoint is discarded ETS-8-172.1 Solubility Test: Slmke Flask Method FACT -TCROOP (LIMS #E00-1716),Page 33 of 67 Page 7 of 11 BACK TO MAIN via the Q-Test, then the non-compliansthake flask data is not tobe used for the timepoint. Include justification for dropping the shakfelask (e.g. q-test results). 14.1.3 And finally, the %RSD betweenthe three days shouldbe 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 timeintervals are required to report the final solubility. If three consecutivetimepoints do notagree within 15%, it may be necessary to repeathe entire test,or re-dilutehe-shoot samples to checkfor error. 14.2 Limit of Quantification.For this study, the LOQ will be equal to the lowest calibration standard used in the calibrationcurve containing more than twictehe 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 levelsof analyte exist inthe initial methodblanks, but subsequent blanks prior to the calibration standard curve are clean, the data may be accepted (provided that there are no other indicators that eitherthe samples or the instrument contain significantbackground levels of analyte). 15.0 POLLUTION PREVENTIONAND WASTE MANAGEMENT 15.1 Dispose of sample waste by placing in highor low BTU containersas appropriate. Use broken glass containers to disposeof 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 AttachmentA-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 ToxicSubstances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-042: pp. 1-17, 1996 18.3 United States Environmental Protectipn Agency. OPPTS 830.7840 Water Solubility: Column Elution Method;Shake Flask Method. Prevention, Pesticides andToxic ETS-8-172.1 Solubility Test: Shake Flask Method FACT -TCR002 (LIMS #E00-1716), Page 34 of 67 Page 8 of 11 BACK TO MAIN Substances: Fate, Transport and Transformation Test Guidelines. EPA 712-C-96-041: pp. 1-12,1996 18.4 ETS-8-170.0, Solubility Screen Test: ApproximateSolubility Determination of a Test Substance in Various Solvents. 18.5 Harris, Daniel C. Statistics. Quantitative Chemical Analysis. 4`h ed.; W.H. Freeman and Company:NewYork1, 982;p70-71. ' 18.6 Natrella, Mary Gibbons. The Treatment of Outliers. Experimental Statistics, National Bureau of StandardsHandbook 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.'' Revision Reason Number 1 For Revision Section 1.2,referencesto specifictestsubstanceswereremoved. Section 3.6, removed"intemal standard blank" fromdefinition section. Section 12.2.5, an optional high-speed centrifbgationstep was added. Fixed the incorrect numberingof sections 13.5 through 14.3. Section 14.3, more clearly defined the acceptance criterion for method and solvent blanks. Revision Date 0 3 z O1 ETS-8- 172.1 Solubility Test: Sl~nkeFlask Method FACT -TCR002 (LIMS #E00-1716), Page 35 of 67 Page 9 of 11 BACK TO MAIN vent: ATTACHMENTA: Example Sample Preparation Sheet (page 1 of 2). Temperature of Study: 0 3M F n v i r o w ntal Laboratory S .o. l u b ~ h ~ Study # prwSepoaarmkrsaphtIieeoent Analyst(s): Test LIM Source: Date: Test IM Vessel: Test 15ml cenliifuge tubelOther(spedfyf hr: s!mf Source: Was the solubility screen test performed prior to start? Y/N If so, what is the approximatceoncentration? . SFM vials I W I : "samplelD" = adDdaedSdsdac:ermidpp:tiloen Balance ID: weight test substance Thermometer ID: Total mass (solute + solvent) . ughl Temp: Room 'Internal Standard addition uu d a +pin IS addedto dilutOiobnsfeawctaotri:onslComments: "C DatelTime/lnitials: samplelD-1.1.1 thru -1.1.4 samplelD-1.2.1 thru -12.4 samplelD-1.3.1 thru -1.3.4 samplelD-1.4.1 thru -1.4.4 samplelD-2.1.1 thru -2.1.4 samplelD-2.2.1 thru -22.4 samplelD-2.3.1 thru -2.3.4 samplelD-2.4.1 thru -2.4.4 samplelD-3.1.1 thru -3.1.4 samplelD-3.2.1 thru -3.2.4 samplelD3.3.1 thru -3.3.4 samplelD-3.4.1 thru -3.4.4 Dayl, Smpl Dayl, SmpP Dayl, Smp3 adDdaeydl. Smp4 blank Day2, Smpl Day2. Smp2 Day2. Smp3 adDdaeyd2, Smp4 blank Day3, Smpl Day3, SmpP Day3. Smp3 adDdaeyd3. Smp4 blank DatelTime: Initials: none none none TesVt esselssealedwt/ape? Y/N TesVt esselsshakenhortex-mixed? Y/N vTeesspstlaeclsed on orbital incubatonY/N IncubatorID Speed 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: Standard: 'Internal Int. Std. Conc.: @aInTietiamlperature: ID#: 0 b a v I: Four "Day 1' samplesarepulledandallawedto be placed in atemperaturecontrolledenvironment for approx.24 hourswith no agitation. LoTceamtiopneoraf tsuarme ples: of environment 0 ID: Visual observations: DaWnmeAnlt!aaL: ETS-8-172.1 Solubility Test:Shake Flask Method Page 10 of 11 BACK TO MAIN ATTACHMENTA CONTINUED @age2 of 2). Day 2: Four"Day 2" samplesarepulledandallowed to be placedina temperaturecontrolledenvironmentforapprox. 24 hourswith noagitation. Oatefrimdlnitiab: mreples: of Location Visual observations: of environment: 0 ID II I I I 1 D: CCeenVntrtei.fsusg1ee"lsdT:est"Day durarteioand:outR: CFRPM or mh. Visual observations: each of the four samples are to be made into glasdplastic autovials (also, check autovials for correct labelinYge).s/No Datefrimdlniliib: the solution-1: with methanol. MethanoTl N-A: Dalflirndlnitials: AdditiodnilaultioDnislu:ftaiocntor (test solution:methanol): 1: X's hmodawinluytio=nfdisnil:aufltaiocntor: 1: methanol, UL used: AmDouiluntser ID used: timeuntil exatrtacoctosleinr a of aCnoaolylesris. ID TeCmopo:ler c ul samples Dalflimdnitials: Day 3: Four"Day 3"samplesarepulledandallowed to be placedina temperaturecontrolledenvironmentforapprox. 24 hourswith no agitation. Datflimdlnitials: m ples: of Location of environment: 0 tet I D I Visuaol bservations: I I 1ID: C"DeCVaneyetnrsi2ftsu."egTleseds:t Visual observations: RPM or RCdFureaatiodno:ut: mia Oatefrimdinitiik. Four aliquots from each of the four replicates atorebe made into glasdplastic autovials (also, check autovials for correct labeling). YeslNo the diluteImmediately Methanoml eth1-:anol. with TN-A: AdditiodnilaultioDnilsu:ftaio(cstnteoos1rtution:rnethanol): 1: X's hmodawinluytio=nfdisnil:aufltaiocntor: Imethanol, UL used: AmDouiluntser ID used: Datflirndlnitils: 1: ul samples -Stosreampleasnedxtracts caoinoulaentritmialonef alysis. Cooler ID: CooTleer mp: 0 Datefrimdlnitiab: lay 4 U approx. 24 hoursoef quilibrationt,he"Day : CCenetn.triVfuegsesde:ls Test 3"samplesare to be aliquoted/extractedE. nvironmenTtemp.: RPM orduRrCaFtiorena: dout: ('2Therm. ID: mh. Datflimdlnitials: I Visual observations: I I each of the four samples are to be made into glasdplastic autovials (also, check autovials for correct labYeleinsgM).o the solution-1: with methanol. Methanol TN-A: AdditiodnilaultioDnislu:ftaio(cstnteos1rtution:methanol): 1: X's hmoadwniluytio=nfdisni:laulftaiocntor: Diluter ID used Amounts used: methanol, uL -StosraemplaenesdxtrcaoacionutsalenttimratiolnefalysCiso. oIlCeDro: oTleermp: 0 Datflimdlnitials: 1: ul samples Datwlimdlnitiab: ETS-8- 172.1 Solubility Test: Shake Flask Method Page 11of 11 BACK TO MAIN 3M ENVIRONMENTLAALBORATORY METHOD ANALYSISOF POTASSIUM PERFLUOROOCTANESULFONATEOR OTHER FLUOROCHEMICIANLWSASTE STREAMOR WATER EXTRACTUSSING HPLC-ELECTROSPRAYMASPSES CTROMETRY Method Number: ETS-8-155.0 Author: Mark L. Anderson, Mark E. Ellefson Approved By: AdoptionDate: / ( / / O / O O Revision Date: Laboratory Manager Date Word 97 ETS-8-155.0 Page 1 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Streamor Water Extracts Using HPLC-Electrospray/Mass Spectrometry FACT -TCR002 (LIMS #E00-1716), Page 38 of 67 BACK TO MAIN 1.0 SCOPE AND APPLICATION 1.1 Scope: This method describes the analysiosf waste stream or water extracts using HPLCelectrospray/mass spectrometry. 1.2 Applicable Compounds:Fluorochemicalsor other electrospray ionizable compounds. 1.3 Matrices: Tap water, ground water, wastewater and other aqueous solutions. 2.0 SUMMARY OF METHOD 2.1 This method describes the analysis of fluorochemicals or other electrospray ionizable compounds extracted from water, using HPLC-electrospray mass spectrometry (HPLCES/MS). The analysis is performed by the mass selectionof a single ion characteristic of a particular compound, suchas the perfluorooctanesulfonate (PFOS) anion,m/z = 499 or perfluorooctanoate (PFOA),m/z = 413. 3.0 DEFINITIONS 3.1 Atmospheric Pressure Ionization(API): The Micromass Platform LCZ single quadrupole system and other commercially available LC/MS systems aflloorwvarious methods of ionization by utilizing a varietyof sources, probes, and interfacesT. hese include butare not limited to: Electrospray Ionization(ESI), Atmospheric Pressure chemical Ionization (ApcI), Thermospray, etc. The ionization in these processes occurs at atmospheric press(uLre., not under a vacuum). 3.2 Electrospray Ionization(ES, ESI): A methodof ionization performedat atmospheric pressure, wherebyions in solution are transferred tothe gas phase via tiny charged droplets. These droplets are produced by the application of a strong electrical field. 3.3 Mass Spectrometer (MS)T: he Platform LCZ and other commercially manufactuEreSdMS systems are equipped with a single quadrupole mass selective detector. Ions are selectively discriminated by mass tocharge ratio ( d z ) and subsequently detected. 3.4 Conventional vs. Z-spray probe interface:The Micromass PlatformLCZ system utilizes a "Z-spray" conformation. The spray emitted from the probise orthogonal to the cone aperture. In the conventional conformationit is aimed directlyat the cone aperture,after passing through a tortuous pathwayin the counter electrode. Though the configurationis different, the methods of operation, cleaning, and maintenancaere the same. However,Zspray components and conventional componentsare not compatible with one another, but only with similar systems (i.e., Z-spray componentasre compatible with some otheZr spray systems, etc.). Other commercially manufacturedES/MS systems may have similar features. 3.5 Mass Lynx Software:System softwaredesigned for the specific operationof Micromass LCZ Mass spectrometer. Currently MassLynx has Window95s and WindowsNT4.0 versions. All versions are similar. For more details see the manual specific to the instrument (MassLynx NT User's Guideor Micromass Platform LCZ User's Guide). ETS-8-155.0 Page 2 of 9 Analysis of Potassium Perfhorooctanesulfonate or Other Fluorochemicalsin Waste Streamor Water Extracts Using HPLC-Electrosprayh4ass Spectrometry FACT -TCROO2 (LIMS #E00-1716), Page 39 of 67 BACK TO MAIN 4.0 WARNINGS AND CAUTIONS 4.1 HealthandSafetyWarnings: 4.1.1 Use cautionwith the voltage cables forthe probe. When engaged, the probe employs a voltageof approximately5000 Volts. 4.1.2 When handling samplesor solvents wear appropriateprotective clothing, gloves, and eyewear. 4.2 Cautions: 4.2.1 Operate solvent pumps below a backpressureof 400 bar (5800psi). If the backpressure exceeds400 bar, theHpl100 will initiate automatic shutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 Interferences 5.1 To minimize interferenceswhen analyzing samples,Teflon should not be usedfor sample storage or any partof instrumentation that comeisn contact with the sample or extract. 6.0 EQUIPMENT 6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications inthe raw dataas method deviations. 6.1.1 Micromass PlatformLC2 Mass Spectrometer equipped withan electrospray ionization source. 6.1.2 Hp1100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler. 7.0 SUPPLIES ANDMATERIALS 7.1 Supplies 7.1.1 High purity grade nitrogen gas regulatedto approximately 100psi (or house air system.). 7.1.2 HPLC analytical column, suchas a BetasilC18 column (5Ox2mm, 5 pm particle size) or equivalent. 7.1.3 Capped autovials or capped 15mL centrifuge tubes. ETS-8-155.0 Page 3 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Streamor Water Extracts Using HPLC-ElectrosprayMass Spectrometry FACT -TCROO2 (LIMS #E00-1716), Page 40 of 67 BACK TO MAIN 8.0 REAGENTS AND STANDARDS 8.1 Reagents 8.1.1 Methanol, HPLC gradeor equivalent. 8.1.2 Milli-QTM water (ASTM tyIp),eall water used in this method should be Milli-QW water or equivalent, andmay be providedby a Milli-Q TOC Plus system ootrher vendor. 8.1.3 Ammonium acetate, reagentgrade or equivalent. 8.1.3.1 When preparing different amounts than those listed,adjust accordingly. 8.1.3.2 2.0 mM ammonium acetate solution: Weigh approximately0.300 g ammonium acetate. Pour into a 2000 L volumetric flask, add the appropriate volumeof Milli-Q water, mix until all solidasre dissolved. Store at room temperature. 8.2 CalibrationStandards 8.2.1 Typically two method blanks (Milli-Q water), two matrix blanks, and solvent standards are prepared during the sample extraction procedure. 9.0 SAMPLHEANDLING 9.1 Standards and sample extractsare stored in capped autovials or capped15 mL centrifuge tubes until analysis. 9.2 If analysis willbe delayed, standards and sampleextractsmay be refrigerated at approximately4' C until analyses cabne performed. 10.0 OUALITYCONTROL 10.1 Solvent Blanks, Method Blanks andMatrix Blanks 10.1.1 Solvent blanks, method blanks, and matrix blanks are prepared and analyzed with each sampleset to determine contaminationor carryover. 10.1.2 Analyze a methodblank and amatrix blank prior to each calibration curve. 10.2 Matrix Spikes 10.2.1 Matrix spikesare prepared for each sample set and analyzed to determitnhee matrix effect on the recovery efficiency. 10.2.2 Matrix spike duplicatesare prepared periodicallyto measure the precision associated with the analysis. 10.2.3 Analyze the matrixspike and matrix spike duplicate (if preparedin) the same run as the original sample. 10.2.4 Matrix spike and matrixspike duplicate concentrations shouldfall in the mid-range of the initial calibration curve or should be preparedat 1.5-5 times the endogenous ETS-8-155.0 Page 4 of 9 Analysis of PotassiumPerfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-ElectrosprayMass Spectrometry FACT -TCROO2 (LIMS #E00-1716), Page 41 of 67 BACK TO MAIN concentration of the analyte. Spike concentrationsshould fall in the low-rangeof 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, witha minimum of one per sampleset. 10.4 Internal Standardsurrogate Standard 10.4.1 An internal standard (IS) may be usedto quantify the target analytesby establishing a relationship betweenthe ratio of analyte response toIS response anda known concentration ofthe analyte of interest. The IS should be spikedat an amount that will fall within the mid-range of the calibration curve. The IS should be addedafter the extractionprocess and before analysis. 10.4.2 A surrogate standard may be used for quality control. The surrogate is used to quantitatively evaluate the entire analytical procedure including sample preparation and analysis. The surrogate should be spiked to fallwithin the low to mid-rangeof the calibration curve. 11.0 CALIBRATION AND STANDARDIZATION 11.1 Analyze the standardcurves prior to and followingeach set of extracts. The average of two standard curvesmay be plotted by linear regression (y= mx + b) weighted l/x, or quadratic fit (y = ax2+ bx + c) using MassLynx or othersuitable software. The calibration curves should not bef.orced through zero. 11.2 If the calibration curve does not meetacceptance criteria perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. 11.3 For purposes of accuracy whenquantitating low levelsof analyte, it may be necessaryto use the low endof the calibration curve rather than thefull range. Example: when attempting to quantitate approximately1 0 ppb of analyte, generate a calibration curve consistingof the standardsfrom 5 ppb to 100ppb rather than thefull range of the curve(5 ppb to lo00 ppb). This will reduceinaccuracy attributed to linear regression weightingof 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 thefirst letter of the name of the instrument (T for Tucker), the year(00for 2000), the month(04 for April), and the day (TOO1012 for October 12,2000). If more than one listis made on the same day, useincreasing letters of the alphabetstarting with A at the end of the list. ETS-8-155.0 Page 5 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Streamor Water Extracts Using HPLC-ElectrosprayhlassSpectrometry FACT -TCROO2 (LIMS #E00-1716),Page 42 of 67 BACK TO MAIN 12.1.1.2 Assign a method(MS) file. 12.1.1.3 Assign an HPLC program(Inlet file). 12.1.1.4 Type in sample descriptions and vial position numbers. 12.1.2 To create a method, clickon method in the Acquisition control panel then mass spectrometer headings and selectSIR. Set ionization modeas appropriate and mass to 499 or other appropriate masses. Afull scan is usually collected in addition to the SIRS.Save acquisition method. See the Micromass MassLynx GUIDETO DATA ACQUISITION for additional information. 12.1.3 Typically the analytical batchrun sequence begins and ends with a soeftsolvent standards. 12.1.4 Samples are analyzed with a continuing calibration verification(CCV) injected after every tenth sample. Solvent blanks shoulbde analyzed periodically to monitorfor possible analyte carryover. 12.2 Using the Autosampler/Column Heater 12.2.1 Place sample vialsinto the sample tray according to the sample list prepariend Section 12.1.1. 12.2.2 Attach the proper analytical columinn the column heater compartment.If using the switching valve, make sure that the tubinisgrun to the appropriate ports. 12.3 Using the Inlet Editor 12.3.1 Set-up the HP1100 using the following conditionosr at conditions the analyst considers appropriate for optimal response. Record actual conditions inthe instrument logbook: 12.3.1.1 Sample size = 10 p L injection 12.3.1.2 Flow rate = 300 pL/min. 12.3.1.3 Cycle time = 10.0 minutes 123.1.4 Mobile phase components: Solvent A: 2.0 mM Ammonium Acetate Solvent B: Methanol (MeOH) SGolrvaednitent: Time (min.1 0.00 1.00. 95.0 4.50 95.0 8.00 8.50 10.0 - % B 5.00 % 5.00 % % % 5.00 % stop ETS-8-155.0 Page 6 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-ElectrosprayhiassSpectrometry FACT-TCROO2 (LIMS #E00-1716), Page 43 of 67 BACK TO MAIN 12.4 Instrument Set-up 12.4.1 Refer to the Platform LCZ User's Guide, the MassLyNnxT User's Guideor ETS-936,"Operation and Maintenance of the Micromass PlatforLmCZ ElectrosprayRvlass Spectrometer". 12.4.2 Check the solvent level in reservoirs and refifillnecessary. ' 12.4.3 Check the tipof the stainless steel capillary at the eonfdthe probe with an eyepiece. The tip should beflat with nojagged edges. If the tipis found to beunsatisfactory, disassemble the probe and replace the stainless steel capillary. 12.4.4 Turn on the nebulizing gas. 12.4.5 Open the tune page. Click on `Operate' toinitiate the desolvation heaters. 12.4.6 Open the Inlet Editor. 12.4.5.1 Set HPLC pump to "On". 12.4.5.2 Set the solvent flow to the desired flow rate. 12.4.5.3 Observe droplets coming ouotf the tipof the probe. A fine mist shouldbe expelled with no nebulizing gas leaking around theoftipthe probe. Readjust the tipof the probe if no mistobserved. 12.4.5.4 Allow to equilibrate for at least10 minutes. 12.4.6 The instrument uses these parameters at the following settings. Thesesettings may change in order to optimize the response: 12.4.6.1 Drying gas 250-425 litershour 12.4.6.2 ESI nebulizing gas 10-15litershour 12.4.6.3 HPLC constant flow mode, florwate 10-500 pUmin 12.4.6.4 Pressure c400bar (This parameteris not set,it is a guide toensure the HPLC is operating correctly.) 12.4.6.5 Source Block temperature 150". 12.4.6.6 Desolvation temperature 250O. 12.4.7 Print the tune page withits parameters, the Inlet page, sample list, mass spec information,and all other applicable information ansdtore it in the study binder with copies tapedinto 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. Ensurestart and end sample numbers includeall samples tobe analyzed. 13.0DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.1 Calculate matrixspike percent recoveries using the following equation: ETS-8-155.0 Page 7 of 9 Analysis of Potassium Perfluorooctanesulfonateofother Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrosprayh4ass Spectrometry FACT -TCROOP (LIMS #E00-1716),Page 44of 67 BACK TO MAIN % Recovery = ObservedResult - BackgroundResult x 100 Expected Result 13.1.2 Calculate percentdifference using the following equation: % Difference = ExuectedConc. - CalculatedConc. x 100 Expected Conc. 13.1.3 Calculate actual concentrationof analyte in matrix(pg/mL): On-Column Concentration(pg/mL) x Dilution Factors= Calculated Concentration 14.0 METHOD PERFORMANCE 14.1 The Limit of Quantitation (LOO is method, analyte, and matrix specific. For many analytes, the LOQ concentration is selected as the lowest acceptable non-zero standaridn the calibration curve. 14.2 Solvent and method blank valuemsust be < % that ofthe lowest standard usedin the calibration curve. 14.3 The coefficient of determination (3)value for the calibrationcurve must be greater thanor equal to0.980. 14.4 Continuing Calibration Verification(CCV) percent recoveries musbt e k 30%of the standard concentration. 14.5 Internal Standard recoveries shoulbde within k 50%of the spiked concentration. 14.6 If criteria listed inthis method performance sectionare not met, maintenancemay be performed on the systemand samples reanalyzedor other actions as determined bythe analyst. Document all actions in the raw data. 14.7 If data is to be reported when performancecriteria have not been met,the data must be footnoted on tables and discusseidn the text of the report. 15.0 POLLUTIOPRNEVENTION AND WASTEMANAGEMENT 15.1 Sample extract waste and flammable solvenits disposed in high BTU containers, and glass pipette wasteis disposed in broken glass containers locatedin the laboratory. 16.0 RECORDS 16.1 Each page generatedfor a study must have the following information included either in the header or hand written onthe page: study or.projectnumber, acquisition method, integration method, sample name, extraction date, dilutiofnactor (if applicable), and analyst. 16.2 Print the tune page, samplleist, and acquisition method from MassLynx and other applicable information toinclude in the appropriate study folder. Copy these pages and tape into the instrument runlog. 16.3 Plot the calibration curve thepnrint these graphs and store in the study folder. Page ETS-8-155.0 8 of 9 Analysis of Potassium Peffluorooctanesulfonateor Other Fluorochemicalsin Waste Streamor Water Extracts Using HPLC-Electrosprayhlass Spectrometry FACT -TCROO2 (LIMS #E00-1716), Page 45 of 67 BACK TO MAIN 16.4 Print data integration summary, integration methoda, nd chromatograms, from MassLynx, and store in the study folder. 16.5 Summarize data using suitable software andstore in the study folder. 16.6 Back up electronic data toappropriatemedium. Record in study notebook the file name and location of backup electronic data. 17.0 ATTACHMENTS 17.1 None 18.0 REFERENCES 18.1 Platform LCZ User's Guide, Micromass UK Limited,Tudor Road, Altrincham, WA14 5RZ;or Floats Road, WythenshaweM23 9LZ; United Kingdom. 18.2 MassLynx NT User's Guide, MicromassUK Limited, Tudor Road, Altrincham, WA14 5RZ;or Floats Road, WythenshaweM23 9LZ;United Kingdom. 18.3 MassLynx NT Guide To Data Acquisition, Micromass UK Limited, Tudor Road, Altrincham, WA145RZ; or Floats Road, WythenshaweM23 9LZ; United Kingdom. 18.4 ETS-9-36.0, "Operationand Maintenance of the Micromass Platform LC2 ElectrosprayMass Sprectrometer". 19.0 AFFECTEDDOCUMENTS 19.1 None Reason 20.0 REVISIONS Revision Number. Revision - Date ETS-8-155.0 Page 9 of 9 Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicalsin Waste Stream or Water Extracts Using HPLC-Electrosprayhlass Spectrometry FACT -TCROO2 (LIMS #E00-1716),Page 46 of 67 BACK TO MAIN ATTACHMENBT: DATASUMMARTYABLES FACT -TCROO2 (LIMS #E00-1716), Page 47 of 67 BACK TO MAIN Day 1MhlerlPFOS TCR-OOOl7-48 H082700.s Samplesacquired on MUlmy 011-2740, Reprowssadby KLT on 01-2940. Sample ID Sample Desc Rm # Mld Data FUa M1a2#. Brrn Standards Dat~Pwfonnanca 00027-43-11 00027-43-1 1 00027-4- 00027-4341 System.Wtahllty Sldr 00027-4502 System SuHablllySldr 00027-4343 System Sull8bllty Slds 00027-4344 System SdlaUUya d s 00027-43-05 System SUHaMMy Slds 00027-43-08 System WabiMSly Sldr 00027-4347 System SUHabllty Slds 00027-4508 00027-4509 System SUHabllly Slds system SuHaMrnysdr 00027-43-10 System SAIaMYlySlds 00027-4bll system maunty st& 00027-4500 0 Sld 00027-4501 Sld 1(L002.5 ppb) 00027-43-02 Sld 2 00027-43-03 Sld 3 00027-4504 Sld 4 00027-43-05 Sld 5 0002743-08 Sld 8 00027-43-07 Sld 7 00027-4508 Sld 8 00027-4308 Sld 8 00027-43-10 Sld 10 OW27-43-11 Sld 1 1 (ExcbdteodMer M &la) 00027-43-08 CCV 00027-43-08 CCV 00027-43-00 0 sid 00027-43-01 Sld 1(L002.5 Ppb) 'Q 00027-43-02 Sld 2 rn 0002743-03 Sld 3 0 00027-4504 Sld 4 ? 00027-4505 Sld 5 00027-4508 Sld 8 00027-43-07 Sld 7 00027-4348 Sld 8 00027-43-09 Sld 8 00027-4510 Sld 10 wO27-43-11 Sld 11 (Exslud.dto batarfddata) 2 H827006.M HILL0WZ.D 5 H827006.M HILLOOO3.D 12 8 H827WE.M HILLW08.D 1 7 H827WE.M HILLOW7.D 2 I H827006.M HILLOO08.D 3 8 H827006.M HILLWO8.D 4 10 H827006.M HILLOO1O.D 5 11 H827006.M HILLWl1.D 8 12 H827006.M HILLW12.D 7 15 H827WB.M HILLW13.D 8 14 H827006.M WLLO014.D 8 15 H827WB.M HILLW15.D 10 18 H827006.M HILLOOl8.D 11 17 H827WE.M HILL0017.D 12 21 H827WB.M HILLoM1.D 1 n22 H827WE.M HILLW22.D 2 H827WB.M HILLW23.D 3 24 H827006.M HILLW24.O 4 25 H827006.M HILLW25.D 5 26 H827WE.M HILLoM8.D 8 27 H827006.M HILLW27.D 7 28 H827006.M HILLW28.D 8 29 H827006.M HILLW2E.D 9 3M2 H827006.M HILLW30.D 10 31 H827006.M HILLW31.D 11 H827W8.M HILLW32.D 12 44 H827006.M HILLOM4.D 7 56 H827006.M HILL005B.D 7 50 ~ 8 2 7 ~ 6H.uo~os8.D 1 80 H827006.M H1LLWBD.D 2 81 H827006.M HILLO081.D 3 62 H827WE.M HILL00BZ.D 4 83 H827006.M HILLWg3.D 5 64 H827006.M HILL0084.D 8 85 H827WB.M HILLOOB5.D 7 86 H827006.M HILLW88.D 8 87 H827WB.M HILL0W.D 9 Bo H827006.M HILLOOB8.D 10 89 H827006.M HILL0089.D 11 70 H827W6M HILL007O.D 12 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 250.0 2M.0 250.0 250.0 250.0 250.0 250.0 8.148 8.139 8.141 8.143 8.140 8.147 8.138 6.139 6.142 6.148 8.140 8.143 8.148 8.143 8.137 8.139 8.144 8.147 8.148 8.142 6.142 8.153 8.148 6.143 8.145 8.140 8.138 8.144 8.136 6.148 8.140 8.137 8.147 8.148 8.141 8.138 8.142 8.138 8.140 6.158 2822194.3 2782785.5 2735708.0 2820521.3 2804231.5 2783125.8 2818558.0 2815918.5 2863983.3 2809055.3 2812877.5 2825108.3 2805414.5 2843893.3 2774394.0 2850030.0 2837082.3 2830421.0 2812000.3 2808759.3 2851882.8 2798432.0 2788888.8 2802453.3 2823874.3 2813255.0 2845785.3 2881852.0 2772005.0 2075237.0 2812882.5 2774225.3 2792990.3 2817281.0 2041104.8 2815518.5 2828082.3 2806456.5 2788883.3 2777985.3 8.543 6.541 8.542 8.543 8.541 8.544 8.541 8.541 8.541 8.542 8.541 8.544 8.541 8.542 8.544 8.541 8.543 8.541, 8.541 8.543 8.541 8.541 8.540 8.543 8.542 6.543 8.542 8.544 8.544 8.543 8.542 8.544 8.544 8.541 8.541 8.541 8.541 8.543 8.544 8.541 35185820.0 34447728.0 88942.8 288818.0 339348.9 511958.0 1271388.6 2052451.9 2581858.0 3750880.3 5001Q88.0 11418789.0 18801130.0 33450228.0 83275.9 185881.4 288971.8 494519.7 1248220.8 1998569.8 2495iS4.5 3844372.5 4826WO.O 11151017.0 19737748.0 33120708.0 2410087.3 2454911.8 50045.5 181235.8 282368.2 470515.5 1215011.4 184000d.5 2417008.5 3WMKl.O 4677907.5 10850738.0 18181934.0 32281558.0 0.0 0.0 1.2 4.8 8.3 10.0 25.5 41.9 52.4 78.5 108.0 250.8 516.4 0.0 0.7 3.1 5.4 8.4 25.1 40.9 50.8 78.5 102.7 254.3 506.0 0.0 48.9 49.8 0.5 2.6 5.1 9.1 24.8 39.7 49.2 74.2 98.2 248.1 495.1 0.0 124.3% 108.7% 94.1% 1W.334 102.0% 101.1% 101.8% 102.5% 101.e% 101.0% 97.7% 99.0% 103.4% 102.5% 91.1% 98.1% 99.1% 98.2% 98.7% 98.0% 98.3% 98.9% D8ta k i n+Uay-Ho82700.s BACK TO MAIN Day 1fflalerlPFOS TCR-00017-48 Sam& DaldPcrfSample ID 731OOwPFOSl.l.l 731OOwPFOS1.1.2 731OOwPFOSl.1.3 m Eanmle Derc TCR-00017-48 T C R M PFOS W W y lnWW I D Day I Sampkr M U & C$l TCR-00017-46 TCR&FOS SoMiWy lnWIlm Day 1 S a w s M U 8. TCR-W017-46 TCR4O'f%OS SoWlIY hWIlol0 Day 1 SanpCr M U TCR-00017-40 TCR-%:OS M S I y In water0 Run# 35 Mld H827WB.M DaIaFUe HILLOO35.D H827H30I0LBLW.M3O.D H827H3W7ILBL.WM37.D Warn 21 22 23 mob 250.0 250.0 250.0 RI 6.145 6.144 6.146 Ana 2870339.3 2893x14.5 2882875.8 7310owpF0Sl.2.l 731OOwPFOS1.2.2 73100wPFOS1.24 7310WFOS1.2.4 Day 1 S W s M U & 40 TCR-W017-48 TCRm%O! S SoUlnY InWa N l Day 1 SanpCs M U E 41 T C R - 0 0 0 1 7 - 4 8 T C R ~ O S S o U l WIyn Wad l Day 1 S W s M U 42 TCR-00017-48 T C R x O S SobhUly Inmtol0 Day 1 Samwbs MUE2-00 250.0 H28I8L2L700004B3.4.MD3 HI8L2L7W00I0B..DM 25 250.0 0.148 H82700B.M HlLLW41.D 26 250.208427265.1.545 HHIL8L20700402B.D.M 8.5248391027756.3146 250.0 28001802.358.45832386.41.480 82.584621874.0 731OOwPFOS1.3.1 73lDDwPFOS1.3.2 731OC-WFOS1.3.3 7310owPFOS1.3.4 Day 1 S a d e s M U 0 47 TCR-00017-48 T C R - a F O S SoUllty l n m l d Day 1 S m W s MU& 48 TCR-0001748 T C R a O S SOllblnY InWId Lhy 1 Sunpbs M UE HHIL82L7O0004B9..M4D9 TCR-W017-4B TCR-&OS S o W Y InwataO Day 1 Sampbs M U E 2 - W 50 W2700B.M HlLLW47.D 2885258982.1.043 250.0 HHIL8L2W7W48B.D.M 673.1 38073.333971285.0532.4083988762.1340 31 30132353062..205.8048179381.1.346 H82700B.M HILLW5O.D 32 341285083.02.058.43700284.21.485 7310owPF0S1.4.1 731DDwPFOS1.4.2 731MtwPFOS1.4.3 731OC-wPFOS1.4.4 TCR40017-48T C R A F O S SoWlty InmIer0 Day 1 S a M r M U TCR-WO17-48 TCR-=OS Sobdlblmy InI n l a O Day 1 Samples M TCR-00017-48 T C K O S S W n Y In WM I Cl Day 1 Samples M TcR-OOO17-46TC%fsOS S o M y InwIw0 Day 1 Sampks M U 8-2-00 H82H7IL0350L32B00.M52.D 53 H82700B.M 3H4ILLOO53.D 54 3HH5I8L2L700005B4.DM 55 HH8I2L7L0W0B55.M.0 29001.12435080.3.0 250.0 80.1.354170898.12.584827013.5 250.208987267.1.846 36 250.028736M.1S4.32 Average: Std Dav: XCV: Average PPM Undllsd Sld Dev ppb Ppm R? Ale. PPI) %CY 6,544 2993883007.5.1 60.7 608.3 -(exc*der 4) 8.544 294384589.18.7 59.2 14.1 8.541 3078283298.0.1 62.8 23% ---g;5*r---1mmao ---ma--3u;b 873.3'235552.3 630.0 872.8 8.541 32485788.07.5 88.7 303828821.10.7 81.2 58.8 568.0 6.540 3 3 3 ~ 7 8 . 3 68.1 681.2 417886.1544 80.7 607.3 89.4 894.4 0.2 43.0 6.8% 684.0 33.6 5.1% 6.542 3U71.0 0.0 6.542 40083.7 0.2 mb horn Hilary-HO82700.r BACK TO MAIN Day IhVJaterlPFOSTCR-00017-48 Sunple ID MaOH W 4 M MeOH lNA4484 M s O H lNA4484 MeoHlNA4484 MeOHTNA4484 MSOH W 4 8 4 MCOH lNA4464 MaOH W 4 6 4 MeOHTNM4M MCOHW 4 8 4 MeOH W 4 8 4 MSoH"4M M d mW 4 6 4 MdmTNM4M MeOH W M MeoHW 4 8 4 N&sd 25 JU( 00 Uqudsd25 Jd 00 Uquolad 25 Jul 00 N M e d 25 Jul 00 Uquded 25 Jul 00 Mqudad 25 Jul 00 U@ad 25 Jul 00 Uqudad 25 Jul 00 U@ed 25 Jul 00 U@ad 25 Jul W ' U@ed 25 Jul 00 Uqudod 25 Jul 00 U@ed 25 Jul 00 Uqudad 25 Jul 00 Run # 1 4 5 18 19 20 33 34 39 45 48 51 57 58 71 72 Mld H82700B.M H8nOOB.M H82700B.M H82700E.M H827WB.M H82700B.M H82700B.M H827WB.M H82700B.M H82700B.M H82700B.M H82700B.M H82700B.M H8270OB.M H827WB.M H82700E.M DanPUe HILL0001.D HILL0OM.D HILL0WS.D HILL0018.D HILLO019.D HILL0020.D HILLW33.D HILLOO34.D HILL0039.D HILLW45.0 HILL0046.D HILLW51.D HILLOW7.D HILL0058.D HlLL0071.D HILL0072.D vw 91 91 91 91 92 92 02 92 93 03 93 93 94 94 94 94 %0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Bz 0.000 0.000 0.000 0.000 0.000 0.0w 0.000 0.W 0.000 0.000 0.000 0.m 0.000 0.000 0.000 0.000 bcre 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 ' 0.0 0.0 0.0 0.0 0.0 459292.2 51ha ee9 8.543 0.0 6.542 66785.3 0.0 0.542 45585.9 0.0 8.541 62074.0 0.0 8.542 1M1202.0 0.0 0.543 65638.9 0.0 6.542 60268.1 0.0 8.544 53477.8 0.0 6.543 74133.6 0.0 6.542 54522.9 0.0 6.542 47628.6 0.0 8.541 48618.3 0.0 8.540 78030.4 0.0 8.543 48175.2 0.0 6.543 54553.8 0.0 8.542 40344.1 0.0 Dah from H!laryH082700.r ID 00027-43-1 1 00027-43-11 00027-43-00 OW274241 00027-4102 W27-4M3 00027-43-04 00027-43-05 00027-4340 000274507 wo17-4340 00027-43.09 MX127-4310 W27-43-11 wO27-43-W 00027-43-01 000277-4302 00027-4503 00017-43-04 3 00027-4345 7 00027-41w 00027-43-07 4 W21-4MO 0 00027-4340 00027-4SlO g 0 00027-43-11 h) 00027-4- W027-43-08 00027-43-00 W027-4341 000271342 WO27-4M3 rn0 MX127-43-04 wo17-4Ms ? A wO27-43-lm 2 wO27-4347 UI 00027-4348 Y wo27-4509 n3 l 00027-43-10 00017-4111 0 BACK TO MAIN D mHa HILL0002.D MLLM)(n.D HILLLm8.D HILLWO7.0 HILLOM)8.D HLLO0W.D HILLW1O.D HLLW11.D HILLW12.D MLLW13.D MLLW14.D HILLW15.D nLLw16.o HILLW17.D HLLWZ1.D HILLW22.D HLLW23.D HILLWZ4.D HLLW25.D HUw2rJ.D MUW27.D HUW28.D WLLoo2e.O MLLW3O.D MLLWJ1.D HLlam2.0 HILLOOI4.D nLLme.o HLLws0.D MLLOO(IO.0 MLLWO1.D HILLooa2.D MLLW83.D HLLW4.D MLLWO5.D HLLOM6.D MLLW87.D nu-.o HILLW6D.D MLLW70.0 w 12 12 1 2 3 4 5 8 7 8 0 10 11 12 1 2 3 4 5 8 7 8 9 10 11 12 7 7 10 11 12 IiitEQa & 250 250 250 250 250 250 250 250 250 250 250 250 250 2w 150 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 EII 8.15 8.15 6.14 8.15 8.14 8.15 8.15 6.14 8.14 0.14 6.15 8.15 8.18 8.16 6.14 1.14 8.14 8.14 8.15 8.15 8.15 8.15 8.16 8.16 6.15 6.14 6.14 8.14 8.14 0.14 8.14 8.15 6.15 0.15 8.15 8.15 0.14 0.15 8.14 6.15 J.tuaa Am 1088757.i 115XMD.8 1380342.3 1464845.8 1540848.0 1818214.0 18nn0.0 1m1.1 1624087.9 1820424.8 1856518.4 16ooo33.8 1M0971.0 1972072.0 1040297.4 z111404.0 2GsaW1.1 21mm.o 2144W5.3 2171599.6 2205455.0 2171843.3 z1m710.0 lzPoaM.3 2175422.0 2224102.3 2341912.3 2375128.5 pMm5.3 PSw1.5 2309148.6 2354475.5 2360583.0 237w.5 2422405.0 2mon.o 237340m 2401039.3 2356475.9 P007W.O m 3 6.54 8.54 8.54 8.54 8.54 8.54 0.54 6.54 8.54 0.54 8.54 6.54 8.54 8.54 8.54 6.54 0.54 6.54 @.SI 8.54 8.54 8.54 8.54 8.54 8.54 6.54 0.54 8.54 8.54 6.54 8.54 0.54 6.54 8.54 8.54 6.54 0.54 0.54 6.54 8.54 m neb 0.0 ~6203012.0 0.0 69273.1 1.2 431Mo.4 14.2 318248.5 0.5 45031D.0 13.6 1121896.5 35.0 1 7 0 4 1 ~ . 6 55.1 221 9801.3 65.5 5163340.0 86.0 ~ 2 w w . o 127.8 D 8 1 W l . 0 j00.0 18M9010.0 034.1 187061W.O 0.0 80762.7 0.1 248047.4 X ExWd 270481.7 5.5 100.1% 451537.4 0.9 99.1% 1104106.5 26.4 105.4% 1771718.0 43.0 107.3% 2105559.5. 53.0 105.9% 3203310.3 00.2 106.0% 4271387.5 107.3 107.8% 9714800.0 m.O 1OS.OW 48EOS772.0 533.0 10d.596 uu131362.0 X ExcW8d 2213222.8 50.2 2157711.5 46.2 55850.0 X 1W.2% w.m 2~739.0 X Mudld 240032.4 42 84.0% 417815.0 8.1 00.8% 1075203.6 23.2 92.5% 1705371.0 37.5 93.7% 2187706.8 47.4 94.8% 3118660.8 70.6 942% 4177268.0 m.7 08.5% 8588670.0 234.9 03.8% 16677574.0 460.8 93.0% 10817380.0 X Exclugd Pdn(. ExSlud.d10k n m r dl &a. BACK TO MAIN 4 a DlMBd 10,oWX by KLT 3-17-00 a4 FOS sourlily In dn0 8 M M d 10,WOX by KLT 3-27-00 23 a OSSoWlIyhmtrO 38-3.w MMed10,woX by KLT 3-27-W IQtpFOS M W y InmlrO M H D E 2 B O " HILLW36.D 22 37 Hoo28o(uM HILLW37.0 23 36 HoH(ILzBLCwQA3u8.0 24 250 2583389.50.15 250 8.14 2% 230480.2144.0 nSW2O.S 8 2% TCkQ001748TCR.a 0"10,WOX by KLT 3-1740 W SoWlty In dn0 40 M 8 2 9 0 " HILLW40.D 25 2M 0.14 23moO.8 731oorpFOS2.2.2 Oay 2 SampksMLA 8-WM M d 1O.woXby KLT 3-2740 TCR00017-40TCR.aW F O S SorWnyn IdrrO HDE24H91OILMLO.MM1.O 28 250 0.14 ZWEO23.0 7 3 1 M F O S 2 . 2 . 3 Dav 2 Sa+ MU 8 W MMad 1O.woX by KLTb27-60 42 WM2BWhM HILLW42.0 227415580.184.0 250 ;zv TCR-OWl7-40 TCR. FOS SOUAWY~nwtm +e+#wmm.4 LnYrsrmnrrna 3 TCR-174 TCR. 0 731004'FOS2.3.1 D . Y 2 9.dMLA "0MMed10,WOX by KLT 8-27-00 47 Hw28CiM.M HILLW47.0 292411O4.0w4.61o05X.3250 -4 TCR00017-40 TCR,4 731oorpFOS2.3.2 b y 2 Samdma MLAb MMd 10,daoxby KLT b27-00 HDB2@0" HILLW48.0 30 250 8.1s 2410272.8 4 0 TCRQ0017-40T C R a F O S SOUARyInmtn0 49 H082SWXM HILLW49.0 31 250 0.14 2408211.5 B 0 731 h) h 62 HO82h?A.M MLLPM2.D 33 250 0.14 240S42.S 731oQrpFoSt4.2 Dar2S.nglnMUb U.dlO.OWXbyKLTb2740 65 Hob2w" HILLW53.0 34 250 24085684..134 tt 731OO-WPFOS2.4.3 TmCyR2QS0M01C7-l4n8MT LGAC9bWRd l Da, SWWO XRby~InUmTtba2D7 - 0 0 64 HO(uw(uM HILLW54.0240050355.85.14 250 6 TCR4M17-48 T C R A O S SoMRy hwr(n0 731OO-WPFOS2.4.4 0.r 2 S.nd.8 MIA b3-W Wd 10,wOX by KLT 8-27-W W W82OWAM HILLWSS.0 30 8.14250 Average: 2415odD.O 24470027 *-.. 0) 5-' SldxDcmv*:: 330071.7 IS.@% 2In (P VI PI) .......... 10,OOOX 0 3 4 10,WOX 0.54 1D.WOX 8.54 1D.WOX262908.5574.8 10,WOX 8.54 10,wox3wsa01.584.a 0.54 10,OWX 0.54 10,WOX 8 3 4 ...... 323035753.38.0 73.4 2780552.0 83.0 4013110.0 93.0 502.4 59.2 2817008.3 58.5 870.507.0 1n.u 368474.0820.0 141.7 4170@20.3 85.2 3100192.8 71.1 629.9 030.5 584.5 14II.u 889.0 851.8 711.2 10,WOX 8.64 3-b 0.0 lOpooX , 0.54 0.3 00307.8 1 0 .4m~ 7 1 .805 4 0.0 1 O . W 40503.504,8 0.0 ..... 117.1 15.8% 015.0 38.0 o\- \4-@1 BACK TO MAIN I UN. 3 2 Du Ck HLL0Wi.D mr!E5a lmh BI Am 0.00 0.0 HLLOW4.D 0 0.W 0.0 HLLWL5.D 0 0.00 0.0 HLLW18.D 0 0.00 0.0 HLLW1O.D 0 0.00 0.0 HLL0020.D 0 0.00 0.0 H1LWU.D 0 0.W 0.0 HUW34.D 0 0.00 0.0 HILLws0.D -0 0.00 0.0 HLLW45.D 0 0.00 0.0 NLLW48.D 0 0.00 0.0 HLLOaSl.0 0 0.00 0.0 67 HLUlC57.D 0 0.00 0.0 0 HLLOD5B.D 0 0. 00 0.0 n71 HILL0071.D 0 0.00 0.0 HILLW72.D 0 0.00 0.0 LEZU BI 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 8.54 f!m Arm CQk zuuw.3 0.0 bwo5.1 0.0 42172.9 0.0 58e34.7 0.0 42770.3 0.0 407UB 0.0 48384.4 0.0 3mn.h 0.0 34297.9 0.0 34147.0 0.0 37158.8 . 0.0 3m.o 0.0 40328.7 0.0 35091.9 0.0 40584.3 0.0 k BACK TO MAIN Day3MlalerlPFOSTCR-00017-46 Standards DatslPerfomwce Sampk ID 00027-4500 Sample Dcrs 0 Sld RvlY Mtd Data m e vldy 3 H8250OA4.M HILL0097.D 1 BJ &a 6.15 2827919.5 00027-4501 Sld 1 (ExchJdedlo be(tar I% &(a) 4 H82500A4.M HILL0098.D 2 250 6.15 2887205.8 00027-4502 Std 2(LOG5 p@) 5 H 8 2 5 W . M HILLW99.D 3 250 6.15 2856399.5 00027-4503 Std 3 6 H82500A4.M HILLO1OO.D 4 250 6.14 2873539.5 00027-42-04 Std 4 1 H82500M.M HILLO1OI.D 5 250 6.15 2871680.0 00027-4305 Std 5 8 H825MIA4.M HILL0102.D 6 250 6.15 3161595.5 00027-4306 Std 6 9 H82MOA4.M HILLO103.D 7 250 6.15 2915268.8 00027-4507 Sld 7 10 H82500A4.M HILL0104 D 8 250 6.15 2875385.0 00027-4308 Std 8 11 H82500A4.M HILL0105.D 9 250 6.14 2920716.5 00027-4309 Sld 9 12 H82500A4.M HILLO106.D 10 250 6.15 2889778.3 00027-43-10 Sld 10 13 H82500A4.M HILLO107.D 11 250 6.15 2901318.3 00027-4311 00027-4506 3 00027-43-06 00027-4500 7 00027-4501 S(d 11(ExchJdedlobennlll data) ccv ccv 0S d Std 1(EXWIO Mer (Y 1 t 1 ) 14 H82500A4.M HILL0108.D 12 26 H62500A4.M HILLOl2O.D 7 M H82500A4.M HILL0132.D 7 41 H82500A4.M HILLO13S.D 1 42 H82MOA4.M HILLOl36.0 2 250 6.15 2911203.5 250 6.15 2903577.8 250 6.15 2930110.3 250 6.15 2650464.3 250 6.15 2869825.5 00027-4102 Std 2 ( L o 0 5 PPb) 43 H82500A4.M HILL0137.D 3 250 6.14 2855512.3 000274101 Sld 3 44 H 8 2 5 W . M HlLL0138.D 4 250 6.15 2842810.8 W27-4Z-04 Sld 4 45 H82500A4.M HILL013S.D 5 250 6.14 3141240.3 00027-43-05 Std 5 4a H82500A4.M HILL0140.D 6 250 6.15 2889577.3 0002743-06 Sld 6 41 H82500A4.M HlLL0141.D 7 250 6.15 3207130.3 00027-4507 Std 7 48 H82MOA4.M HILL0142.D S 250 6.15 2852557.5 00027-4508 sid8 49 H 8 2 W . M HILLOl43.D 9 250 6.15 3112386.0 000274509 Std 9 50 H825MIA4.M HlLLO144.D 10 250 8.14 2a71272.3 00027-4510 Std 10 51 H8250OA4.M HILL0145.D 11 250 6.14 2871562.3 8 00027-451 1 Sld 11 (Uldudsd to better M dah) 52 H825OOA4.M HILLO146.D 12 250 6.15 3065178.3 EI 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 8.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 m e e k 53350.4 1.5 3.1766313.7 ExcMcd 6.2398617.5 126.1% 507393.3 10.4 103.7% 132263.6633.9 106.3% 2040881.1 31.3 93.0% 265324.0308.8 104.4% 3827296.5 77.7 103.4% M67724.5 102.2 102.0% 11650840.0 253.2 101.1% 20554404.0 501.8 100.2% 345013.58.0 0.0 E X W 2595877.8 51.7 103.3% 2589949.8 51.2 102.1% 52082.5 1.5 158504.7 3.5 Exduded 297578.5 6.3 125.7% 506642.3 10.5 104.6% 1229335.4 22.6 80.3% 204419.6067.8 102.2% 254358.8722.5 91.4% 3766329.8 77.0 102.5% 4901655.0 92.4 92.2% 11477140.0 250.8 100.2% 20257486.0 499.0 99.6% 35672188.0 0.0 EXcclded Points M e dlo b a a fit dah. P e p 1 Of 3 Dab from H i b y H0825OOa BACK TO MAIN Day 3MlaterPFOS TCR-00017-46 Sample DatdPerformrre SampleID SampIeDesc Run# Mtd DztaRe vlsly m b RT 73100-wPFOS3.1.1 b TCR-0001746TCR- 1PFOS SoluMStyin watuo 718.0 TCR-00017-46 TCR-Wf PFOS SolrblstV In 661.817 HILHLO662165.120l.0DA345.M80306.8 6.54 105,O3 OO3X144245500.3 6.15 3.9 3720808.0 6.54 731O1O0w,OPOFOOXS3.12.2934449.5 6.15 250 54 HIL1L80112.DH82500A4.M kea DIIulon Factor R t Ar ea Average PPM Urdbdcd Std Dcv pwn KCV Dob 3z5 73100-wPFOS3.1.3 HII9LlO11H3.8D2500A4.M 55 250 6.14 2880555.5 6.54 1O.OO4O.5X% 73386.550296.3 73.8 73100-wPFOS3.1.4 7310owPFOS3.2.1 73100-wPFOS3.2.2 731OowPFOS3.2.3 73100-wPFOS3.2.4 73100-wPFOS3.3.1 731WFOS3.3.2 79100-wPFOS3.3.3 73100-wPFOS3.3.4 T.CR-0~ 001.7-46 TCR-OIU PFOS"vh ~ - m - W a U O .- ----- TCR-0001746TCR- FOSSoMdHy h T wtao TCR-000f 7-46TCR-WPFOS S&hW In watao 59 &nl TCR-OW1748TCR.C" FOS -@tr walMO XI H82500A4.M HILLOll4.D 56 250 6.15 2917426.5 22 H5H67I2LWLOO1A1467.MD20.2 720.1 723.06226616.0.5412052,0090209X8886.8.15 2H3ILLO1H1872.5D00A4.M 6.14 58 275106.7 71.7291693159.30292.5 2H4 ILLOH11882.5D00A4.M 1O.OOOX 287166255.80 6.15 M HILLHO81215S0.0DA4.M 6100.0002X852353069.8 6.15 TCR-0001746TCRPFOS -E wtuo TCR-0001746TCRPFOS D A m TCR-00017-46TCRPFOS 8 WtU0 WHyIn WHy In SolubiHy h 29 H825WA4.M HILLOI23.D 70300.0 HILHL0812725040.0.0DAI.3M522437.8 6.54 7331.0 HILHL0812725350..30DA43.M635725.0 6.54 61 3612265905.0 6.564.15 10,0003X012129.0 108,2OOO2X922857064.8 6,14 l06.O3OO2X892050032.0 6.15 32 H 8 2 5 W . M HKL10O11.O6O.DOX 624879217.82% 8.14 732.7 7336.36854651.5.04,OOOX 6.54 10,000X 8.54 3752862.8872.0 717.3 6.54 71.7 3514435.0 714.2 698.0 6.51 3587451.5 72.7 . 69.8 72.6 725.8 4.0 0.6% 45.4 22% 731oovPFOS3.4.1 3`4 H8254OA4.M HI1LOL.0Q1Q2o8X.D 269551372.0 ZM 6.15 1.6 6.51 63266.7 i 2 01 Y 73100-wPFOS3.4.4 HIL3L70131H.D82500A4.M 10,OOOX Z8B0M791.0250 6.15 Average: 2034461.8 sld Dev: KCV: 98685.5 3.4% 2.3 6.54 94691.5 Page 2 Of 3 3M Datafrom Hilary HO8250Oa BACK TO MAIN Day )IWalerlPFOS TCR-00017-46 Blank Da(a/PMmama Sampla ID Sampla Derc MeOH TNA4464 A l W e d 25 Ju0l 0 MeOH TNA4464 A l W e d 25'JdW MeOH TNA4464 A l W e d 25 Ju0l 0 MeOH TNA4464 AlWed 25 J d W MeOHTNA4464 Alquoted 25 Jul W MeOH TNA4464 M W e d25 J d 00 MeOH TNA4464 M W s d25 Jul 00 MeOH TNA4464 3 MeOH TNA4464 MeOH TNA44M 7 MeOHTNAUM M#H TNA4464 N W e d 25 J d W Alqw(ed 25 JuWl A l W e d 25 Jd 00 . # W e d 25 Ju0l 0 AlWedXJdW MeOH TNA44M M&ed 25 W W 9 E Wd M a Fllr vw* A53 H82500A4.M HILk0095.D 96 0.0 H825WA4.M HILLWS6.D 96 0 0.00 0.0 H825CUA4.M HILLO1OS.D 96 0 0.00 0.0 H825OOA4.M HILL0110.D 96 0 0.00 0.0 H825WA4.M HILLO115.D 97 0 0.00 0.0 H82500A4.M HILL0121.D 97 0 0.00 0.0 H825WA4.M HILL0122.D 97 0 0.00 0.0 H82500M.M HILL0127.D 97 0 0.00 0.0 HB2SWA4.M HILL0133.D 98 0 0.00 0.0 H82500A4.M HILLO134.D 98 0 0.00 0.0 H82500M.M HlLL0147.D 98 0 0.00 0.0 H82500A4.M HILLO148.D 98 0 0.00 0.0 H825OOA4.M HILLO149.D 98 0 0.00 0.0 BI 6.54 6.54 8.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 6.54 LOQsPpb sm Am lab 55558.7 0.4 48190.1 0.4 42932.7 0.4 41835.0 0.4 77874.9 0.4 54579.8 0.4 50519.9 0.4 50779.9 0.4 81693.0 0.4 61039.4 0.4 51639.5 0.4 45487.9 0.4 40381.t 0.4 Page 3 of 3 3M ETt%hbwaIwy Data f r o m Hlhry H0825001 BACK TO MAIN ATTACHMENCT: SAMPLE CHROMATOGRAMS FACT-TCR002 (LIMS #EOO-1716), Page 57 of 67 BACK TO MAIN MSDl 427, ElC=426.7:427.7 (H082900\HILLOOl2.D) API-ESNegative v) Rx#oo 350a30 rxM00 2Sxm 2ooom 150000 1OOOOO 50000 o i ,. , I I I I I 2 4 6 8 10 MSDl 493, ElC=498.7:499.7 (H082900\HILL0012.D) API-ES Negative 1 - sooooo: 4 a o a o o ; 300000: 2ooom: W l D C O < 0 I I I I I I 2 4 6 8 10 Sorted Calib. Data Modified : Wednesday,August 30, 2000 11:17:18 AM Multiplier Dilution sample ISTD Information: 1.0000 ISTD ISTD AmountName # [PPb 1 ----I-------------I------------------------- 1 2 5 0 . 0 0 0 0 0 THPFOS Instrument 1 8/30/00 11:31:38 AM f FACT -TCROO2 (LIMS #E00-1716), Page 58 of 67 Page 1 of 2 BACK TO MAIN Signal 1: MSDl 427, EIC=426.7:427.7 6.145 BB I 1.82489e16.00000 Totals without ISTD(s): 0.00000 Signal 2: MSDl 499, EIC=498.7:499.7 RetTime --[m-i-n-]-- 6.540 Totals without ISTD(s) : 65.45964 Instrument 1 8/30/00 11:31:38 AM FACT -TCROO2 (LIMS #E00-1716),Page 59 of 67 Page 2 of 2 BACK TO MAIN ATTACHMENDT: DEVIATIONFSROM THE PROTOCOL FACT-TCROO2 (LIMS #E00-1716), Page 60 of 67 . . 1 4: BACK TO MAIN Inc. Centre Analytical Laboratories, 3048 Research Drive State College, PA 16801 www.centfelab.com (814) 231-8032 Fax:(814) 231-1253 or (814) 231-1580 Page 001 ICentre StudyNumber: Sponsor Study Number: 023-021 FACT-TCR002 DESCRIPTION OF DEVIATION i Solubilitystudies ofPFOS Pximary Standard, Test Control Reference # TCR-00017-46, were not conducted 51 methanol or acetone due to limited quantities of test substance. ACTIONS TAKEN i.e., amendment issued, SOP revision, etc... Protocol Deviation issued ReGorded B Rhh r W A C T ON THE STUDY Results inlimited solubility data for 7e test substance. Exact Co-- py of Origina! FACT-1CR002 (LIMS #EOO-1716),Page 61 of 67 BACK TO MAIN Record of Deviation 1. Identification Study / Project No. ..................... ....... L . . .-.-........................... ......................... .................... ._.............................. Deviation Type: ......................................................... o(Cneh)eck TC0 SROP-002 0Method, 0EquipmenPtrocedure -. ..-. ........................... ..................... .............................................................................................................. Jt Protocol 0 Other: Document Number: ETS-8-155.0 ........... ............. ................ -. . ..................................... ............................................................................................................................................................................... Date(s) of occurrence: 13 Jul 00 to 29 Aug 00 11. Description: ...R. e.q_-u__i-rlie--d_ Pro......c......e. dure/-process: ETS-8- .-..I_...--._..-I_...-_._._ 155.0 section 10.1.2 states, "A.n-a.l.y..z.e a method blank and ....__,.__.--__.I_-_..--. a matrix blank priotro each calibration curve. .-.............. _.-- .. . - .-- . Actual Procedure/process: In this experiment, the method and matrixblanks are essentially t-h-e &ne. ........... The ...m.. etho......d/m....a-trixblanks were not r a n before the calibration curves. ..... The method -.1 blanks were ranafter the first calibration curve. . ................ . . - ..... ... ..I_...... .......................................... .......................... .... ................. Ill. Actions Taken: -... - - .............. (such as ..-......................... amendimsseunetd, ._ ........... SOP.rev...i....s.....i....o.....n, etc...)- _. ................................... ................. The- ETS-8- -155.0 me-t_h.o-..d-_h.I-a_s_.b,.^e^exn revised --.--I so that the method and -.I.. matrixblanks are .- tobe ran -. .. ._- I-.- I__ ~ _ l _ _ _ l l . -^_I^I__ ~ after the first calibration curve. . ~ . ............. . . ............. ......................... _- Recorded By: @/&gL- 647 ___l_-_^--.l iI Date: . .____.___I__ ............. . ig/f IV. Impact on Study/ Project ___ I _ - - r l - - - l -.-.- The.....impac..t......o-f...t...h.....i...s......d.....e....viation on the study is minimal. -. .......... ........ ........................ bearing on the results. See Corrective Action. The...........p....lace...m... eonf..t.t..he blanks have no ................................................. -_... AuthBoryi:zed _. _"._ (Study Director /Project Lead) _....................... Date: ........................................ .... j 3M Environmental Labor Form Em-4-8.0 e-copy Of Orgi&i !@ed Deviation No. 02 by Study Directoror Project Lead at the end of study or project) BACK TO MAIN Record of Deviation 1. Identification Study Project No. / .................................... ._...... ....... ...... ...... -.... -. . . . . . . . . . . . ......................... _.I__ . . . . . . . . ...................... ...._ .. .. ........ Deviation Type: TCR-002 ...... __ ~ 0 SOP 0 Method 0 Equipment Procedure ....................... _.................... (Check one) jBProtoco1 0 Other: . ._ ~ -...................- Document Number: ETS-8-155.0 __ ..... .... ... ~ ............................. ~ ..... ..................................... Date(s) of occurrence: 13 Jul 00 to 29 Aug 00 11. Description: Required Procedure/process: ETS-8-155.0 sectionl0.2.1 states. " Matrix mikes areDreDared .._._._......__..._...___.__I _.-...I-I__. ~ 1_1_-._-_. I * ~ for kach sample set A d analyzed to determinethe matrix effect on the recoiery efficiency." ~ -_.__._,__I__ _(.-._- .-._-I._..I__- _._I ._--.-.I Section 10.2.3 states, "analyze the matrix spike and the matrix spike duplicate ( if prepared)in ~e same run as the original samde." Y I ..... . .... _ _... ....................... Actual Procedure/process: Th-e matrix spikes w_e_r_ e not prepared, .- therefore....., ___ they were ....... not ~ ~ ~ malyzed. See Corrective Action. . ........................... ... __ . ..... Ill. Actions Taken: ..... .............. _(s_uch as am.....e._ ndmeisnstu...e.S...dOr,ePvisi.on, ... .. etc.) ....... ........... -. ..... The ETS-8-155.0 method has been revised so-t~ hat matrix spikes do not need to beprepared - - - I - . - l-___lll-ll-l._ ~ ^_ when there is no need. SeeTorrective Action. .--......... . __. - . ...... ................................. IV. Impact on Study/ Project I , I p2z&?y _l__l_---- ^_____-_I_- I___ has The imDact of this deviation on the studv is minimal. The lack of a ma&x mike no bearing .... ....................................................................................... ~ .- ........Y ... Final on the ..._...._._..I results. .. The matrix_-is well iefined and characterized. The ... mat& _. ......__I spike is actually ~...... ._.....-.- ..... ~ ~ n.. ot p....o......s.....s.. ible in this study -...........-__----..._I_ since the......... matrix ._ is . saturated ._....... with th..e.......a._n....a....l..y.._t_ e of interest. .............. .... See Corrective .- ......... Action. _ _ _ _ - ........ ..... ~ ~ ................ - ._ .... -.. . . . AuthBoyri:zed (Study Director /Project Lead) j Date: 3M EnvironmentalLaboratory Form ETS-4-8.0 Deviation No. 0.3 (assigned by Study Directoror Project Lead at the end of study or project) FACT -TCR002(LIMS #E00-1716), Page 63 of 67 BACK TO MAIN Record of Deviation stu.dy /.proj.ect ................ ............. 1. Identification ........................................................................................... Deviation Type: .................................................. TCb R~ o-0p02 -............... 0 Method ....... -. .............-..................... ...... ......... . 0 EquiPprmoceendt ure .......................... .................... (Check one) `BProtocol 0 Other: D o c ~ e n t ~ . ~ ~: ................. b - e ....... r ... - ........................ ............. .......... .... ........ Date(s) of occurrence: 13Jul 00 to 29 Aug 00 11. Description: (2) _____ Required Procedure/process: In dealing with the ..s...t...a...n. dardcurve, the curve must havea .. ....... _.-._-I-. _ _ . - I - - ~ coefficient of determination value greater than or equ.a....l to 0.980 and individual curve .._..-._....I.______.--__._.l__l -_.I__..._ ._.-______-_ __--.---I.- -_____I.______. points must have `Percent Deviations' equal to or less than +/- 30%. .............. ....... . . ~ .... __,.__I__ ... ..... .... .- .......... Actual ................ Pr..oced- ure/...p...-r-.ocess: In some cases, hi..gh andor low curv.....e.........points were -e-l-im-.inatedso.... 3 that the value would meet the criteria. In other cases. high and/or low curve Doints were .................... .................................... ......... ...........-. .................._....... ..- ..I... Y ... ._...... .......... I.................. ..... ......... eliminated, as they were .................. .......... ._ not within the +/-30% ._ .. accurac-y re-quirements. Whenthis was done, - ...... ~ .. .. the ____ .... curve point(s)afTected ..... and the action(s) taken were annotatedon the `Data S u m m a r y .- - .- - ~ ~ ~ Sheet'. - - - - ~ - - ~ ---_____I .--I _~-"--11-~---- The .. ..... follow. ing......... ias list ofall curve points affected and the reasons why. ... ....... .. . ...... .......... Day 1, (H08?700), points Hill003iand Hi110070. .... ___--__-..I__-_ .- I _ - D ~.y ~ 2 ~.... , ~- ~ _ o ~ 8 ~-2 g ~Hoio11~0)0;2~2,_opo~3o2,~o.o~6~0ta_ sn_ d oo70..... Day- 3-,(H. 082500al points L Hi110098.0108.0136 and 0146. I- ..... . - . appropriateto the data.' Ill. Actions Taken: r I 6 (such as amendment issued, SOP revision, etc.) Th.e 3.M Environme.n... tal___M_. ethod ETS-8-155.03as .amended to take into account eltihmein.ation .-.___-_I_.-.___., of curve points. Curve. po_in_ts_may be omitted..for the following reasons: ............_......__.__._._I-_._._._......._____.... I __-I ..(....C.. om..m....... ents are b.ulleted as they would ap..pear on the ................... s u m m a r y sheets.) .... (1) "HigMow curve points were excluded to provide a better fit over the range appropriateto .... the data.'' _ ... .......................... ...................................................................................... . .. .. ... . . . - .... ...^........-.I _ l ^ _ I -... .... ............. . I (..2) "LOWlevel point(s) ............. ............................... wneroet........2x high..e....r.....t..h.....a....n the ext.r....a.. ction.blan......k......;.....t. hese p......o.. ints were .......... .. . exclud.ed from th.e curve to disqualifla data range that may have been significantlyaffected by ._ ^I__ .backogf..round... levels th...e analyte." . . ... . ..... (3) "Highllow ... ...... curve ... point(s) were ... excluded. as they were not within ...................... the +/- 30% accura.cy . . . ._.._l,_____._._ll__..-__--. requirements ...... .- .... of...... the_ m_ et-h_ .o. _ d. .whe...n.....t. he-c-u. .r...v....es were . evaluatedover a range appropriateto the .- ................ _.................... -. . . . . .- I data." 3M Environmental Laboratow Form En-4-8.0 (assigned FACT -TCROOZ (LIMS #E00-1716),Page 64of 67 BACK TO MAIN Recorded By: Record of Deviation ; Date: I IV. Impact on Study/ Project .. - . , .. .. - ._ . . ._ The ....._- .. elimination of curve . ...... points had a ...... ....... positive impact .... tohni s st.u.....d....-y.. " eliminated, the curves would not baes accurate. ....._ ................................. than __. ~ ^... -..-~._._~..__.,_~._.._____.____.__I__ Authorized By: fJj!/T&d ky If curve . ..^ . points ... .. . were not .- .,. . . ... .. . , .......... ~ ........ . . .. ......... i ~ Date: I__ .. . r I 0 3M Environmental Laboratory Form ETS-4-8.0 (assigned by Study Director or Project Leadat the end of study or project) FACT -TCROO2 (LIMS #E00-1716), Page 65 of 67 h b ATTACHMENET: SAMPLE CALCULATIONS BACK TO MAIN FACT-TCROOZ (LIMS #E00-1716), Page.66of 67 BACK TO MAIN 3M Environmental Laboratory Report NO. FACT-TCROOP Mean and standard deviation wercealculatedusing fundions providedIn MicrosoftExcd software. Standard deviation population was usteodmeasurethe scatter about the meanof a data thus, it can be used to estimate the precisioonf a method. Relativestandard deviationpmsentsa measure of the magnitude of the standard deviataionnd is calculated by dvid i n i gthestandard deviation populatianby the mean. Means are calculatedby adding individuael ntitiesand dvid i n i g the resultant sumby the numberof individual entities. Standard deviation populatiownas calculated using the following equation: Average/Standard Deviation PopulationPhRSD examplecalculatiom: I Sample ID Concentratlon WmL 647.9 Calculations Average (647.9+723.4+723.0+717.3)/4= 702.9 723.4 Std Dev Pop (seeabove equation)= 31.8 723.0 %RSD (31.8RO2.9)= 4.5% I Rep 4 717.3 I I I I Data that did not meet acceptance criteria as desicnritbheedOPPTS and OECD guidelineswas excluded using statistical justification providbeydDxon'sQ-test A data pointmay be excluded if "Q-' is greater than"Q-' with 90% confidence. Q- = gaplrange where "gap"is the difference between the questionable data point and the closest ovfatlhue dataset, and "range" is the differencebetween the highest andlowestvalue of the dataset. Q-tabulated(90% confidence) for 4 observations (or data points) is 0.679 and for 3 observations is 0.886. Q-Test Example (Shaded number represents #le suspected outiietj: 1 Description1 Sample Conc., Rep1I SamDle Conc.. Rm2l 607.1 591.7 I I . - -. 1 Range(RePQ-RepZ): Gap (RePQ-Rep3): 2057.14 2022.66 No. observations: 4 Retain? no Q-Test example conclusion: Since Qexcluded. ,0.983, was greater thanQtkwd,0.679, rep4 may be FACT-TCROO2 (LIMS #E00-1716), Page67 of 67