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BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Study Title ' HydrolysisReactions of 2-(N-ethylperfluoroctanesulfonamido)-EthyAl lcohol (N-EtFOSE Alcohol) Data Requirement: Based on OPPTS: 835.210 Author Thomas L. Hatfield, Ph.D. Study CompletionDate March 23,2001 Performing Laboratory 3M Environmental Laboratory Building 2-3E-09,935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Laboratory Report No: W1872 Total Number of Pages 83 Page 1 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 This page has been reservefodr specific country requirements. Page 2 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 Statement of Non-Compliance StudyTitle:HydrolysisReactionsof 2-(N-ethylperfluoroctanesulfonamido)-Ethyl Alcohol (N-EtFOSE Alcohol) Study Identification Number: W1872 This study does not fully comply with the requiremofetnhtes US EPA Good Laboratory Practices (GLP) Standards a4t0 CFR Part792 (TSCA). However, many GLP standards were usedin the development of the analytical method (Appendix A), and the quality assurance procedures followedin this study were based on the practices described in the GLP documentation. This is a revised reporitn that the statistics on the study data and the discussion were changed from the initial study report. Changtoesthese interpretive sections were made to better represent the experimental results of the study. Date Page 3 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 ~~~~ ~ Quality Assurance Statement StudyTitle:HydrolysisReactionsof2-(N-ethylperfluoroctanesulfonamido)-Ethyl Alcohol (N-EtFOSE Alcohol) Study Identification Number: W1872 The following table provides detaiolsf the audits performed by the 3M Environmental Laboratory Quality Assurance Unit (QAU). I I I Inspection Dates Phase I II 9/21,252/,2000 DaaDntRradaeftport Date Reported to Management I Study Director I 10/03/00 1 10/03/00 1 10/2/2000 10/03/00Da1ta0/a0n3d/00Draft Report 3/14, 15/2001 Draft Report 3/15/0 1 3/ 51/01 Page 4 of 83 BACK TO MAIN 3M Environmental Laboratory Report No.W1872 Table of Contents Statement of Non-Complianc..e.......................................................................................... 3 Quality Assurance Statemen..t............................................................................................ 4 List of Tables ........................................................................................................................ 6 List of Figure..s..................................................................................................................... 6 Study Personnel and Contributo.r..s.................................................................................... 6 Location of Archive..s........................................................................................................... 7 Summary.............................................................................................................................. 8 Introduction .......................................................................................................................... 9 Summary of Kinetics Mod.e..l............................................................................................ 10 Materials and Method.s...................................................................................................... 1'I Chemical Characterizations.......................................................................................... 11 Sample Preparation...................................................................................................... 11 Sample Analysis............................................................................................................ 12 Deviations...................................................................................................................... 12 Results and Discussion..................................................................................................... 13 Data Quality Objectives (DQO.'.s..).............................................................................. 13 Anomalous Analytical Resu.l.t.s.................................................................................... 13 Statistical Methods and Calculatio.n..s......................................................................... 14 Data Summary and Discuss.i.o..n................................................................................. 14 Conclusions........................................................................................................................ 18 References......................................................................................................................... 19 Signatures .......................................................................................................................... 20 Appendix A: Analytical Method.......................................................................................... 21 Appendix B:Kinetics Mode.l.............................................................................................. 40 Appendix C: Selected Analytical and Kinetics Result.s.................................................... 50 Appendix D: Selected Chromatogram..s........................................................................... 61 Page 5 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 List of Tables Table 1. Summary of Results Based on N-EtFOSE Alcohol Concentratio..n..s................8 Table 2. Summary of Results Basedon PFOS Limit of Quantification............................. 8 Table 3. Characterizations of Test and Reference Substance...s................................... 11 Table 4. Observed (50' C) Degradation Rates of N-EtFOSE Alcoholin Aqueous Buffered Solutions and at Various pH Lev..e...l.s............................................... 14 Table 5. Degradation Rate and Half Life of N-EtFOSE AlcohinoAl queous Buffered Solutions Using Data Pooled Over pH Le..v..e..l.s............................................. 15 Table 6. Degradation Rate and Half Life of N-EtFOSE Alcohol in Aqueous Buffered Based on PFOS Limit of Quantificat.i.o..n.......................................................... 17 List of Figures Figure 1. Structures of N-EtFOSE Alcohol and the Potassium Salt of PFO...S...............9 Figure 2. Observed N-EtFOSE Alcohol Degradation for Various pH lev.e..ls.................15 Figure 3. Pooled N-EtFOSE Alcohol Data and Slope Regressi.o...n.............................. 16 Study Personnel and Contributors Study Director Thomas L. Hatfield, Ph.D. 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN55106 (651) 778-7863 Sponsor 3M Corporation 3M Environmental Laboratory andProfessional Services Contributing Personnel Kuruppu Dharmasiri, Ph.D Mark T. McCann Anthony E. Scales Joseph J. S. Tokos, Ph.D (Pace Analytical Services, Inc., 1700 Elm St., Minneapolis, MN 55144) Gregory Maisel Jill Maloney (Braun lntertec Corporation, 6875 Washington Ave. South, Minneapolis, MN 55439) Page 6 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 Location of Archives The 3M Environmental Laboratory will retain the original data documents and digital copies ofthe original data relatetdo this work for at lea1st0 years following the effective date of any related final ruling. Information may be obtained through written inquiry addressed as follows: 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Page 7 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Summary We report here the results of our study of the hydro2ly-s(Nis-eothf ylperfluorooctanesu1fonamido)-ethyl alcohol (hereafter, N-EtFOSE alcohol). Our methods are described below andin Appendix Ato this work; our results are basoendthe observed concentrations of N-EtFOSE alcohol and its potential hydrolysis product perfluorooctane sulfonate (PFOS)in buffered aqueous solutions as a function of time. The chosen analytical technique was high performance liquid chromatography with mass spectrometry detection (HPLWMS). Table1sand 2 summarize the results of the study. During this study, we prepared and examined samples at six different pH leve1l.s5 from to 11.O over a period o4f9 days, and our results indicate no dependence of the degradation rate of N-EtFOSE alcohol on the sample pH level. Our results baosnetdhe N-EtFOSE alcohol concentrations, pooled over the observed pH levels, are presinented Table 1. I Table 1. Summary of ResultsBasedonN-EtFOSEAlcohol Concentrations I Constant at 50" c (day-') 0.00262 Constant at 25" c 0.000262 at 250 c (years) 7.3 Half Life Range at 25" C (years) 5.3 to 11.5 We also monitored the concentration of one of the potential hydrolysis products (perfluorooctane sulfonate, PFOS), but never observed this compound at levels above its limit of quantification(LOQ, equal to 12.8 ng/mL). AssumingPFOS to be the only hydrolysis product of N-EtFOSE alcohol, thLOisQ (and other experimental data) provide a second estimate of the N-EtFOSE alcohol half-life, presented in2T. able Table 2. Summary of Results Basedon PFOS Limitof Quantification Maximum Maximum Possible Rate CalcuClatlecdulaHtaeldf LRiafete Constant at 50" C Constant at 25" C I (.day. -'.) I (.day.-'.) I 6.1 X1o4 I 6.1 XIO-~ I at 25" C (years) 2 31 According to the data available from this study, the half-life estimate of Table 2 represents the minimum possible half-life of the compound N-EtFOSE alcohol under the assumption thatit hydrolyzesto form only the compound PFOS. Page 8 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 introduction Three primary chemical routeosf environmental degradation are hydrolysis, photolysis, and biodegradation. Studiesof these routes provide information on the environmental persistence of both the "parent" compounds and their reaction products, and are ideally carried out over the range of chemical conditions pertinent to both environmental and metabolic processes. The hydrolysis of N-EtFOSE alcohol (or, more generally, its degradation in the presence of H20)is addressed in this report. Structures of the "parent" compound N-EtFOSE alcohol and the potassium saolftpossible hydrolysis product perfluorooctane sulfonate (PFOS) are illustrated in Figure1. Figure 1. Structures of N-EtFOSE Alcohol and the Potassium Salot f PFOS N-EtFOSE Alcohol OH Potassium Saltof PFOS FFFFFFFFO II F S-0- K" II F F F F F F F F O Page 9 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Summary of Kinetics Model A full mathematical description of the kinetics model emplionytehdis studyis presented in Appendix B. The study data allotwwo independent estimates of the hydrolytic half-life of N-EtFOSE alcohol. The first estimate (see Tab1le) is based on the observed degradation of the "parent" compound N-EtFOSE alcohol in dilute, appropriately buffered aqueous solutions. Equation 1 describes the estimatedhalf-life('fq;)l in terms of the estimated total parent hydrolysis rateip(see Appendix B, Equation B10): We determined the quantitiypfrom the experimental data as described in AppenBd.ix The data correspondingto "Day 0" (t = 0) were usedto determine the relative concentration ratios (see Equations B8 and B9). The measured concentrations of the potential hydrolysis proPdFuOctS (also obtained during the experiments described here) provide a second estimate (see Table 2) of the parent half-life. During the course of this study, we did not detect PFOSitasbliomviet of quantitation (LOQ), and related studies' show thaPt FOS is itself hydrolytically stable. Assuming also thaPt FOS is the only hydrolytic product of the parent compound N-EtFOSE alcohol, these PFOS analyses provide the following esti(mTaytte)2of the minimum N-EtFOSE alcohol half-life (see Equatio8n3s2 and B33): Eq. 2 where [Po]= the initial N-EtFOSE alcohol molar concentration, A t = the time interval over which the study was conducte(4d9 days), and = the molar limit of quantitation for the compound PFOS. All the samples usedin this study were maintained at a reaction temperatu5re0"oCf . The quoted results, valid for the reaction temperature25o"fC, were calculated from our experimental results accordingto methods described in Appendix B (Eq. B38 and 839). Page 10 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No.W1872 Materials and Methods Details of the characteristics of the test materials, sample preparation techniques, and analytical methods are presenteindAppendix A (ETS-8-179.0, "Preparation o2f -(Nethylperfluoroctanesulfonamido)Ethyl Alcohol (N-EtFOSE Alcohol) Hydrolysis Samples and Analysis by High Performance Liquid Chromatography with Mass Spectrometry Detection.") A summary of these itemiss provided below, as well aasdescription the known deviations from the proceduroefsAppendix A. 3M prepared and analyzed the samples includedin this study between March 13 and Octob5e,r 1999. Chemical Characterizations Table 3 describes the sources and properties of the materials used in this work. These materials were used to prepare both the samples and the quantitative standatords used quantify them. For this reason, and because Equations 36ainnvdolve only ratios of the parent and product concentrations, the resulting rate and half-life estimates are largely independent of the material purity levels. I Table 3. Source Chemical Lot Numbee -Characterizationsof Test and ReferenceSubstances N-EtFOSE Alcohol PFOS (Potassium Salt) THPFOS N-MeFOSE Alcohol 3M Specialty Chemistry 3M Icp'pcp Division ICN Biomedicals 3M Specialty Chemistry S398-332 Batch # 171 Batch # 53406 S398-331 Physical Description Off-white powder , I I Molecular Weight lam mole"\ 570.9 Lighptow"loderer d 538 Brown waxy solid White powder I 428 I 556.9 a The "S"and 'TNA" designations are based on reference numbers tinwo redundant databases maintained by3M. Sampie Preparation We prepared three .O1 -mL aqueous buffer samples (a sample, a duplicate, and a "matrix spike") at each of six pH lev(eals5 , 3, 5,7, 9 and 11) for analysis at eight time intervals (0, 7, 14, 21, 28, 35,42 and 49 days). Buffered solutions containing455 ng/mL of the analyte N-EtFOSE alcohol and 403 ngo/mf TLHPFOS (3,3,4,4, 5,5, 6, 6, 7, 7, 8, 8-8, tridecafluorooctane sulfonic acid), the latter serving as a surrogate for the compound PFOS, formed the basis aolfl these samples. The chosen buffer solutions are described fully in Appendix A. All the samples were prepared simultaneously,alal nbdut the "Day0 samples were placed in an orbital incubatodshaker maintained 5a0t " (* 3") C. After at least three minutes of agitation, the "Da0y samples were spiked (as required) with the PFOS solution, diluted 1O:l with methanol containing the internal standard THPFOS, and refrigerated. After the appropriate incubation times, subsets of the sample vials were removed from the incubator and then spiked, diluted, and stored as described Page 11 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo.W1872 immediately above. Except during the relatively short periods of time required to prepare them, the samples were shielded from light. Eight calibration standards containing N-MeFOSE alcohol (31 6 ng/ml), N-EtFOSE alcohol (155 to 932 ng/ml), THPFOS (403 ng/ml) and PFOS (1to.364 ng/ml) served as the quantitative basis of the study. All these standards were prepared at the appropriate pH levels using the buffer solutions described above. Sample Analysis The equipment we used for the HPLC/ITMS analysis was a Hewlett Packard 1m1o0d0el equipped with a Dionex lonPac@ NG-1 HPLC column (aqueous ammonium acetate/methanol solvent gradient) and an ALS ModGeIl322A degassing module. An ALS Model G1315A column heater maintained the column temperature "aCt ,4a0 quaternary pump supplied a columflnow rate of 0.3mumin, and an ALS Model G1313A auto-sampler provided 5 pL sample injections. The detector was a Hewlett Packard MSD mass spectrometer, operatedin negative-mode electrospray ionization mode; anions of PFOS, THPFOS, and acetate adducts of N-MeFOSE alcohol and N-EtFOSE alcohol were detected at the charge-to mass ratios 499,4267,, a6n1d 630 respectively. We processed the resulting data using the computer progHrPamChernSfationforLC (Rev.A.06.0). Further analytical details, including the gradient elution program, instrument and detector parameters, and performance specifications, are preseinted Appendix A. Deviations No deviations from the procedures definiendthe analytical method (Appendix A) were noted duringthe study. As noted in the following sections, some calibration and sample data failedto meet data quality objectives and were rejected. Page 12of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 Results and Discussion Data QualityObjectiies(DCWs) Below is a brief description of the data quality objectives ainptphliisedstudy, a full description is presented in Appendix A. With the exceptions of the anomalous results noted below, all the DQO's were met. Appendix C presents the results for each sample set, organized bypH level. Calibrations. The minimum acceptable coefficienotf determination(P) for linearfits to calibration data is 0.990. The acceptance criterion for individual calibration points is that their values fall wiItfhr 2in5% of the linear fit value; data outside this range are excluded andthe linear fitis recalculated. No more thantwo points maybe rejected from a calibration data set. Data for the ohriglohw calibration standards mabye rejected, though this results in a smaller effective calibration range. Continuing Calibration Verification (CCV)I.dentical calibration samples are examined at the beginning and end of each sample run. Results of the second calibration run may not deviate by morekth2a5n% of the first run for any analyte. The average results of the calibration runs are utsoecdalculate the analyte concentrations. Matrix Spikes. The acceptable percent spike recovery range 7is5% to 125%. Analyte specificity is demonstrated by acceptable analyte spike recoveries. Sample Duplicates. Duplicate pairs with relative percent deviation (RSD) greater than 25% may be accepted at the analyst's discretion, but must be noted. Solvent Blanks. Concentration results for solvent blanks may exceed neith5e%r of the highest calibration standard nor 25o%f the lowest calibration level. System Suitability. Suitability was demonstrated by either an abbreviated mass-tocharge (mh) check-tuneor performanceof a fullauto-tune routine. Anomalous AnalyticaRl esults Calibrations. Of the 288 calibration results obtained, 22 individual values failed to meet the stated DQO and were rejected.No more thantwo values for any compound were rejected for any particular calibration run. Spike Recoveries. Results for three sample pairs (EFA -136 and -137, at pH= 7.0; EFA-31 and -32 at p=H9.0; and EFA -139 and -140, at pH= 9.0) failedto meet the related DQO and were rejected. Page 13 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1872 Statistical Methods and Calculations Using functions providedin Microsoft@ Excel@ software, we calculated means, standard deviations, and first-order rate constants (see AppenBd,iExquation B8)for various subsets of the acquired data. Our linear regressions included the determinationf constant terms,that is, the regression fits were not forcteodpass throughthe origin. As describedin Appendix B (Equations B38 and B39), rates measured50atCwere extrapolated to 25Cby dividing by a factorf 10; this approximation is valifdorz reactions, such as these, with Arrhenius heats of activation near 18 Kcallmole. Data Summary and Discussion The LOQ is defined asthe concentration of the lowest (accepted) standard in the calibration set for which the known concentration exc4e0e0d%s of the indicated solvent blank level (see Appendix A). During this study, tLhOe Q's for N-EtFOSE alcohol and PFQS were 155 ng/mL and12.8ng/mL, respectively Results for the surrogate compoun(TdHPFOS) and the internal standard (N-MeFOSE alcohol) were very consistent throughout the study. The percent relative standard deviations of the measured values, calculated for each pH level, ranged f1ro.8m% to 7.4%. Table 4 presents the results of the rate determinations atpsHixlevels and 50C. Table 4. Observed (SO" C) Degradation Ratesof N-EtFOSEAlcohol in I AqueoBuusfferSeodlutiVoaanntrsdious pH Levels. I These degradation rates are generally only poorly determined; their percent re2la0tive (95% confidence) uncertainties range from36% to 115%. The data do not indicate any dependence of the degradation rate on the sample pH. Page 14 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 In the absenceof a clear trend relating the degradation troatseample pH,it is appropriate to "pool" the data from all pH levelstoanddetermine the degradation rate using the entire data set. Figur3e illustrates the results of this pooled analysis according to Equation 1, and Table5 summarizes the results ofthe analysis. Table 5. Degradation Rate and Half Life of N-EtFOSE AlcohinolAqueous Buffered Solutions Using Data PooledOver pH Levels. Figure 2. Observed N-EtFOSE Alcohol Degradation for Various pH levels. 0.05 0.00 -0.05 - -0.10 ---L -0.15 E -0.20 1 -0.25 0 10 20 30 40 Time (days) 1 ...... pH 1.5 pH 3.0 .,.,.. pH 5.0 - - - .pH 7.0 - . - - pH 9.0 I - _ -p_H 11 50 Page 15 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 Figure 3. Pooled N-EtFOSEAlcohol Data and SlopeRegression. 0.05 1 0.00 - -0.05 p" -0.10 % B-c -0.15 -0.20 - -0'25 { -0.30 8 - Solid Line: y = -2.62E-03~ 2.30E-02 R2= 4.10E-01 '0 DashedLines2: olimits (slope and intercept) I , i I 0 10 20 30 40 50 time (days) We also monitored the concentration of the hydrolysis product pemuorooctane sulfonate (PFOS), but never observed this compound at levels above its limit of quantification (LOQ, equal to 12.8 ng/mL). The initial N-EtFOSE alcohol concentratio(n455 ng/ml) and the PFOS LOQ provide a second estimatoef the N-EtFOSE alcohol half-life (seine Appendix B, Equations 832 and B33). The maximum degradation rateis given by Equation 3: N k, I (kp)- = -Z A F Q Po A t m=l and the minimum half-life is given by Equa4tion Eq. 3 Eq. 4 Page 16 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 We note thaitn both Equations3 and 4,the initial N-EtFOSE alcohol concentratio(nPo) and the PFOSLOQ (AiFzs) are molar quantities. Table 6 presents the resouflttshe calculation. ITable 6. At (days) 49 Degradation Rate QuanotfLiPfiimFcQaiBottSinaosned and Half Life of N-EtFQASlEcohol in Aqueous I Buffered [P,1 A% Maximum Observed Rate at 50" C Maximum Calculated 25" C Calculated Half Life at 25" C Rate at (nm/ml) (nm/ml) (day-') (daY-') (years) 0.80 0.024 6.1 x IO-" 6.1 x 2 31 Page 17 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 ~~~ ~ Conclusions We have performed a study of the aqueous hydrolytic degrad2a-t(ioNn-ethylperfluorooctanesu1fonamido)-ethyl alcohol (N-EtFOSE alcohol). Six different pH levels were incluinded the study, which were carried out at 50C and extrapolated to 25C. Our results based on direct observation of the N-EtFOSE alcohol concentration indicate no clear dependenthcee of degradation rate of N-EtFOSE alcohol on pH. From the data pooled over thpeHsliexvels, we estimate that the hydrolytic half-life of N-EtFOSE alcohol at 25C lies b5.e3twaenedn 1I.5 years, with the most likely value7.o3fyears. The concentration of the compound PFOS, a likely hydrolytic product of N-EtFOSE alcohol, was monitored throughout the study, but remained undetected above its limit of quantification (LOQ = 12.8 ng/mL). Using the LOQ for PFOS and the initial N-EtFOSE alcohol concentration (455 ng/ml), and assuming PFOiSs the only hydrolytic product of N-EtFOSE alcohothl,e data indicate that the hydrolytic half-life of N-EtFOSE alcohol at 25C is greateorr ethqaunal to 25 years. Page 18 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 References ' "Fate, Transport and Transformation Test Guidelines: 835I.O21: Hydrolysis as a Function of pH," U.S. EPA Officeof Prevention, Pesticides and Toxic Substances, publication number712-C-98-057, January 1998. * "Experimental Physical Chemistry", F. Daniels, et al., McGraw HBiollok Co. (New York), p. 131, 1962. Page 19 of 83 Signatures BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 Page 20 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W 1872 Appendix A: Analytical Method ETS - 8-179.0, "Preparation of 2-(N-ethylperfluorooctanesdfonamido)Ethyl Alcohol (N- EtFOSE Alcohol) Hydrolysis Samples and AnalysisHibgyh Performance Liquid Chromatography with Mass Spectrometry Detection." This Appendix presents the analytical method employiendthis study. Page 21 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 3M ENVIRONMENTAL LABORATORY METHOD PREPARATIONOF 2-(N-ETHYL PERFLUOROOCTANESULFONAMIDO)-ETIIYLALCOHOL (N-ETFOSE ALCOHOL) HYDROLYSIS SAMPLESAND ANALYSIS BY HIGH PERFORMANCELIQUIDCHROMATOGRAPHY WITH MASS SPECTROMETRY DETECTION Method Number: ETS-8-179.1 Approved by: Laboratory Manager Adoption Date: 9/14/00 Effective RevisionDate: 3/19/01 Date ETS-8-179.1 Method Page 1 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLCMS Page 22 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 1.0 SCOPE AND APPLICATION 1.1 This procedure defines the steps for analysis of 2-(N-ethyl peffluorooctanesulfonamido)- ethylalcohol(N-EtFOSE alcohol) hydrolysis samplebsy high performance liquid chromatography (HPLC)with mass spectrometry (MS) detection and quantitatioInt .is based on EPA OPPTS:835.2110 (Reference 18.1). N-EtFOSE alcohol and the potential hydrolysis product perfluorooctane sulfona(tPeFOS anion) are detected and quantifibeyd this method. N-EtFOSE alcohol is quantified using N-MeFOSE alcohol (2-(Nmethylperfluorooctanesulfonamido)-ethyl alcohol) as an internal standard. The surrogate THPFOS (the anion o3f,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctane sulfonic acid) is used to quantifyPFOS. Representative structures are shownin Attachment A. 1.2 Compatible analytes. 2-(N-ethyl perfluorooctanesulfonamido)-ethyl alcohol (N-EtFOSE alcohol),perfluorooctanesulfonate(PFOS anion),perfluorooctanesulfonamide(FOSA), 2-(N-methylperfluorooctanesulfonamide)-ethyl alcohol (N-MeFOSE alcohol) and the anion of3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctane sulfonic acid(THPFOS). 1.3 Compatible matrices foarnalysis. Aqueous solutions at various buffered pH levels. 1.4 This is a performance-based method. Target analytoer surrogate matrix spike recoveries (100 25%) are used for each sample matorixevaluate method performance. Refer to Section 10for the frequency of quality control parametetorsbe performedin this method. Refer to Section 14 for the quality assurance evaluation criteriathfoisr method. 2.0 SUMMARY OF METHOD 2.1 Aliquots of N-EtFOSE alcohol stock solution containing THPFOS surrogate are added to vials that contain bufferspHat 1.5,3.0,5.0,7,0,9.0and 11.O. The vials are then placed in an orbital incubatorhhaker set 5a0t.0 f 3 OC. Sets of vials are removed at designated intervals and the date and time recorTdheed.aqueous samplefrom the hydrolysisof N- EtFOSE alcoholis diluted tenfoldwith methanol (h4eOH) and spiked with N-MeFOSE alcohol internal standard. The parent compounNd-,EtFOSE alcohol, and thePFOS hydrolysis product are separated on a Dionex IonNPGa1c@reversed-phase HPLC column usingan ammonium acetateMeOH solvent gradienwt,ith detectiodquantitation by electrospray ionizationmass spectrometry inthe negative mode. 3.0 DEFINITIONS 3.1 Calibration Standard. A dilution of various amounts of a stock, intermediate or purchased standard to achieve standard solutinonasconcentration rangeof interest. Hydrolytic half-lives resulting from these analyses are calculatedobnaasneadlytical ratios and not absolute numbers. Therefore, results do not depend pounrityheofthe standards used. 3.2 Calibration Curve. The graphical relationship between knownvalues, such as concentration of a series of calibration standards and their instrumental response. 3.3 Internal StandardCalibration. Process of establishing a relationship between theratio of the target analyte(s) response to internal standard or surrogate responskenoanwdn a concentration ofthe target analyte(s). The ratio of analyte to internal standard respoinsse used to generatethe calibration curve and determiunneknown concentrations. ETS-8- 179.1 Method Page 2 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLCMS Page 23 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 3.4 3.5 3.6 3.7 3.8 3.9 3.10 3.11 3.12 3.13 3.14 Correlation Coefficient (r)A. m e m e of the degreeof correlation betweentwo variables. This term is generally usteodevaluate the linearity of a Least Squares Linear regression. An r value of0.98 is at the lower boundosf whatis considered linear. Values of r may range from-1 to +l. A value of +1 denotes perfect direct functional relationship between two variables. A value of -1 also denotes a perfect inverse relationship. When r = 0, there is no effectof one variable upotnhe other variable. Coeffrcient of Determination (8).The squareof the correlation coefficient. Itis the proportion of the variation in the dependent variabilseacthcoatunted for btyhe independent variable. Internal standard. A known amount of a compound or element similianr analytical behavior to the compound(s) or element(os)f interest, added to all samples and standards, and carried through the entire measurement process (post-hydrolysis, faifntearl dilution). It provides a reference for evaluating and controlling the precision andofbtihase applied analytical method. Surrogate. An organic compound whichis similar to the target analyte(isn)chemical composition and behavior in the analytical process, but iws nhoicthnormally foundin the sample(s). In hydrolysis studies, surrogate is addedto CCVs, samples, sample duplicates, and matrix spike samples along with the test analyte (pre-hydrolysis). Continuing CalibrationVerification (CCV). Standards analyzed duringan analytical run to veri@ the continued accuracoyf the calibration curve.This solutionmay or may not be prepared from a different source or lot number than the calibration curve standards. Solvent Blank. A sample of analyte-fiee medium(for example, methanol) thaits not taken through the sample preparation proceTshsi. s blank is used to evaluate instrument contamination. Limit of Quantitation (LOQ). The lowest concentration that can be reliably measured within specified limitsof accuracy during routine laboratory operating conditions. The LOQ is generally 5 to 10times theminimumconcentration witha 99% confidence limit that the concentration is greater than zero. However, it may be nominally chosen within these guidelinesto simplifjr data reporting. For many analytes,LtOheQ is selected as the lowest non-zero standard in the calibrationcurve that is greaterthan 4 times thelevel of the solvent blanks. Sample LOQs are highly matrix-dependent. Sample Duplicates. Two samples taken from and representativoef the same sample source and separately carried through all sotfetphse extraction and analytical procedures in an identical manner. Duplicate samples are used to assess variaonfctehe total method, including sampling, extraction, and analysis. Relative Percent Difference(RPD). A measure ofprecision definedas the absolute value of the differenceof two values divided by the averaogfethe two values and multiplied by 100. Matrix Spike(MS). Prepared by addinga known mass of target analyteto a specified amount of a sample matrix prior to analysiTs.his assumes that an independent estimate of target analyte concentratioins available. Matrix spikes areusedto determine the effect of the matrix on method recovery efficiency. Accuracy. The closeness of agreement betweaennexperimentally determined valueand an accepted reference value. When applied to a set of observed values, accuirsaacy ETS-8-179.1 Method Page 3 of 18 Prep. of N-EtFOSE Alcohol HydrolysisSamples and Analysis by HPLCMS Page 24 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 combination of a random (precision) and a common systematic (bias) component. For purposes of the study, the acceptance criterio7n5%isto 125% of the nominal value. 3.15 Dilution. A step in the hydrolysis study procedurien which a solventis added to the test malytehuffer solution to prepariet for instrumental analysis.This step occurs afterthe vials areremoved fiom incubation and before the samples are analyzeIdf.the solvent used is miscible with the test analytehuffer solution, the diluting soilsvemnetrely added and mixed. If the diluting solvenits non-miscible, a liquid-liquid extractionis performed. 4.0 WARNINGS AND CAUTIONS 4.1 Health andsafetywarnings 4.1.1 Wear the proper lab attire for all parotsf this procedure. Wear gloves and proper eyewear at all times. 4.1.2 Handle all solventsin a hood for all parts of the described sample preparation procedure. Whenever possible and practical, dilute samples with solveinnta hood. 4.13 For potential hazards of each chemical used, rteofemr aterial safetydata sheets, packing materials,and the 3M Environmental Laboratory Chemical Hazard Review. 4.2 Cautions 4.2.1 All glassware in whichstandards are prepared should be triple-rinsed with:1 acetone/MeOHto reduce the possibilityof contamination. 4.2.2 Ensure that theHPLC mobile phases are prepared priotro beginning arun sequence, and that there is sufficient quanttoitcyomplete therun. Do not allow the pump torundry. 4.2.3 Ensure that before starting threunsequence thereis ample hard disk spaceon the runs. computer to save arlulndata. 4.2.4 Ensure that there is enough nitrogeinnthe supplytankto complete sequence 5.0 INTERFERENCE 5.1 Contaminants in solvents,reagents, glassware, and other sample processing or analysis hardware may cause interference. Use the routine analysiosf laboratory method blanks to demonstrate that thereis no such interference. 5.2 Contamination fiom columns, HPLC tubing, and detector components may cause interference atlow detection levels.The routine analysisof solvent blanks must be used to demonstrate that thereis no such interference. 6.0 EQUIPMENT 6.1 Analyticalbalancesensitiveto 0.1 mg 6.2 Incubatodshaker capableof maintaining temperature at 50.0 f 3C 6.3 Hewlett-Packard (HP) 1100HPLCSystem,orequivalent 6.3.1 Pump, binary, Model G1312, or equivalent 6.3.2 Solvent degasser, Model G1322A or equivalent 6.3.3 Autosampler, ALS ModelG1313A, variable injection volume capable 6.3.4 Column heater, Model G1316A ETS-8-179.1 Method Page 4 of 18 Prep. ofN-EtFOSE Alcohol Hydrolysis Samples and Analysisby HPLCMS Page 25 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1872 6.4 Dionex IonPac"NGl Guard column,4 x 35 mm, or equivalent 6.5 Mass spectrometer. Hewlett-PackardMSD Model G1946A, or equivalent, operatingin Electrospray negativeSIM mode 6.6 Clock, digital.Only one clock should buesed,to insure unambiguous documentationof the correct performanceof procedures. 6.7 pH meter. Corning Model308 pWTemperature Meterwith 3-in-1 gel-filled combination electrode (pWreference/temperatut.e),or equivalent 6.8 Refligerator capableof maintaining 4 f 3 "C 6.9 Data system. A personal computer capableof controlling the HPLC systemas well as recording and processing signalsfrom the detector, Hewlett-Packard ChemStation@ Version A.06.01 or later 7.0 SUPPLIESAND MATERIALS 7.1 Vials, 40 mL, VOA (I-Chem or equivalent) 7.2 Crimp cap autoyials, 1.m8 L 7.3 Labels 7.4 Graduated pipets, glass, disposable, 1mL to 10 mL 7.5 Pasteur pipets, glass, disposable 7.6 Hamilton Gastight* syringes (precisiofn 1%of total volume),10 VI-1000 pL 7.7 Volumetric flasks, various sizes 7.8 Beakers, glass, varioussizes 7.9 Automatic pipettor, capabloef dispensing 10-5000 pL 8.0 REAGENTS AND STANDARDS 8.1 Methanol (MeOH). HPLC/SPEC/GC grade fromEM Science, or equivalent 8.2 Acetone. HPLC/SPEC/GC gradefrom EMScience, or equivalent 8.3 18.0 Mi2 water. Water with lower resistance must not be used. 8.4 Ammonium acetate,2 mMin water, This solution is chromatographic solvent A (see Section 12.3.1). (Example:An acceptable eluent solutionis made by adding 0.15 g ammonium acetate crystals to a I-Lvolumetric flask containing about 500 mL water, adding 10 mL of methanol, diluting to the mawrkith 18.0 MQ water andmixing.) 8.5 Stock, internaI standard, surrogate and calibration solutions All weights should be recorded to the nearest 0g.0i0n0a1 standards preparation log: 8.5.1 N-EtFOSE alcohol prepared in acetone. (Example: A stock solution is preparedat a concentration of approximately 30,000 pg/mL by weighing 0.3 g of N-EtFOSE alcohol in a 10-mL volumetric flask and bringing to the mwairtkhacetone. This solution is diluted iMn eOH to make additional, appropriate standards.) 8.5.2 N-MeFOSE-alcohol internal standard preparedinacetone. (Example: A stock solution is prepared at a concentratioonf approximately 30,000 pg/mL by weighing 0.3 g of N-EtFOSE-OH in a10-mLvolumetricflask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) ETS-8-179.1 Method Page 5 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLCMS Page 26 of 83 BACK TO MAIN 3M Environmental Laboratory Report No.W1872 8.5.3 Perfluorooctanesulfonate(PFOS) prepared in methanol. (Example: A stock solution is prepared at a concentration of approximate3l0y00 pg/mL by weighing 0.06 g of PFOS in a20-mL ,volumetric flask and bringitnogthe mark with methanol. Thissolution is dilutedin MeOH to make additional, appropriate standards.) 8.5.4 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctanesulfonic acid(THPFOS) surrogate prepared inMeOH. (Example: A stock solutionis prepared at a concentration of approximately 20,0p0g0/rnL by weighing 0.2 g of THPFOSin a 10-mL volumetric flask and bringing tothe mark with methanol. This solution is diluted in methanol to make additional, appropriate standards.) 8.6 Buffers for calibrationof pH meter Purchased pH calibration standards poHf 4.0,7.0,and 10.0 (suppliers vary). 8.7 Buffer solutions for hydrolysis study. Prepare buffer solutions of pH 1.5,3.0,7.0,9.0 and 11.Ousing guidelinesfiom CRC Handbookof Chemistry and Physics (Reference 18.2). Prepare buffer solution of p5H.0 using guidelines from Fate, Transport and Transformation Test Guidelines (Referenc1e8.2). Prepare the buffer solutionsin 1-liter quantities. Calibrate a portable pWtemperature metuesring purchased pH calibration standardsof pH 4.0,7.0, and 10.0, and measure the pHof all buffer solutions. Prepare buffer solutionsof pH 1.5,3 .O,5,0,7.0,9,a0nd 11.0 at ambient mom temperatureT. he concentrations are given below. Record final pH measuremoefndtsl buffers. Store buffers in sealed glass containers. 8.7.1 pH 1.5 8.7.1.1 207 mL of 0.1 N HCl (reagent grade) 8.7.1.2 125 mL of 0.2 M KC1 (reagent grade) 8.7.1.3 Add 18.0 Mi2 water to about 900mL total volume 8.7.1.4 Adjust pH to1.5 with additional1N HC1 8.7.1.5 Bring to a final volumoef 1 L with 18.0 MQ water 8.7.2 pH 3.0 8.7.2.1 223 mL of 0.1 M HCl (reagent grade) 8.7.2.2 500 mL of 0.1 M potassium hydrogen phthalate (reagentgrade) 8.7.2.3 Add 18.0MQ water to'about900 mL total volume 8.7.2.4 Adjust pH to3.0 with 1N HC1 or 1N NaOH 8.7.2.5 Bring to a final volume of 1wLith 18.0 MS2 water 8.73 p H 5.0 8.7.3.1 Add 3.8777g ammonium acetate (reagent gradet)o 250 mL 18.0 Mi2 water 8.7.3.2 Add 250 mL 0.052 M acetic acid (reagent grade) 8.7.3.3 Add 18.0MQ water to abou9t00 mL total volume 8.7.3.4 Adjust topH of 5.0 withglacial acetic acid (approximately0.5d) 8.7.3.5 Bring to afinal volume of 1 L with 18.0 MR water 8.7.4 p H 7.0 8.7.4.1 500 mL 0.1 M KH,PO, buffer (reagent grade) 8.7.4.2 291 mL 0.1 N NaOH (reagent grade) 8.7.4.3 Adjust topH 7.0 with either 1N HCl or 1 NNaOH ETS-8-179.1 Method Page 6 of 18 Prep. ofN-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 27 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 8.7.4.4 Bring to a final volumeof 1L with 18.0 MR water. 8.7.5 pH 9.0 8.7.5.1 500 mL 0.025 M sodium borate decahydrate (reagent grade) 8.7.5.2 46 mL of 0.1 N HC1 (reagent grade) 8.7.5.3 Add 18.0 Ma water to approximately900 mL 8.7.5.4 Adjust topH 9.0 with either 1N HCl or 1N NaOH 8.7.5.5 Bring to a final volume.of 1 L with 18.0MR water. 8.7.6 pH 11.0 8.7.6.1 500 mL 0.05 M NaHCOz (reagent grade) 8.7.6.2 227 mL 0.1 N NaOH (reagent grade) 8.7.6.3 Add 18.0 M a water to approximately 900 mL 8.7.6.4 Adjust pH to 11.0 with 1N NaOH 8.7.6.5 Bring to a final volumeof 1L with 18.0 MR water 8.8 Test analyte and spike solutions: 8.8.1 N-EtFOSE alcohol test analyte soIutionwith THPFOS surrogate. [Example: An analyte solutionof N-EtFOSE alcohol at 500 pg/mL and THPFOS at 400 pg/mL is used (a dilutionin MeOH of the solutions preparedin Sections 8.5.1 and 8.5.4). A 10-pL aliquotof this solution added to m1 L buffer (the step performed in Section 12.1.6) resulitns a f d concentration of 500 ng/mL N-EtFOSE alcohol and 400 ng/mL THPFOS after MeOH dilution (the step perfoirnmSeedction 12.1.13)]. 8.8.2 N-MeFOSE alcohol internal standard solution. pxample: An analyte solution of N-MeFOSE alcoholat 30 pg/mL is used (a thousand-fold dilutionin MeOH of the solution preparedin Section 8.5.2). A 100-pL aliquot of this solution addedto 1mL buffer (the step performeidn Section 12.1.14) resultsin a finalN-MeFOSE . alcohol concentration of approximatel3y00 ng/mL. 8.8.3 Spiking solution. pxample: A spiking solutionis prepared by adding 70 pL of the N-EtFOSE alcohol stock solution (Section 8.5.1t)o a 10-mL volumelricflask and diluting to themark with methanol. A 10-pL aliquot of this solutionadded to the 1.O mL sample (the step performed in Sect1io2.n1.I5) results in a final spike concentration of 217 ng/mL of N-EtFOSE alcohol after MeOH dilution. 9.0 SAMPLHEANDLING 9.1 Record times of initial preparation and dilution on the fluorochemical degradation (hydrolysis) analysis sample preparation sheet (AttachmBe).nt 9.2 For Time 0 samples, aliquot only the 1mLof buffer intothe vials. DO NOT spike with test analyte. Store the vialsat room temperature until ready to analyze. Then proceed from Section 12.1.12. 9.3 Once the 9.0 mL of diluting solventhas been added to the hydrolysis mixtures,the samples are ready to be analyzed. Alternatively, aliquoof tshe methanol-diluted samples should be refrigerated at 4 k 3 OC or frozen until analysis can be performed. ETS-8- 179.1 Method Page 7 of 18 Prep. of N-EtFOSE Alcohol HydrolysisSamples and Analysis by HPLC/MS Page 28 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 10.0 QUALITCYONTROL 10.1 Calibration Standards,Calibration standards (Section 11)used to generate a calibration curve should be prepared in the same type of solvent oarsminatthriex study samples. The numberof calibration standards and the concentration levels shboeusludfficientto encompass the expected concentratioonfs the study samples.In general, a minimumof five calibration standards is required foroffliitnear regression. Broad calibration ranges (greater that three orders of magnitude between lohwighasntdandards), may require use of a quadraticfit of the data and requires more potiontasdequately representhe calibration range. '10.2 Internal standard(IS). N-MeFOSE alcohol internal standardis added in a constant concentration to all standards, samples, and matrix spikes. 10.3 Surrogate. THPFOS surrogate is addedin a constant concentration tCoCVs, samples, sample duplicates, and matrix spike Samples along withN-tEhteFOSE alcohol test analyte (pre-hydrolysis). 10.4 Continuing CalibrationVerification (CCV). A standard analyzed periodically during an analyticalrunto verify the continued accuracy tohfe calibration curve anids run in tandem with the solvent blank. This solution may be preparfreodm a different source or lot number than the calibration curves standards. 10.5 Solvent blank. Solvent blanks (or clean methodblanks) should berunbefore and after every calibration curveC, CV,method blank(ifcontamination is noted), anadfter batches of no more than20 injections. Acceptable values for the blanks are values betlhoew limit of quantitation(LOQ)of the instrument (Section3.10). If analyte carryoveirs a problem (see Section14.5), use back-to-back solvent blanks. 10.6 Sample Duplicates. Prepareand analyze all samples in duplicate to provide a measure of the precision of analysis. 10.7 Matrix spikes. Prepare a post-hydrolysis matrix spike sample (the step perforimn ed Section 12.I.15) for each interval and pH leuvseeld in the study. Concentrations of the spike should be approximately equal to a mid-range calibration standard. The matrix spike sample should be analyzed immediately following the sample'duplicates toit which corresponds. The analyst shall accept percent spike recoveriesof 100 f 25%. Spike recoveries outsideof this range should be noted. Appropriate steps must be taken to correct the problem before analysisisallowed to proceed. Before the analysisisallowed to proceed, consult with tTheeam Leader or designee for direction and final acceptance or rejectionof the analyticalrun. 11.0. CALIBRATION AND STANDARDIZATION 11.1 Standard preparation. Prepare six calibration standards containing N-EtFOSE alcohol, N-MeFOSE alcohol, PFOS and THPFOS in 9:l Me0H:buEer for each pH level. Standards from approximately 150n g / d to 1000ng/mL ofN-EtFOSE alcohol and1 n g / d to 100 ng/mL of PFOS are suggested. 11.2 Calibration Standards. Analyze the calibration standardsat the beginning and endof the run. Individual calibration data poinftrsom both setsof calibration standards are used to generate a calibration curve. The calibration cuirsvtehen used by the data reduction software programfor linear regression calculationsto relate the analyte peak area ratio ETS-8- 179. I Meihod Page 8 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 29 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 versus amount ratio, using internal standard calibration. Use N-MeFOSE alcaoshthoel internal standardfor N-EtFOSE alcohol andTHPFOS surrogateas the internal standard for PFOS quantitation. Quadratic regression may be used if data review sthioswtos be a consistent and more accurate representaotifotnhe instrument response. Consult with the Team Leader for direction prior to performing the quadratic calibration methodology. 12.0 PROCEDURES 12.1 Sampleandspikepreparation 12.1.1 Before spiking with any of the stock standards, transfer approxim1mateLlyof the solution toan autovial and cap the vial. Utsheis smaller volume for spiking to minimize the effectosf evaporation from stock solutionasnd to prevent contamination of the larger volume of stock solution. 12.1.2 Determine the numbeor f time intervals thawt ill be analyzed. Each intervawlill have three vials for each pH, multiplied by the onfumpHbseranalyzed. One vial ateach level will be labeleads sample, duplicate, and spike. 12.1.3 Obtain the appropriate number of 40-mVOLA vials with caps and cardboard boxes. Prepare appropriate sample preparation worksheets, create labels, and affix them to the vials. The labels should include the sample numbIe.rD.a,nd temperature, pH, time interval, test analyanted, date of preparation. Record the pH of each buffer solution. 12.1.4 Remove the capof the VOA vial and add1mL of the appropriate buffer soIution to allof the pre-labeled vials. Always replace the cap immediately after any addition to minimize evaporation. 12.1.5 Put `(Time0" samples aside at this point. For all other samples, continue on to Section 12.1,6. 12.1.6 To all of the vials, add10 pL of the mixedN-EtFOSE alcohol analyte and THPFOS surrogate solution (Sectio8n.8.1) with a 25-pL Hamilton Gastight@ syringe. 12.1.7 Make sure that the cahpas been firmly tightened and placethe samples backin the cardboard case. 12.1.8 Place the case into a pre-warmed incubator/shaker tfhoer appropriate time. Record the time, temperature, and raotef shaking. The temperatureis determined by the conditions of the experiment. Continue to manually monitor the incubator temperature daily during the entire incubation. Recthored temperature on the sample preparation sheet (AttachmBe).nt 12.1.9 Store "Time On samples at room temperature unttihl e time of analysis. 12.1.10 Remove each case from the incubaatot trhe designated preset time. 12.1.11 Remove the vials from the case and place in racks. tAhellovwials to cool for approximately 15 minutes to room temperature. 12.1.12 While vials are cooling, spike the store`dT i e 0" samples with test analyte solution (Section12.1.5) on Day 7. Then continueon to (Section 12.1.13)with all samples. 12.1.13 Add 9 mL of methanol to each vial. ETS-8-179.1 Page Method 9 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLClMS Page 30 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 12.1.14 12.1.15 12.1.16 Using a 100-pL gas tight syringe, add 10p0L of N-MeFOSE alcohol internal standard solution (Section 8.8.2) to each sample and spike vial. Using a25-pL gas tight syringe, add 10pL of spiking solution (Section8.8.3) to the sample spike vials. Shake the vials for three minbuytehsandor Vortex mixer tomix the contents and extract any analytes that may have adtsoorbed the vial. Aliquot approximately1mL of each sampleto the appropriately labeled autovial, cap,and reiiigerateat 4 k 3 "C until analysis. 12.2 Instrument set up 12.2.1 Check that the appropriateHPLC column isin the instrument for analysis. run. 123.2 Check that the correct eluent solutions ainrebottles to be usedand that enough is availabIe to complete the sequence 12.2.3 Place the samples in the autosampler tray and construct a seqtaubelnecwe ith appropriate calibration standards, calibration check standards and sobllvaennkts. 12.2.4 Verify thatall samples and standardsare positioned.correctly. Enter sequence information (sample or standarIDd , method name). Use one injectionper sample. 12.2.5 Save sequenceas analysis date and instrument lett(eer.g. on March14,1999, save sequence tableas 031499,s). Saveall data to a subdirectory labeledwith analysis date. (e.g. 031499). 12.2.6 Set post-sequence commandmacro to shut down system after trhuenis completed (Example": STANDBY"on HP11OOMSD systems). 12.3 HPLC set up: 12.3.1 Analysisof N-EtFOSE alcohol hydrolysis sampleisn buffers at pH levels 1.5,3.0, 5.0,7.0,9.0and 11.0. Column: Dionex IonPac" NG1Guard column, 4x 35 mm, or equivalent Solvent A: Ammonium Acetate2mM in water (with 1% MeOH). Solvent B: Methanol Recommended Solvent Gradient: TIME(MIN) 0.0 %A %B I FLOWRATE 60 40 0.3 mumin 1.o 60 40 0.3 mUrnin I I I I 1 4.0 ..~.5 ~ 95 0.3 mtlmin ~~ 11.0 5 95 0.3 rnllrnin Post timc: 6 nhtttss, column temperature:35C. ETS-8- 1 79. t Method Page 10 of 18 Prep. of N-EtFOSE AlcohoI Hydrolysis Samples and Analysis bHyF'LChiS Page 31of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 12.4 Recommended mass spectrometer set up': MSD: ionization mode API-ES Polarity Negative Acquisition mode SIM Gain 1.o Fragmentor 70 Dwell time 183 msec Capillary voltage 3500 I gas I Drying 1 Nitrogen I ~ ~~~~ ~ ~ Nebulizer pressure 30 psig Drying gasflow 8 Umin Drying gastemp 300"C *Example conditions are applicableto HPl IOO/MSD equipment only. 12.5 Auto-samplesretup*.: AUTO-SAMPLER: ALS Model G1313A AUTO-SAMPLERPROGRAM: None I INJECTION VOLUME: I 5.0 pL I *Example conditionsare applicableto Hewlett Packard 1100 only 12.6 Ions used for identification and quantification: APPROX. RETENTION TIME(MIN) COMPONENT NAME 8.3 MeFOSE-OH I I I 8.6 ' EtFOSE-OH DESCRIPTION Internal Standard AnTaleystte QUANTIFICATION ION 6 I 6 (M+OAc-) I 630 (M+OAc') I MONITOR ION 617 631 I I I I I I I 6.2 PFOS PotDenegtiraaldation Product 499 (M-IT) 500 I I I I I I 5.9 THPFOS Surrogate 427 (M-H*) Nomonitor I 6.3 I FOSA I QualitPautirvpOeonsleys I 498 (M-I?) I No monitor I 12.7 Sample analysis 12.7.1 Enter the standard, sample,and QC informationinto the sequence table. Analyze calibration standardsfust,then up to 20 injections, followed btyhe calibration standards. If more than 20 injections areto be run,analyze a continuing calibration standard(CCV)after every 20 injectionsand runthe calibration ETS-8-179.1 PageMethod 11 of 18 Prep. of N-EtFOSE AlcohoHlydrolysis Samples and Analysis by HPLCMS Page 32 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 standards again at the end of the sequence. Run solvent (or method) blanks after the highest calibration standard, before and after CthCeV,and after the seotf samples to check for any analyte carryover. 12.7.2 Place standards, samples, anQd C (matrix spikes, sample duplicates, and blanks) into the autosampler tray according to the order they areinltihsetesdequence. 12.7.3 Identify the electronic acquisition filewsith an appropriate prefix (e.gE.tFOS). Do not exceed five characters if the sequence contains tmhaonre99 lines. 12.7.4 Save sequence as analysis date (e.g. on March 14,1999, save sequenceatsable 031499.s). Save all data to a subdirectory labeled with analysis date (e.g. 031499). 12.7.5 Start the sequence. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Peak Evaluation. Peaks must be symmetricin shape and identified by extracting compound-specific ions. Peaks considered for Calibration must have peak heights greater than 4 (four) times the baseline noise for that region of the chromatogram.arPeeaak integration is from baseline to baseline using automatic or manual integratiNon-. EtFOSE alcohol concentrations are calculateudsing N-MeFOSE-OH as the intend standards. THPFOS surrogateis used as the "internalstandard" to calculatePFOS concentrations. External standard calibration may be acceptable. Consult wtihteh team leader for directionpriorto performing the external calibration methodology. Document change in raw data and final report. 13.2 Calculation of Rate Constant(k). Calculate the test analyte concentratioinnseach of the pH matricesusing the curves obtained fiom the calibrations. Assuming first-order ( ~ ~ ~ ~ ~ ~ ~ kinetics, a rate constan(tk)can be determined by plotting: Ln versus minus elapsed time(-0. The subscriptst and 0 refer to andyte concentrations determinedat some elapsed timet and att =0, respectively. The slope of the resulting line is k. 13.3 Target analyte concentrations. Calculate theEtFOSE-OH and PFOS concentrationsin each ofthe pH matrices using the curves obtained fiom the calibrations 13.4 Matrix spikes. Calculate the percent recovery for each tohfe matrix spikes. Calculate the matrix spike percent recoveries usintghe following equation: YORecovery = (observed spikedsamde result - observed samule result)x 100 Actual amount spiked Using the observed matrix spike recoveries, calculate the average spike recovery. ETS-S- 179.1 Method Page 12 of 18 Prep. of A'-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 33 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 13.5 Sample Duplicates. Calculate the relative percent deviation(%FtPD) for the duplicate sam*ples: RPD= IA-BI x 100% (A+B)/2 Where A =the concentration measuredin the sample B =the concentration measuredin the duplicate 14.0 METHOPDERFORMANCE 14.1 Coefficient of Determination (9).The coeffkientof determination(8)for the calibration curves should b0e.990 or greater. The curves shouldbe examined closely for linearity and intercept, particularly for accuorafcqymtitation at the low anhdigh ends of the curve. The accuracy oafll standards usedfor calibration mustbe within 75125%. On occasion it may be necessary to use exponeonrtiqauladratic fits of the data, usually when broad range curves (greatthearn 3 orders of magnitude between the lowand high concentration standards) are used. Documeinnthe raw dattahe technical justification for using quadratic equations. Consult with Ttheeam Leader or designee for direction and for fmal acceptancoer rejection forthe data. 14.2 Calibration Standards. The acceptance criterion for the calibratiosntandards is that the accuracy of each standard 7is5% to 125% of the nominal value. Calibration standards outsidethis range are to be considered outliers and exclufdioedm the linear regressionI.t may be necessary to use exponential or quadfirtastoicfthe data,usually when broad range curves (greatetrhan 3 orders of magnitude between thleow and high concentration standards) are used. Document in the raw data the technical justification for using quadratic equations. 14.3 Internal Standard (IS)and Surrogate. Review of the internal standard and surrogate performance is performed by averagingthe area response throughotuhte analytical run and calculating%RSD. Inconsistencies in the internasltandardpeak area may indicate instrumental changes over time. Inconsistenciiensthe surrogate peak area may indicate instrumental changes, changes in the test-system, or hydroflythsies surrogate over time. Consult with the Team Leader or designee for direction and final acocrerpetjaenccteion of the analytical run. 14.4 Continuing CalibrationVerification. If the percent difference for the amooufnt quantitated analyteis greater than25% fiom the true value relative tohe initial standard curve, the Team Leader should be consulted. Only those samples analyzed before the last acceptable calibration check standard will be used. ConsultthweitTheam Leader or designee for direction and for final acceptoarnrceejection forthe data. 14.5 Solvent Blanks. Solvent blanks should show no mtohraena 5% carryover from a high standard or calibration check standardI.f so, two solvent blanks may be necessary to rule out instrumental contaminationI.f peaks with greater tha2n5% of the peak areaof a low standard value are observed in sequential solvent blanksr,utnheshould bestopped. This ETS-S- 179.1 PageMethod 13 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLCMS Page 34 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 14.6 14.7 14.8 14.9 14.10 is indicative of instrument contamination. The instrument shall be sebryvitcheodroughly cleaning the electrospray souracned, replacingkleaning columns, tubing, etc. Limit of Quantitation (LOQ).The LOQ is equalto the lowest standardin the calibration curve thaist greaterthan 4times the level of the solvent blanks. Sample Duplicates. The analyst shall accep%t RPD (See Section 13.5) value<s 25%. %RPD values 25% should be noted. Appropriatesteps must be taketno correctthe problem before analysis allowed to proceed (e.g. sample re-runs, additional blanks, etc.). Consult with the Team Leaderor designee for direction, and fionral acceptanceor rejectionof the analyticalrun. Matrix Spikes. The analyst shall accept percent spike recovery values10of0f 25%. Spike recoveries outside tohfis range should be noted. Consult with the Team Loerader designeefor direction, and for final acceptance or rejecotfiothne data. Datathat are used in finalreport thatis deemed outof control willbe requiredto have a technical justification forwhy the data are being used, documentiendthe finalreport and raw data. Specificity. Analyte specificityis demonstratedby acceptable post-hydrolysis analyte spike recoveries. System Suitabiiity. Without performinga method validation, system suitabilitcyan be demonstratedby acceptable instrumental checks (e.g. abbreviamted/z check-tune, or full auto-tune routines. Consult the appropriate instrumenmtaalnuals (Reference 18.3). 15.0 POLLUTION PREVENTIONAND WASTE MANAGEMENT 15.1 Disposeof sample wasteby placing in high or lowBTU containers as appropriate. Use broken glass containertso disposeof glass pipettes. 15.2 Collect HPLC solvent waste in the satellite accumulation can. Empty into the flammable storage drum in the hazardous waste collection area on the 2nd floor. 15.3 Use smaller borecolumns when possibleto minimize waste generation. 16.0 RECORDS 16.1 Print hard copies of all graphics and data analysis summaries for archiving. 16.2 Sign and dateall graphics and label with instrumeIDnt. 16.3 Fill out the hydrolysis sample preparation worksheet completely, makingtosiunrcelude all initials and dates. 16.4 Print out the sample sequence table, reduce twheithsipzheotocopying andtape the photocopy intothe instrument log. Keep the original copy ftohre raw data files package. 16.5 Print chromatograms and quantification reports for all analyses. 16.6 Print calibration tablesand curve information and store in the raw data file. 16.7 Storehydrolysis sample preparation worksheientsthe raw data file. 16.8 Enter all standard preparation information in the standards preparation logbook. Make a photocopy of the logbook pageand include the copyin the raw data file. 16.9 Archive electronic datao appropriate mediawhen necessary. ETS-8-179. I Method Page 14 of 18 Prep. of ,V-EtFOSE Alcohol HydrolysisSamples and Analysisby HPLClMS Page 35 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 17.0 ATTACHMENTS 17.1Attachment A. Representativechemicalstructures 17.2 Attachment B. Hydrolysissamplelogsheet 18.0 REFERENCES 18.1 Fate, Transport and Transformation Test Guidelines Office of Prevention, Pesticides and Toxic Substances (OPPTS) 835.210 Hydrolysis as a Function of pH, EPA 712-C-98057, January 1998. 18.2 CRC Handbook of Chemishy and Physics, 1st Student Edition, "Buffer Solutions Operational Definitionsof pH," Robert C. Weast, Ph.D., 1988, p. D-87. 18.3 Hewlett Packard 1100MSD instruction CD/ROM 19.0 AFFECTED DOCUMENTS 19.1 None. ETS-8-179.1 Method Page 15 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis byHPLCMS Page 36 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 20.0 REvrSIONS Revision number 001 RevisioRneason for revision Resizing of graphics and re-formattingto remove page orphans Date of 03/19/0 1 ETS-8- 179.1 Method Page 16 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samplesand Analysis by HPLCMS Page 37 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1872 Attachment A. Representative Chemical Structures N-MeFOSE alcohol (FW = 55n THPFOS F W = 427 (anion)) PFOS (FW =499 (anion)) N-EtFOSE alcohol FW = 526) ,I, F F d/ Go Counter cation = K+in this study, butmay also be Li', Ca", or DEA (diethanolamine) ETS-8-179.1 Method Page 17 of 18 Prep. of N-EtFOSE Alcohol Hydrolysis Samples and Analysis by HPLCMS Page 38 of 83 5 z 0 I1 iia EE CC BACK TO MAIN a t P II BACK TO MAIN 3M Environmental Laboratory Report NoV. V1872 Appendix B: Kinetics Model This Appendix includes a mathematical descriptiofnthe kinetics model employedin the study. Page 40 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 Kinetics Model B I . Reaction Components and Rates The arguments below are based on the following idealized set of reactions representing the hydrolysisof a parent compoundP and its hydrolysis productAs, which numberN. The actual hydrolysis reactions that occur under neutral, acidic, and basic conditions are subsumed in these equations, and are assumed to proceed with pseudo-first order rates k,, (for the parent) ankd,, (for the parent's hydrolysis products). P + H20 kP, @ n, A,+Y, (m=ltoN) (m= 1toN) where the general symbolYs,, and Ym2representall the other hydrolysis products. B2. Parent Compound Concentrations Equation B1 indicates that the pseudo-first order differential change in the parent concentration P is given by which is equivalent to the separable differential equation -P?p=-n,(,).,$ Equation B4 may be directly integrated to obtain the general solution With the initial conditionP(t = 0) = Po,the specific solutionto Equation B4 is P =Po exp (-8n, k,, t ] P~oe-kpt using the additional definition of the total parent hydrolysis rate Page 41 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 m=l Equation B6 can be re-writteinna form that allowsa least-squares estimateof the total parent hydrolysis rate: kpt=-In [i) Using the initia(lt =0)measured valueof the parent concentrationPoand later values P measured at later timest one can calculate and plot the (linear) quantity cp [- In (P/P,)] versus time and obtaianleast -squares estimate of the slope of the line. The resulting slope is the least-squares estimate of the total parent hydrolysis rate. Equation 56 indicates that ovear period of timeTI'; (the parent hydrolysis half-life) the parent concentrationP is reduced through hydrolysis abyfactor oftwo, where A least squares estimatefqiof the parent hydrolysis half-life is therefore available from 83. ProductCompoundConcentrations The pseudo-first order differential changes in the product concentraA,tio(nussing Equations B2 and B6) are dA, = ( n,kpmP - kA,Am)dl:= ( n,kPmP,, e-kp- kA,Am)dt (B11) and the (first order, non-separable) differential equation governing the product concentrations is %+ k,A, = n,k,,P, dt The "standard formo" f Equation 512 i s e-kpt. Page 42 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 A`, + S (t) A, = Q(t) where the "functionS" (t) is actually a constant: (t)= kAm and Q(t>=nmkPmPeO-kpc. The general solutionA, to EquationB12 is contained in where and /Q(t) ejs(t')ddtt' + C = s nmkPmPOekbte-kptdt+ C There aretwo cases of Equation B18to consider. In the circumstance that kA, = k, , which occurs only whetnhe hydrolysis rateof the mthproduct is identical to the total parent hydrolysis rate, the general solution to Equat8io1n8 is (for k, = k,) A, ekp=t nmkPmPtO+ C and, usingthe initial conditionAm(t= 0) = A, ,the specific solutionto Equation18 is (for k, = k,) A, = (nmkPmPto+ Amoe)-kp . We note that whenk, = k, = 0 (that is, when both the parent and potential product are hydrolytically stable), EquationB7 requires (also)that k,, =0 , so Equation B20 becomes Page 43 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 indicating, as required, that the product concentration ndootecshange with time. The circumstancek, = k p is highly improbable, andis neglectedin the remainderof this discussion. However, the reader should beainr mind that the expressions derived below do not hold when the parent hydrolysis krapteand the product hydrolysis ratek, approach each other. In the more probable case, for whickh,, # kp (i.e. that the hydrolysis rate of thme th product is different from the total parent hydrolysis rate), the general solution to Equation B18 is and the specific solution to Equatio8n18 with the initial conditioAn ,(t = 0 )= Am, is Of greatest interest here is the casinewhich the product compounds are known to be hydrolytically stable, that is, whekn, = 0 for all m. In this case, EquationB23 becomes (for hydrolytically stable products) + A, =AmO nmkPmPO kP (1- e-kp t ) . B4. Relationships Between the Parent and Compound Concentrations Equations B7 and B24 can be combined to obtain (for hydrolytically stable products) Page 44 of 83 so that or BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 (for hydrolytically stable products) (for hydrolytically stable products) If the changesin the product concentrations are all small compared to the original parent concentration, thatis, if we may use the expression (valid for -1 I X I 1 h ( l + X ) = X - -1x2 1+-x-3-P x 4 +..... 2 3 4 and Equation B23 becomes (for hydrolytically stable products andC A m-Amo << Po1 m Page 45 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W4872 or (for hydrolytically stable products cand rn A, -Amo << Po) k,t E m=l 85. Parent Half-Life Estimates Basedon Limits of Quantification of the Products In every experimental determination okf, , there is some set of valuesA F Q(the "limits of quantitation") below which the product concentratioAns, cannot be reliably measured. If during an experiment carried out over the period of timteadl,l the product concentrations A, remain below their limits of quantitation, then the maximum possible value of the ratek, is obtained by assuming (for all the products) th1a)t A,, = 0 and 2) at time =t A t , the product concentrations have increasetdo the values A, = A P Q . With these assumptions, the experimental data indicate that the reaction kra,teis less than some maximum value(kp)- as follows: (for hydrolytically stable products at concentrations below the limiotsf quantitation) 1 N k, I(kP)- = - C A k o Q . Po A t m=l Underthe same circumstances and assumptionsth, e experimental data indicatethat the parent half-lifeT''; (see EquationB9)is greater than the value(T!$) , as follows: mm (for hydrolytically stable products at concentrations betlohwe limits of quantitation) The reader should note that Equatio8ns32and B33 are valid only when bot1h) the products are hydrolytically stable an2d)the concentrationsof all the potential products are measured. Otherwise, the quantity (kp),, in Equation B32 may not actually represent the maximum possible valuoef the rate constantk, ,and the related resultin Equation 833 for (TyE)rm.n is also questionable. Page 46 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo.W1872 B6. Parent Half-Life Estimates Based on Limiotsf Quantification and Experimental Precision of Product Concentrations In certain experiments, some hydrolysis products are present at quantifiable but essentially constant concentrations over the ti(mAet ) of the experiment.In this case, it is the experimental precision of the measured product concentrations, rather than the limits of quantitation, which contribute to the estimate of the maximum value of the parent hydrolysis ratek, . If the set of concentrations measured for mthtehproduct have the mean valuep, and standard deviationO, ,the datado not exclude the possibility that the product concentration increased from the initial0v,al-upe, to the value O, + p, at timeft = A t . Taking this possibility to be the actual case for the measured products, the maximum value of the quant(itAym-Amo)is 20,. This reasoning suggests that the following estimate of the maximum parent hydrolysisis rate appropriate: (for hydrolyticallystable products at either1) constant measured concentrations with standard deviationa, or 2) concentrations below the limitsof quantitation) r 1 Under these circumstances and assumptions, the experimental data indicate that the parent half-life TI': is greater than the value(Tv;) , as follows: mm (for hydrolytically stable products at either1) constant measured concentrations with standard deviationa, or 2) concentrations below the limitsof quantitation) (Ti':)min C ~ JO , TYt 2 r 1-1 =-(kIPn(12- ) = At P, ~n2() A ~ ; P+~ Below LOQ Constant . (B35) The reader should note that Equatio6ns34 and 635 are valid only when boIth)the products are hydrolytically stable a2n)dthe concentrationsof all the potential products are measured. B6. Parent Half-Life Estimates Basedon the Experimental Precisionof Parent Concentrations In certain experimentst,he hydrolytic parent remains at an essentially constant concentration over the tim(eA t ) of the experiment. In this caseit, is the experimental precision of the measured parent concentrations that determines the maximum value of the parent hydrolysis ratke p. If the set of concentrations measured for the parent have the mean valueppand standard deviationO,,the data do not exclude the possibility Page 47 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo.W1872 that the product concentration increased from the initial valupep - G, to the value pp +CY,at timet = A t . This reasoning suggests that the following estimate of the maximum parent hydrolysis rate is appropriate: (for essentially constant parent concentrations with mean valpuepand standard deviationCY,) Under these circumstances and assumptions, the experimental data indicate that the ) parent half-life T1'i is greater than the value(Tf . as follows: rmn (for essentially constant parent concentrations with mean valupepand standard deviation0,) B8. Temperature Dependence of the Reaction Rate and Half-Life In order to increase the speed of the reactionosf interest, we conducted this experimental study using samples maintained at the temperatu5r0eC = 323 K. Of greater interest are the corresponding results for the environmentally important temperature 25C = 298 K. When the Arrhenius activation energy for a reaction isAH,, Equation B38 provides the following relationship between the hydrolysis ra(kte, sand k2)for that reaction at two different absolute temperatures(TIand T2): where R = 1.99 x Kcal mole" K-' is the ideal gas constant. Using the valueB2 AH, = I 8 Kcal/mole, the rate ratiokl/k2 at the corresponding temperaturesTI =298 K and T2=323 K is = exp{ 18 [L-I]=} exp(-2.35) = 0.095 k2 1.99~10" 323 298 Page 48 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 Equation B39 indicates that the hydrolysis reactioonfsinterest proceed approximately ten times more slowly 2a5t C than atthe chosen experimental temperature o5f0C. Accordingly, the rate reactions reported here for the temperature 25Cteanretimes lower than those measured at 50"@, and the hydrolysis half-life estimates reported here for 25Csamples areten times longer than those calculated from t5h0eC experimental data. References to Appendix B: 61 I. N Levine, "Physical Chemistry," McGraw-Hill (New York), pp. 498-501 (1978). F. Daniels, et al., "Experimental Physical Chemistry", McGrawHill (New York), p.131 ( I962). Page 49 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Appendix C: Selected Analytical and Kinetics Results This Appendix includes selected sample data and their related kinetics results. Page 50 of 83  BACK TO MAIN : i |e tt [E48 bi ; i P2 H iOR tbl Ele LH| ClalEboilksei ELEnNI _ -- 1 Bod [1,1 [foes T, J | 4 : eee Log $1 Les IN i ezzzzay i llc i5 4 i See FETT | He czaanes i J Jraanaaas | SSgesee | ole ~=mraes f "lle 2222222 | FRRExFY | BACK TO MAIN . a E 2 0 (D 1 O e h% E t G 0 J BACK TO MAIN 1 L BACK TO MAIN s f BACK TO MAIN m 03 a" z IC 0 m m 0 a, m I BACK TO MAIN P d' -I 1 BACK TO MAIN I . .._. 3 z 0 mmmwwm rdddddd BACK TO MAIN s c m n 8 m -1 BACK TO MAIN CD v) z 0 a, w a m BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Pooled N-EtFOSE Alcohol Data and Slope Regression 0.05 0 0.00 -0.05 2- -0.10 2 E -0.15 Line: Solid 0 -0.20 y = -2.62E-03~- 2.30E-02 R2= 4.1OE-01 0 -0.25 Dashed Lines: 20 limits (sloapinendtercept) . -0.30 0 10 20 30 time (days) 0 - . . _-- -. *. -. 40 50 L SUMMARY OUTWT Regression Statistics Multiole R 0.640148735 R Sduare 0.409790403 Adjusted R Sq 0.396064598 Standard Err0 0.049634563 Observations 45 ANOVA df ss MS 1 Regression 0.074145584 2.21908.80057854E417-4045605584 840,1067898 43 Residual 44 Total 0.180935383 0.098035439 -1.691199513 Intercept X Variable 1 CoefficiSetnatnsEdt rarrodr Stat -0.0230159405.013609243 -0.0002.060-1507.4647562924.1008-1086.02809-0E33.5-0-08064.10060-650341.5063044106654164 % 2a Slope Uncertainly 37% F P-value Signiricance F Lower 95% Upper95% Lower95.0% , . ! Upper 95.0% 0.00442969 Page 60 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Appendix D: Selected Chromatograms A representative seot f chromatograms from the present studisy included in this Appendix. Page 61 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 Batch Run # 2 o f 56 Data File C: \HPCHEM\l\DATA\O92899\EtFOSOO2.D Sample Name: MeOH ----------=----------------------------------------------------------========== InjectioDnate : 9/28/89 3:10:02 PM SeqL.ine : 2 Sample Name : MeOH Blank Vial : 91 Acq. OperaItnojr : MTM : 1 A c q . Instrument : Instrument 1 Acq. Method : c : \HPCHEM\I\METHO&\FOSESIM.M Last changed : 9/28/99 2:49:57 PM'by MTM AnalysisMethod C:\HPCHEM\l\METHODS\0824_1IS.M Last,changed : 9 / 5 / 0 0 8:28:58 AM by kej (Results arefr0m.a previously saved Batch) SIM Analysis (ES-j f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. Blank 400 350 Page 62 of 83 Batch Run # 2 o f 56 DaFtiale C:\HPCHEM\l\DATA\O92899\EtFOS002.D BACK TO MAIN 3M EnvironmentalLaboratoryReport No.W1872 Sample'Name:MeOH Blank Signal 1: MSD1 427, EIC=426.5:427.5 Signal 2 : MSD1 499, EIC=498.5:499.5 Signal 3: MSD1 616, EIC=615:5:616.5 ' Signal 4: MSD1 630, EIC=629.5:630.5 RetTimSeig Type AreAamt/Are&amount G r p , Name ratio ---[--m--1i--n1-]-----1----------~----------~----------~--~------~---,----- ' - 5.940 . 1 6.221 2 - - - - - THPFOS , -- PFOS MeFOSE-OH 8.355 3 8.586 4 - I - -. EtFOSE-OH Totals without ISTD(s1 : 0. aoooo 1. Warnings or Errors : . Warning : ISTD compound(s1 not found Sorted By TiRmeetention Calib. Data Modified : , Tuesday, September 05, 2000 8:28:56 AM Multiplier 1.0000 Dilution I.0000 Peak RetTimSeiTgypAerea ----1#-------1---[1-m---i--nI-l----------l--------l------------------------- 1 5.940 1 2 6.221 2 3 8.355 3 4. 8,.586 4 I 0.00000 0.00000 I 0.00000 0.00000 Totals : 0.00000 1 Warninus or Errors &ea Name % '0.0000 THPFOS 0.0000 PFOS 0.0000 MeFOSE-OH 0.0000 EtFOSE-OH Page 63 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 Batch R u n # 3 of 56 Data File C:\HPCHEM\l\DATA\O92899\EtFOS003.D Sample Name: L1 1.5, 155.40 -.-.--.-.--.-.--.-.--.-.--.-.--.-.--.--.-.-=..--.--.-.--.-.--.-.--- --------------e'=lcIr2rr===-ppe-I= I n j e c t i o n Date : 9/28/99 3:28:12 PM LSeinqe. : 3 Sample Name : L1 1 . 5 , 1 5 5 . 4 0 Vial : 1 Acq. Operator : MTM Inj : 1 Acq. Instrument : Instrument 1 AcqM.ethod : C:\HPCHEM\I\METHODS\FOSESIM.M Last changed : 9/28/99 2:49:57 PM by MTM \ Analysis Method : C:\HPCHEM\1\METHODS\O824 lIS.M Last changed : 9/5/00 8:28:58 AM by kej- (Results are from a previousiy saved Batch) , SIM Analysis (ES-) f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. .. [TM 50000 30000 20000 10000 a ' 0 , N I s D l - R ? i U .t r t i ; = ~ ~ T P l - t s 30000 20000 15000 5000 4". 6' NeQanve 4' 40 mir Page 64 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1872 Signal'l: MSDl 427, EIC=426.5:427.5 Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3 : MSDl 616, EIC=61.5.5:616.5 Signal 4: MSDl 630, EIC=629.5:630.5 RetTime Sig Type Area Amt/Area ' Amount Grp Name ----[-r--n1-i-n1-l-----1----------1----------1----------1--1------------'--- ratio ppb (ng/ml 5.925 1 PB 1: 4.65165e5 1.00000 402.60000 THPFOS 6.207 8.313 8.538 2Mb' 3 PB 4 PB 5973.06250 3.07266e-1 1.58847 r 8.80788e5 1.00000 315.60000 5.05275e5 8.83244e-1, 159.90937 PFOS MeFOSE-OH EtFOSE-OH Totals without IS!CD(s) : 161.49784 Page 65 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo. W1872 .D Batch Run# 4 of 56 Data File C:\HPCHEMYl\DATA\O92899\EtFOS004 ' Sampie Name: LZ 1.5, ..................... -- ---- ---_ -----c ------ ----- ---- ---- ==- ---- ---- ---- ---- ---- ----- --=I =_ ------ ---- ---- ----------- ---------------qt Injection Date : 9/28/.39:946:24 PM Seq. Line : 4 Sample Name Acq. Operator : L2 1.5, 310.80 : MTM Vial : 2 Inj : 1 Acq. Instrument : Instrument 1 Acq. Method : C:\HPCHEM\I\METHODS~FOSESIM.M Last changed ' : 9/28/99 2:49:57 PM by MTM Analysis Method : C:\HPCHEM\l\METHODS\0824~~IS.M Last changed :.9/5/00 8:28:58 AM by kej (Results.arefrom a previouslysaved Batch) SIM Analysis (ES-1 f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35m Dionex IonPac NG1 column, S/N 12879. < yTI 310.80 I. i Page 66 of 83 Batch RUn # 4 Of 56 Data File C:\HPCHEM\l\DATA\O92899\EtFOS004.D BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 Sample Name:. 212 1.5, 310.80 Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD Amount Name ----I#---------p--p--Ib-----(---n--g---/--m--l;-------- 1 402.60000 THPFOS 315.260000 MeFOSE-OH Retention Time Tuesday, September 1.0000 1.0000 05,'2000 8:28:56 AM JJf.y/~yl'. Signal 1: MSDl 427, EIC=426.5:427.5 Si.gna1 2: MSDl 499,'EIC=498.5:499.5 Signal 3: MSDl 616, EIC=615.5:6i6.5 Signal 4: MSD1 630, EIC=629.5:630.5 RetTime Sig Type [minl , I --1--L---I 1 BV I 6.218 2 PB 8.320 3 BB I 8.545 4 PB Area Amt/Area Amount G r p Name - - - .. - - I -- - ,r-- a-t-i- -o- -, - -- I -p-p-b- - -(-n-g-/-mlI - +.- - - - - - - - - - - - - - 4.56303e5 1,00000 402.60POO W P F O S 4.24807e4 3.52850e-1 13.22520 PFOS 8.73131e5 1.00000 315.60000 MeFOSE-OH 9.57739e5 9.'01077e-1 311.93664 EtFOSE-OH wTIioSttThaDol(usSt) : 325.16184 Page 67 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No. W1872 Batch Run # 5 of 56 Data F i l e C:\HPCHEM\l\DATA\O92899\EtFOS005.D Sample Name: L3 1.5, 466.20 -.-.--.-.--.-.--.-.--.--.-.--.-.--.-.--.-.--.-.--.--.-.--.-.--.-.--.-.--.--.-.--.-.--.-.-=.=..--.--.-.- ............. - - I _ _ - _ _ _ _ _ _ I n j e c t i o n Date : 9/28/99 4:04:35 PM Sample Name : L3 1.5, 466.20 Acq, Operator : MTM Acq. Instrument : Instrument 1 Seq. Line : 5 Vial : 3 Inj : 1 Acq. Method : C:\HPCHEM\~\METHODS\FOSESIM.M Last changed : 9/28/99 2:49:57 PM by MTM Analysis Method : C:\HPCHEM\l\METHODS\O824-1IS.M Last changed : 9/5/00 8:28:58 AM by kej (Results arefrom a previously saved Batch) SIM Analysis (ES-) for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4 m m 3 5 m m Dionex IonPac NG1 column, S/N 1,2879. ,l Page 68 of 83 Batch Run # 5 of 56 Data File C:\HPCHEM\l\DATA\O92899\EtFOS005.D BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Sample Name: L3 1.5, 466.20 Signal 1: MSDI 427, EIC=426.5:427.5 Signal 2: MSDl 499, EIC=4.98.5:499,5 Signal 3: MSDl 616, EIC=6.15.5:616.5 Signal 4: MSDl 630, EIC=629.5:630;5. RetTime Sig Type. ArAemat/Area Amount Grp' Name --[-m-in-l-- I--f-,-----l----------~----------~----------~--~-----r--a---t--i-~o- ppb (ng/ml , , 5.928 1 BB I'4.47867e5 1.00000 402.60000 THPFOS 6.217 2 BB 8'.87226e4 3.56716e-1 28.44993 8.322 3 'PB I 8.68287e5 1.00000 315.60000. PFOS MeFOSE-OH 8.546 4 PV 1.39693e6 9.07298e-1 460.67730 EtFOSE-OH ' Totals without ISTD(s) : 489.12723 Page 69 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No.W1872 Batch Run # 6 of 56 DataFileC:\HPCHEM\l\DATA\O92899\EtFOSOO6.D Sample Name: L4 1 3 , 621.60 --------- ---------rlPPIPI=tP==E===I=lt=PI-P==== Injection Date Sample Name A c q . Operator Acq. Instrument A c q . Method Last changed Analysis Method : 9/28/99 4:22:47 PM 2 L4 1.5, V6i2a1l.60 : MTM : Instrument 1 : C:\HPCHEM\I\METHODS\FOSES.IM.M : 9/28/99 2:49:57 PM by MTM : C:\HPCHEM\l\METHODS\O824-11S.M Seq. Line' : : Inj : 6 4 1 3;w ' Last changed : 9/5/00 8:28:58 AM by kej (Results are from a previously saved'Batch) TJdW SIM Analysis (ES-} f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex fonPac NG1 column, S/N 12879. .. mom^I00000 60000 ' 3 40000-j 20000 Page 70 of 83 Batch Run # 6 of 56 Data File C:\HPCHEM\l\DATA\O92899\EtFOS006.D BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Sample Name: L4 1.5, 621.60 Sorted By Retention Time Calib. Data Modified : . . Tuesday, September 05, 2000 8:28:56 AM . , Multiplier 1.0000 ion Bi lut 1.0000 Sample ISTD Information: ISTD ISTD Amount Name - - -#- I - - -p-p-b- - - -(-n-g-/- [m,l- - - - - - - - - - - - - - - - - - - - - - - - - .1 ' 402.60000 THPFOS 2 315.60000 MeFOSE-OH Signal 1: MSDl 427, EIC=426.5:427.5 Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3: MSDl 616, EIC=615.5:6.26.5 Signal 4: MSDl 630,'EIC=629.5:630.5 RetTime Sig TyApreea ---[---m-1i--n1-l-----1------,----1----------1----------1 5.926 1'BB I 4.51811e5 6.215 2 VB 1.120.72e5' 8.534 3 BB I 3.00592e4 8.546 4 PP 1.86971e6 .Amt/Area Amount Grp' Name r a t i o ppb (ng/tnl - - I - - - - - - ' - - - - - - - - - 1.00000 402.60000 THPFOS , 3.57406e-1 35.69254 PFOS 1.00000 315.60000 MeFOSE-OH 9.20283e-1.1.80657e4 EtFOSE-OH , Tokals without ISTD(s)' : 1.81014e4 Page 71 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo.W1872 Batch Run # 7 of 56 Data File C:\HPCHEM\l\DATA\O92899\EtFOSOO7 .D' Sample Name: L5 2.5, ---_---_---_---_---_---_------_---_------_--_---_------_---_---_---_--------=-=-----------_--_-----------_-------_-_-_-_-_-_-_-_-_-_-_-_ Injection Date : 9/28/99 4:41:01 PM Seq. Line : 7 Sample Name : L5 1.5, 777.00 Vial : 5 Acq. Operator : MTM Inj : I Acq. Instrument : Instrument 1 Acq. Method : C:\HPCHEM\I\METHODS\FOSESIM.M Last changed : 9/28/99 2:49:57 PM by'MTM Analysis Method : C:\HPCHEM\l\METHODS\0824-lIS.M Last changed : 9/5/00 8:28:58 AM by kej , ' (Results are from. apreviously saved Batch) SIM Analysis (ES-) for Et-FOSE-OH, MeFOSE-OH,. ,THPFOS, andPFOS usihg 4mmx35mm Dionex IonPac N G l column, S/N,l2879. 777.00 Page 72 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Sorted By TiRmeetention Calib. Data Modified : Tuesday, September 05, 2000 8:28:56 AM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name _ _ _ _#1 _ _ _ _ _ _ _p_ _p_ _b_ _ 1 _ _(_ _n_ _g_ _/_ _m_ _l_ _ _ _ _ _ _ _ _ - - - - 1 402.60000 THPFOS 2 315.60000 MeFOSE-OH Signal 1: MSD1'427, EIC=426.5:427.5 ' Signal 2: MSDI. 499, EIC=498.5:499.5 Signal 3: MSD1.616, EIC=615.5:616.5 Signal 4: MSDl 630, EICs629.5:630.5 RetTi,Smieg Type, Area Amt/ArAemaount G r p Name - - -[- m_ -i- n1 -]- 1.- - - -, - - 1 - - - - - - - - - - ~ - - - - _ - - - - - ~ - - - - - - - - -r- a~ ~t-i~ o- - - - - - - -p- -p- -b- - - (ng/ml' 5.940 1 - - - THPFOS ' 6..213, 2 BB 1.67565e5 ' 0.00000 0.00000 PFOS 8.318 3 PB 1 8.56724e5 1.00000 315.60000 , MeFOSE-OH 8.544 4 PB 2.34711e6 9.12800e-2 789.23438 EtFOSE-OH Totals without ISTD(s1 : 789.23438 Page 73 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No. Wl872 Batch Run # 8 o f 56 Data File C: \HPCHEM\l\DATA\O92899\EtFOSOO8;D Sample Name: L6 1.5, 932.40 ............................................................................. Injection Date : 9 / 2 8 / 9 9 4:59:12 PM Seq. Line : Sample Name Acq. Operator : L69312..54,0 : MTM Vial : Inj : ACq. Instrument 2 Instrument 1 Acq. Method C:\HPCHEM\I\METHODS\FOSESIM.M . , Last changed 2 9/28/99 2:49:57 PM by M T M Analysis Method .: C:\HPCHEM\l\METHODS\0824_1IS .M Last changed ' : 9/5/00 8 : 2 8 : 5 8 AM by kej (Results are from a previously savedBatch) SIM Analysis (ES-1 for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS 4 m m x 3 5 9 Dionex IonPac NG1 column, S/N 12879. 8 6 1 5J4 y/yb using MTM Page 74 of 83 BACK TO MAIN 3M Environmental Laboratory ReportNo.W1872 Signal 1: MSDX 427, EfC=426.5:427.5 Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3: MSDl 616, EIC=615.5:616.5 Signal 4: MSD1'630, EIC=629.5:630.5' ' RetTime S'ig Type Area ----[--m-Ii--n[--]----I----------l---------- Amt/Area Amount Grp ratio IP- -P- -~-f-nI g-/-,-ml 5.927 6.215 8.323 8.546 1 BB 2 VB 3 PB 4 BV I 4:60976e5 . 1:OOOOO 2.10524e5 3.58656e-1 I 8.87024e5 ' 1,00000 2.91155e6 . 9..14094e-1 402.60000 65.94394 315.60000 946.92850 Totals without ISTD( s ) : 1012.87244 Name Page 75 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No.W1872 Batch Run # 9 of 56 Data FileC:\HPCHEM\l\DATA\O92899\EtFOSOO.9.D Sample Name: MeOH Blank - --- --- -- - -- --- -II I- Il_ ===- ==PPIPC=01=E======Lr=I===~========================== I n j e c t i o n Date : 9/28/99 5:17:26 PM Seq. Line : 9 SampleName ' : MeOH Blank Vial : 91 Acq. Operator : MTM Inj : 1 Rcq. Instrument : Instrumexit 1 A c q . Method : C:\HPCHEM\~\METHODS\FOSESIM.M Last changed : 9/28/99 2:49:57 PMMTMby Analysis Method : C:\HPCHEM\l\METTIODS\0824 1IS.M L a s t changed : 9/5/00 8:28:58 AM by kej- 13-+f lb (Results are from a previously 'saved Batch) SLM Analysis (ES-) f o r Et-FOSE-OH, MeFOSE-OH,,THPFOS,and PFOS using . 4mmx35mm Dionex IonPac NG1 column, S/N 12879. Page 76 of 83 Batch Run # 9 .of 56 Data F i l e C:\HPCHEM\3,\DATA\O92899\EtFOSOO9.D BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Sample Name:MeOH Blank TiRmeetentionBy Sorted Calib. Data Modified : Multiplier Dilution Sample ISTD Information: ISTD ISTD AmountName I Tuesday, September 0 5 , 2000 8:28:56 AM 1 ..oooo 1.0000 , Signal 1: MSDl 427, 3IC=426.5:427.5 Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3: MSDl 616, BIC=615.5:616.5 Signal 4 : MSDl 630, EIC=629.5:630.5 G r p Name .TwoittaIhlSosTuDt(.S) : 1 Warriings or Errors : . Warning : ISTD compound(5) not .found . 0.00000 TiRmeetentionBy Sorted ' . Calib. DataModified : Multiplier ,Dilution Tuesday,September OS, 2000 8 : 2 8 : 5 6 : A M 1.0000 1.0000 Totals : 1 Warninas or Errors 0.00000 Page 77 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 Batch Run #'23 of 5 6 Data File C:\HPCHEM\l\DATA\O92899\EtFOS023.D Sample Name :. EFA-073 - - - - _ - - _ _ _ _ _ _ _ =====--- ---1- - -- - -- - -- --- --- --- --- - -I-----p ----- --I -p ----r ---= ----= -r-= ----1 ---1 ----= - 1---------- 1njec.tion Date : 9/28/99 9:32:40 PM Seq. Line : .. 23 NameSample : EFA-073 Vial : 23 Acq. Operator : MTM Inj I ' 1 Acq. Instrument : Instrument 1 Acq. Method ' : C:\HPCHEM\I\METHODS\FOSESXM.M ,' Last changed ' . : 9/28/99 2:49:57 PM by M T M Analysis Method : C:\HPCHEM\l\METHODS\0824 lIS.M Lacshtanged : 9/5/00 8 :28 :58 AM by kej- Pf ' SpYI" , (Results are from a previously saved Batch) SIM Analysis (ES-1 f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. MTM 40000 35000 30000 25000 20000 15000 1 woo 5000 . . . 4 ' 8 - 8 , ' I 0, MYU-l49l8, t 1' Wl-kb N%am $0 m Page 78 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Batch Run # 23 of 56 Data File C:\HPCHEM\l\DATA\092899\EtFOSO23.D Sample Name: EFA-073 Sorted By Calib. Data Modified ' : Multiplier Retention Time Tuesday, September 05, 2000 8:28:56 AM 1.0000 Signal 1: MSDl 427, EIC=426.5:427.5 Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3.: MSDl 616, EIC=615.5:616.5 Signal 4: MSDl 630, EIC=629.5:630.5 RetTi~eSig Type Area' - -I-m-in-l- - 5.926 6.221 I----------I---------- 4 - 647- 74e5 8.326 3 v B I 8.48397e5 8.550 4 BB 1.33227e6 Amt /Area Amount Grp ratio Ip_p-b- _ -(- _d- _m. .1 I.-00000 4.02-.60000 1.00000 315.60000 9.06971e-1 449.49185 Totals without ISTD(s) : Name 1'Wariiings or Errors : . . . Page 79 of 83 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1872 Batch R u n # 24 of 56 Data F i l e C:\HPCHEM\l\DATA\o92899\EtFOS024.D Sample Name: EFA-074 -_ -----_ ------ ----_ -----_ ------_ -----_ ------- ---_ -----_ ------I ---_ -----_ ------ -=-- -- __-_-____ _ I _ _ _____ Injection Date : 9/28/99 9:50:54 PM Seq. Line ; 24 Sample Name : EFA-074 Vial : 24 Acq. Operator : MTM Inj : 1 Acq. Instrument : Instrument 1 A c q . Method : C:\HPCHEM\~\METHODS\FOSESIM;M Last changed : 9/28/99 2:49:57 PM by MTM Analysis Method : C:\HPCiEM\l\METHODS\0824-11S.M Last. changed : 9/5/00 8:28:58 AM by kej (Results are from a previously saved Batch) SIM Analysis (ES-1 f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4 m 3 5 m Dionex'IonPac NGI column, SjN 12879.. 40000 35000 30000 ' 25000 20000 . 15ooo I0000 5000 0, h 5 - - .1.. . ' S 4. N I S' egawe 8' .Ib mb Page 80 of 83 Batch Run # 24 of 56 Data File C:\HPCHEM\l\DATA\092899\EtFOSO24.D BACK TO MAIN 3M Environmental LaboratorRyeport No. W1872 Sample Name: EFA-074 Sorted By Retention Time . . Calib. Data Modified : Multiplier ' . ' Tuesday, September 05, 2000 8:28:56 AM I. 0000 Dilution 1.0000 Sample ISTD'Information: ISTD ISTD Amount Name ---#-1-p-p--b--(-n-g--/-m-l-I--------- ---------------- 1 402.60000, , THPFOS 2 315.60000 MeFOSE-OH Signal l:.MSDl 427, EXC=426.5:427.5 Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3: MSDl 616, EIC=615.5:616.5 Signal 4: MSDl 630, EIC=629.5:630.5 RetTime Sig Type Area ----[--m-l-i-In--l----I----------l---------- 5.925 1 BB 6.221 2 I - 4.56350e5 8.329 3'BB I 8.55992e5 8.555 4 PB 1.28265e6 Amt/Area ratio 1.- 00000 1.00000 9.06316e-1 Totals without ISTD(s1 : Grp 428.60235 Name Page 81 of 83 BACK TO MAIN 3M Environmental LaboratoryReport No.W1872 Itch Run # 25 of 56 / Data FileC:\HPCHEDl\l\DATA\O92899\EtFOS025.D Sample Name: EFA-075 ======LXI==t=IQDC=3IIflP==t==-DI=PIIX InjectioDnate : 9/28/99 10:09:06 PM Seq. Line : 25 Sample Name : EFA-075 Vial : 25 Acq. Operator : MTM A c q - Instrument : Instrument 1 Acq. Method ; C:\HPCHEM\~\METHODS\FOSESIM.M Last changed : 9/28/99.2:49:57 PM by MTM ' Analysis Method : C : \ H P C H E M \ l \ M E ~ O D S \ 0 8 2 4 _ 1 I S . M Inj : 1 >Yf. . ' #Ji. Last changed : 9/5/00 8:28:58 AM by kej (Results are from a previously savedBatch) SIM Analysis (ES-1 f o r Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS .using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. 50000 moo . . 30000 20000 1Oooo 220000 iowoo 60000 40000 20000 I Page 82 of 83 BACK TO MAIN 3M Environmental Laboratory Report No. W1872 Batch Run # 25 o f . 5 6 Data File C:\HPCHEM\l\DATA\092899\EtFOSO25.D Sample Name: EFA-075 Sorted By Calib. Data Modified : Multiplier Dilution Sample ISTD Information: Retention Time Tuesday, September 05, 2000 8:28:56 AM 1.0000 1.0000 ratio 2 315.60000 . MeFOSE-OH 'I Signal 1: MSDl 427, EIC=426.5:427.5 . Signal 2: MSDl 499, EIC=498.5:499.5 Signal 3: MSDl 616,. EIC=615.5:616.5 Signal 4: MSDl 630, 31C=629.5:630.5 RetTiSmieg Type ArAemat/ArAemaount ----[--m-1i--n1--l----1----------1----------1----------1--1--------------- 5.923 1 BB 6.221 2 I - 4.64731e5 1.00000 - 8.332 3.PB I 8 . 4 6 0 7 2 e 5 1.00000 8 . 5 5 9 4 MM 1.80524e6 9.10577e-1 G r p Name 402.- 60000 315.60000 613.16914 THPFOS PFOS MeFOSE-OH EtFOSE-OH Totals without "ISTD( 9 ) : 613.16914 1 Warnings or Errors. : . Warning : Calibrated compound(s1 notfound Page 83 of 83