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BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Study Title Hydrolysis Reactions of 2-(/V-Methylperfluoroctanesulfonamido)-Ethyl Alcohol (A/-MeFOSE Alcohol) Data Requirement: Based on OPPTS: 835.2110 Author Thomas L. Hatfield, Ph.D. Study Completion Date March 30, 2001 Performing Laboratory 3M Environmental Laboratory Building 2-3E-09, 935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Laboratory R eport No: W 1880 Total Number of Pages 71 Page 1 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 This page has been reserved for specific country requirements. Page 2 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Statement of Non-Compliance Study Title: Hydrolysis Reactions of 2-(A/-Methylperfluoroctanesulfonamido)-Ethyl Alcohol {N-MeFOSE Alcohol) Study Identification Number: W1880 This study does not fully comply with the requirements of the US EPA Good Laboratory Practices (GLP) Standards at 40 CFR Part 792 (TSCA). However, many GLP standards were used in the development of the analytical method (Appendix A), and the quality assurance procedures followed in this study were based on the practices described in the GLP documentation. This is a revised report in that the statistics on the study data and the discussion were changed from the initial study report. Changes to these interpretive sections were made to better represent the experimental results of the study. Study Director P 5" Sponsor Representative Date Page 3 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Quality Assurance Statement Study Title: Hydrolysis Reactions of 2-(/V-Methylperfluoroctanesulfonamldo)-Ethyl Alcohol (N-MeFOSE Alcohol) Study Identification Number: W1880 The following table provides details of the audits performed by the 3M Environmental Laboratory Quality Assurance Unit (QAU). Inspection Dates 10/4-6/2000 3/19, 20/2001 Phase Data and Draft Report Date Re ported to Management Study Director 10/06/00 10/06/00 Draft Report 3/21/01 3/121/01 QAU Representative 2 , 2001 Date Page 4 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Table of Contents Statement of Non-Compliance..............................................................................................3 Quality Assurance Statement................................................................................................4 List of Tables...........................................................................................................................6 List of Figures......................................................................................................................... 6 Study Personnel and Contributors.........................................................................................6 Location of Archives............................................................................................................... 7 S u m m ary.................................................................................................................................8 Introduction.............................................................................................................................9 Summary of Kinetics Model................................................................................................. 10 Materials and Methods......................................................................................................... 11 Chemical Characterizations............................................................................................ 11 Sample Preparation........................................................................................................ 11 Sample Analysis.............................................................................................................. 12 Deviations.........................................................................................................................12 Results and Discussion....................................................................................................... 14 Data Quality Objectives (DQO's )....................................................................................14 Anomalous Analytical Results.........................................................................................14 Statistical Methods and Calculations..............................................................................15 Data Summary and Discussion.......................................................................................15 Conclusions...........................................................................................................................19 References........................................................................................................................... 20 Signatures............................................................................................................................ 21 Appendix A: Analytical Method............................................................................................22 Appendix B: Kinetics Model.................................................................................................39 Appendix C: Selected Analytical and Kinetics Results......................................................49 Appendix D:: Selected Chromatograms.............................................................................. 60 Page 5 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 List of Tables Table 1. Summary of Results Based on N-MeFOSE Alcohol Concentrations.................. 8 Table 2. Summary of Results Based on PFOS Limit of Q uantification............................. 8 Table 3. Characterizations of Test and Reference Substances.......................................11 Table 4. Observed (50 C) Degradation Rates of N-MeFOSE Alcohol in Aqueous Buffered Solutions and at Various pH Levels..................................................... 15 Table 5. Degradation Rate and Half Life of A/-MeFOSE Alcohol in Aqueous Buffered Solutions Using Data Pooled Over pH Levels....................................................16 Table 6. Degradation Rate and Half Life of A/-MeFOSE Alcohol in Aqueous Buffered Solutions Based on PFOS Limit of Quantification.............................................. 18 List of Figures Figure 1. Structures of N-MeFOSE Alcohol and the Potassium Salt of PFOS................9 Figure 2. Observed N-MeFOSE Alcohol Degradation for Various pH Levels................16 Figure 3. Pooled N-MeFOSE Alcohol Data and Slope Regression................................. 17 Study Personnel and Contributors 3M Environmental Laboratory Building 2-3EE-09 935 Bush Avenue St. Paul, MN 55106 (651) 778-7863 3M Environm ental Laboratory and Professional Services C ontributing 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 Intertec Corporation, 6875 Washington Ave. South, Minneapolis, MN 55439) Page 6 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Location of Archives The 3M Environmental Laboratory will retain the original data documents and digital copies of the original data related to this work for at least 10 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 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Summary We report here the results of our study of the hydrolysis of 2-(A/-methylperfluorooctanesulfonamido)-ethyl alcohol (hereafter, N-MeFOSE Alcohol). Our methods are described below and In Appendix A to this work; our results are based on the observed concentrations of A/-MeFOSE 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 (HPLC/MS). Tables 1 and 2 summarize the results of the study. During this study, we prepared and examined samples at six different pH levels from 1.5 to 11.0 over a period of 49 days, and our results indicate no dependence of the degradation rate of A/-MeFOSE alcohol on the sample pH level. Our results based on the W-MeFOSE alcohol concentrations, pooled over the observed pH levels, are presented in Table 1. Table 1. Summary o f Results Based on N-MeFOSE Alcohol C o n ce n tra tio n s Observed Rate Constant at 50 C (day1) Calculated Rate Constant at 25 C (day1) Calculated Half Life at 25 C (years) Calculated (2a) Half Life Range at 25 C(years) 0.0030 0.00030 6.3 3.8 to 19.4 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). Assuming PFOS to be the only hydrolysis product of /V-MeFOSE Alcohol, this LOQ (and other experimental data) provide a second estimate of the A/-MeFOSE alcohol half-life, presented in Table 2. Table 2. Summary o f Results Based on PFOS Lim it o f Q u a n tifica tio n Maximum Possible Rate Constant at 50 C (day1) Maximum Calculated Rate Constant at 25 C (day1) Calculated Half Life at 25 C (years) 7.0 x104 7.0 x10b >27 According to the data available from this study, the half-life estimate of Table 2 represents the minimum possible half-life of the compound A/-MeFOSE alcohol under the assumption that it hydrolyzes to form only the compound PFOS. Page 8 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Introduction Three primary chemical routes of environmental degradation are hydrolysis, photolysis, and biodegradation. Studies of 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 A/-MeFOSE alcohol (or, more generally, its degradation in the presence of H2O) is addressed in this report. Structures of the "parent" compound NMeFOSE alcohol and the potassium salt of possible hydrolysis product perfluorooctane sulfonate (PFOS) are illustrated in Figure 1. Figure 1. Structures o f W-MeFOSE Alcohol and the Potassium Salt o f PFOS W-MeFOSE Alcohol Potassium Salt o f PFOS FFFFFFFFO FFFFFFFFO Page 9 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Summary of Kinetics Model A full mathematical description of the kinetics model employed in this study is presented in Appendix B. The study data allow two independent estimates of the hydrolytic half-life of A/-MeFOSE Alcohol. The first estimate (see Table 1) is based on the observed degradation of the "parent" compound W-MeFOSE alcohol in dilute, appropriately buffered aqueous solutions. Equation 1 describes the estimated half-life ( f ^ ) in terms of the estimated total parent hydrolysis rate k p (see Appendix B, Equation B10): Eq.1 We determined the quantity k pfrom the experimental data as described in Appendix B. At each pH level, we used the earliest study data meeting the data quality objectives to determine the relevant concentration ratios (see Equation B8). The measured concentrations of the potential hydrolysis product PFOS (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 PFOS above its limit of quantitation (LOQ), and related studies1show that PFOS is itself hydrolytically stable. Assuming also that PFOS is the only hydrolytic product of the parent compound NMeFOSE alcohol, these PFOS analyses provide the following estimate (t ^ ) of the minimum N-MeFOSE alcohol half-life (see Equations B32 and B33): A t [P ,,]h(2) a l q -^P F O S E q.2 where [P0]= the initial A/-MeFOSE alcohol molar concentration, A t = the time interval over which the study was conducted (49 days), and A pfos = the molar limit of quantitation for the compound PFOS. All the samples used in this study were maintained at a reaction temperature of 50 C. The quoted results, valid for the reaction temperature of 25 C, were calculated from our experimental results according to methods described in Appendix B (Eq. B38 and B39). Page 10 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Materials and Methods Details of the characteristics of the test materials, sample preparation techniques, and analytical methods are presented in Appendix A (ETS-8-178.0, "Preparation of 2-(NMethylperfluoroctanesulfonamido) Ethyl Alcohol (A/-MeFOSE Alcohol) Hydrolysis Samples and Analysis by High Performance Liquid Chromatography with Mass Spectrometry Detection.") A summary of these items is provided below, as well as a description the known deviations from the procedures of Appendix A. 3M prepared and analyzed the samples included in this study between March 13 and October 5,1999. Chem ical C haracterizations 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 standards used to quantify them. For this reason, and because Equation B8 (see Appendix B) involves only ratios of parent concentrations, the resulting rate and half-life estimates are largely independent of the material purity levels. Table 3. Characterizations o f Test and Reference Substances Source Chemical Lot Number8 N-EtFOSE Alcohol8 3M Specialty Chemistry TN-A-1284 Physical Description Off-white powder Molecular Weight (gm mole'1) 570.9 PFOS 3M Specialty Chemistry TN-A-2130 SD108 White powder 498.9 THPFOSb ICN Biomedicals TN-A-1339 (Lot #59909, SD028) Light brown powder JV-MeFOSE Alcohol 3M ICP/PCP Division TN-A-1282 SD 015 Amber waxy solid 428.0 556.9 aThe "S" and "TNA" designations are based on reference numbers in two redundant databases m aintained by 3M . b3, 3, 4, 4, 5, 5, 6, 6, 7, 7, 8, 8, 8-tridecafluorooctane sulfonic acid. 0 2-(N-ethylperfluoroctanesulfonamido)-ethyl alcohol. Sam ple Preparation We prepared three 1.0-mL aqueous buffer samples (a sample, a duplicate, and a "matrix spike") at each of six pH levels (1.5, 3, 5, 7, 9 and 11) for analysis at eight time intervals (0 ,7 ,1 4 ,2 1 ,2 8 ,3 5 ,4 2 and 49 days). Buffered solutions containing the analyte NMeFOSE alcohol and THPFOS (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 of all these samples. The chosen buffer solutions are described fully in Appendix A We prepared all the samples simultaneously, and placed all but the "Day 0" samples in an orbital incubator/shaker maintained at 50 ( 3) C. After at least three minutes of agitation, we diluted the "Day 0" samples 10:1 with methanol, added solutions of the internal standard 2-(A/-ethylperfluoroctanesulfonamido)-ethyl alcohol (A/-EtFOSE Page 11 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 alcohol), spiked the samples (as required) with a solution of A/-MeFOSE alcohol, and then refrigerated them. After the appropriate incubation times, subsets of the sample vials were removed from the incubator and then diluted, spiked, and stored as described immediately above. Except during the relatively short periods of time required to prepare them, the samples were shielded from light. In all the samples, the resulting THPFOS and A/-EtFOSE alcohol concentrations were 403 and 311 ng/ml, respectively. In the samples spiked with the anlatye N-MeFOSE alcohol, the resulting analyte levels were 221 ng/ml above those In the unsplked sam ples. Six calibration standards containing N-MeFOSE alcohol (78.9 to 789 ng/ml), A/-EtFOSE alcohol (311 ng/ml), THPFOS (403 ng/ml) and PFOS (3.1 to 94 ng/ml) served as the quantitative basis of the study. All these standards were prepared at the appropriate pH levels using the chosen buffer solutions (see Appendix A.) Sam ple Analysis The equipment we used for the HPLC/MS analysis was a Hewlett Packard model 1100 equipped with a Dionex lonPac NG-1 HPLC column (aqueous ammonium acetate/methanol solvent gradient) and an ALS Model G1322A degassing module. An ALS Model G1315A column heater maintained the column temperature at 40 C, a quaternary pump supplied a column flow rate of 0.3 mL/min, and an ALS Model G1313A auto-sampler provided 5 pL sample injections. The detector was a Hewlett Packard MSD mass spectrometer, operated in negative-mode electrospray ionization mode; anions of PFOS, THPFOS, and acetate adducts of A/-MeFOSE alcohol and A/-MeFOSE alcohol were detected at the charge-to mass ratio (m/z) values 499,427, 616, and 630 respectively, We processed the resulting data using the computer program HP ChemStation for LC (Rev.A.06.0). The calibration curves and analytical results are based on the measured area ratios for the analyte/internal standard pairs; A/-EtFOSE alcohol served as the internal standard for A/-MeFOSE alcohol, and THPFOS served as the internal standard for PFOS. Additional analytical details, including the gradient elution program, instrum ent and de te cto r param eters, and perform ance specificatio ns, are presented in Appendix A. D ev iatio n s The gradient parameters and column temperature were incorrectly entered on the instrument parameters spreadsheet dated 12/17/99. The correct parameters are listed in the data acquisition method file "FOSESIM.m." Through either human or mechanical error, samples for "Day 42," pH 1.5 were not analyzed. The mass spectrometer detector gain was set to 2.0 for samples analyzed on 12/20/99. Earlier sets were analyzed with gain = 1.0. The pH 3 data were reanalyzed using the correct quantitative ion at m/z = 616; an earlier analysis incorrectly employed the "monitor" ion value m/z = 617. Page 12 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 The actual concentration of the THPFOS solution added as a surrogate (21,030 |ig/mL) was incorrectly entered as 20,130 pg/mL in some documents. This error has no effect on the accuracy of the analytical results, but has been noted on the reagent preparation logs. The analytical results for one set of samples, intended for the pH 1.5, "Day 49" analysis, indicated much lower concentrations of N-MeFOSE than for any other similar sample prepared for this study. All other analytical results were quite consistent, so it is highly likely that these samples were not properly prepared. Incorrect amounts of either the surrogate or internal standard solutions were added to the following pH = 1.5 samples: M FA-002,127,128, and 129. The same Is true of the following pH = 11 samples: MFA-016, 017, 018, 034, 035, 036,124,125, and 126. In many cases, these samples were rejected on the basis of the data quality objectives (see the section immediately below). The resulting analytical values for the surrogate and internal standards were also excluded from evaluations of the consistency of the related results (see the "Data Summary and Discussion" section below). Page 13 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Results and Discussion D ata Q uality O bjectives (DOC^s) Below is a brief description of the data quality objectives applied in this study. 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 by pH level. Calibrations. The minimum acceptable coefficient of determination (r2) for linear fits to calibration data is 0.990. The acceptance criterion for individual calibration points is that their values fall within 25% of the linear fit value; data outside this range are excluded and the calibration curve is recalculated. No more than two points may be rejected from a calibration data set. Data for the high or low calibration standards may be rejected, though this results in a smaller effective calibration range. Continuing Calibration Verification (CCV). Identical calibration samples are examined at the beginning and end of each sample run. Results of the second calibration run may not deviate by more than 25% of the first run for any analyte. The average results of the calibration runs are used to calculate the analyte concentrations. M atrix Spikes. The acceptable percent spike recovery range is 75% 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 neither 5% of the highest calibration standard nor 25% of the lowest calibration level. System Suitability. Suitability was demonstrated by either an abbreviated mass-tocharge (m/z) check-tune or performance of a full auto-tune routine. Anom alous A nalytical Results Calibrations. Of the 144 calibration results obtained, 13 individual values failed to meet the stated DQO and were rejected. We rejected fewer than two values for any particular calibration run and compound. S p ik e R ecoveries. Results for the following sample pairs failed to meet this data quality objective and are excluded from the following analyses: MFA-127-128 ("Day 49", pH = 1.5); MFA-004-005 ("Day 0", pH = 3.0); MFAr016-017 ("Day 0", pH = 11); MFA034-035 ("Day 7", pH = 11); and MF/V124-125 ("Day 42", pH = 11). Sample Duplicates. The following sample pairs failed to meet this data quality objective and are excluded from the following analyses: MFAr001-002 ("Day 0", pH = 1.5) and MFAr040-041 ("Day 14", pH = 3.0). Solvent Blanks. PFOS results for two of the 12 solvent blanks (the initial blanks performed at pH levels 3.0 and 7.0) exceeded 25% of the calibration standards values below 15.7 ng/ml. As a result, the detection limit for PFOS quoted in this study is 15.7 ng/ml (see Appendix A). Page 14 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 S tatistical Methods and C alculations Using functions provided in Microsoft Excel software, we calculated means, standard deviations, and first-order rate constants (see Appendix B, Equation B8) for various subsets of the acquired data. Our linear regressions included the determination of constant terms, that is, we did not force the regression fits to pass through the origin. As described in Appendix B (Equations B38 and B39), rates measured at 50C were extrapolated to 25C by dividing by a factor of 10; this approximation is valid for reactions with Arrhenius heats of activation near 18 Kcal/mole.2 D ata Sum m ary and Discussion The LOQ is defined as the concentration of the lowest (accepted) standard in the calibration set for which the known concentration exceeds 400% of the indicated solvent blank level (see Appendix A). During this study, the LOQ's for A/-MeFOSE alcohol and PFOS were 78.9 ng/mL and 15.7 ng/mL, respectively. Excluding those samples noted above in the "Deviation" section, our results for the surrogate compound (THPFOS) and the internal standard (A/-EtFOSE alcohol) were quite consistent throughout the study. The percent relative standard deviations of the measured THPFOS values, calculated for each pH level, ranged from 2.1% to 10.2%. The corresponding values for A/-EtFOSE alcohol ranged from 4.7% to 11.3%. Figure 2 illustrates the observed N-MeFOSE alcohol concentrations at the six pH levels (at 50C), and Table 4 presents the results of the slope determinations (see Equation B8) for the same data. Table 4. Observed (50 C) Degradation Rates o f M-MeFOSE Alcohol in Aqueous Buffered Solutions and at Various pH Levels. PH Observed Rate (day1) Percent (2a) Rate Uncertainty 1.5 4.26 x 10'3 3.0 -4.42 x 10-4 5.0 5.68 x 10'3 7.0 4.83 x 10'3 9.0 3.97 x 10'3 11 2.98 x 10'3 67 582 25 24 34 25 These degradation rates, with the exception of the pH = 3.0 data, are generally well determined, with percent relative 2a (95% confidence) uncertainties in the range 24% to 67%. The data do not indicate any clear dependence of the degradation rate on the sample pH. Page 15 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 In the absence of a clear trend relating the degradation rate to sample pH, it is appropriate to "pool" the data from all pH levels and to determine the degradation rate using the entire data set. Figure 3 illustrates the results of this pooled analysis according to Equation 1, and Table 5 summarizes the results of the analysis. Table 5. Degradation Rate and Half Life o f M-MeFOSE Alcohol in Aqueous Buffered S olutions Using Data Pooled Over pH Levels. Observed Rate Constant at 50 C (day1) 0.0030 Percent (2c) Rate Constant Uncertainty at 50 C (day1) 66 Calculated Rate Constant at 25 C (day1) 0.00030 Calculated Half Life at 25C (years) 6.3 Calculated (2c) Half Life Range at 25 C (years) 3.8 to 19.4 Figure 2. Observed N-MeFOSE Alcohol Degradation fo r Various pH Levels p H 1.5 pH 3.0 ......pH 5.0 - - p H 7.0 - - p H 9.0 - - - p H 11 Page 16 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Figure 3. Pooled N-MeFOSE Alcohol Data and Slope Regression. We also monitored the concentration of the hydrolysis product perfluorooctane sulfonate (PFOS), but never observed this compound at levels above Its limit of quantification (LOQ, equal to 15.7 ng/mL). The initial A/-MeFOSE alcohol concentration (473 ng/ml) and the PFOS LOQ provide a second estimate of the N-MeFOSE alcohol half-life (see in Appendix B, Equations B32 and B33). The maximum degradation rate is given by E quation 3: (k p )_= 0 m=l and the minimum half-life is given by Equation 4 A t [P0]ta (2 ) a l q PFOS Eq. 3 E q .4 Page 17 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 We note that in both Equations 3 and 4, the initial A/-MeFOSE alcohol concentration (P0) and the PFOS LOQ (Aps) are molar quantities. Table 6 presents the results of the calculation. Table 6. Degradation Rate and Half Life o f W-MeFOSE Alcohol in Aqueous Buffered Solutions Based on PFOS Lim it o f Quantification At (days) 49 [P j (nm/ml) 0.85 a loq ^ P FO S (nm/ml) 0.029 Maximum Observed Rate at 50 C (day'1) 7.0 x 10^ Maximum Calculated Rate at 25 C (day'1) 7.0 x1 0 s Calculated Half Life at 25 C (years) >27 Page 18 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Conclusions We have performed a study of the aqueous hydrolytic degradation 2-(A/-ethylperfluorooctanesulfonamido)-ethyl alcohol (A/-MeFOSE Alcohol). Six different pH levels were included in the study, which were carried out at 50C and extrapolated to 25C. Our results based on direct observation of the N-MeFOSE alcohol concentration indicate no clear dependence of the degradation rate of A/-MeFOSE alcohol on pH. From the data pooled over the six pH levels, we estimate that the hydrolytic half-life of N-MeFOSE alcohol at 25C lies between 3.8 and 19.4 years, with the most likely value of 6.3 years. The concentration of the compound PFOS, a likely hydrolytic product of A/-MeFOSE Alcohol, was monitored throughout the study, but remained undetected above its limit of quantification (LOQ = 15.7 ng/mL). Using the LOQ for PFOS and the initial A/-MeFOSE alcohol concentration (473 ng/ml), and assuming PFOS is the only hydrolytic product of N-MeFOSE Alcohol, the data indicate that the hydrolytic half-life of N-MeFOSE alcohol at 25C is greater than or equal to 27 years. Page 19 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 References 1"Fate, Transport and Transformation Test Guidelines: 835.2110: Hydrolysis as a Function of pH," U.S. EPA Office of Prevention, Pesticides and Toxic Substances, publication number 712-C-98-057, January 1998. 2"Experimental Physical Chemistry", F. Daniels, et al., McGraw Hill Book Co. (New York), p. 131,1962. Page 20 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 ______________________________O 3 /^ /0 / William K. Reagen, Ph.D., Laboratory Management Date Page 21 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix A: Analytical Method ETS - 8-178.0, "Preparation of 2-(N-Methylperfluorooctanesulfonamido) Ethyl Alcohol (N-EtFOSE Alcohol) Hydrolysis Samples and Analysis by High Performance Liquid Chromatography with Mass Spectrometry Detection." This Appendix presents the analytical method employed in this study. Page 22 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 3M Environmental Laboratory Method Preparation of 2-(N-M ethylperfluorooctanesuufonamtoo)-ethyl alcohol (N-MeFOSE Alcohol) Hydrolysis Samples and Analysis by High Performance Liquid Chromatography with Mass Spectrometry Detection M ethod N um ber: ETS-8-178.0 Adoption Date: ^fl^ o o Approved by: Exact C opy o f O riginal j y r q/wjw In itia l D ate Revision Effective Date: Page 23 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 1.0 Scope and Application_________________________________ 1.1 This procedure defines the steps for analysis o f 2-(Nmethylperfluorooctanesulfonamido)-ethyl alcohol (N-MeFOSE alcohol) hydrolysis samples by high performance liquid chromatography (HPLC) with mass spectrometry (MS) detection and quantitation. It is based on EPA OPPTS: 835.2110 (Reference 18.1). N-MeFOSE alcohol and the potential hydrolysis product perfluorooctane sulfonate (PFOS anion) are detected and quantified by this method. Internal standards for the method are 2-(N-ethylperfluorooctanesulfonamido)ethyl alcohol (N-EtFOSE alcohol) and 3 ,3 ,4 ,4 , 5, 5 ,6 , 6 ,7 , 7, 8, 8, 8-tridecafluorooctane sulfonic acid anion (THPFOS). This method may also be used to screen for the presence o f FOSA (perfluorooctanesulfonamide). Representative structures are shown in Attachment A. 1.2 Com patible analytes. 2-(N-methylperfluorooctanesulfonamido)-ethyl alcohol (NMeFOSE alcohol), perfluorooctanesulfonate (PFOS anion), 2-(Nethylperfluorooctanesulfonamide)-ethyl alcohol (N-EtFOSE alcohol), 3 , 3 , 4 , 4 , 5 , 5 , 6 , 6 , 7,7, 8, 8 ,8-tridecafluorooctane sulfonic acid anion (THPFOS) and perfluorooctanesulfonamide (FOSA). 1 3 This is a performance-based method. Target analyte or surrogate matrix spike recoveries (100 25%) are used for each sample matrix to evaluate method performance. (Refer to Section 10 for the quality control parameters to be analyzed by this method. Refer to Section 14 for the quality assurance evaluation criteria for this method.) 2.0 Summary of Method______________________________________________________ 2.1 Aliquots o f N-MeFOSE alcohol stock solution are added to vials that contain buffers at pH 1.5, 3.0, 5.0,7.0, 9.0 and 11.0. The vials are then placed in an orbital incubator/shaker set at 50.0 3 C. Sets of vials are removed at designated intervals and the date and time recorded. The aqueous sample from the hydrolysis o f N-MeFOSE alcohol is diluted tenfold with methanol (MeOH). The parent compound, N-MeFOSE alcohol, and the PFOS hydrolysis product are separated on a Dionex IonPac NG1 reversed-phase HPLC column using an ammonium acetate/MeOH solvent gradient, with detection/quantitation by electrospray ionization mass spectrometry in the negative mode. 3.0 Definitions 3.1 Solvent blank. A sample o f analyte-free medium (for example, methanol) that is not taken through the sample preparation process. This blank is used to evaluate instrument contamination. 3.2 Sample duplicates. Two samples taken from and representative o f the same sample source and separately carried through all steps o f the extraction and analytical procedures in an identical maimer. Duplicate samples are used to assess variance o f the total method, including sampling, extraction, and analysis. 3.3 M atrix spike (MS). Prepared by adding a known mass o f target analyte to a specified amount o f a sample matrix. This assumes that an independent estimate o f target analyte concentration is available. Matrix spikes are used to determine the effect o f the matrix on method recovery efficiency. ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 24 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 3.4 C alibration standard. A dilution o f various amounts o f a stock, intermediate or purchased standard to achieve standard solutions in a concentration range o f interest. Hydrolytic half-lives resulting from these analyses are calculated based on analytical ratios and not absolute numbers. Therefore, results do not depend on the purity o f the standards used. 3.5 In tern al standard. A known amount o f a compound or element similar in analytical behavior to the compound(s) or elem ents) of interest, added to all samples and standards, and carried through the entire measurement process (post-hydrolysis, alter final dilution). It provides a reference for evaluating and controlling the precision and bias o f the applied analytical method. 3.6 Surrogate. An organic compound which is similar to the target analyte(s) in chemical composition and behavior in the analytical process, but which is not normally found in the sample(s). In hydrolysis studies, surrogate is added to CCVs, samples, sample duplicates, and matrix spike samples along with the test analyte (pre-hydrolysis). 3.7 C ontinuing calibration verification. (CCV). Standards analyzed during an analytical run to verify the continued accuracy o f the calibration curve. This solution may or may not be prepared from a different source or lot number than the calibration curve standards. 3.8 D ilution. A step in the hydrolysis study procedure in which a solvent is added to the test analyte/buffer solution to prepare it for instrumental analysis. This step occurs alter the vials are removed from incubation and before the samples are analyzed. If the solvent used is miscible with the test analyte/buffer solution, the diluting solvent is merely added and mixed. I f the diluting solvent is non-miscible, a liquid-liquid extraction is performed. 3.9 L im it o f quantitation (LOQ). The lowest concentration that can be reliably measured within specified limits o f accuracy (see Sections 14.1 and 14.2) and precision (see Sections 14.3 and 14.4) during routine laboratory operating conditions. The LOQ is generally 5 to 10 times the minimum concentration with a 99% confidence lim it that the concentration is greater than zero. However, it may be nominally chosen within these guidelines to simplify data reporting. For many analytes, the LOQ is selected as the lowest non-zero standard in the calibration curve that is greater than 4 times the level o f the solvent blanks. Sample LOQs are highly matrix-dependent. 3.10 A ccuracy. The closeness o f agreement between an experimentally determined value and an accepted reference value. When applied to a set o f observed values, accuracy is a combination o f a random (precision) and a common systematic (bias) component. For purposes o f the study, the acceptance criterion is 75% to 125% o f the nominal value. 4.0 W arnings and Cautions___________________________________________________ 4.1 Health and safety warnings 4.1.1 Wear the proper lab attire for all parts o f this procedure. Wear gloves and proper eyewear at all times. 4.1.2 Handle all solvents in a hood for all parts of the described sample preparation procedure. Whenever possible and practical, dilute samples with solvent in a hood. 4.1.3 For potential hazards o f each chemical used, refer to material safety data sheets, packing materials, and the 3M Environmental Laboratory Chemical Hazard Review. ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 25 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 4.2 Cautions 4.2.1 All glassware in which standards are prepared should be triple-rinsed with 1:1 acetone/MeOH to reduce the possibility o f contamination. 4.2.2 Ensure that the HPLC mobile phases are prepared prior to beginning a run sequence, and that there is sufficient quantity to complete the run. Do not allow the pump to run dry. 4.2.3 Ensure that before starting the run sequence there is ample hard disk space on the computer to save all run data. 4.2.4 Ensure that there is enough nitrogen in the supply tank to complete sequence runs. 5.0 Interference________________________________________ ____________ 5.1 Contaminants in solvents, reagents, glassware, and other sample processing or analysis hardware may cause interference. Use the routine analysis of laboratory solvent blanks to demonstrate that there is no such interference. 5.2 Contamination from columns, HPLC tubing, and detector components may cause interference at low detection levels. The routine analysis o f solvent blanks must be used to demonstrate that there is no such interference. 6.0 Equipment_______________________________________________________ 6.1 Analytical balance sensitive to 0.1 mg 6.2 Incubator/shaker capable o f maintaining temperature at 50.0 3 C 6 3 Hewlett-Packard (HP) 1100 HPLC System, or equivalent 63.1 Pump, binary, Model G l 312, or equivalent 6 3 .2 Solvent degasser, Model G l 322A or equivalent 6 3 3 Autosampler, ALS Model G1313A, variable injection volume capable 6 3 .4 Column heater, Model G1316A 6.4 Dionex IonPac NG1 Guard column, 4 x 3 5 mm, or equivalent 6.5 Mass spectrometer. Hewlett-Packard MSD Model G l 946A, or equivalent, operating in electrospray-negative SIM mode 6.6 Clock, digital. Only one clock should be used, to insure unambiguous documentation o f the correct performance o f procedures. 6.7 pH meter. Coming Model 308 pH/Temperature Meter with 3-in-l gel-filled combination electrode (pH/reference/temperature), or equivalent 6.8 Refrigerator capable o f maintaining 4 3 C 6.9 Data system. A personal computer capable o f controlling the HPLC system as well as recording and processing signals from the detector, Hewlett-Packard ChemStation Version A.06.01 or later 7.0 Supplies and Materials__________________________________________________ 7.1 Vials, 40 mL, VOA (I-Chem or equivalent) 7.2 Crimp cap autovials, 1.8 mL 7 3 Labels 7.4 Graduated pipets, glass, disposable, 1 mL to 10 mL 7.5 Pasteur pipets, glass, disposable ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 26 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 7.6 Hamilton Gastight syringes (precision 1% o f total volume), 10 pL-lOOOpL 7.7 Volumetric flasks, various sizes 7.8 Beakers, glass, various sizes 7.9 Automatic pipettor, capable o f dispensing 10-5000 pL 8.0 Reagents and Standards_______________________________ 8.1 M ethanol (MeOH). HPLC/SPEC/GC grade from EM Science, or equivalent 8.2 Acetone. HPLC/SPEC/GC grade from EM Science, or equivalent 8 3 18.0 M Q w ater. Water with lower resistance must not be used. 8.4 Ammonium acetate, 2 mM in water. This solution is chromatographic solvent A (see Section 12.3.1). (Example: An acceptable eluent solution is made by adding 0.15g ammonium acetate crystals to a 1-L volumetric flask containing about 500 mL 18.0 MQ water, adding 10 mL o f methanol, diluting to the mark with 18.0 MQ water and mixing.) 8.5 Stock, internal stan d ard and calibration solutions A ll weights should be recorded to the nearest 0.0001 g in a standards preparation log: 83.1 N-MeFOSE alcohol prepared in acetone. (Example: A stock solution is prepared at a concentration o f approximately 30,000 pg/mL by weighing 0.3 g o f N- MeFOSE alcohol in a 10-mL volumetric flask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) . 8.5.2 N-EtFOSE-alcohol internal standard prepared in acetone. (Example: A stock solution is prepared at a concentration o f approximately 30,000 pg/mL by weighing 0.3 g o f N-EtFOSE-OH in a 10-mL volumetric flask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) 8.5.3 Perfluorooctanesulfonate (PFOS) prepared in acetone. (Example: A stock solution is prepared at a concentration o f approximately 3000 pg/mL by weighing 0.06 g o f PFOS in a 20-mL volumetric flask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) 8.5.4 3, 3 ,4 ,4 , 5, 5, 6, 6, 7 ,7 , 8, 8, 8-tridecafIuorooctane sulfonic acid (TH PFO S) internal standard prepared in MeOH. ( Example: A stock solution is prepared at a concentration of approximately 20,000 pg/mL by weighing 0.2 g o f IH PFO S in a 10-mL volumetric flask and bringing to the mark with MeOH. This solution is diluted in MeOH to make additional, appropriate standards.) 8.6 Buffers for calibration of pH m eter Purchased pH calibration standards of pH 4.0,7.0, and 10.0 (suppliers vary). 8.7 Buffer solutions for hydrolysis study. Prepare buffer solutions o f pH 1.5,3.0, 7.0,9.0 and 11.0 using guidelines from CRC Handbook o f Chemistry and Physics (Reference 18.2). Prepare buffer solution o f pH 5.0 using guidelines from Fate, Transport and Transformation Test Guidelines (Reference 18.2). Prepare the buffer solutions in 1-liter quantities. Calibrate a portable pH/temperature meter using purchased pH calibration standards o f pH 4.0,7.0, and 10.0, and measure the pH of all buffer solutions. Prepare buffer solutions o f pH 1.5,3.0, 5.0,7.0,9.0 and 11.0 at ambient room temperature. The ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 27 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 concentrations are given below. Record final pH measurements o f all buffers. Store buffers in sealed glass containers. 8.7.1 pH 1.5 8.7.1.1 207 mL o f 0.1 N HC1 (reagent grade) 8.7.1.2 125 mL o f 0.2 M KC1 (reagent grade) 8.7.13 Add 18.0 MQ water to about 900 mL total volume 8.7.1.4 Adjust pH to 1.5 w ith additional 1 N HC1 8.7.13 Bring to a final volume o f 1 L with 18.0 MQ water 8.7.2 pH 3.0 8.73.1 223 mL o f 0.1 MHC1 (reagent grade) 8.7.2.2 500 mL o f 0.1 M potassium hydrogen phthalate (reagent grade) 8.7.2.3 Add 18.0 MQ water to about 900 mL total volume 8.73.4 Adjust pH to 3.0 with 1 N HCI or 1 N NaOH 8.73.5 Bring to a final volume o f 1 L with 18.0 MQ water 8.73 pH 5.0 8.73.1 467 mL o f 0.1M NaOH (reagent grade) . 8 .7 3 3 500 mL o f 0.1 M monopotassium citrate (reagent grade) 8 .7 3 3 Add 18.0 M il water to about 900 mL total volume 8.73.4 Adjust to pH o f 5.0 with IN NaOH or IN HCI 8.73.5 Bring to a final volume o f 1 L with 18.0 M Q water 8.7.4 pH 7.0 8.7.4.1 500 mL 0.1 M KH2P 0 4buffer (reagent grade) 8.7.43 291 mL 0.1N NaOH (reagent grade) 8.7.43 Adjust to pH 7.0 with either 1 N HCI or 1 N NaOH 8.7.4.4 Bring to a final volume o f 1 L with 18.0 MQ water. 8.7.5 pH 9.0 8.7.5.1 500 mL 0.025 M sodium borate decahydrate (reagent grade) 8.7.53 46 mL o f 0.1N HCI (reagent grade) 8.7.53 Add 18.0 MQ water to approximately 900 mL 8.7.5.4 Adjust to pH 9.0 with either 1 N HCI or 1 N NaOH 8.7.5.5 Bring to a final volume o f 1 L with 18.0 MQ water. 8.7.6 pH 11.0 8.7.6.1 500 mL 0.05 M NaHC02(reagent grade) 8.7.63 227 mL 0.1 N NaOH (reagent grade) 8.7.63 Add 18.0 MQ water to approximately 900 mL 8.7.6.4 Adjust pH to 11.0 with IN NaOH 8.7.6.5 Bring to a final volume of 1 L with 18.0 MQ water 8.8 Test analyte and spike solutions: . 8.8.1 N-M eFOSE alcohol test analyte solution w ith THPFOS surrogate. [Example: An analyte solution o f N-MeFOSE alcohol, TN-A-1282, at approximately 500 pg/mL and THPFOS at approximately 400 pg/mL is used (a dilution in MeOH o f the solutions prepared in Sections 8.5.1 and 8.5.4). A 10-pL aliquot o f this solution added to 1 mL buffer (step found in Section 12.1.5) results in a final concentration o f approximately 500 ng/mL N-MeFOSE alcohol and ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 28 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 8.8.2 8 .8 3 approximately 400 ng/mL THPFOS after MeOH dilution (step found in Section 12.1.13)]. N-EtFO SE alcohol internal standard solution. [Example: An analyte solution o f N-EtFOSE alcohol, TN-A-1284, at approximately 30 pg/mL is used (a thousand-fold dilution in MeOH o f the solution prepared in Section 8.5.2). A 100-pL aliquot o f this solution added to lm L buffer (step found in Section 12.1.5) results in a final concentration o f approximately 300 ng/mL after MeOH dilution (step found in Section 12.1.13)]. Spiking solution. [Example: A spiking solution is prepared by adding 10-pL o f the N-MeFOSE alcohol stock solution (Section 8.5.1) to a 10-mL volumetric flask and diluting to the mark with methanol. A 70-pL aliquot o f this solution added to the 1 mL sample (Section 12.1.15) results in a final spike concentration o f approximately 210 ng/mL o fN-MeFOSE alcohol]. 9.Q Sample H andling__________________'___________________________ ___________ 9.1 Record times o f initial preparation and dilution on the fluorochemical degradation (hydrolysis) analysis sample preparation sheet (Attachment B). 9.2 For Time 0 samples, aliquot only the lm L o f buffer into the vials. DO NOT spike with test analyte. Store the vials at room temperature until ready to analyze. Then proceed from Section 12.1.12. 9 3 Once the 9.0 mL o f diluting solvent has been added to the hydrolysis mixtures, the samples should be analyzed as soon as possible. Alternatively, aliquots o f the methanoldiluted samples should be refrigerated at 4 3 C until analysis can be performed. 10.0 10.1 10.2 103 10.4 Quality Control______________________________________ __________________ Sam ple D uplicates. Prepare and analyze all samples in duplicate to provide a measure o f the precision o f analysis. M atrix spikes. Prepare a post-hydrolysis matrix spike sample (Section 8.8.3 and 12.1.12) for each interval and pH level used in the study. Concentrations o f the spike should be approximately equal to a mid-range calibration standard. The matrix spike sample should be analyzed immediately following the sample duplicates to which it corresponds. The analyst shall accept percent spike recoveries o f 100 25% . Spike recoveries outside o f this range should be noted. Appropriate steps must be taken to correct the problem before analysis is allowed to proceed. Before the analysis is allowed to proceed, consult with the Team Leader or designee for direction and final acceptance or rejection o f the analytical run. Solvent blank. Solvent blanks should be run before and after every calibration curve, CCV, and after no more than 20 injections. Acceptable values for the blank are values less than 25% o f the LOQ standard. If analyte carryover is a problem, use back-to-back solvent blanks. C ontinuing C alibration V erification (CCV). A standard analyzed periodically during an analytical run to verify the continued accuracy o f the calibration curve and is run in tandem with the solvent blank. This solution may be prepared from a different source or lot number than the calibration curves standards. ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 29 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 10.5 In tern al standard. Internal standard is added post-hydrolysis (after final dilution) to all standards, samples, and matrix spikes at a constant concentration. 11.0 Calibration and Standardization_________________________________________ 11.1 S tandard preparation. Prepare six calibration standards containing N-MeFOSE alcohol, N-EtFOSE alcohol, PFOS and THPFOS in 9:1 MeOH:bufFer for each pH level. Standards from approximately 75 ng/mL to 750 ng/mL o f N-MeFOSE alcohol and 3 ng/mL to 100 ng/mL o f PFOS are suggested. A level o f approximately 400 ng/mL THPFOS surrogate and 300 ng/mL o f N-EtFOSE alcohol internal standard in each calibration standard is suggested. 11.2 C alibration standards. Analyze the calibration standards at the beginning and end o f the run. Average the peak area response from both curves. Use the data reduction software program for linear regression calculations to relate the analyte peak area ratio versus amount ratio, rising internal standard calibration. Use N-EtFOSE alcohol as the internal standard for N-MeFOSE alcohol quantification, and THPFOS surrogate as the "internal standard" for PFOS quantitation. Quadratic regression may be used if data review shows this to be a consistent and more accurate representation o f the instrument response. Consult with the Team Leader for direction prior to performing the quadratic calibration methodology. 12.0 Procedures________________ ______________________________________________ 12.1 Sam ple and spike preparation 12.1.1 Before spiking with any o f the stock standards, transfer approximately 1 mL of the solution to an autovial and cap the vial. Use this smaller volume for spiking to minimize the effects o f evaporation from stock solutions and to prevent contamination o f the larger volume of stock solution. 12.1.2 Determine the number of time intervals that will be analyzed. Each interval will have three vials for each pH, multiplied by the number o f pHs analyzed. One vial at each level will be labeled as sample, duplicate, and spike. 12.13 Obtain the appropriate number of 40-mL VOA 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 number and i.d., temperature, pH, time interval, test analyte, and date o f preparation. Record the pH o f each buffer solution. 12.1.4 Remove the cap o f the VOA vial and add 1 mL o f the appropriate buffer solution to all o f the pre-labeled vials. Always replace the cap immediately after any addition to minimize evaporation. 12.1.5 P u t "Time 0" samples aside at this p o in t For all other sam ples, continue on to section 12.1.6. 12.1.6 To all o f the vials, add 10 pL o f the combined N-MeFOSE alcohol and THPFOS analyte solution (Section 8.8.1) with a 25-pL Hamilton Gastight syringe. 12.1.7 Make sure that the cap has been firmly tightened and place the samples back in the cardboard case. ETS-8-I78.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 30 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 12.1.8 Place the case into a pre-warmed incubator/shaker for the appropriate time. Record the time, temperature, and rate o f shaking. The temperature is determined by the conditions o f the experiment. Continue to manually monitor the incubator temperature daily during the entire incubation. Record the temperature on the sample preparation sheet (Attachment B). 12.1.9 Store "Time 0" samples at room temperature until the time o f analysis. 12.1.10 Remove the case from the incubator at the designated preset time. 12.1.11 Remove the vials from the case and place in racks. Allow the vials to cool for approximately 15 minutes to room temperature. 12.1.12 While vials from the first time point are cooling, spike the stored `T im e 0" samples with test analyte solution (Section 12.1.6). Then continue on to section 12.1.13 with all samples. 12.1.13 Add 9 mL o f methanol to each vial. 12.1.14 Using a 100-pL gas tight syringe, add 100 pL of N-EtFOSE alcohol internal standard solution (Section 8.8.2) to each sample and spike vial. 12.1.15 Using a 100-pL gas tight syringe, add 70 pL o f N-MeFOSE alcohol spiking solution (Section 8.8.3) to the sample spike vials. Shake the vials for three minutes by hand or Vortex mixer to mix the contents and extract any analytes that may have adsorbed to the vial. 12.1.16 Aliquot approximately 1 mL o f each sample to the appropriately labeled autovial, cap, and refrigerate at 4 3 C until analysis. 12.2 Instrument set up 12.2.1 Check that the appropriate HPLC column is in the instrument for analysis. 12.2.2 Check that the correct eluent solutions are in bottles to be used and that enough is available to complete the sequence run. 12.2.3 Place the samples in the autosampler tray and construct a sequence table with appropriate calibration standards, calibration check standards and solvent blanks. 12.2.4 Verify that all samples and standards are positioned correctly. Enter sequence information (sample or standard ID, method name). Use one injection per sample. 12.2.5 Save sequence as analysis date and instrument letter (e.g. on March 14,1999, save sequence table as 031499.S). Save all data to a subdirectory labeled with analysis date. (e.g. 031499). 12.2.6 Set post-sequence command macro to shut down system after the run is completed (Example: "STANDBY" on HP1100/MSD systems). 12.3 H PLC set up: 12.3.1 Analysis o f N-MeFOSE alcohol hydrolysis samples in buffers at pH levels 1.5, 3.0,5.0,7.0,9.0 and 11.0. Column: Dionex IonPac NG1 Guard column, 4 x 3 5 mm, or equivalent Solvent A: Ammonium Acetate 2mM in water (with 1% MeOH). Solvent B: Methanol Recommended Solvent Gradient: ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 31 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Tim e (m i n ) %A 0.0 60 1.0 60 4.0 5 11.0 5 Post time: 6 minutes, column temperature: 40C. 12.4 Recommended mass spectrometer set up*: %B 40 40 95 95 MSD: Ionization mode API-ES Polarity Negative Acquisition mode SIM SIM resolution High Time filter Enabled Peakwidth 0.25 min Gain 1.0 Fragmentor 70 Dwell time 183 msec Capillary voltage 3500 Drying gas Nitrogen Nebulizer pressure 30 psig Drying gas flow 8L/min Drying gas temp 300* C Example conditions are applicable to HP1100/MSD equipment only. 12.5 Auto-sampler setup*: A uto-sa m p le r : A uto-sam pler Program: ALS Model G1313A None In je c t io n v o l u m e : 5.0 pL Example conditions are applicable to Hewlett Packard 1100 only Flow rate 0.3 mL/min 0.3 mL/min Q.3 mL/min 0.3 mL/min 12.6 Ions used for identification and quantification: A pprox. R e t e n t io n T im e ( m in ) 8.4 C o m p o n e n t Na m e MeFOSE-OH D e s c r ip t io n Test Analyte Q u a n t if ic a t io n Ion 616 8.6 EtFOSE-OH Internal Standard 630 62 PFOS' Potential Degradation 499 Product . ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS M o n it o r Io n 617 631 500 Page 32 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 5.9 THPFOS' Surrogate 427 None * FOSA Qualitative use only None 498 *No calibration standards analyzed, so no retention times available. 12.7 Sample analysis 12.7.1 Enter the standard, sample, and QC information into the sequence table. Analyze calibration standards first, then up to 20 injections, followed by the calibration standards. If more than 20 injections are to be run, analyze a continuing calibration standard (CCV) after every 20 injections and run the calibration standards again at the end o f the sequence. Rim solvent (or method) blanks after the highest calibration standard, before and after the CCV, and after the set o f samples to check for any analyte carryover. 12.7.2 Place standards, samples, and QC (matrix spikes, sample duplicates, and blanks) into the autosampler tray according to the order they are listed in the sequence. 12.73 Identify the electronic acquisition files with an appropriate prefix (e.g. MeFOS). Do not exceed five characters if the sequence contains more than 99 lines. 12.7.4 Save sequence as analysis date (e.g. on March 14,1999, save sequence table as 031499.s). Save all data to a subdirectory labeled with analysis date (e.g. 031499). 12.7.5 Start the sequence. 13.0 13.1 133 133 13.4 D a t a A n a l y s is a n d C a l c u l a t io n s ____________________________________________ P eak Evaluation. Peaks must be symmetric in shape and identified by extracting compound-specific ions. Peaks considered for calibration must have peak heights greater than 5 (five) times the baseline noise for that region o f the chromatogram. Peak area integration is from baseline to baseline using automatic or manual integration. C alculation of Rate C onstant (A). Calculate the test analyte concentrations in each o f the pH matrices using the curves obtained from the calibrations. Assuming first-order kinetics a rate constant (k) can be determined by plotting: Ln H N -M eFO SE - d- --h! ") versus minus elapsed time (-t). The subscripts t and 0 refer " (^[N-MeFOSE- to analyte concentrations determined at some elapsed time t and at / = 0, respectively. The slope o f the resulting line is k. The r2value for this plot should be > 0.80. For r2 values less than this, consult the Team Leader or designee. T arget analyte concentrations. Calculate the Me-FOSE alcohol and PFOS concentrations in each o f the pH matrices using the curves obtained from the calibrations. M atrix spikes. Calculate the percent recovery for each o f the matrix spikes. Calculate the matrix spike percent recoveries using the following equation: % Recovery = (observed spiked sample result - observed sample result-! x 100 Actual amount spiked Using the observed matrix spike recoveries, calculate the average spike recovery. ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 33 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 13.5 Sam ple D uplicates. Calculate the relative percent deviation (RPD) for the duplicate samples: RPD = lA-Bl x 100% (A+B)/2 Where A= the concentration measured in the sample B= the concentration measured in the sample duplicate 14.0 14.1 14.2 14.3 14.4 14.5 14.6 Method Performance A ccuracy. For purposes of the study, the acceptance criterion is 75% to 125% o f the nominal value. Coefficient o f determ ination (r2). The coefficient o f determination (r2) for the calibration curves should be 0.99 or greater. The curves should be examined closely for linearity and intercept, particularly for accuracy o f quantitation at the low and high ends o f the curve. The accuracy o f all standards used for calibration must be within 75-125%. On occasion it may be necessary to use exponential or quadratic fits o f the data, usually when broad range curves (greater than 3 orders o f magnitude between the low and high concentration standards) are used. Document in the raw data the technical justification for using quadratic equations. M atrix spikes. The analyst shall accept percent spike recovery values o f 100 25%. Spike recoveries outside o f this range should be noted. Consult with the Team Leader or designee for direction, and for final acceptance or rejection o f the data. Data that are used in final report that is deemed out o f control will be required to have a technical justification for why the data are being used, documented in the final report and raw data. Sam ple duplicates. The analyst shall accept %RPD (See Section 13.5) values < 25%. %RPD values > 25% should be noted. RPD values o f 25% or greater should be noted. Appropriate steps must be taken to correct the problem before analysis is allowed to proceed (e.g. sample re-runs, additional blanks, etc.). Consult with the Team Leader or designee for direction, and for final acceptance or rejection o f the data. Continuing calibration verification (CCV). If the percent difference for the amount o f quantitated analyte is greater than 25% from the true value relative to the initial standard curve, stop the run. Only those samples analyzed before the last acceptable calibration check standard w ill be used. Consult with the Team Leader or designee for direction, and for final acceptance or rejection o f the data. In tern al standard and Surrogate. Review o f the internal standard and surrogate can be performed by averaging the area response throughout the analytical run and calculating the relative standard deviation (%RSD). %RSD values >10% should be noted. Inconsistencies in the internal standard peak area may indicate instrumental changes over time. Inconsistencies in the surrogate peak area may indicate instrumental changes, changes in the test-system, or hydrolysis of the surrogate over time. Consult with the Team Leader or designee for direction and final acceptance or rejection o f the analytical run. ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 34 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 14.7 Limit of Quantitation (LOQ). The LOQ is equal to the lowest standard in the calibration curve that is greater than 4 times the level o f the solvent blanks. Ifblank contamination is present, it may he necessary to delete the lowest standard o f the calibration curve. 14.8 Solvent blanks. Solvent blanks should show no more than a 5% carryover from a high standard or calibration check standard. If so, two solvent blanks may be necessary to rule out instrumental contamination. If peaks with greater than 25% o f the peak area o f a low standard value are observed in sequential solvent blanks, it is indicative o f instrument contamination. The instrument shall be serviced by thoroughly cleaning the electrospray source, and replacing/cleaning columns, tubing, etc. 14.9 Specificity. Analyte specificity is demonstrated by acceptable post-hydrolysis analyte spike recoveries. 14.10 System Suitability. Without performing a method validation, system suitability can be demonstrated by acceptable instrumental checks (e.g. abbreviated ra/z check-tune, or full auto-tune routines). Consult the appropriate instrumental manuals (Reference 18.3). 15.0 Pollution Prevention and Waste Management___________________________ 15.1 Dispose o f sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose o f 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 bore columns when possible to minimize waste generation. 16.0 R ecords_________________________________________________________________ 16.1 Print hard copies o f all graphics and data analysis summaries for archiving. 16.2 Sign and date all data packages and label with instrument ID. 16.3 Fill out the hydrolysis sample preparation worksheet completely, making sure to include all initials and dates. . 16.4 Print out the sample sequence table, reduce the size with photocopying and tape the photocopy into the instrument log. Keep the original copy for the raw data files package. 16.5 Print chromatograms and quantification reports for all analyses. 16.6 Print calibration tables and curve information and store in die raw data file. 16.7 Store hydrolysis sample preparation worksheets in the raw data file. 16.8 Enter all standard preparation information in the standards preparation logbook. Make a photocopy o f the logbook page and include the copy in the raw data file. 16.9 Archive electronic data to appropriate media when necessary. 17.0 Attachments__________________________ _________________________________ 17.1 Attachment A. Representative chemical structures 17.2 Attachment B. Hydrolysis sample logsheet ETS-8-178.0 Prep, o f N-MeFOSE Alcohol Hydrolysis Samples and A nalysis by HPLC/MS Page 35 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 18.0 References______ ___________________________ _____________ 18.1 Fate, Transport and Transformation Test Guidelines Office o f Prevention, Pesticides and Toxic Substances (OPPTS) 835.2110 Hydrolysis as a Function o f pH, EPA 712-C-98057, January 1998. 18.2 C R C Handbook o f Chemistry and Physics, 1st Student Edition, "Buffer Solutions Operational Definitions o f pH," Robert C. Weast, Ph.D., 1988, p. D-87. 18.3 HP 1100 Series LC/MSD Reference Collection, Rev. A.00.01 June 1997 CD/ROM 19.0 Affected Documents 19.1 None. 2 0 .0 REVISIONS Revision number Reason for revision Date o f Revision ETS-8-178.0 Prep, o f N-M eFOSE A lcohol Hydrolysis Samples and Analysis by HPLC/MS BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Attachment A. Representative Chemical Structures N-MeFOSE Alcohol FW = 557.23 N-EtFOSE Alcohol FW = 571.25 FOSA FW=499.14 CgOlefin FW=382.07 i 1 PFOS FW=500.13 I II C8Hydride FW=420.07 j ?VH F F , r p f F il 1 1 . . . . THPFOSFW=428.17 A ttachm ent A ETS-8-178.0 Prep, o f MeFOSE A lcohol Hydrolysis Samples and Analysis by HPLC/MS Page 37 of 71 3M Environmental Laboratory Report No. W1880 Attachm ent B. H ydrolysis Sample Logsheet Fluoroctiemical Degradation (Hydrolysis)Analysis TEST ANAYLTE: Lai R ia u u t N um btr. N o m in al In ta n a i: Nomina! Temperatur*: C Incubator ID: Start Date: Stop Date: Incubation interval: days Time: Time: hours Freezer ID: Start Oate: Stop Date: min Sample No. Date: Time: Initials: Rep k it Tomp pH n 1.5 n 1.5 r3 1.5 ms 1.5 blk 1.5 rl 3 r2 3 i3 3 ms 3 blk 3 rt 5 r2 5 r3 5 ms 5 blk 5 rl 7 x2 7 r3 7 ms 7 blk 7 n9 r2 9 t3 9 ms 9 blk 9 rl 11 n 11 r3 11 ms 11 blk 11 B u ffe r Teat A n ahrte S p ik e S o lu tio n D ilu tio n S o lv e n t In te rn a l S ta n d a rd Solution ID Amount (ML) S o lu tio n - Am ount 0A-) - S o lu tio n .. - _ _ - Amount ft* .) ~ - S o lu tio n Am ount (m u S o lu tio n Amount (DU _ - Component ~ - ~~ _ .. .. .. .. .. Cone. (pg/mL) Component _ _ _ - Cone. (pg/mL) Component Cone. (pg/hiLJ Component Cone. (pg/mL) Comments: Time: Time: Com m ent I Attachment B ETS-8-178.0 Prep, o f MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS h- ^4o-- 00 CO 0O(00.) BACK TO MAIN BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix B: Kinetics Model This Appendix includes a mathematical description of the kinetics model employed in the study. Page 39 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 K in e tics Model B1. Reaction Components and Rates The arguments below are based on the following idealized set of reactions representing the hydrolysis of a parent compound P and its hydrolysis products A m, which number N. 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 Pm(forthe parent) and k Am(forthe parent's hydrolysis products). P + H 20 n mA m+ Y ml (m = l to N) (B1) A m+ H 20 Y,, (m = 1 to N) (B2) where the general symbols Y ml and Y m2 represent all 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 Equation B4 may be directly integrated to obtain the general solution ln [P ]= " X nmk Pm t +C v m=l y With the initial condition P(t - O) = P0, the specific solution to Equation B4 is (B3) (B4) (B5) using the additional definition of the total parent hydrolysis rate Page 40 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 N k p = X n " kpTM m=l (B7) Equation B6 can be re-written in a form that allows a least-squares estimate of the total parent hydrolysis rate: k pt = -In (B8) Using the initial (t = 0) measured value of the parent concentration P0and later values P measured at later times t , one can calculate and plot the (linear) quantity [- In (P/P0)] versus time and obtain a least -squares estimate of the slope of the line. The resulting slope is the least-squares estimate k p of the total parent hydrolysis rate. Equation B6 indicates that over a period of time T 1^ (the parent hydrolysis half-life) the parent concentration P is reduced through hydrolysis by a factor of two, where (B9) A least squares estimate of the parent hydrolysis half-life is therefore available from HZ) kP (B10) B3. Product Compound Concentrations The pseudo-first order differential changes in the product concentrations An (using Equations B2 and B6) are dAm= ( nmkPmP - kAmAm)dt = ( nmkPmP0 e-kp' - kAmAm)dt (B11) and the (first order, non-separable) differential equation governing the product concentrations is HA ^ + k AmA m= nmkPniP0 e~kpt . (B12) The "standard form" of Equation B12 is Page 41 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 A m+ S ( t ) A m = Q (t) where the "function" S (t) is actually a constant: S(t)=k*m and Q ( t ) = n mk PmP0e _ l f l . The general solution A mto Equation B12 is contained in (B13) (B14) (B15) A m e 's (,)d ,= j Q ( t ) e ' s(,')d ' d t + C (B16) where g J S (t)d t _ e J S (t')d t' _ gkAm Jd t _ g k ^ t (B17) and j Q ^ ^ ^ ' d t + C = n m k PmP 0J e k ^ e - k p ,d t + C (B18) There are two cases of Equation B18 to consider. In the circumstance that k Am = k p , which occurs only when the hydrolysis rate of the mth product is identical to the total parent hydrolysis rate, the general solution to Equation B18 is (forkAm= k P) A m ekpt = nmk PmP0 t + C (B19) and, using the initial conditionAm(t = 0) = A ^ , the specific solution to Equation18 is (for k ^ = k P) A m= (nmkPmP0 t + A,*, ) e'kp' . (B20) We note that when k ^ = k p = 0 (that is, when both the parent and potential product are hydrolytically stable), Equation B7 requires (also) that k Pm= 0, so Equation B20 becomes Page 42 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 A,, = AirO (B21) indicating, as required, that the product concentration does not change with time. The circumstance k Am = k p is highly improbable, and is neglected in the remainder of this discussion. However, the reader should bear in mind that the expressions derived below do not hold when the parent hydrolysis rate k pand the product hydrolysis ra te k ^ approach each other. In the more probable case, for which k Am * k p (i.e. that the hydrolysis rate of the mth product is different from the total parent hydrolysis rate), the general solution to Equation B18 is ^ Qk Amt _ | n k p m Po ( k Am- k p ) t I kAn, - k P (B22) and the specific solution to Equation B18 with the initial condition A m(t = 0) = A m0 is A,, = A,,n+ n mkpn,P0 kp - k Am t n m kp..m..P..Q c-kP t kp - km (B23) Of greatest interest here is the case in which the product compounds are known to be hydrolytically stable, that is, when k ^ = 0 for all m. In this case, Equation B23 becomes (for hydrolytically stable products) A m~ A m0 + nm^ FmP (l --e~kpt) . kP (B24) B4. R elationships Between the Parent and Compound Concentrations Equations B7 and B24 can be combined to obtain (for hydrolytically stable products) kP v K - A j (B25) Page 43 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 so that (for hydrolytically stable products) V ( A n , - A m0) ( l - e `kp' ) = Xif p0 or (for hydrolytically stable products) (B26) -Ikpt = -In ( A m ~ A mo) m=l P 0 (B27) If the changes in the product concentrations are all small compared to the original parent concentration, that is, if y Am- Am0 1, m-1 P 0 we may use the expression (valid for -1 < X < 1 ) (B28) ln(l + X) = X - - X 2 + - X 3 - - X 4 + . 234 and Equation B23 becomes (for hydrolytically stable products and ^mO <<P0) ( -- A -A k pt = - - r p V m-1 ^ J (B29) (B30) Page 44 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 or (for hydrolytically stable products and m =l Po) (B31) B5. Parent Half-Life Estimates Based on Lim its o f Q uantification o f the Products In every experimental determination of k p, there is some set of values A ^OQ (the "limits of quantitation") below which the product concentrations A mcannot be reliably measured. If during an experiment carried out over the period of timeA t all the product concentrations A mremain below their limits of quantitation, then the maximum possible value of the rate k p is obtained by assuming (for all the products) that 1) A m0 = 0 and 2) at time t = A t , the product concentrations have increased to the values A m= A ^0Q. With these assumptions, the experimental data indicate that the reaction rate k pis less than some maximum value (kp^ as follows: (for hydrolytically stable products at concentrations below the limits of quantitation) k P < ( kpL * = ^ 7 t X A "0Q ' * 0 A 1 m=l (B32) Under the same circumstances and assumptions, the experimental data indicate that the parent half-life T 1^ (see Equation B9) is greater than the value (t ^ ) . as follows: (for hydrolytically stable products at concentrations below the limits of quantitation) T 1/2 H 2 ) A t P0 ln(2) a !:oq P (kp ima (B33) The reader should note that Equations B32 and B33 are valid only when both 1) the products are hydrolytically stable and 2) the concentrations of all the potential products are measured. Otherwise, the quantity ( k p ) ^ in Equation B32 may not actually represent the maximum possible value of the rate constant k p, and the related result in Equation B33 for (t ^ ) is also questionable. Page 45 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 B6. Parent Half-Life Estimates Based on Lim its o f Quantification and Experimental Precision o f Product Concentrations In certain experiments, some hydrolysis products are present at quantifiable but essentially constant concentrations over the time (A t ) 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 rate k p. If the set of concentrations measured for the mth product have the mean value p mand standard deviation c m, the data do not exclude the possibility that the product concentration increased from the initial value o m- p mto the value o m+ | imat time t = A t . Taking this possibility to be the actual case for the measured products, the maximum value of the quantity (A m- A m0) is 2 o ra. This reasoning suggests that the following estimate of the maximum parent hydrolysis rate is appropriate: (for hydrolytically stable products at either 1) constant measured concentrations with standard deviation am or 2) concentrations below the limits of quantitation) kp ^ (k p L * = 1 A r + 2' P0 A t Below LOQ Cons tan t (B34) Under these circumstances and assumptions, the experimental data indicate that the parent half-life T 1^ is greater than the value (t ^ ) as follows: " ' P 'm in (for hydrolytically stable products at either 1) constant measured concentrations with standard deviation om or 2) concentrations below the limits of quantitation) t !/2 > (t ^2 P - V P 'm Jn(2) ( k P )m a At P0 2) X A - Q+ 5 > m Below LOQ Cons tan t (B35) The reader should note that Equations B34 and B35 are valid only when both 1) the products are hydrolytically stable and 2) the concentrations of all the potential products are measured. B6. Parent Half-Life Estimates Based on the Experimental Precision o f Parent C o n ce n tra tio n s In certain experiments, the hydrolytic parent remains at an essentially constant concentration over the time (A t) of the experiment. In this case, it is the experimental precision of the measured parent concentrations that determines the maximum value of the parent hydrolysis rate k p. If the set of concentrations measured for the parent have the mean value |j.pand standard deviation a p, the data do not exclude the possibility Page 46 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 that the product concentration increased from the initial value p p - o P to the value p P + c P at time t = A t . This reasoning suggests that the following estimate of the maximum parent hydrolysis rate is appropriate: (for essentially constant parent concentrations with mean value p pand standard deviation Gp) k p * (k p L x = "f-A^pTAATl - (B36) Under these circumstances and assumptions, the experimental data indicate that the parent half-life T 1^r is greater than the value ( T ^ ) as v P 7min follows: (for essentially constant parent concentrations with mean value p pand standard deviation a P) -p/2 > (t V21 _ 2) _ M-p A t ln( 2) P ' P /min (\r \ (k p)n 2. (B37) B8. Tem perature Dependence o f the Reaction Rate and Half-Life In order to increase the speed of the reactions of interest, we conducted this experimental study using samples maintained at the temperature 50C = 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 is A H a, Equation B38 81 provides the following relationship between the hydrolysis rates (lq and k2) for that reaction at two different absolute tem peratures (T, and T 2): (B38) where R = 1.99 x 10'3 Kcal mole-1 K-1is the ideal gas constant. Using the value82 A H a=18 Kcal/mole, the rate ratio k , / k 2at the corresponding temperatures T, =298 K and T2=323 K is --k, = expf^-----1-8------- 1 k2 [1.99X10-3 323 = exp(-2.35) = 0.095 (B39) Page 47 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Equation B39 indicates that the hydrolysis reactions of interest proceed approximately ten times more slowly at 25C than at the chosen experimental temperature of 50C. Accordingly, the rate reactions reported here for the temperature 25C are ten times lower than those measured at 50C, and the hydrolysis half-life estimates reported here for 25C samples are ten times longer than those calculated from the 50C experimental data. References to Appendix B: 81 I. N Levine, "Physical Chemistry," McGraw-Hill (New York), pp. 498-501 (1978). 82 F. Daniels, et al., "Experimental Physical Chemistry", McGraw Hill (New York), p.131 (1962). Page 48 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix C: Selected Analytical and Kinetics Results This Appendix includes selected sample data and their related kinetics results. Page 49 of 71 N-MeFOSE Alcohol Buffer Hydrolysis Study at 50 C. 3M Environmental Laboratory Report No. W1880 All concentrations in ng/ml pH T im e (D ays) 1.5 0 1.5 42 1.5 49 1.5 7 1.5 14 1.5 21 1.5 28 1.5 34 Cone. 441 398 402 390 385 ln([P ]t/[P]o) (A ) (A ) (A ) 0.000 -0 .1 0 2 -0 .0 9 2 -0 .1 2 2 -0 .1 3 4 N-MeFOSE Alcohol @ 50 C pH 1.5, Rate constant g raph Time (days) SUMMARY OUTPUT R eg ression Statistics Multiple R R Square Adjusted R Square Standard Error Observations 0.86541 0.74893 0.66524 0.03064 5 ANOVA ___________________________________________________d f Regression Residual Total 1 3 4 0.00840 0.00282 0.01122 ____________________________________________ C o e ffic ie n ts Intercept -0.00143 X Variable 1 -0.00426 Standard Error 0.03264 0.00142 % 2a Slope Uncertainty 67% pH T im e (D ays) Cone. ln([P ]t/[P]o) 3.0 0 (A ) 3.0 14 (A ) 3.0 7 348 0.000 3.0 21 338 -0.027 3.0 28 333 -0.042 3.0 34 373 0.069 3.0 42 353 0.014 3.0 49 343 -0.014 0.05 -, 0 -0.05 - 2b. -0.15 - -0.25 -0.35 N-MeFOSE Alcohol @ 50 C pH 3J litote. constant g rap h .. y = 4 .4 2 E -0 4 x - 1.34E-02 R 2 = 2 .8 7 E - 0 2 _________ 20 30 40 50 Time (days) SUMMARY OUTPUT R eg ression Statistics Multiple R R Square Adjusted R Square Standard Error Observations 0.16934 0.02868 -0.21415 0.04329 6 ANOVA Regression Residual Total d f _______________ S S 1 0.00022 4 0.00750 5 0.00772 Intercept X Variable 1 % 2 a Slope Uncertainty 582% C o e ffic ie n ts -0.01338 0.00044 Standard Error 0.04261 0.00129 (A) Data in Italics have been rejected from the fits on the basis of the data quality objectives (see text). BACK TO MAIN Page 50 of 71 N-MeFOSE Alcohol Buffer Hydrolysis Study at 50 C. 3M Environmental Laboratory Report No. W1880 All concentrations in ng/ml pH T im e (D ays) 5.0 0 5.0 7 5.0 14 5.0 21 5.0 28 5.0 34 5.0 42 5.0 49 Cone. 482 439 407 394 398 381 358 357 ln([P ]t/[P]o) 0.000 -0 .0 9 4 -0 .1 7 0 -0 .2 0 2 -0.191 -0 .2 3 6 -0 .2 9 9 -0 .3 0 2 0.00 -, O -0.10 aBn. -0 .2 0 - -0.30 -0.40 t N-MeFOSE AJcohol @ 50 C pH 5.0 Rate constant graph y = -5.68E-03X - 4.82E -02 \ R2 = 9 .12E-01 . > s# '1 20 40 Time (days) 1 60 pH T im e (D ays) Cone. ln([P ]t/[P]o) 7.0 0 467 0 .000 7.0 7 446 -0.047 7.0 14 410 -0.130 7.0 21 399 -0.159 7.0 28 393 -0.173 7.0 34 396 -0.164 7.0 42 375 -0.220 7.0 49 360 -0.262 0 10 20 30 40 50 60 Time (days) SUMMARY OUTPUT R eg ression Statistics Multiple R R Square Adjusted R Square Standard Error Observations 0.95481 0.91167 0.89695 0.03261 8 ANOVA Regression Residual Total df 1 6 7 SS 0.06584 0.00638 0.07222 Intercept X Variable 1 % 2o Slope Uncertainty 25% C oefficients -0.04820 -0.00568 Standard Error 0.02104 0 .0 0 0 7 2 SUMMARY OUTPUT R eg ression Statistics Multiple R R Square Adjusted R Square Standard Error Observations 0.96050 0.92256 0.90965 0.02578 8 ANOVA Regression Residual Total df 1 6 7 SS 0.04749 0.00399 0.05147 Intercept X Variable 1 % 2 a Slope Uncertainty 24% C o e ffic ie n ts -0.02680 -0.00483 Standard Error 0.01664 0.00057 BACK TO MAIN Page 51 of 71 N-MeFOSE Alcohol Buffer Hydrolysis Study at 50 C. 3M Environmental Laboratory Report No. W1880 All concentrations in ng/ml pH T im e (D ays) 9.0 0 9.0 7 9.0 14 9.0 21 9.0 28 9.0 34 9.0 42 9.0 49 Cone. 412 437 394 393 383 382 355 349 ln ([P ]t/[P ]o ) 0.000 0 .0 6 0 -0.043 -0.046 -0.072 -0.075 -0.148 -0.164 N-MeFOSE Alcohol @ 5# C pH 9.0 rate constant graph SUMMARY OUTPUT R eg ressio n Statistics Multiple R R Square Adjusted R Square Standard Error Observations 0.92392 0.85363 0.82924 0.03027 8 ANOVA ___________________________________________________d f Regression Residual Total 1 6 7 0.03207 0.00550 0.03757 ____________________________________________ C oefficien ts Intercept 0.03566 X Variable 1 -0.00397 Standard Error 0.01954 0.00067 % 2 c Slope Uncertainty 34% pH 11.0 11.0 11.0 11.0 11.0 11.0 11.0 11.0 Tim e (D ays) 0 7 42 14 21 28 34 49 Cone. 480 450 413 413 398 395 392 369 ln([P]t/[Plo) 0.000 -0 .0 3 7 -0 .0 4 4 -0.053 -0.113 0 .0 5 -i N-MeFOSE Alcohol @ 50 C ft. 2. -0.15 St - -0.25 - y = -2.98E-03X + 3.77E -02 R2= 9.55E -01 0 20 40 60 Time (days) SUMMARY OUTPUT R eg re s s io n Statistics Multiple R R Square Adjusted R Square Standard Error Observations 0.97704 0.95460 0.93947 0.01002 5 ANOVA Regression Residual Total df 1 3 4 SS 0.00634 0.00030 0.00664 Intercept X Variable 1 % 2 o Slope Uncertainty 25% C o e ffic ie n ts 0.03775 -0.00298 Standard Error 0.01183 0.00037 (A) Data in Italics have been rejected from the fits on the basis of the data quality objectives (see text). BACK TO MAIN Page 52 of 71 3M Environmental Laboratory Report No. W1880 MeFOSE A lcoh ol H ydrolysis Study 50C; pH 1.5 R ete ntion tim es (RT) in m inu te s A ll c o n c e n tr a tio n s In n g / m l ________ N-MeFOSE A lc o h o l Sam ple t.0. M eOH Blank 99039-30-1 99039-30-2 99039-30-3 99039-30-4 99039-30-5 99039-30-6 MeOH Blank MeOH Blank M FA-001 MFA-002 M FA -003 M FA -019 M FA -020 M FA-021 M FA -037 M FA-038 M FA -039 M FA -055 MFA-056 MFA-057 MFA-073 MFA-074 MFA-075 MeOH Blank M eOH Blank M FA-091 M F A-092 M FA-093 F ile M e F O S 0 0 4 .D M e F O S 0 0 6 .D M e F O S 0 0 7 .D M e F O S 0 0 8 .D M e F O S 0 0 9 .D M 0FOSOIO.D M eFOSOH.D M e F O S 0 1 1 .D E IFO S 0 10 .D M e F O S 0 1 1 .D M 0F O S O I2 .D M e F O S 0 1 3 .D M 0FOSO14.D M 0 F O S O 1 5 .D M 0 F O S O 1 6 .D M0FOSO17.D M 0FOSO18.D M e F O S 0 1 9 .D M eF O S 02 0.D M e F O S 0 2 1 .D M 0 F O S O 2 2 .D M 0 F O S O 2 3 .D M e F O S 0 2 4 .D M e F O S 0 2 5 .D M e F O S 0 2 8 .D M 0FO S O 3O .D M e F O S 0 3 1 .D M 0 F O S O 3 2 .D M e F O S 0 3 3 .D vw # 92 1 2 3 4 5 6 92 91 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 92 92 26 27 28 R i Tifi* 0.0 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.3 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 A re* 0 159912 302118 583920 882215 1143693 1499047 0 0 901027 76714 1272654 818000 830680 1196650 824922 804837 1204286 818907 823891 1209639 796832 803728 1213336 0 0 793217 793658 1194136 Cone. 0 87 159 303 466 610 794 0 0 794 0 660 437 444 644 393 403 596 400 404 577 392 388 586 0 0 381 390 580 % Standard or RSO 110% 101% 96% 98% 97% 101% 169% 2% 3% 1% 1% 2% M FA-127 M FA-128 M FA-129 M 0 F O S O 3 4 .D M 0 F O S O 3 5 .D M 0 F O S O 3 6 .D 29 30 31 8.6 20502 8.6 22336 8.4 410622 186 M eOH Blank M eOH Blank M 0FOSO4 2 .D M e F O S 0 4 3 .D 92 92 8.4 8.4 0 0 0 0 99039-30-1 MeFOS044.D 1 8.4 160028 69 99039-30-2 M 0 F O S O 4 5 .D 2 8.4 296600 159 99039-30-3 M eFO S 046.D 3 8.4 569502 310 99039-30-4 99039-30-5 99039-30-6 M eFO S 047.D 4 8.4 862113 472 M e F O S 0 4 8 .D 5 8.4 1153879 614 M 0FOSO49.D 6 8.4 1507978 826 M eth od ID; In ternal Standard quant: =0.999 O riginal cone, (ng/ml) = 473 C urve A ve ra ge d, Linear, include origin Spike cone, (ng/ml) = 221 C alibration range; 79-789 ng/mL {A ) D ata excluded on th e basis o f data quality objectives; see text. (B ) S a m p les not analyzed; see text. (C ) In tern al standard om itted; see text. 113% 100% 98% 100% 97% 105% PFOS_________________________________________________ T H PF O S_____________________________ N -E tF O S E A lc o h o l ttS p ilw R e co ve ry T im e P o in t Ret Tim e 6.3 A re * 0.0 Cone. 0.0 % Standard 0.0 R et Time 6.0 A fe a 0 Cone. 0 Ret Tim e 8.6 Area 0 6.3 10709 6.2 20812 6.2 39143 6.2 93452 6.3 168595 6.3 260180 4.0 7.5 14.3 34.2 62.1 96.6 128% 96% 91% 109% 99% 103% 6.0 327940 403 8.6 629180 6.0 335820 403 8.6 635756 6.0 329167 403 8.6 637107 6.0 325653 403 8.6 623482 6.0 323376 403 8.6 616127 6.0 320840 403 8.6 619682 6.3 0 6.3 0 0.0 0.0 6.0 0 6.0 0 0 8.6 0 0 8.6 0 92% 90% 79% 89% (A) (A.C) (A) Day 7 Day 7 Day 7 D ay 14 D ay 14 D ay 14 D ay 21 D ay 21 D ay 21 Day 28 Day 28 Day 28 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 0 9367 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.7 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 6.0 340395 6.0 1843930 6.0 334722 6.0 315749 6.0 319962 6.0 320529 6.0 321851 6.0 311489 6.0 313734 6.0 321434 6.0 321231 6.0 323500 6.0 323192 6.0 315968 6.0 321240 403 403 403 403 403 403 403 403 403 403 403 403 403 403 403 8.6 631887 8.6 21628 8.6 633337 8.6 616995 8.6 615930 8.6 610314 8.6 692682 8.6 658945 8.6 663813 8.6 675939 8.6 672256 8.6 689292 8.6 670521 8.6 683560 8.6 679976 6.3 0 6.3 0 0.0 0.0 6.0 0 6.0 0 0 8.6 0 0 8.6 0 88% Day 34 D ay 34 Day 34 (B) (B) (B) (C) (C) (C) 6.3 6.3 6.3 6.3 6.3 6.3 0 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 0.0 6.0 317397 6.0 316037 6.0 322198 403 403 403 8.6 687397 8.6 671514 8.6 677205 6.0 0 6.0 0 6.0 0 403 8.6 732926 403 8.6 772348 403 8.6 737726 6.3 0 6.3 0 0.0 0.0 6.0 0 6.0 0 0 8.6 0 0 8.6 0 6.3 11627 4.4 6.3 20367 7.5 6.3 40566 14.6 6.3 80242 30.1 6.3 163639 61.1 6.3 251082 92.3 internal S tandard quant; =0.999 Curve Averaged, Linear, include origin Calibration range; 7.8-94 ng/m L 141% 95% 93% 96% 98% 98% 6.0 6.0 6.0 6.0 6.0 6.0 mean S.D. %S.D. 321139 329327 332980 318130 319076 323806 323030 6782 2.1% 403 8.6 615234 403 8.6 626824 403 8.6 608282 403 8.6 601758 403 8.6 617694 403 8.6 598749 mean 643498 S.D. 30538 %S.D. 4.7% Cone. 0 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 0 0 311 311 311 311 311 311 Page 53 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 MeFOSE A lcoh ol H ydrolysis Study 50C; pH 3 R s ts n tio n to ne s (RT) In m inu te s . . S : : .............. MeOH Blank O a teF e M eFOS002.D N-MsFOSE A lcohol Ret T im * Ana 91 0.0 0 Cone. 0 99039-31-2 99039-31-4 99039-31-6 M eFOS003.D M eFO S 0O 4 .D M eFOS005.D MeFOSOOS.D M eFOS007.D M eFOS008.D 1 2 3 4 5 6 8.3 151736 8.3 341079 8.3 660722 8.3 1127126 8.3 1442949 8.3 1796489 82 155 301 484 639 778 MeOH Blank MeOH Blank M eFOS009.D MeFOSOIO.D 91 91 8.3 8.3 0 0 0 0 M FA-004 M FA-006 M FA-022 M FA-024 M FA-040 MFA-041 MFA-042 MFA-059 MFA-076 MFA-077 MFA-078 M eFOS011.D M eFOS012.D M eFOS013.D M eFOS014.D M e F O S 0 1 5 .D M eFOS016.D M e F O S 0 1 7 .D M eFOS018.D M eFOS019.D M eFOS020.D M eFOS021.D M eFOS022.D M e F O S 0 2 3 .D M e F O S 0 2 4 .D M e F O S 0 2 5 .D 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 8.3 1134989 8.3 390170 8.3 1589221 8.3 829229 8.3 764651 8.3 1403195 8.3 866899 8.3 597135 8.3 1297314 8.3 825047 8.3 869628 8.3 1317427 8.3 915705 8.3 738271 8.3 1351582 483 951 683 361 334 555 355 247 520 327 349 537 367 300 553 MeOH Blank MeOH Blank MeFOS026.D 91 8.3 MeFOS028.D 91 8.3 0 0 0 0 MFA-094 MFA-096 MFA-112 MFA-113 MFA-114 MFA-130 MFA-131 MFA-132 MeFOS029.D 26 8.3 901464 371 MeFOS030.D 27 8.3 908936 375 MeFOS031.D 28 8.3 1346477 561 MeFOS032.D 29 8.3 952582 359 MeFOS033.D 30 8.3 907374 346 M eF O S 034.D 31 8.3 1435453 545 M e F O S 0 3 5 .D 32 8.3 948868 346 M e F O S 0 3 6 .D 33 8.3 919227 339 MeFOS037.D 34 8.3 1428869 525 MeOH Blank MeOH Blank MeFOS043.D 91 8.3 MeFOS044.D 91 8.3 0 0 0 0 99039-31-1 MeFOS045.D 1 8.3 168468 79 99039-31-2 MeFOS046.D 2 8.3 336376 158 99039-31-3 MeFOS047.D 3 8.3 654789 308 99039-31-4 M e F O S 0 4 8 .D 4 8.3 1038617 487 99039-31-5 M e F O S 0 4 9 .D 5 8.3 1350045 640 99039-31-6 MeFOSOSO.D 6 8.3 1678778 781 M ethod ID: M FO 0828. M Internal Standard qu an t r2 = 0.998 Original co n e (ng/m l) * 473.4 C urm averaged, Linear,indude origin. Spike cone, (ng/ml) * 221 Calibration range: 155-932 ng/ml (A) Data excluded on the basis of data quality objectives; see te x t U S ttO drnJw RSO 104% 98% 95% 102% 101% 99% 65% 8% 36% 7% 20% 1% 4% 2% 100% 100% 97% 103% 101% 99% % S P *. Recovery ; th *** Ret Tim * 6.2 Ar*a 12328 Cone. 0.0 6.1 20171 6.1 34709 6.1 53143 6.2 106282 6.2 195877 6.2 286910 3.2 7.8 13.6 31.2 59.4 91.2 6.2 13023 6.2 10891 0.0 0.0 -15% 94% 99% 90% 100% (A) (A) (A) (A) (A) (A) (A) (A) (A) Day 20 Day 20 Day 20 Day 28 Day 28 Day 28 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 14132 23260 11570 21187 14049 13409 13922 11887 14064 12959 13631 13975 12740 12296 13770 0.7 0.0 0.0 3.5 1.0 0.4 1.0 0.3 1.1 0.7 0.8 1.2 0.7 0.6 1.1 6.2 12588 6.2 11439 0.0 0.0 85% 87% 82% Day 34 Day 34 Day 34 Day 42 Day 42 Day 42 Day 49 Day 49 Day 49 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 12253 16034 12839 12120 13021 13729 13388 13132 12775 0.6 2.0 0.9 0.5 0.8 1.1 0.9 0.8 0.7 6.2 12279 6.2 12022 0.0 0.0 6.2 21343 3.8 6.2 33543 8.2 6.2 53137 15.4 6.2 101279 33.3 6.2 189417 64.5 6.2 272461 97.1 Internal Standard quant: r2 * 0.997 Curve averaged, U near.indude origin. Calibration range: 15.7-94 ng/ml % S ten da rd 0.00 Ret Thu* 5.8 Area 0 101% 100% 87% 100% 95% 97% 5.8 419594 5.8 430655 5.8 439792 5.9 438422 5.9 446792 5.9 435601 5.8 0 5.8 0 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9. 5.9 5.9 5.9 5.9 457701 2178603 439722 418052 425475 468367 417398 416159 412958 420234 426878 408760 414108 407935 404854 5.9 0 5.9 0 5.9 406516 5.9 402971 5.9 396695 5.9 406709 5.9 410407 5.9 405375 5.9 416004 5.9 410564 5.9 408186 5.9 0 5.9 0 121% 105% 98% 107% 103% 103% 5.9 5.9 5.9 5.9 5.9 5.9 mean S.D. %S.D. 406294 403340 400256 393427 399772 389466 417298 18014 4.3% N-EtFOSE A lc o h o l uonc. 0 403 403 403 403 403 403 0 0 403 403 403 403 403 403 403 403 403 403 403 403 403 403 403 0 0 403 403 403 403 403 403 403 403 403 0 0 403 403 403 403 403 403 Ret T im * 8.5 Area 0 8.5 636812 8.5 744336 8.5 738471 8.5 780745 8.5 757128 8.5 773265 8.5 0 8.5 0 8.5 788849 8.5 137293 8.5 779045 8.5 771699 8.5 768972 8.5 847709 8.5 820997 8.5 815246 8.5 837107 8.5 847509 8.5 B36796 8.5 822995 8.5 839193 8.5 827596 8.5 819079 8.5 0 8.5 0 8.5 817359 8.5 814894 8.5 804552 8.5 890510 8.5 881985 8.5 882466 8.5 920660 8.5 911546 8.5 912B01 8.5 0 8.5 0 8.5 8.5 8.5 8.5 8.5 8.5 mean S.D. %S.D. 730702 723068 715896 715694 706581 719573 800052 66857 8.4% Cone. 0 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 BACK TO MAIN Page 54 of 71 3M Environmental Laboratory Report No. W1880 Mb FOSE A ko bo J H yd ro lysis Study 50C; pH 5 R etention tim es (RT) in m inutos A ll concentrations ir '- MeOH Blank " f r i t t i FHu M eFOS051.D 92 99039-32-1 99039-32-2 99039-32-3 99039-32-6 M eFOS052.D M eFOS053.D M eFOS054.D M eFOS055.D M eFOS056.D M eFOS057.D 41 42 43 44 45 46 MeOH Blank MeOH Blank M eFOS058.D M eFOS059.D 92 92 MFA-O07 MFA-007 MFA-009 MFA-025 MFA-026 MFA-027 MFA-043 MFA-044 M FA-04 5 M FA-061 M FA-062 M FA-063 MFA-079 M FA-080 MFA-081 M eFOS060.D M eFOS061.D M eFOS062.D M e F O S 0 6 3 .D M eFOS064.D M eFOS065.D MOFOS066.D M eFOS067.D M eFOS068.D M eFOS069.D M eFOS070.D M eFOS071.D M eFOS072.D M eFOS073.D M eFOS074.D 51 51 53 54 55 56 57 58 59 60 61 62 63 64 65 M -O H Blank MeOH Blank M eFOS075.D M eFOS077.D 92 92 MFA-097 MFA-098 MFA-099 MFA-115 MFA-116 M FA-117 MFA-133 MFA-134 MFA-135 M eFOS078.D M eFOS079.D MeFOSOBO.D M eFOS081.D M eFOS082.D M eFOS083.D M e F O S 0 8 4 .D M eFOS085.D M eFOS086.D 66 67 68 69 70 71 72 73 74 MeOH Blank MeOH Blank M eFOS092.D M eFOS093.D 92 92 99039-32-1 99039-32-2 99039-32-3 99039-32-4 99039-32-5 99039-32-6 M ethod ID: Original cone. (ng/nV) a Spike cone. (ng A nl)1 M eFOS094.D M eFOS095.D M eFOS096.D M eFOS097.D M eFOS098.D M eFOS099.D 41 42 43 44 45 46 473.4 221 n e i Tgli* 8.4 Aa 0 8.4 149060 8.4 280351 8.4 544208 8.4 858881 8.4 1154462 8.4 1368379 Cone. 6 82 159 306 485 628 785 Standard o r RSC .V R ecovery 104% 101% 97% 103% 99% 100% Tim Point Ret Time 6.3 6.3 6.3 6.3 6.3 6.3 6.3 Area 0 9155 18855 69313 81289 178117 240605 Cone. 0.0 3.5 7.4 27.2 31.6 66.6 93.4 8.4 0 8.4 0 0 0 6.3 0 6.3 0 0.0 0.0 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 . 8.4 8.4 8.4 8.4 8.4 966445 972256 1262465 764741 801784 1155792 778763 818063 1165951 791698 798843 1174315 773986 768748 1157303 482 483 697 428 450 641 406 408 607 394 394 601 401 396 597 0% 5% 0% 0% 1% 97% 92% 91% 94% 90% Day 0 Day 0 Day 0 Day 7 Day 7 Day 7 Day 14 Day 14 Day 14 Day 20 Day 20 Day 20 Day 28 Day 28 Day 28 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 84 0 8.4 0 0 0 6.3 0 6.3 0 0.0 0.0 8.4 768874 8.4 750087 8.4 1142873 8.4 750291 8.4 771988 8.4 1161257 8.4 765373 8.4 779909 8.4 1157828 383 379 586 359 358 562 354 359 559 1% 1% 1% 93% 92% 91% Day 34 Day 34 Day 34 Day 42 Day 42 Day 42 D ay 49 Day 49 Day 49 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 6.3 0 0 0 0 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 8.4 0 8.4 0 0 0 6.3 0 6.3 0 0.0 0.0 8.4 146569 82 8.4 276797 158 8.4 535672 306 8.4 839851 483 8.4 1133812 627 8.4 1354192 793 Internal Standard qu an t 1 ^0 90 9 Curve A veraged, Linear, include origin Callbradon range: 79-789 ng/mL 104% 100% 97% 102% 99% 100% 6.3 8210 3.1 6.3 18161 7.0 6.3 67607 26.4 6.3 77275 29.9 6.3 168159 63.7 6.3 241085 91.5 Internal Standard qu an t *=0.999 Curve Averaged, Linear, include origin Calibration range: 3-94 ng/mL ` S ta nd ard 0.0% Ret Time 6.0 Area 0 111% 94% 174% 101% 106% 99% 6.0 316040 6.0 313687 6.0 315295 6.0 318179 6.0 331279 6.0 319232 6.0 0 6.0 0 6.0 369039 6.0 358611 6.0 330072 6.0 318812 6.0 321150 6.0 304415 6.0 318966 6.0 327562 6.0 316845 6.0 318539 6.0 321130 6.0 326528 6.0 316457 8.0 314675 6.0 314393 6.0 0 6.0 0 6.0 315754 6.0 313136 6.0 315673 6.0 308799 6.0 323683 6.0 317017 6.0 319465 6.0 320409 6.0 314997 6.0 0 6.0 0 99% 90% 169% 96% 102% 97% 6.0 6.0 6.0 6.0 6.0 6.0 mean S.D. %S.D. 317732 316658 315957 319036 326660 326403 321175 11910 3.7% Cone. 0.00 402.60 402.60 402.60 402.60 402.60 402.60 0.00 0.00 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 0.00 0.00 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 402.60 0.00 0.00 402.60 402.60 402.60 402.60 402.60 402.60 Ret Time 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 mean S.D. %S.D. Aras 0 598824 589170 597555 596598 620432 588439 0 0 676077 678936 611827 602188 600482 608185 645859 676091 647772 676708 681943 659289 650045 653653 654428 0 0 676650 666126 658463 702846 729469 697385 727348 731061 699055 0 0 590591 584803 588651 585887 610173 577012 642778 46797 7.3% eonc. 0 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 BACK TO MAIN Page 55 of 71 3M Environmental Laboratory Report No. W1880 MeFOSE A lc o h o l H y dro ly l* Stud y 50C; pH 7 Retention tim os (RT) in m inutes All con c e n tra tio n s In ng/m l________________________ N jjte F O S E A lc o h o l _______________________________________________ PFOS_______________________________________________ THPFOS___________________________ N-EtFOSE A lco h o l S -n & te 'tb . MeOH Blank D stsPS * M eF O S 052.D 2 91 8.3 Arsa VI 0 Cone. 0 %Stendard or RSC ::;Rscvsrir':.' lm P o in t Ret Tim s 6.2 Arsa 12060 Cone. 0.0 '/S ta nd ard 0.00 RetTlm e 5.9 Area 0 Cone. 0 R e i Tim e 8.5 Area 0 99039-33-1 99039-33-2 99039-33-3 99039-33-4 99039-33-5 99039-33-6 M eFOS053.D 41 8.3 163904 74 M eF O S 054.D 42 8.3 353900 162 M eFOS055.D 43 8.3 672851 313 M eF O S 056.D 44 8.3 1043250 487 M eF O S 057.D 45 8.3 1848028 863 M eF O S 058.D 46 8.3 1678345 781 94% 103% 99% 103% 137% 99% 6.2 21926 6.2 33291 6.2 54104 6.2 120587 6.2 197863 6.2 283971 3.4 7.3 14.6 37.5 64.7 93.6 110% 94% 93% 120% 103% 100% 5.9 413061 403 8.5 722548 5.9 415745 403 0.0 726841 5.9 414516 403 0.0 721094 5.9 416734 403 0.0 721008 5.9 414375 403 0.0 722268 5.9 418593 403 0.0 724772 MeOH Blank MeOH Blank M eFOS059.D 91 8.3 M eFOS060.D 91 8.3 0 0 0 0 6.2 10884 6.2 8447 0.0 0.0 5.9 0 5.9 0 0 0.0 0 0 0.0 0 MFA-010 M eF O S 061.D 51 8.3 1059849 463 MFA-011 M eF O S 062.D 52 8.3 1035099 471 MFA-012 MFA-028 M eF O S 063.D 53 8.3 1493105 693 M eFOS064.D 54 8.3 950167 440 MFA-Q29 M eFOS065.D 55 8.3 971370 451 M FA-030 M eFOS066.D 56 8.3 1421931 668 M FA-046 M eFOS067.D 57 8.3 945561 409 MFA-047 M eFOS068.D 58 8.3 955303 412 MFA-048 M eFOS069.D 59 8.3 1390135 604 MFA-064 M 6FOS070.D 60 8.3 960628 398 MFA-065 M eF O S 071.D 61 8.3 966872 400 MFA-066 M eF O S 072.D 62 8.3 1429859 597 MFA-082 M eF O S 073.D 63 8.3 938910 387 M FA-083 M eFOS074.D 64 8.3 938077 400 M FA-084 M eF O S 075.D 65 8.3 1373974 590 2% 2% 1% 1% 3% 102% 100% 88% 90% 89% Day 0 Day 0 Day 0 Day 7 Day 7 Day 7 Day 14 Day 14 Day 14 Day 20 Day 20 Day 20 Day 28 Day 28 Day 28 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 13752 13431 12470 12076 11626 12182 11943 10864 11746 10395 9801 11029 10525 10803 10416 0.4 0.3 0.0 0.1 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 5.9 431433 403 0.0 769705 5.9 427482 403 0.0 738896 5.9 424804 403 0.0 726047 5.9 406484 403 0.0 725630 5.9 412600 403 0.0 724750 5.9 408432 403 0.0 717606 5.9 410627 403 0.0 777412 5.9 411793 403 0.0 780071 5.9 405115 403 0.0 775133 5.9 415819 403 0.0 812219 5.9 425350 403 0.0 812637 5.9 416204 403 0.0 806256 5.9 414343 403 0.0 816223 5.9 406804 403 0.0 789126 5.9 406568 403 0.0 784737 MeOH Blank MeOH Blank M eFOS076.D 91 8.3 M eFOS078.D 91 8.3 0 0 0 0 6.2 10001 6.2 10980 0.0 0.0 5.9 0 5.9 0 0 0.0 0 0 0.0 0 MFA-100 MFA-101 MFA-102 MFA-118 MFA-119 M F A-120 M FA-136 M F A-137 M FA-138 M eFOS079.D 66 8.3 917975 398 MeFOSOBO.D 67 8.3 917551 395 M eF O S 081.D 68 8.3 1364592 618 M eFOS082.D 69 8.3 908151 380 M eF O S 083.D 70 8.3 875016 369 M eF O S 084.D 71 8.3 1355262 565 M eFOS085.D 72 8.3 894105 366 M eFOS086.D 73 8.3 871427 353 M eF O S 087.D 74 8.3 1360766 544 1% 3% 3% 100% 86% 84% Day 34 Day 34 Day 34 Day 42 Day 42 Day 42 Day 49 Day 49 Day 49 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 10737 10892 10866 10383 11979 10102 10966 10552 11994 0.0 0.0 0.0 0.0 0.2 0.0 0.0 0.0 0.1 5.9 396679 403 0.0 774961 5.9 401501 403 0.0 781448 5.9 395834 403 0.0 744222 5.9 387918 403 0.0 802155 5.9 389550 403 0.0 796321 5.9 397888 403 0.0 808103 5.9 392635 403 0.0 821314 5.9 392646 403 0.0 828208 5.9 404326 403 0.0 841482 MeOH Blank MeOH Blank M eFOS093.D MeF OSO94. D 91 91 8.3 8.3 0 0 0 0 6.2 9877 6.2 10936 0.0 0.0 5.9 0 5.9 0 0 0.0 0 0 mean 0 99039-33-1 99039-33-2 99039-33-3 99039-33-4 99039-33-5 99039-33-6 Method ID: Original cone, (ng/ml) * S pire cone, (ng/m l) = M eF O S 095.D M eF O S 096.D MeFOS097.D M eF O S 098.D M eF O S 099.D M eF O S IO O .D 41 42 43 44 45 46 473.4 221 8.3 162533 75 8.3 338524 161 8.3 651730 310 8.3 1013215 480 8.3 1816530 853 8.3 1634822 788 Internal Standard q u a n t r2 = 0.998 Curve averaged. Linear, Include origin. Calibration range: 155-932 ng/ml 95% 102% 98% 101% 135% 100% 6.2 17446 1.9 6.2 29991 6.4 6.2 58016 16.2 6.2 112160 35.3 6.2 190397 61.9 6.2 272629 90.2 Internal Standard q u a n t r2 = 0.997 Curve averaged, Linear,Inelude origin. Calibration range: 15.7-94 ng/ml 62% 82% 103% 113% 99% 96% 5.9 5.9 5.9 5.9 5.9 5.9 mean S.D. %S.D. 407992 407169 409832 409148 415661 416714 409510 10423 2-5% 403 mean 707642 403 S.D. 700669 403 mean 705419 403 S.D. 709949 403 mean 718262 403 S.D. 699829 mean 759304 S.D. 43356 %S.D. 5.7% uonc. 0 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 BACK TO MAIN Page 56 of 71 MaFOSE Alcohol Hydrolysis Study 50C; pH 9 Rstsntlon times (RT) In mlnutss AH concentrations In ng/ml N-MaFOSE Alcohol MeOH Blank MeFOS011 D 92 8.4 A ... 0 . Cane. 0.0 99039-34-1 99039-34-2 99039-34-3 99039-34-4 99039-34-5 99039-34-6 M eFOS012.D 1 8.4 163460 80 M eF O S 013.D 2 6.4 309997 146 M eF O S 014.D 3 6.4 680067 304 M eF O S 015.D 4 6.4 1006367 464 M eF O S 016.D 5 6.4 1546780 692 M eF O S 017.D 6 8.4 1664630 761 MeOH Blank MeOH Blank MeFOS018.D 92 0.0 MeFOS019.D 92 0.0 0 0 MFA-013 MFA-014 MFA-015 MFA-031 MFA-032 MFA-033 MFA-049 MFA-050 MFA-051 MFA-067 MFA-068 MFA-069 MFA-085 MFA-086 MFA-087 M eF O S 020.D 11 8.4 1028452 463 M eFOS021.D 12 8.4 847651 360 M eF O S 022.D 13 8.4 1569790 666 M eFOS023.D 14 8.4 996448 440 M eFOS024.D 15 8.4 964111 434 M eF O S 025.D 16 8.4 1413056 654 M eF O S 026.D 17 8.4 1033316 396 M eFOS027.D 18 8.4 940771 392 M eF O S 026.D 19 6.4 1387762 586 M eF O S 029.D 20 6.4 972226 393 M eF O S 030.D 21 6.4 965677 394 M e F O S 0 3 1 .D 22 8.4 1405487 570 MeFOS032.D 23 8.4 944313 383 MeFOS033.D 24 8.4 926910 384 M eF O S 034.D 25 8.4 1407593 576 MeOH Blank MeOH Blank M eFOS035.D 92 0.0 M eFOS037.D 92 0.0 0 0 0 0 MFA-1Q3 MFA-104 MFA-105 MFA-121 MFA-122 MFA-123 MFA-139 MFA-140 MFA-141 M eFOS038.D 26 8.4 942289 378 M eF O S 039.D 27 8.4 923555 386 M eF O S 040.D 28 8.4 1390550 561 M eF O S 041.D 29 6.4 929686 361 M eF O S 042.D 30 8.4 903803 349 M eF O S 043.D 31 8.4 1418215 541 M eF O S 044.D 32 8.4 918039 345 M eF O S 045.D 33 6.4 930094 353 M eF O S 046.D 34 8.4 1380184 536 MeOH Blank MeOH Blank MeFOS052.D 92 8-4 MeFOS053.D 92 8.4 0 0 0 0 99039-34-1 99039-34-2 99039-34-3 99039-34-4 99039-34-5 99039-34-6 M ethod ID Original cone. (ng/mO = Spfce cone, (n g /m l) M e F O S 0 5 4 .D M eF O S 055.D MeFOS056.D M eF O S 057.D M eF O S 058.D M eF O S 059.D 1 2 3 4 5 6 473.4 221 8.4 171724 8-4 314956 8.4 693304 81 150 311 8.4 1012133 466 8.4 1499739 8.4 1623048 685 744 Internal Standard q u a n t i3 0.995 Curve averaged, linear. Include origin Calibration ra n g e : 155-932 ngAnl %Standard or RSD 102% 92% 96% 98% 110% 96% 25% 1% 1% 0% 0% 2% 3% 2% 103% 95% 99% 99% 109% 94% % 3p*e Recovery 115% 98% 88% 80% 87% 81% 84% 85% 3M Environmental Laboratory Report No. W1880 PFOS______________________________________ THPFOS_____________________ N-EtFOSE Ateohot T k iM P tfitt Ret Time 6.2 Area 0 Cone. 0.0 6.2 15569 6.2 22667 6.2 35291 6.2 76745 6.2 166823 6.2 248041 5.5 8.7 13.9 31.3 67.2 100.3 6.2 0 6.2 0 0.0 0.0 Day 0 Day 0 Day 0 Day 7 Day 7 Day 7 Day 14 Day 14 Day 14 Day 20 Day 20 Day 20 Day 28 Day 28 Day 28 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 6.2 0 6.2 0 0.0 0.0 Day 34 Day 34 Day 34 Day 42 Day 42 Day 42 Day 49 Day 49 Day 49 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 0 0 0 0 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 6.2 0 6.2 0 0.0 0.0 6.1 19200 4.9 6.2 25918 7.1 6.2 43090 12.5 6.2 106004 32.4 6.2 192716 59.1 6.2 278633 87.0 Internat Standard q u a n t r2 = 0.995 Curve averaged, linear, Indude origin Calibration range : 7.8-94 ng/tnl ' S ta n d a rd 0% 175% 111% 89% 100% 107% 107% 156% 91% 80% 104% 94% 93% Ret Tim 5.9 5.9 5.9 5.9 5.9 5.9 5.9 0.0 0.0 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 Mean S.D. %S.D. Area 0 237029 232506 235554 236190 243630 244027 0 0 260732 213698 278506 257996 260367 251637 281850 262960 257391 268694 271896 268042 269583 268663 272969 0 0 287634 288790 283426 285239 283638 291538 293710 296728 293597 0 0 321749 316093 317775 315713 319381 315350 274020 27955 10.2% Cone. 0.0 402.6 402.6 402.6 402.6 402.6 402.6 0.0 0.0 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 0.0 0.0 4Q2.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 402.6 0.0 0.0 402.6 402.6 402.8 402.6 402.6 402.6 Ret Time 8.6 Area 0 8.6 699032 8.6 706051 8.6 726783 8.6 700715 8.6 719387 8.6 703692 8.6 0 8.6 0 8.6 717735 8.6 764466 8.6 759367 8.6 733413 B.6 718160 B.6 696132 8.6 845429 8.6 777318 8.6 761035 8.6 802228 8.6 .795082 8.6 794941 8.6 800204 8.6 783174 8.6 788327 8.6 0 8.6 0 8.6 807638 8.6 776325 8.6 800018 8.6 835761 8.6 841667 8.6 846199 8.6 863218 8.6 855340 8.6 831130 8.6 0 8.6 0 8.6 8.6 8.6 8.6 8.6 8.6 Mean S.D. %S.D. 725918 698921 724207 701526 704933 701733 764089 54352 7.1% Cone. 0.0 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 311 311 311 0 0 311 311 311 311 311 311 Page 57 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Me FOSE A lcoh ol H ydrolysis Study 50C; pH 11 R ete ntion tim e s (RT) in m in u te s MeOH Blank O rnaras M eFOS061.D 0.0 0Rei i uns 92 Arsa 0Cone. 99039-35-1 99039-35-2 99039-35-3 99039-35-4 99039-35-5 99039-35-6 MeOH Blank M eFOS062.D 41 8.4 162927 81 M eFOS063.D 42 8.4 319287 145 M eFOS064.D 43 8.4 647404 308 M eFOS065.D 44 8.4 1035120 490 M eFOS066.D 45 8.4 1363926 650 M eFOS067.D 46 8.4 1661391 764 0.0M e F O S 0 6 8 .D 0.0MeFOS069.D 92 92 0 0 00 MFA-016 MFA-017 M FA-034 M FA-035 M FA-052 MFA-053 MFA-054 MFA-070 MFA-071 MFA-072 MFA-088 MFA-089 MFA-090 MeOH Blank MeOH Blank M e F O S 0 7 0 .D 51 8.4 1015166 473 M e F O S 0 7 1 .D 52 8.4 1001588 487 MeFOS072.D 53 8.4 1454983 M e F O S 0 7 3 .D 54 8.4 932168 448 M eFOS074.D 55 8.4 933193 451 M eFOS075.D 56 8.4 1349755 M eFOS076.D 57 8.4 1333450 410 MeFOS077.D 56 8.4 916402 418 MeFOS078.D 59 8.4 1330022 609 M e F O S 0 7 9 .D 60 8.4 924714 396 M e F O S 0 8 0 .D 61 8.4 933652 . 400 M e F O S 0 8 1 .D 62 8.4 1341660 598 M e F O S 0 8 2 .D 63 8.4 923952 397 M e F O S 0 8 3 .D 64 8.4 886677 393 MeFOS084.D 65 8.4 1320368 590 0.0M eF O S 08 5.D 0.0M eF O S 08 7.D 92 92 00 0 0 MFA-106 M FA-107 MFA-108 MFA-125 MFA-126 MFA-142 MFA-143 MFA-144 MeOH Blank MeOH Blank MeFOS088.D 66 8.4 895537 389 M eFOS089.D 67 B.4 889957 395 MeFOS090.D 68 8.4 1330772 588 MeFOS091.D 69 8.4 900341 374 MeFOS092.D 70 8.4 866673 543 M e F O S 0 9 3 .D 71 8.4 1299388 546 M e F O S 0 9 4 .D 72 8.4 876044 367 M e F O S 0 9 5 .D 73 8.4 906488 370 MeFOS096.D 74 8.4 1302131 544 0.0MeFOS102.D 0.0MeFOS103.D 92 92 00 00 99039-35-1 99039-35-2 99039-35-3 99039-35-4 99039-35-5 99039-35-6 Method ID: M e F O S 1 0 4 .D M e F O S 1 0 5 .D M e F O S 1 0 6 .D M eF O S 107.D M e F O S 1 0 8 .D M e F O S 1 0 9 .D 41 8.4 160080 82 42 8.4 305109 146 43 8.4 621321 310 44 8.4 964994 496 45 8.4 1292328 636 46 8.4 1570588 782 Internal Standard quant x2 0.999 Original cone, (ng/ml) - 473.4 Curve averaged. Linear,Include origin. Spike cone, (ng/ml) = 221 Calibration range: 155-932 ng/ml (A) Data excluded on the basis of data quality objectives; see te x t % Standard o r RSD 103% 92% 98% 103% 103% 97% 3% 1% 2% 1% 1% 1% 37% 1% 104% 93% 98% 105% 101% 99% % S p R te Recovery T h * P o in t Ret Tim e 6.2 0A r e a 0.0Cone. -217% -203% 89% 90% 88% 89% 39% 79% (A) (A) (A) (A) (A) (A) Day 14 Day 14 Day 14 Day 20 Day 20 Day 20 Day 28 Day 28 Day 28 Day 34 Day 34 Day 34 (A) (A) (A) Day 49 Day 49 Day 49 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 8592 24325 43465 97689 203483 297189 00 00 0 00000 00000 00 00 0000 0 000 0 00 2.4 7.8 14.2 32.5 67.1 97.4 00..00 000000000000000...............000000000000000 00..00 000000000.........000000000 00..00 6.2 8830 6.2 24409 6.2 44065 6.2 122620 6.2 199720 6.2 282960 2.2 7.1 13.1 37.8 61.8 86.1 Curve averaged, Linear,Include origin. Calibration range: 7.8-94 ng/ml 0.0% S te n d a rd Ret Tim e 5.9 0Area 76% 99% 91% 104% 107% 104% 5.9 334451 5.9 336584 5.9 340261 5.9 341410 5.9 347910 5.9 350784 00..00 00 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 00..00 361052 358929 365070 350786 346018 351584 518509 349233 339650 356993 362219 345470 360186 352051 345627 0 0 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 00..00 355343 353220 356886 361820 354024 348654 357923 372805 358209 00 70% 90% 84% 121% 99% 92% 5.9 364922 5.9 368496 5.9 372249 5.9 369974 5.9 370582 5.9 377502 S.D. %S.D. N-EtFOSE A lco h o l Cone. 0 403 403 403 403 403 403 00 403 403 403 403 403 403 403 403 403 403 403 403 403 403 403 00 403 403 403 403 403 403 403 403 403 00 403 403 403 403 403 403 R et Tim e 8.6 0A r e a 8.6 693381 8.6 748590 8.6 706006 8.6 707232 8.6 701518 8.6 726509 08.6 08.6 8.6 719209 8.6 688438 6.6 23244 8.6 696227 08.6 693022 8.6 8.6 1090670 8.6 738040 8.6 729875 8.6 783229 8.6 781943 8.6 750255 8.6 780059 8.6 755853 8.6 747940 08.6 08.6 8.6 772071 8.6 755961 8.6 756782 8.6 806499 8.6 533536 8.6 796217 8.6 799439 8.6 820989 8.6 800015 08.6 08.8 8.6 673354 8.6 707289 8.6 673339 8.6 664378 8.6 679464 8.6 670949 S.D. %S.D. 84954 11.3% 0Cone. 311 311 311 311 311 311 00 311 311 311 311 0311 311 311 311 311 311 311 311 311 311 00 311 311 311 311 311 311 311 311 311 00 311 311 311 311 311 311 BACK TO MAIN Page 58 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Pooled N-MeFOSE Alcohol Data and Slope Regression Solid Line: Dotted Lines: 2s Uncertainty Limits (slope and intercept) SUMMARY OUTPUT Regression Statistics Multiple R 0.438823641 R Square 0.192566188 Adjusted R Sq 0.17131793 Standard Erro 0.09318559 Observations 40 ANOVA Regression Residual Total d f SS 1 0.078696282 :38 0.329975058 39 0.408671341 Intercept X Variable 1 Coefficients Standard Error -0.0125162.55 0.029261069 -0.002996307 0.000995309 %2 s slope uncertainty 66.4 MS 0.078696282 0.008683554 F Significance F 9.062681099 0.004617044 tStat -0.427744267 -3.010428723 P-value 0.671251755 0.004617044 Lower 95% -0.071752194 -0.005011205 Upper 95% 0.046719685 -0.000981409 Lower 95.0% -0.071752194 -0.005011205 Upper 95.0% 0.046719685 -0.000981409 Page 59 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix D: Selected Chromatograms A representative set of chromatograms from the present study is included in this Appendix. Page 60 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Method C:\HPCHEM\l\METHODS\1227 701.M Calibration Table P H ^o, Calib. Data Modified : Calculate Based on Rel. Reference Window Abs. Reference Window Rel. Non-ref. Window Abs. Non-ref. Window Multiplier Dilution Sample Amount Uncalibrated Peaks Partial Calibration Correct All Ret. Times Curve Type Origin Weight Wednesday, December 29, 1999 9:20:34 AM Internal Standard Peak Area iyrl l 10.000 % 0 . 0 0 0 min 10.000 % - J it e T s & l / i ' <L 0.000 min fS /I Copy Of 1.0000 1.0000 3/tf 0.00000 not reported Yes, identified peaks are recalibrated No, only for identified peaks Linear Included Equal `S p p w tQ -- : Ci/' __ . Recalibration Settings Average Response Average Retention Time Average all calibrations SAMPL Floating Average New 75%" l / p U L f f & i Calibration Report Options : Printout of recalibrations within a sequence: Calibration Table after Recalibration Normal Report after Recalibration If the sequence is done with bracketing: Results of first cycle (ending previous bracket) ^^ Sample ISTD Information: ISTD ISTD Amount Name # tppb] I --------------------- I --------------------------- X 4 0 2 .6 0 0 0 0 THPFOS 2 310.80000 EtFOSE-OH Signal 1: MSD1 427, EIC=426.7:427.7 Signal 2: MSD1 499, EIC=498..7:499.7 Signal 3: MSD1 630, EIC=629.7:630.7 Signal 4: MSD1 616, EIC=615.7:616.7 RetTime Lvl Amount [min] Siq [ppb] Area Amt/Area Ref Grp Name 5.870 11 2 3 4 5 6 11 402.60000 4.13061e5 402.60000 4.15745e5 402.60000 4.14516e5 402.60000 4.16734e5 402.60000 4.14375e5 402.60000 4.18593e5 402.60000 4.07992e5 9.74674e-4 9.68383e-4 9.71252e-4 9.66084e-4 9.71584e-4 9.61793e-4 9.86784e-4 II THPFOS Page 61 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Method C:\HPCHEM\l\METHODS\1227 701.M RetTime 6.162 8.306 8.535 Lvl Amount Area r [ppb] II- - 1! 12 402.60000 4.07169e5 13 402.60000 4.09832e5 14 402.60000 4.09148e5 15 402.60000 4.15661e5 16 402.60000 4.16714e5 : 1 3.13000 2.19264e4 11 3.13000 1.74464e4 2 7.83000 3.32915e4 12 7.83000 2.99914e4 3 15.65000 5.41038e4 13 15.65000 5.80162e4 4 31.30000 1.20587e5 14 31.30000 1.12160e5 5 62.60000 1.97863e5 15 62.60000 1.90397e'5 6 93.90000 2.83971e5 16 93.90000 2.72629e5 : 1 78.90000 1.63904e5 11 78.90000 1.62533e5 2 157.80000 3.53900e5 12 157.80000 3.38524e5 3 315.60000 6.72851e5 13 315.60000 6.51730e5 4 473.40000 1.04325e6 14 473.40000 1.01321e6 6 789.00000 1.67835e6 16 789.00000 1.63482e6 i 1 310.80000 7 .22548e5 2 310.80000 7.26841e5 3 310.80000 7.21094e5 4 310.80000 7.21008e5 5 310.80000 7.22268e5 6 310.80000 7.24772e5 11 310.80000 7.07642e5 12 310.80000 7.00669e5 . 13 310.80000 7.05419e5 14 310.80000 7.09949e5 15 310.80000 7.18262e5 16 310.80000 6.9982 9e5 Amt/Area 9.88780e-4 9.82353e-4 9.83996e-4 9.68579e-4 9.66131e-4 1.42750e-4 1.79406e-4 2.35195e-4 2.61075e-4 2.89259e-4 2.69752e-4 2.59563e-4 2.79065e-4 3.16381e-4 3.28787e-4 3.30667e-4 3.44424e-4 4.81379e-4 4.85439e-4 4.45889e-4 4.66141e-4 4.69049e-4 4.84249e-4 4.53774e-4 4.67226e-4 4.70106e-4 4.82621e-4 4.30145e-4 4.27604e-4 4.31012e-4 4.31063e-4 4.30311e-4 4.28824e-4 4.39205e-4 4.43576e-4 4.40589e-4 4.37778e-4 4.32711e-4 4.44109e-4 12 sessessex It II II H II II II Rn R Peak Sum Table ***No Entries in table*** PFOS MeFOSE-OH EtFOSE-OH Page 62 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 1 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS051.D Sample Name: MeOH Blank Injection Date 10/7/99 5:09:30 AM Seq. Line : 51 Sample Name MeOH Blank Vial : 91 A c q . Operator Acq. Method Last changed MTM . C:\HPCHEM\l\METHODS\FOSESIM.M 10/6/99 1:56:00 PM by MTM Inj : 1 pH Analysis Method C:\HPCHEM\1\METHODS\1227_70I.M Last changed 12/29/99 9:23:15 AM by MTM (modified after loading) (Results are from a previously s SIM Analysis (ES for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. Internal Standard Report Sorted By Signal Calib. Data Modified Multiplier Dilution 12/29/99 9:23:15 AM 1.0000 1.0000 Sample ISTD Information: ISTD ISTD Amount Name # _______ I [ppb] 1_____ 1 402.60000 'THPFOS 2 310.80000 EtFOSE-OH Page 63 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Method C:\HPCHEM\1\METHODS\1227_70I.M Calibration Curves THPFOS at exp. RT: 5.870 MSDl 427,, EIC=426.7:427.7 Correlation: 1.00000 Residual Std. Dev.: 0.00000 Formula : y = mx + b m: 1.00000 b: 0.00000 x: Amount Ratio y : Area Ratio PFOS at exp. RT: 6.162 MSD1 499, EIC=498.7:499.7 Correlation: 0.99654 Residual Std. Dev.: 0.02056 Formula: y = mx + b m: 2.79093 b: 2.92806e-2 x: Amount Ratio y: Area Ratio MeFOSE-OH at exp. RT: 8.306 MSD1 616, EIC=615.7 :616.7 Correlation: 0.99973 Residual Std. Dev.: 0.01989 Formula: y = mx + b m: 9.19192e-l b: . 6.85505e-3 x: Amount Ratio y: Area Ratio EtFOSE-OH at expl RT: 8.535 MSD1 630, EIC=629.7:630.7 Correlation: 1.00000 Residual Std. Dev.-: 0.00000 Formula: y = mx + b m: 1.00000 b: 0.00000 x: Amount Ratio y: Area Ratio Page 64 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 1 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS051.D Sample Name: MeOH Blank Signal 1: MSD1 427, EIC=426.7:427.7 RetTime Type Area Amt/Area [min] ratio ------ |______ I_________ I 5.870 Totals without ISTD(s) : Amount Grp Name [ppb] - . . THPFOS 0.00000 Signal 2: MSD1 499, EIC=498.7:499.7 RetTime Type Area Amt /Area [min] ratio ______ |______ |__________ | 6.213 MM 1.21236e4 o.ooooo' Totals without ISTD(s) : Amount Grp Name [ppb] 0.00000 PFOS 0.00000 Signal 3: MSD1 630, EIC=629.7:630.7 RetTime Type [min] _____ |______ 1 8.535 Area Amt/Area Amount ratio [ppb] 1_________ 11 -. Grp Name ` 'EtFOSE-OH Totals without ISTD(s) : O.QOOOO Signal 4: MSD1 616, EXC=615.7 :616.7 RetTime Type Area Amt/Area [min] ratio ______ |..... . I__ _______| __ _ ___ | 8.306' Amount [ppb] - Grp Name 1~~ 1 . MeFOSE-OH Totals without iSTD(s) : 0.00000 2 Warnings or ]Errors : Warning : ISTD compound(s) not found Warning : Negative results set to zero (cal. curve intercept), (PFOS) *** End of Report *** Page 65 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 11 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS061.D Sample Name: MFA-010 Injection Date : 10/7/99 8:11:41 AM Seq. Line : 61 Sample Name :MFA-010 Vial 51 Acq. Operator :MTM Acq'. Method Last changed :C:\HPCHEM\l\METHODS\FOSESIM.M :10/6/99 1:56:00 PM by MTM `yfj/flPi Analysis Method : C:\HPCHEM\1\METHODS\1227_70I.M Last changed ' : 12/29/99 9:23:15 AM by MTM bo (modified after loading) (Results are from a previously s SIM Analysis (ES-) for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. Internal Standard Report Sorted By : Calib. Data Modified : Multiplier : Dilution . : Sample ISTD Information: ISTD ISTD Amount Name # [ppb] Signal 12/29/99 9:23:15 AM 1.0000 1.0000 1 402.60000 THPFOS 2 310.80000 EtFOSE-OH Page 66 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 11 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS061.D Sample Name: MFA-010 Signal 1: MSD1 427, EIC=426.7:427.7 ` RetTime Type Area Amt/Area Amount Grp Name [min] ratio [ppb] ' ---------- I ............I ---------------- I ........... - " - I ........... -- - I " l ----------- 5.874 BB I 4.31433e5 1.00000 402.60000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2: MSD1 499, EIC=498.7:499.7 RetTime Type Area Amt/Area Amount Grp Name [min] . ratio [ppb]' - -- -- I.........I------------- I------------- I........... l - l - ....... 6.203 PBA 1.37520e4 2.9l660e-2 3.74287e-l PFOS Totals without ISTD(s) : 3.74287e-l Signal 3: MSD1 63 0, EIC=629.7 :630.7 RetTime Type Area Amt/Area Amount Grp Name --[-m-i-n-]-- I-........I-------------I---r-a-t--i-o--- -I---[-p--p-b-]---- l - l ---------------------- 8.538 VB I 7.69704e5 1.00000 310.80000 EtFOSE-OH Totals without ISTD(s) : 0.00000 Signal 4: MSD1 616, EIC=615.7 :616.7 RetTime Type Area Amt/Area Amount Grp Name --[-m-i-n-]-- I-- -- 1-- ........- | --r--a-t-i-o----- I--- --[ppb] - l - l ------- --------------- 8.310 BV 1.05985e6 1.08250 463.26250 MeFOSE-OH Totals Without ISTD(s) : 463.26250 *** End of Report *** Page 67 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 12 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS062.D Sample Name: MFA-Oll Injection Date : 10/7/99 8:29:54 AM Seq. Line : 62 Sample Name : MFA-011 Vial : 52 Acq. Operator : MTM Inj : 1 Acq. Method C:\HPCHEM\1\METH0DS\F0SESIM.M Last changed 10/6/99 1:56:00 PM by MTM Analysis Method C :\HPCHEM\1\METHODS\1227_7 01 .M O ofU L m , ! . Last changed 12/29/99 9:23:15 AM by MTM yyf (modified after loading) (Results are from a previously s SIM Analysis (ES-) for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. Internal Standard Report Sorted By : Signal Calib. Data Modified : 12/29/99 9:23:15 AM Multiplier : 1.0000 Dilution : 1.0000 Sample ISTD Information: ISTD ISTD Amount Name # [ppb] ___ 11 1___ _____ 1 402.60000 THPFOS 2 310.80000 EtFOSE-OH Page 68 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 13 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS063.D Sample Name: MFA-012 Injection Date 10/7/99 8:48:10 AM Seq. Line : 63 Sample Name MFA-012 Vial : 53 Acq. Operator Acq. Method Last changed MTM C:\HPCHEM\l\METHODS\FOSESIM.M 10/6/99 1:56:00 PM by MTM Inj : 1 t P ij Analysis Method C:\HPCHEM\1\METHODS\1227_70I.M ----< vj,,/ Last changed 12/29/99 9:23:15 AM by MTM /YifrftVyL (modified after loading) (Results are from a previously SIM Analysis (ES-) for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879, MTM Internal Standard Report Sorted By : Calib. Data Modified : Multiplier : Dilution : Sample ISTD Information: ISTD ISTD Amount Name # [ppb] Signal 12/29/99 9:23:15 AM 1.0000 1.0000 1 402.60000 THPFOS 2 310.80000 EtFOSE-OH Page 69 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 12 of 50 Data File C:\HPCHEM\l\DATA\l00699\MeFOS062.D ' Sample Name: MFA-011 Signal 1: MSD1 427, EIC=426.7-.427.7 ` RetTime [min] Type Area Amt/Area Amount ratio ' [ppb] Grp Name - ................. I ---------------- I ....................................... I ....................-- " I - ...................- 1 - 1 --------------------------- ------------- -- 5.873 BB I 4.27482e5 1.00000 402.50000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2: MSD1 499, EIC=498.7:499.7 RetTime [min] Type Area Amt/Area ratio Amount [ppb] Grp Name . ------------------I ........................I ........................- - I ---------------------------I -- - -- -- I -- I t ----------------------------------------------- S.207 MM 1.34305e4 2.43726e-2 3.08284e-l PFOS Totals without ISTD(s) : 3.08284e-l Signal 3: MSD1 630, EIC=629.7:630.7 RetTime Type Area [min] Amt/Area ratio Amount [ppb] Grp Name ------------------| ---------------- | ----------- -- ------------- ] --------------------------- | - ....................-- - 1 - 1 - ---------------- --------------------------- 8.538 BB I 7.38896e5 1.00000 310.80000 EtFOSE-OH Totals without ISTD(s) : 0.00000 Signal 4: MSD1 616, EIC=615.7:616.7 RetTime Type Area Amt/Area Amount Grp Name [min] ratio [ppb] -----------1-- -- | ----------------- | -----------------| - ------------- | - | ---------- -------------------- 8.310 BV 1.03510e6 1.08259 471.34921 MeFOSE-OH Totals without ISTD(s) : 471.34921 *** End of Report *** Page 70 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 13 of 50 Data File C:\HPCHEM\l\DATA\100699\MeFOS063.D Sample Name: MFA-012 Signal 1: MSD1 427, EIC=426.7:427.7 ` RetTime Type Area Amt/Area Amount Grp Name [min] . ratio [ppb] ---------- 1----------1.............- -- I--------- 1---------------------- 1 ~ | ----------- 5.872 PB I 4.24804e5 1.00000 402.60000 THPFOS Totals without ISTD(s) : 0.00000 Signal 2: MSD1 499, EIC=498.7:499.7 RetTime Type [min] Area Amt/Area ratio Amount Grp Name [ppb] .......... - I ----------- I ----------------7 - 1 ....................... I " ..................... 1 - 1 ................ 6.206 PBA 1.24702e4 9.10909e-4 1.07655e-2 PFOS Totals without ISTD{s) : 1.07655e-2 Signal 3: MSD1 630, EIC=629.7:630.7 RetTime Type Area Amt/Area Amount Grp Name [min] ratio [ppb] -----------I ---------- I -- - ........... I -------------- I - ....................I " ! ------------- - 8.535 PB I 7.26047e5 1.00000 310.80000 EtFOSE-OH Totals without ISTD(s) : 0.00000 Signal 4: MSD1 616, EIC=615.7:616.7 RetTime Type Area Amt/Area Amount Grp Name [min] ----------- | -----------| ............................. | -----r-a--t--i--o------| .........[..p..p...b..]...........| _ _ | ----------------- 8.307 BV 1.49310e6 1.08429 693.02691 MeFOSE-OH Totals without ISTD(s) : 693.02691 *** End of Report *** Page 71 of 71