Document 6bdJ9NGweZ21gkee7eek7kQad

BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Study Title Screening Studies on the Aqueous Photolytic Degradation of Perfluorooctanoic Acid (PFOA) Data Requirement Consistent With: OPPTS: 835.5270 "Indirect Photolysis Screening Test" -and- OECD Draft Document "Phototransformation of Chemicals in Water - Direct and Indirect Photolysis", August 2000 Author Thomas L. Hatfield, Ph.D. Study Completion Date April 20th, 2001 Performing Laboratory 3M Environmental Laboratory Building 2-3E-09, 935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Laboratory Report No: E00-2192 Total Number of Pages 148 Page 1 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 This page has been reserved for specific country requirements. Page 2 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Non - Compliance Statement Study Title: Screening Studies on the Aqueous Photolytic Degradation of Periluorooctonic Acid (PFOA) Study Identification Number: E00-2192 By design, this study does not comply with the requirements of the US EPA Good Laboratory Practices Standards at 40 CFR Part 792 (TSCA). However, the 3M Environmental Laboratory Quality Assurance Unit has performed audits of all data, related documentation and final report. Test and reference substance receipt and use, dosing and incubation of the test system, and analyses were conducted and documented according to procedures developed by 3M, based on References 1 and 2. Page 3 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Quality Assurance Statement Study Title: Screening Studies on the Aqueous Photolytic Degradation of Perfluorooctanoic Acid (PFOA) Study Identification Number: E00-2192 This study has been inspected by the 3M Laboratory Quality Assurance Unit as indicated in the following table Inspection Dates 03/30/01-03/31/01 04/18/01-04/19/01 Phase Data Draft Report Date Reported to Management Study Director 03/31/01 03/31/01 04/19/01 04/19/01 Quality Assurance Unit C/JX ojQ \ Date Page 4 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Table of Contents Non - Compliance Statement........................................................... ,....................................... 3 Quality Assurance Statement................................................................................................... 4 Table of Contents......................................................................................................................5 Study Personnel and Contributors...........................................................................................7 Summary................................................................................................................................... 8 Introduction................................................................................................................................ 9 Materials and Methods............................................................................................................. 11 Chemical Characterization..................................................................................................11 Method Summaries............................................................................................................. 11 Deviations............................................................................................................................. 13 Results and Discussion........................................................................................................... 14 Data Quality Objectives....................................................................................................... 14 Analytical Results................................................................................................................ 14 Data Summary and Discussion......................................................................................... 15 Conclusions............................................................................................................................... 18 References................................................................................................................................19 S ig n a tu re s ................................................................................................................................. 20 Appendix A: Analytical Methods............................................................................................... 21 Appendix B: Chemical Characterization.................................................................................105 Appendix C: Kinetics Model and Kinetics Calculations........................................ 109 Appendix D: Representative Chromatograms........................................................................119 Appendix E: Soil Types and Characterizations...................................................................... 138 Appendix F: Light Intensity Measurements at 45 South Latitude (Miami F L)..................... 140 Appendix G: Characteristics of the Spectral Output of the Suntest Instruments................ 142 Page 5 of 148 BACK TO MAIN IKIKKKIKKKIKKIIKMI 3M Environmental Laboratory Report No. E00-2192 List of Tables Table 1. Typical Sample Preparation Scheme Used in the Present Investigation...............12 Table 2. Observed Products and Mass Balance Determinations for pH 7 Buffer, Synthetic Humic Water and Iron Oxide Containing Water...................................................................16 List of Figures Figure 1. Structures of the Compounds Targeted by LC/MS Analysis.................................9 Figure 2. Pooled concentration data from the iron oxide rich matrix.................................... 17 Page 6 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Study Personnel and Contributors Study Director Thomas L. Hatfield, Ph.D. 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 (651) 778-7863 Sponsor 3M Corporation 3M Environmental Laboratory Contributing Personnel Kent Lindstrom Anh Dao Vo 3M Environmental Laboratory Professional Services Contributing Personnel Anthony (Tony) Scales Debra Wright Jan Schutz Rufat Mischiev (Pace Analytical Services, Inc., 1700 Elm St., Minneapolis, MN 55144) Kristin Terrell Jill Maloney Karen Johnson (Braun Intertec Corporation, 6875 Washington Ave. South, Minneapolis, MN 55439) Location of Archives Digital copies of original data, and all original paper data have been archived and will be retained in the 3M Environmental Laboratory archives for at least 10 years Page 7 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Summary We report here the results of studies performed to determine the aqueous photolytic behavior of perfluorooctanoic acid (PFOA) and to identify its primary degradation products. Our techniques are based on both EPA and OECD guidance documents.1,2 In this study, both direct photolysis (the interaction of light with the target molecule leading to a chemical change) and indirect photolysis (the Interaction of light with the sample matrix to produce radical species that subsequently react with the target material) were studied using a synthetic light source. Neither direct nor indirect photolytic decomposition of PFOA were observed based on loss of starting material, nor were any of the predicted degradation products detected above their limits of quantitation. The rates of photolytic degradation are highly dependent on the experimental conditions. However, using an iron oxide (Fe203) photoinitiator matrix model, we estimate the PFOA half-life to be greater than 349 days. Page 8 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Introduction Photolysis reactions, hydrolysis reactions and biodegradation are the primary routes of degradation of chemical compounds in the environment. Studies of photo-induced reactions yield information on the persistence of the parent material as well as information on the identity and stability of products formed. Photolytic reactions occur by two types of mechanisms. The first mechanism, direct photolysis, can be defined as the direct absorption of a photon by the target species that leads to a chemical change. The second mechanism, indirect photolysis, can be defined as a chemical or electronic excitation transfer from a light absorbing species to the test substance, which then undergoes some type of chemical change. In the present investigation, an artificial light source was used to study both the direct and indirect photolysis reactions of PFOA. The test material, PFOA, was dissolved in pH 7 buffered water and then exposed to simulated sunlight to test for direct photolysis.3,4 To test for indirect photolysis, PFOA was dissolved into three separate matrices and exposed to simulated sunlight for periods of time from 69.5 to 164 hours. These exposures tested how each particular matrix would effect the photolytic decomposition of PFOA. The first test matrix was a pH 7 buffered aqueous solution to which H2O2 was added as a well characterized source of .OH radicals.5,6 This was used to test for the propensity of PFOA to undergo indirect photolytic decomposition. The second matrix contained Fe20 3 in water, as this matrix has been shown to generate hydroxyl radicals via a Fenton-type reaction in the presence of both natural and artificial sunlight.7,8 The third matrix contained a standard humic material, diluted to levels that have been shown to approximate an environmental environment.1,2 To effectively determine photolytic decomposition, the concentration of parent material must be monitored over time. Further, it is also important to understand what the degradation products are and how much of each are formed. The present investigation quantified the parent material and two predicted degradation products (PFHpA - perfluoroheptanoic acid and PFPA perfluoropentanoic acid) and monitored for any change in concentration of PFHxA - perfluorohexanoic acid by LC/MS. Structures of the pertinent compounds are illustrated in Figure 1. Figure 1. Structures of the Compounds Targeted by LC/MS Analysis PFOA 0 PFHpA PFHxA PFPA Page 9 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Because it was possible that volatile degradation products could be produced, we decided to monitor for selected C2 through C8 2- or 3- substituted periluoronated olefins (e.g. C8Fi 6) and 1or 2- substituted hydrides (e.g. C8F17H) in both the iron-rich matrix and the pH 7 buffer matrix by dynamic purge and trap gas chromatography/mass spectrometry. The selected target compounds are representative of the types of volatile compounds that could be generated in the photolysis of PFOA. Determination of a maximum kinetic rate constant ( k p ) ^ was based on the data from the iron rich matrix (as the experimental error was lowest in this matrix) using the following first order kinetics equation. (See Appendix C for a complete kinetic derivation and the exact mathematical solution. This equation is valid for essentially constant parent concentrations with a mean value Pp and a standard deviation of c p.) S (k ,)_ = (039) Page 10 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Materials and Methods Chemical Characterization Information on the chemical characterization of both reference substances and control substances is presented in Appendix B. Method Summaries Copies of all analytical methods used in this investigation are attached in Appendix A. Equipment settings, conditions and complete quality control parameters are listed in the pertinent methods. UVA/isible analysis were performed following 3M Environmental Laboratory Method ETS-946.0 "Operation and Maintenance of the Hewlett Packard 8453 UV-Visible Spectrophotometer" An aqueous saturated solution of PFOA was prepared and an initial UVA/IS spectrum recorded. No absorbance above baseline over the range 190-1100 nm was detected. Sample preparation for this analysis followed 3M Environmental Laboratory Method ETS-8177.0 "Indirect Photolysis Screening Test in Synthetic Humic Water" or ETS-8-176.0 "Preparation of Samples for Photolytic Exposure Studies in Aqueous Matrices". A typical sample preparation table for an indirect photolysis screening test is shown in Table 1 on the following page. The general method of sample preparation is as follows. For each time point under each condition shown in Table 1, ten 40-mL sample screw cap VOA vials were prepared: sample, duplicate, triplicate, sample spike, matrix blank, matrix blank spike (assured no accidental contamination of matrix by target compounds), direct photolysis sample, direct photolysis sample spike (assured that degradation observed was due to indirect photolysis and not another process), control blank and control blank spike (assured no accidental contamination of the blank had occurred). All vials contained 5 mL of appropriate matrix: pH 7 buffer, Fe+3 at a 24X molar excess in water, or H20 2 at 1:1 molar equivalent, added every 24 hours. Aliquots of PFOA were added to the vials as indicated in Table 1. The initial time point vials (labeled as "Time Zero" on the sample preparation sheets in Appendix B) were then refrigerated at 42C. These samples served as controls with which to determine what change, if any, occurred during the time the remaining vials were in the photo-reactor. Exposed samples were placed upside down in a custom built holder in the photo-reactor. Unexposed samples were wrapped in aluminum foil, sealed in a plastic bag and placed under the sample rack inside the photo-reactor to assure that any degradation or difference in degradation was due to photolysis and not some other process. During the course of the exposure, a water bath held the temperature of the water surrounding the bottom of the vials (which contained the aqueous samples) at 25 3C. The temperature of the chamber itself was allowed to drift to 70 10C. After exposure, the samples were removed for analysis. Selected portions of the sample setup were modified to accommodate additional samples and controls, to improve the quality control of the analysis or out of experimental necessity. Details of these modifications are shown on the individual sample data sheets shown in Appendix D. For example, the portion of the study employing iron oxide as a radical generating species was set up with both triplicate samples and duplicate sample spikes. Page 11 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 The artificial light chamber used in this analysis was either a Suntest CPS+ or Suntest XLS+, the operation of which followed 3M Environmental Laboratory Method ETS-9-44.0 "Operation and Maintenance of the Sunlight Exposure System, Immersion Unit, and Recirculating Water Chiller System". The output intensity was held at the desired level (680 w/m2) by a continuous feed back loop between an internal radiometer and the variable light source. Table 1. Typical Sample Preparation Scheme Used in the Present Investigation D escription Ind. Photo. Sam ple Rep 1 Ind. Photo. Sam ple Rep 2 Ind. Photo. Sam ple Rep 3 Ind. Photo. Sam ple Spike M atrix Blank M atrix Blank Spike Direct Photo. Sam ple Direct Photo. Spike Control Blank Control Blank Spike T est M atrix (e.g. Fe20 3 in w a te r) + + + + + + 0 0 0 0 C o n tro l M atrix (e.g. w a te r) 0 0 0 0 0 0 + + + + Test S u b s ta n c e (P F O A ) + + + + 0 0 + + 0 0 Post Photolysis T arget A nalyte spike 0 0 0 + 0 + 0 + 0 + Sam ple Type (C onditions) LC/MS* W ith HA W /out H 02 GC/MS* W ith HQ, W /out H 02 Initial Initial Initial Initial . Initial Initial Initial Initial Initial Initial XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X Ind. Photo. Sam ple Rep 1 Ind. Photo. Sam ple Rep 2 Ind. Photo. Sam ple Rep 3 Ind. Photo. Sam ple Spike M atrix Blank M atrix Blank Spike Direct Photo. Sample Direct Photo. Spike Control Blank Control Blank Spike + + + + + + 0 0 0 0 0+ 0+ 0+ 0+ 00 00 ++ ++ +0 +0 0 0 0 + 0 + 0 + 0 + Light Light Light Light Light Light Light Light Light Light XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X ind. Photo. Sam ple Rep 1 Ind. Photo. Sam ple Rep 2 Ind. Photo. Sam ple Rep 3 Ind. Photo. Sam ple Spike M atrix Blank M atrix Blank Spike D irect Photo. Sam ple D irect Photo. Spike Control Blank Control Blank Spike + + + + + + 0 0 0 0 0+ 0+ 0+ 0+ 00 00 ++ ++ +0 +0 0 No Light XX X 0 No Light XX X 0 No Light XX X + No Light XX X 0 No Light XX X + No Light XX X 0 No Light XX X + No Light XX X 0 No Light XX X + No Light XX X + = added to test vial; 0 = NOT added to test vial; ` Duplicate sets, one with H20 2, one without (excludes the Humic material test where H20 2was not added) X X X X X X X X X X GC/MS analysis followed 3M Environmental Laboratory Method ETS-8-182.0 "Analysis of Fluorochemicals by Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent Gas Chromatograph/Mass Spectrometer". Equipment settings, separation conditions and ions monitored are presented in this method. Equipment procedures for the GC/MS system followed 3M Environmental Laboratory SOP ETS-9-49.0 "Routine Maintenance Page 12 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 of Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent Gas Chromatograph/Mass Spectrometer". The HPLC/MS analysis followed 3M Environmental Laboratory Method ETS-8-181.0 "Analysis of Photolysis Samples for Fluorochemicals by High Performance Liquid Chromatography With Mass Spectrometry Detection". Equipment settings, separation conditions and ions monitored are presented in this method. Deviations Calibration points were deleted at both ends of the PFHpA and PFPA curves. The lowest point on the curve was deleted due to poor integration and the two highest points were deleted to improve quantitation. The resulting effective concentration ranges were 5.5-219 ppb for PFPA and 5.2-206 ppb for PFHpA. While setting up additional studies on the photochemical behavior of other compounds in synthetic humic material, it was determined that the commercial humic material (Aldrich) did not contain the appropriate amount of dissolved organic carbon when the solution is prepared as recommend in reference 1. The study director believes that this may have inhibited any possible photodegradation of PFOA and that these data should therefore be considered of screening quality only. Further, the samples from this matrix showed high levels of background ions with the same charge to mass ratio as that of the targeted materials and the manual intergration of some peaks was therefore required. PFHxA was identified as a possible degradation product. However, no suitable standard of this material could be obtained for the LC/MS portion of this study. Area of the chromatographic peak corresponding to the PFHxA anion (m/z = 313) were monitored. No changes greater than 10% relative were observed above the small amounts of PFHxA initially present in the PFOA standard. The PFOA samples numbered 0515-PFOAfe-07, and 30 (LC/MS) failed to meet spike recovery criteria in the iron oxide portion of this investigation and 0515PFOAfe-30 and 90 failed control sample recovery. LC/MS method blanks 0515-PFOAfe-21,31,45,55 showed low recovery. The GC/MS CCV R0602046 failed to pass criteria. The samples numbered 0515PFOAshw-04,14 and 28 failed to meet spike recovery criteria and control sample 0515shw-28 showed low recovery in the synthetic humic portion of this investigation. Samples numbered 0515-PFOA-08, 50,14 and 48 failed to meet spike recovery criteria in the pH 7 buffer portion of this investigation. Control sample 0515-PFOA-48 showed low PFOA recovery while four sample triplicate sets for PFPA analysis had high RSD. The GC/MS samples numbered 051500PFOAfe-067, 077 and 090 failed to meet spike recovery criteria in the iron oxide portion of the investigation. Two samples, 051500-PFOAshw-12 and 051500-PFOAshw-17 leaked. These failures represent only 2% of all samples and QC data and therefore have no significant impact on the results. During the exposure time of the pH 7 buffered matrix, the lamp went out and the temperature drifted. However, this represents less than 3% of the total time the samples were exposed. The conclusions are not effected and the kinetic determinations didn't rely on this data. Page 13 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Results and Discussion Data Quality Objectives The following data quality objectives are summaries of those found in the methods from Appendix A. Calibration curves. An acceptable coefficient of determination (R2) for linear curves is 0.990 or greater. Curve linearity, intercept, and quantitation accuracy should be verified, particularly at upper and lower calibration limits. Residuals generated in curve-fitting must be within 25% of the known standard value. Alternative methods of curve-fitting (e.g., quadratic) require a correlation coefficient (r) of 0.990 or greater. Reasons for the use of quadratic curve-fitting must be documented in the raw data. Solvent blanks, Matrix blanks, Control blanks. Blanks should show no more than 5% of the level of a high standard or CCV and should show less than 25% of the lowest point of the calibration curve. If solvent blanks show more than a 5% carryover, it may be necessary to rule out instrument contamination using duplicate solvent blank injections. If, after duplicate solvent blanks, there is still more than 5% carry-over, or the LOQ is adversely affected, the run should be stopped. This indicates that the instrument is contaminated and should be thoroughly cleaned. Sample spikes, Matrix spikes, Control spikes. Acceptable spike recoveries must be between 75 and 125% for both LC/MS and GC/MS analysis. Values outside these ranges must be documented and evaluated by the Team Leader or designated supervisor. Sample triplicates. All samples are prepared in triplicate (unless otherwise noted). Acceptable RSD precision values are less than or equal to 25%. Values above 25% must be documented and evaluated by the Team Leader or designated supervisor. Continuing calibration verification (CCV). The analyte concentrations must not vary by more than 25% of their expected values, relative to the initial calibration curve. Accept only those samples analyzed before the most recently accepted calibration verification. Reanalyze remaining samples with a new calibration curve. Limit of Quantitation. The limit of quantitation (LOQ) is equal to the concentration of lowest standard in the calibration curve that has an area greater than or equal to four times the solvent blanks and possessing a residual less than 25% of the actual value. Control Samples. Control samples must be within 25% of the nominal concentration. Analytical Results Data quality objectives for this study, outlined in the 3M laboratory method for this study (see Appendix A), were met, with the exceptions noted in the Deviations section. Calibration curves. Calibration curves were prepared according to the particular target at the required levels. For example, calibration curves for PFOA ranged from 25 to 2000 ppb. Calibration curves for degradation products for analysis by LC/MS typically ranged from 2 200 ppb. Calibration curves for GC/MS analysis ranged from 1 ppb to 15 ppb. Using these standards, calibration curves were run before and after every analytical sequence. A correlation coefficient (r) of 0.990 or greater was achieved for all curves. Page 14 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Solvent blanks. All solvent blanks (MeOH) were less than 25% of the method LOQ. In certain cases, as noted in the Deviations section, the LOQ was raised so that the solvent blanks met this criteria. Sample spikes. All spike recoveries for the LC/MS portion of the investigation were between 75 and 125%, except where noted in the Deviations section. All spike recoveries were between 75 and 125% for the GC/MS portion of the investigation, except were noted in the Deviations section. Sample triplicates. All sample RSD values were 25% or less. Continuing calibration verification. All CCV samples were within 25% of the expected value. Limit of Quantitation. The LOQ varied dependant upon target. In some cases, the LOQ was defined as the lowest standard that was greater than 4X the solvent blank level, see deviations section for additional information. Method Blanks. All method blanks were below 25% of the LOQ. Control Samples. All control samples were within 25%. Data Summary and Discussion Direct and indirect photolytic decomposition of PFOA was tested in three separate matrices: a pH 7 buffered water, a synthetic humic water and an iron rich water. The samples were exposed to 680 w/m2 over the wavelength range of 290-800 nm and for time periods of 69.5-164 hours. Results from the quantitation of the parent material and the potential degradation products over time, as well as mass balance determinations, are shown in Table 2. As observed in Table 2a, direct photolytic decomposition of PFOA could not be detected within experimental error. Indirect photolysis was not observed in any of the three matrices (the H2O2 rich pH 7 buffer - Table 2a, the Fe+3 containing matrix - Table 2b and the humic containing matrix - Table 2c). There was a slight increase (4 ppb) in the PFHpA concentration in the Fe+3 and H2O2 containing matrix. However, because of the small size of the increase, it is unclear whether this increase was due to indirect photolysis of PFOA, to photodegradation of an unknown impurity in the PFOA standard, or to experimental error. Page 15 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Table 2. Observed Products and Mass Balance Determinations for pH 7 Buffer, Synthetic Humic Water and iron Oxide Containing Water 2a. Matrix: pH 7 Buffer, With and Without H2O2 S am ple Conditions PFPA PFHxA PFHpA PFOA V o la tile s 164 hour Exposure Ind. Photo. Sam ple1 Initial Tim e Point Dir. Photo. Sam ple2 Initial Tim e Point Ind. Photo. Sam ple1 Exp. to Light Dir. Photo. Sam ple2 Exp. to Light Ind. Photo. Sam ple1 Not Exposed Dir. Photo. Sam ple2 Not Exposed Detection Limits nanom oles nanom oles nanom oles nanom oles nanom oles ND ND ND ND ND ND 0.292 NC NC NC NC NC NC 0 .1 7 8 0.394 0.385 0.404 0.385 0.356 0.356 0.2019 57.1 56.3 55.1 56.8 56.4 55.7 4.06 ND ND ND ND ND ND 0.0095* Mass Balance (percent) 99.3% 97.9% 95.8% 98.8% 97.9% 96.8% Vials initially contained 57.9 nMoles PFOA. 1. Indirect Photloysis Sample: These samples had H 20 2 added as a radical source, results are from triplicate samples. 2. Direct Photolysis Sample: These samples did not have H20 2 added, results are from triplicate samples. *Sum total of all volatiles. ND = non detect. NC = No ChangeDetection limit represents the change in the initial concentration that could be relaiably determined, see deviations section for discussion. 2b. Matrix: Fe20 3 in Water, With and Without H20 2 S am ple Conditions PFPA PFHxA PFHpA PFOA V o la tile s 69.5 hour exposure nanom oles nanom oles nanom oles nanom oles nanom oles Fe20 3 W / H A 1 Initial Tim e Point ND NC 0.846 122 ND Fe20 3 W O /H A 2 Fe20 3 W / H A 1 Initial Tim e Point Exp. to Light ND ND NC 0.808 121 NC 1.27 105 ND ND Fe20 3 W O /H A 2 F e A W /H A 1 Exp. to Light Not Exposed ND ND NC 0.808 116 NC 0.769 121 ND ND F e A W O /H A 2 Not Exposed ND NC 0.789 118 ' ND Detection Lim its 0.729 0 .3 4 8 0.500 2.03 0.0095* Mass Balance (percent) 106% 105% 91.8% 101% 105% 102% Vials initially contained 116 nMoles PFOA. 1. Samples had H20 2and Fe20 3 added as a radical generating species, results are from triplicate samples. 2. Samples contained just Fe20 3as a radical generating species, results are from triplicate samples. *Sum total of all volatiles. ND = non detect. NC = No Change - Detection limit represents the change in the initial concentration that could be relaiably determined, see deviations section for discussion. 2c. Matrix: Synthetic Humic Water S am ple Conditions 164 hour exposure Humic W ater1 Initial Tim e Point W ater2 Initial Tim e Point Humic W ater1 Exp. to Light W ater3 Exp. to Light Humic W ater1 W ater2 Not Exposed Not Exposed Detection Limits PFPA nanom oles ND ND ND NA ND ND 0.729 PFHxA nanom oles NC NC NC NA NC NC 0 .1 7 8 PFHpA nanom oles 0.433 0.469 0.404 NA 0.394 0.391 0.500 PFOA nanom oles 55.7 60.2 5 2 .0 NA 59.3 55.0 2.03 Mass Balance (percent) 96.8% 105% 90.5% NA 103% 95.6% Vials initially contained 57.9 nMoles PFOA. 1. Samples contained Humic Materials, samples are from triplicate analysis. 2. Samples were plain water, results are from a single replicate. 3. Vial leaked, all data listed as NA or not applicable. ND = non detect. NC = No Change - Detection limit represents the change in the initial concentration that could be relaiably determined, see deviations section for discussion. Page 16 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 The Fe203 data were used as an environmental model to generate a half-life estimate for the degradation of PFOA. Because no degradation was observed, data from Table 2b, Fe203 with and without H20 2, was pooled to determine the experimental error of the analysis. This data, shown graphically in Figure 2, yielded an estimated environmental half-life of > 349 days. The exact mathematical solution is shown in Appendix C. Figure 2. Pooled concentration data from the iron oxide rich matrix. Page 17 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Conclusions A quantitative preliminary investigation was undertaken to determine the photolytic stability of perfluorooctonic acid (PFOA) and to identify the primary degradation products. The investigation included experimental conditions that addressed both direct and indirect photolysis. To test for direct photolysis, samples of PFOA in pH 7 buffer were exposed to a synthetic light source for selected periods of time. Direct photolytic decomposition of PFOA was not observed based on loss of starting material, nor were any of the predicted degradation products detected above their limit of quantitation. To test for indirect photolysis, a synthetic light source was used to initiate radical formation in three separate matrices: a synthetic humic matrix, a hydrogen peroxide rich matrix, and an iron containing matrix (as Fe203). Degradation of PFOA was not observed in any matrix outside of the experimental precision of the analytical methodology. Mass balance for the degradation study was 100 10% under all experimental conditions. A minimum half-life for PFOA based upon these preliminary studies was calculated to be > 349 days. Page 18 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 References 1. Fate, Transport and Transformation Test Guidelines, OPPTS 835.5270 Indirect Photolysis Screening Test, E P A 7 1 2 -C -9 8 -0 9 9 ; United States Environmental Protection Agency, U.S. Government Printing Office: Washington, DC, 1998, pp. 1-22. 2. OECD Guideline for Testing of Chemicals, Phototransformation of Chemicals in Water--Direct and Indirect Photoysis, (Draft Document); OECD, 2000, pp. 1-- 59. 3. Scrano, L.; Bufo, S. A.; Perucci, P.; Meallier, P.; Mansour, M. Photolysis and Hydrolysis of Rimsulfuron. Pestic. Sci. 1999, Vol. 55, pp. 955-961. 4. Nubbe, M. E.; Adams, V. D.; Moore, W . M. The Direct and Sensitized Photo-oxidation of Hexachlorocyclopentadiene. Wat. Res. 1995, Vol. 29, No. 5, pp. 1287-1293. 5. Ogata, Y.; Tomizawa, K.; Furuta, K. Chemistry of Peroxides, in S. Patai (ed.). The Chemistry of Peroxides 1983, p. 720. 6. Lunak, S.; Sedlak, P. Photoinitiated Reactions of Hydrogen Peroxide in the Liquid Phase. J. Photochem. Photobiol. A.: Chem. 1992, Vol. 68, pp. 1-33. 7. Kachanova, Z. P.; Kozlov, J. N. Zh. Fiz. Khim. 1973, Vol. 47, p. 2107. . 8. Behar, B.; Stein, G. Science 1966, Vol. 154, p. 1012. 9. Takahashi, N.; Ito, M.; Mikami, N.; Matsuda, T.; Miyamoto, J. Identification of Reactive Oxygen Species Generated by Irradiation of Aqueous Humic Acid Solution. J. Pesticide Sci. 1988, Vol. 13, pp. 4 2 9 -4 3 5 . , Page 19 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Signatures The final draft of this report is a true representation of the data developed in this study. It has been issued by: Page 20 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix A: Analytical Methods This appendix presents the analytical methods and Standard Operating Procedures used in the present study. ETS-9-46.0 Operation and Maintenance of the Hewlett Packard 8453 UV-Visible Spectrophotometer ETS-9-44.0 Operation and Maintenance of the Sunlight Exposure System, Immersion Unit, and Recirculating Water Chiller System ETS-9-49.0 Routine Maintenance of Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent Gas Chromatograph/Mass Spectrometer ETS-8-182.0 Analysis of Fluorochemicals by Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent Gas Chromatograph/Mass Spectrometer ETS-8-177.0 Indirect Photolysis Screening Tests in Synthetic Humic Water ETS-8-176.0 Preparation of Samples for Photolytic Exposure Studies in Aqueous Matrices ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by High Performance Liquid Chromatography With Mass Spectrometry Detection Page 21 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory M ethod Analysis of Photolysis samples for fluorochemicals by High Performance Liquid Chromatography w ith Mass Spectrometry Detection M ethod Number: ETS-8-181.0 Exact Copy of Original ------- Initial D a te - Approved by: Adoption Date: Effective Revision Date: OC7~ Date ^Date^7 Page 22 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application______________________________________________ 1.1 This procedure defines the steps for analysis o f fluorochemicals in photolysis study samples by high performance liquid chromatography (HPLC) with mass spectrometry (MS) detection and quantification. Refer to the HP 1100 HPLC/MSD Standard Operating Procedure ETS-9-34.0 for operating and maintenance procedures related to the instrument. Refer to Standard Operating Procedures ETS-8-176 and ETS-8-177 for information involving sample preparation and photolytic exposure procedures. 1.2 Compatible analytes. Test substance and degradation products for fluorochemicals or other fluorinated compounds, or other ionizable compounds, which includes but is not limited to: Coipnoimd Acronvm Perfluorooctanoic acid Perfluorooctanesulfonate PFOA PFOS Perfluorooctanesulfonamide FOSA N-methylperfluorooctanesulfonamide N-MeFOSA N-ethylperfluorooctanesulfonamide N-EtFOSA 2-(Kmethylpcrfluorooctanesulfonamido) N-MeFOSE-OH ethyl alcohol 2-(N-ethylperfluorooctanesulfonanudo) N-EtFOSE-OH ethyl alcohol Compound Acropvm Perfluorobutanoic acid Perfluorobutanesulfonate PFBA PFBS Perfluorobutanesulfonamide N-methylperfluorobutanesulfonamide FBSA N-MeFBSA N-ethylperfluorobutanesulfonamide 2-(N-methylperfluorobutanesulfonamido) ethyl alcohol N-EtFBSA N-MeFBSE-OH 2-(N-ethylperfluorobutanesulfonamido) ethyl alcohol N-EtFBSE-OH ... and other C4 thru Cio homologues, and polymeric materials based on the above aforementioned compounds. 1.3 Compatible matrices for analysis. Aqueous (Millipore ASTM Type I water), buffered water, lake water, sea water and metal slurries (TiC>2, Fe2 0 3, etc.) that have been diluted with an appropriate analytical solvent such as acetone or methanol. 2.0 Summary of Method____________________________________________________ 2.1 This method describes the analysis of fluorochemicals in a specified matrix, using HPLC electrospray mass spectrometry for chemical separation and detection/quantification. The analysis is performed by separating target analytes on an HPLC analytical column such as a Dionex NG1 (35x 4.6mm, 10pm particle), Betasil C l8 column (50X2 mm, 5 pm particle) or equivalent using an ammonium acetate/MeOH solvent gradient. Detection by electrospray ionization mass spectrometery in either the positive or negative mode is utilized to quantify data. The MSD may be run in Selected Ion Monitoring (SIM) mode, looking for specific, pre-selected and set analyte ions (i.e. m/z 499 for PFOS (deprotonated)), or SCAN mode which collects and stores data for all ions in a specified mass range. Data quantification is then performed using either HP ChemStation or Target Software. ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 2 of 15 Page 23 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.0 Definitions____________________________________________________________ 3.1 Calibration Standard. A dilution of various amounts o f a stock, intermediate or purchased standard to achieve standard solutions in a concentration range of interest. 3.2 Calibration Curve. The graphical relationship between known values, such as concentration of a series of calibration standards and their instrumental response. 3.3 Internal Standard Quantification. Process of establishing a relationship between the ratio of the target analyte(s) response to internal standard or surrogate response and a known concentration of the target analyte(s). The ratio of analyte to internal standard response is used to generate the calibration curve and determine unknown concentrations. 3.4 External Standard Quantification. Process of establishing the concentration o f a target analyte by plotting the theoretical amount (in units of ppb or ppm, etc.) versus the response o f the target analyte(s) on column. The resultant curve(s) shall be used to determine unknown concentrations by comparing the area response of target analyte(s) to the area response and corresponding analyte amount on the appropriate analyte's calibration curve. Differences in sample mass/volume analyzed, if noted, must be compensated for by a factor applied to the value. 3.5 Correlation Coefficient (r). A measure of the degree of correlation between two variables. This term is generally used to evaluate the linearity of a Least Squares Linear regression. An r value of 0.98 is at the lower bounds of what is considered linear. Values o f r may range from -1 to +1. A value o f +1 denotes perfect direct functional relationship between two variables. A value o f -1 also denotes a perfect inverse relationship. When r = 0, there is no effect o f one variable upon the other variable. 3.6 Coefficient of Determination (r2). The square of the correlation coefficient. It is the proportion of the variation in the dependent variable that is accounted for by the independent variable. 3.7 Internal standard. A known amount of a compound or element similar in analytical behavior to the compound(s) or elements) of interest, added to all samples and standards, and carried through the entire measurement process (post-photolysis, after solvent dilution). It provides a reference for evaluating and controlling the precision and bias o f the applied analytical method. Samples are to be quantified using the internal standard. 3.8 Surrogate. An organic compound similar to the target analyte(s) in chemical composition and behavior in the analytical process but is not normally found in the sample(s). A surrogate may be added to samples along with the test analyte (pre and/or post photolysis) to monitor the sample integrity (leaks or matrix effects). The surrogate may be added to the calibration standards to serve as a qualitative reference for the samples. 3.9 Continuing 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. ETS-8-181.0 Analysis o f Photolysis Samples for Fluorochemicals by HPLC/MS Page 3 o f 15 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.10 Solvent Blank. A sample o f analyte-free medium (for example, methanol, 1:7 diluted buffenmethanol) that is not taken through the sample preparation process. This blank is used to evaluate instrument contamination. 3.11 Blank. For photolysis studies, there are multiple blanks to adequately represent the variables o f the study (Exposed, Unexposed and Day 0 samples with/without peroxide addition). These blanks are carried through the sample preparation, photolytic and analytical procedures to monitor for contamination during any step. It is also used to establish a chromatographic baseline/background and monitor for analytical interference or suppression o f target analyte(s) from the matrix. 3.11.1 M atrix Blank: An analyte-free matrix (buffered water, lake water, etc.) to which all reagents are added in the same volumes or proportions as used in sample processing. It is used to document the test system without test analyte present. 3.11.2, Control Blank: An analyte-free matrix (ASTM Type n water) to which all reagents are added in the same volumes or proportions as used in sample processing. It serves as a control for the test matrix to monitor background levels, interferences or suppression of target analyte(s) from the test matrix. 3.12 Lim it of Quantitation (LOQ). The lowest concentration that can be reliably measured within specified limits of accuracy during routine laboratory operating conditions. The LOQ is generally 5 to 10 times the minimum concentration with a 99% confidence limit 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 of the matrix blank. Sample LOQ are highly matrix-dependent. 3.13 Sample Triplicates. Three samples taken from and representative o f the same sample source and separately carried through all steps of the extraction, photolysis and analytical procedures in an identical manner. There are multiple sets o f triplicate samples to adequately represent the photolytic variables o f the study (Exposed, Unexposed and Day 0 with/without peroxide addition). Triplicate samples are used to assess variance o f the photolytic method, including sample preparation, photolysis, and analysis. 3.14 Control Sample. A known matrix (ASTM Type II water) containing the test analyte(s) carried throughout the entire sample preparation, photolytic and analytical procedure. There are multiple sets o f triplicate samples to adequately represent the photolytic variables o f the study (Exposed, Unexposed and Day 0 with/without peroxide addition). This is used to document method performance and matrix effects by comparing recoveries from the different matrices and sample types. 3.15 Relative Standard Deviation (RSD). A measure o f precision defined as the standard deviation o f three or more values divided by the average of the values and multiplied by 100. (Also reported as Coefficient of Variation (CV)). 3.16 Analytical Spike (AS). Prepared by adding a known mass o f target analyte(s) to a specified amount o f a sample or control matrix prior to analysis. This assumes that an independent estimate of target analyte concentration is available. Analytical spikes are used to determine the effect o f the matrix on recovery efficiency. There are multiple ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 4 of 15 Page 25 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 types of spiked samples to adequately represent the photolytic variables of the study (Exposed, Unexposed and Day 0 ; with/without peroxide addition.) 3.16.1 M atrix Spike. The test matrix (buffered water, lake water) sample containing the test analyte or blank to which a known mass of target analyte(s) is added prior to analysis. 3.16.2 Control Spike. The control matrix (ASTM Type n water) sample containing the test analyte or blank to which a known mass of target analyte(s) is added prior to analysis. 3.17 Accuracy. The closeness o f agreement between an experimentally determined value and an accepted reference value. When applied to a set of observed values, accuracy is a combination of 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. 3.18 Dilution. A step in the sample preparation procedure in which a solvent (i.e. methanol, acetone) is added to the test analyte/sample matrix (i.e. water, buffer, etc.) to prepare it for instrumental analysis. 3.19 Atmospheric Pressure Ionization (API): The Agilent Technologies HPLC 1100/MSD system allows for ionization of incoming liquid sample from the analytical column to the mass spectrometer interface by utilizing a source, probe, hot gas, and specific voltages. 3.20 Electrospray Ionization (ES, ESI): A method of ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charge droplets. These charged droplets are produced by the application o f a strong electrical field. 3.21 Mass Spectrometry, Mass Spectrometer (MS), Mass Spectrometer Detector (MSD): The API HP 1100 MSD system equipped with a quadrupole mass selective detector. Ions are selectively discriminated by mass to charge ratio (m/z) and subsequently detected. 3.22 Geometric Mean of the calibration curve: The square root o f the product of the high standard concentration and the low calibration curve standard. When preparing calibration curve standards, the number of calibration standards below the geometric mean shall equal the number o f calibration standards above the geometric mean. Having equal distribution o f calibration standards above and below the geometric mean when analyzing and reprocessing data, effectively weights the curve such that both the high and low ends of the curve are given equivalent significance. 4.0 Warnings and Cautions________________________________________________ 4.1 Health and safety warnings 4.1.1 Wear the proper lab attire for all parts of this procedure. Wear gloves and proper eyewear when performing sample preparation in the laboratory at all times. Wear proper eyewear when working at the instrument in the laboratory. ETS-8-181.0 Analysis o f Photolysis Samples for Fluorocheinicals by HPLC/MS Page 5 of 15 Rage 26 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 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 of each chemical used, refer to material safety data sheets, packing materials, and the 3M Environmental Laboratory Chemical Hazard Review. 4.2 Cautions 4.2.1 All glassware in which standards are prepared should be rinsed with acetone and methanol to reduce the possibility of 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 rims. 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 method 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 Eq u i p m e n t __________________________________________ 6.1 Analytical balance sensitive to 0.1 mg. 6.2 Hewlett-Packard (HP) 1100 HPLC System, or equivalent. 6.2.1 Pump, binary, Model G1312; Quaternary, Model G1311A; or equivalent. 6.2.2 Solvent degasser, Model G1322A or equivalent. 6.2.3 Autosampler, ALS Model Gl 313A, variable injection volume or equivalent. 6.2.4 Column heater, Model G1316A, or equivalent. 6.3 Betasil C 18,50 x 2 mm; Dionex IonPac NG1 Guard column, 4 x 3 5 mm; or equivalent. 6.4 Mass spectrometer. Hewlett-Packard MSD Model Gl 946A, or equivalent. 6.5 Refrigerator capable of maintaining 4 3 C. 6.6 Data system. A personal computer capable o f controlling the HPLC system as well as recording and processing signals from the detector. ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 6 o f 15 Page 27 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 6.7 System control/data analysis software: Hewlett Packard ChemStation, Version A.6.03 or later. 6.8 Data reprocessing software: Thru-Put Systems Target NT, Revision 4.03, Build 157 or later. Hewlett Packard ChemStation, Version A.6.03 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 7.6 Hamilton Gastight syringes (precision 1% of total volume), 10 pL~l 000 pL 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 Methanol (MeOH). HPLC/SPEC/GC grade from EM Science, or equivalent 8.2 Acetone. HPLC/SPEC/GC grade from EM Science, or equivalent 8.3 ASTM Type II Water. Water with lower resistance must not be used. 8.4 Ammonium acetate, 2 mM in water. This solution is chromatographic solvent A (see Section 12.2.1). (Example: An acceptable eluent solution is made by adding 0.15 g ammonium acetate crystals to a 1-L volumetric flask containing about 500 mL water, adding. 10 mL o f methanol, diluting to the mark with 18.0 M il water and mixing.) 8.5 Stock, internal standard, surrogate, post-photolysis spike and calibration solutions All weights should be recorded to the nearest 0.0001 g in a standards preparation log: 8.5.1 Fluorochemical or target analyte prepared in acetonitrile (or suitable analytical solvent). (Example: A stock solution is prepared at a concentration of approximately 30,000 pg/mL by weighing 0.3 g of target analyte in a 10-mL volumetric flask and bringing to the mark with suitable analytical solvent. This solution is diluted in solvent to make additional, appropriate standards. Follow specified guidelines for documenting removal of test analyte and target analyte(s), use o f balance, preparation of diluted solutions and calibration standards in the appropriate log books. Maintain photocopies of the preparation pages and worksheets in a raw data file. ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 7 of 15 Page 28 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 9.0 Sample Handling______________________________________ _______________ 9.1 Standards and diluted samples are stored in capped autovials or capped 40 mL VOA vials until analysis. 9.2 If analysis will be delayed, standards and sample extracts may be stored at 4 C 3 C or room temperature, until analysis can be performed. Document storage conditions on sample prep worksheet with date and initials. 10.0 Quality Control____________________________________________ _________ 10.1 Calibration Standards. Calibration standards (Section 11) used to generate a calibration curve should be prepared in the same type of solvent or matrix as in the study samples. The number o f calibration standards and the concentration levels should be sufficient to encompass the expected concentrations of the study samples. In general, a minirruim o f five calibration standards is required for fit of linear regression. Broad calibration ranges (greater than three orders of magnitude between low and high standards), may require use o f a quadratic fit of the data and requires more points to adequately represent the calibration range. 10.2 Continuing Calibration Verification (CCV). Analyze a mid-range calibration standard after a maximum of every fifteen samples. 10.3 Solvent blank. Solvent blanks are run before and after every calibration curve, CCV, matrix and control blank (if contamination is noted), and after batches of no more than 30 injections. Acceptable values for the blanks are values below 25% of the limit o f quantitation (LOQ) of the instrument. If analyte carryover is a problem, use back-to-back solvent blanks. 10.4 Sample Triplicates. Analyze all sets of triplicate samples to provide a measure o f the precision o f analysis. Study samples will be analyzed in batches of no more than 30 samples. Multiple batches in an analytical sequence will be bracketed by calibration standards at the beginning and end o f each study sample batch. All samples (matrix and control samples, blanks and spikes) from a specified exposure type or time may be analyzed within the same analytical batch. 10.5 Analytical spikes. Prepare analytical spike sample for each sample type as applicable to determine the matrix effect on the recovery efficiency. Concentrations of the spike should be approximately equal to a mid-range calibration standard. The matrix spike sample should be analyzed periodically to measure the precision associated with the analysis. The analyst shall accept percent spike recoveries of 100 25%. Spike recoveries outside of this range should be noted and used with other criteria to evaluate the condition of the analytical run or necessity for repeat analysis. Consult with the Team Leader or designee for direction and final acceptance or rejection of the analytical run. Samples may be spiked at two different concentrations to ensure that the resulting levels of target analyte(s) are within the viable range of the calibration curve. ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 8 of 15 Page 29 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 11.0 Calibration and Standardization_________________________________ ______ 11.1 Analyze standards prior to and following each set of samples. The linear regression will be calculated from the plot of all individual calibration points, including but not forced through zero, using HP ChemStation or Target NT Software. A minimum o f five calibration standards is required to generate linear regression for target analyte(s). If the calibration curve residuals are greater than 25% deviation from the theoretical value, quadratic curve fitting and/or dropping low/high curve points may be required if data review shows this to be a consistent and more accurate representation o f the instrument response. Document in the raw data the technical justification for any deviation and consult with the team leader or designee for direction and for final acceptance or rejection of the data. 11.1.1 Use the following documentation/footnotes maybe used to justify dropping high/low curve points. 1) "High/low calibration points (list points) were excluded to provide a better fit over the linear range appropriate to the measured data." 2) "Low level calibration point(s) were not 4x higher than the extraction blank; these points were excluded from the curve to disqualify a data range that may have been significantly affected by background levels of the analyte." 3) "High/low calibration point(s) (list points) were excluded as they were not within the +/-25% accuracy requirements of the method when the curves were evaluated over a linear range appropriate to the data.' 11.2 If the curve does not meet requirements perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. 12.0 Procedures_________________________________________________________ 12.1 Instrument set up. Within "Method and Run Control" in the HP ChemStation Software window, turn the system "on" to: turn on file drying gas flow; initiate solvent flow through the column and nebulizing needle; equilibrate the column compartment; and equilibrate the MSD spray chamber temperatures and conditions. The system module displays should turn a green color to indicate the instrument is "ready" for analysis. A yellow color indicates that the system is not ready, but is working to "get ready." A red colored module icon indicates a type of systematic failure and should be corrected prior to proceeding. Check the run log for error messages and error codes if the problem is not apparent. 12.2 MSD set-up. Turn the MSD "on" in the software to equilibrate the system. 12.2.1 Check the level o f nitrogen in the tank and ensure there is enough to complete the impending run. 12.2.2 Clean the MSD according to the Equipment Procedure ETS-9-34.0 Operation and Maintenance o f HP LC/MS System. ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 9 of 15 Page 30 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.2.3 Perform a Check Tune or Autotune to ensure system operational qualification and performance verification o f the MSD. Log the Tune results and keep a copy with the analytical raw data. 12.2.4 Load the method file and ensure that the following parameters are appropriately set for the target analyte(s): Example mass spectrometer set up*: MSD: Ionization mode API-ES (or API-APCI) Polarity Negative (or Positive) Acquisition mode SIM (or SCAN) Gain 1.0 (up to 7.0) Fragmentar 70 (may be set to one voltage, or ramped for each ion) Dwell time 183 msec (time is a function of the amount of ions). Capillary voltage 3500, or equivalent Drying gas Nitrogen, or equivalent Nebulizer pressure 30 psig, or equivalent Drying gas flow 8 L/min, or equivalent . Drying gas temp 300 C, or equivalent Example conditions are applicable to HPl 100 LC/MSD equipment only. 12.3 LC Check 12.3.1 Check that the appropriate HPLC column is in the instrument for analysis. 12.3.2 Check that the correct eluent solutions are in bottles to be used and that enough is available to complete the sequence run. Adjust the solvent bottle level electronically within the method and run control window. 12.3.3 Ensure that the method file has the appropriate LC pump parameters for solvent flow/gradient program, column I.D/temperature, injection volume and stop time. Solvent A: Ammonium Acetate 2mM in water (with 1% MeOH) (or equivalent). Solvent B: Methanol (or equivalent). Example Solvent Gradient: T ime (min) %A %B Flow Rate 0.0 60 40 0.3 mL/min 1.0 60 40 0.3 mL/min 4.0 5 95 0.3 mL/min 11.0 5 Post time: 6 minutes, column temperature: 35C. 95 0.3 mL/min The initial solvent composition is set to a higher amount of aqueous solvent so as to achieve sufficient sample retention on the column. The gradient composition ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 10 of 15 Page 31 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 increases to a higher organic content over time to separate the analytes, and elute them off the column in a timely fashion. After all analytes have eluted, the solvent ratio is then switched back to "initial conditions" and held until the column pressure has stabilized ( indicating re-equilibration to initial conditions) prior to the next injection. 12,3.4 Auto-sampler setup: A uto-sam pler: ALS Model G1313A A u to -sa m pler Pro g ram : None In j e c t io n v o l u m e : 5.0 pL, or equivalent * Example conditions are applicable to Hewlett Packard 1100 LC/MSD only 12.3.5 Place the samples in the autosampler tray and construct a sequence table with appropriate calibration standards, calibration check standards and solvent blanks. 12.3.5.1 Verify that all samples and standards are positioned correctly. 12.3.5.2 Enter the identification code for each standard and samples. For solvent blanks, identify the solvent and the traceability number. 12.3.5.3 Use one injection per sample. 12.3.5.4 Ensure the method file is correctly entered for all samples. 12.4 Sequence and electronic storage of data files. 12.4.1 Within the sequence parameters, enter sequence information (brief sample population description and instrument name). 12.4.2 Set post-sequence command macro to shut down system after the run is completed (Example: "STANDBY" on HP1100/MSD systems). 12.4.3 Save all data to a subdirectory labeled with instrument and analysis date (e.g. H100200 for analysis on "Hillary," on 2 October, 2000). 12.4.4 Name data within the subdirectory with instrument ID and injection/run number (e.g. for samples acquired on "Hillary", data files shall be "HILL0001".... "HILL00##"). DO NOT exceed five identification characters for analysis of more than 99 samples since eight characters total are available for sample ID, and the last three digits are for sample numbering purposes (leaving the first five characters for data file identification). 12.4.5 Save sequence as analysis date and instrument letter (e.g. For analysis on instrument "Hillary" on October 2,2000 save sequence table as H100200.s). 12.5 Sample analysis 12.5.1 Enter the standard, sample, blank identification into the sequence table. Analyze calibration standards first, then up to 30 injections, followed by the calibration standards re-injected. Multiple sets o f samples can be set up in the sequence table with each set bracketed by calibration standards. Analyze a single continuing calibration standard (CC\0 after a maximum of 15 injections. Solvent blanks shall be analyzed before and after the CCV and before method and control blanks, if ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 11 of 15 Page 32 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 necessary. Two solvent blanks shall be analyzed at the end of the calibration standards to ensure that there is no carry over from the highest standard concentration. Solvent blanks may also be used to separate groups o f samples and evaluate for carry over problems from actual samples. Ensure standards, blanks, samples, and matrix spikes in the auto-sampler tray vials are in same order as listed in the sequence. 12.5.2 Print a copy o f the tune results, method and sequence to be stored with raw data. 12.5.3 Start the sequence. 12.6 Post Analysis. Prepare a folder identified specifically to the project and save data, method and sequence files. This will be considered the raw electronic data to be archived. 13.0 Data Analysis and Calculations________________ ______________________ 13.1 Peak Evaluation. Peaks must be symmetric in shape and identified by extracting compound-specific ions. Peaks considered for quantification must have peak heights greater than 4 times any baseline level for that region of the chromatogram. Peak area integration is from baseline to baseline using automatic or manual integration. Manual integration is not acceptable for calibration standards and should only be used in extreme cases as designated by the Team Leader. Samples and standards that may need to be manually integrated must be documented in the raw data as to why the peak was manually integrated. 13.2 Integration Codes. The following integration codes may be utilized to document what type o f manual integration was performed. . A: Adjust Left Anchor B: Adjust Right Anchor C: Delete Integration D: Add Integration Additionally, QAU encourages the data reviewer to write comments directly on the chromatogram if there is anything unusual. Date and initial all documentation. 13.3 M atrix spikes. Calculate the percent recovery for each of the matrix spikes. Calculate the matrix spike percent recoveries using the following equation: % Recovery = (observed spiked sample result - observed sample result! x 100 Nominal amount spiked Using the observed matrix spike recoveries, calculate the average spike recovery. 13.4 Accuracy. Calculate the accuracy o f each calculated calibration standard and CCV samples using the following equation. Accuracy = (Measured Conc.l x 100 Nominal Cone. ETS-8-181.0 Analysis o f Photolysis Samples for Fluorochenricals by HPLC/MS Page 12 of 15 Page 33 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 13.5 Sample Triplicates. Calculate the relative standard deviation (%RSD) for the triplicate samples: RSD = Standard Deviation of Sample Set x 100 Average o f Sample Set 14.0 Method Performance_______________________________________________ 14.1 Coefficient of Determination (r2). The coefficient of determination (r2) for the calibration curves should be 0.990 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%. It may be necessary to use quadratic fits of the data, usually when broad range curves (greater than 3 orders of magnitude between the low and high concentration standards) are used. Document in the raw data the technical justification for using quadratic equations. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. 14.2 Calibration Standards. The acceptance criterion for the calibration standards is that the accuracy of each standard is 75% to 125% ( 25 % difference) of the nominal value. Calibration standards outside this range are to be noted. Document in the raw data the technical justification for deviations. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. 14.3 Internal Standard (IS) and Surrogate. Review of the internal standard and surrogate performance is performed by averaging the area response throughout the analytical run and calculating %RSD. Inconsistencies in the internal standard peak area may indicate instrumental changes over time. Inconsistencies in the surrogate peak area may indicate instrumental changes, injection error, or changes in the test-system. Consult with the Team Leader or designee for direction and final acceptance or rejection o f the analytical run. 14.4 Continuing Calibration Verification. If the accuracy for the amount o f measured analyte is greater than 25% from the nominal value relative to the initial standard curve, the Team Leader should be consulted. Only those samples analyzed before the last acceptable calibration check standard may be used. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. 14.5 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 greater than 25% of the peak area o f the designated LOQ value are observed in sequential solvent blanks, this is indicative o f instrument contamination. The instrument shall be serviced by thoroughly cleaning the electrospray source, and replacing/cleaning columns, tubing, etc. (as designated in the Equipment Procedure, ETS-9-34.0) and the analysis restarted. Consult with the Team Leader or designee for direction and final acceptance or rejection of the analytical run. 14.6 M atrix Blanks. Matrix blanks are the basis for determining the LOQ and are monitored at various times in the analytical run. Samples with greater than 25% of the peak area o f the designated LOQ value observed in matrix blanks are indicative of matrix effect, ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 13 of 15 Page 34 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 sample contamination or instrument contamination. Evaluation o f the solvent and control blades may be necessary to determine these effects. Use of solvent blanks prior to the matrix blank may be necessary to rule out instrumental or sample contamination. 14.7 C ontrol Blanks. Control blanks are the basis for determining matrix effect (interference or suppression) and also to monitor for instrumental or sample contamination. Use of solvent blanks prior to the matrix blank may be necessary to rule out instrumental or sample contamination. 14.8 Lim it of Quantitation (LOQ). The LOQ is equal to the lowest acceptable standard (i.e. % accuracy is 25 % nominal value) in the calibration curve that is greater than 4 times the level of the matrix blanks. 14.9 Sample Triplicates. The analyst shall accept %RSD values < 25%. %RSD values > 25% should be noted. Data used in the final report that is deemed out o f control will be required to have technical justification for why the data is used, documented in the final report and raw data. Consult with the Team Leader or designee for direction, and for final acceptance or rejection of the data. 14.10 Control Samples. The acceptance criterion for the control samples is that the accuracy is 75% to 125% o f the nominal value. These will be used as a reference for matrix effect and overall method performance. Control samples outside this range are to be noted. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. Data used in final report that is deemed out of control will be required to have a technical justification for why the data are being used, documented in the final report and raw data. 14.11 Analytical Spikes. The analyst shall accept percent spike recovery values of 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 of the data. Data used in final report that is deemed out of control will be required to have a technical justification for why the data are being used, documented in the final report and raw data. 14.12 System Suitability. Without performing a method validation, system suitability can be demonstrated by acceptable instrumental checks (e.g. abbreviated m/z check-time, or full auto-tune routines. Consult the appropriate instrumental manuals (Reference 18.2). Furthermore, overlaying calibration curves and implementing check standards (CCV), the method shall be self-validating if all data quality objectives are satisfied. 15.0 Pollution Prevention and Waste Management__________________________ 15.1 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 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. ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 14 of 15 Page 35 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 16.0 Records ____________________________________________________ 16.1 Print hard copies of all graphics and data analysis summaries for archiving. 16.2 Sign and date all graphics and label with instrument ID. 16.3 Fill out appropriate preparation worksheets completely, making sure to include all initials and dates, along with the study number and sample identification. 16.4 Print out the sample acquisition sequence table, reduce the size with photocopying and tape the photocopy into the instrument log. Keep the original copy for the raw data files. 16.5 Print chromatograms, reprocessing sequence and batch reports for all analyses. 16.6 Print calibration tables and curve information and store in the raw data file. 16.7 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. 16.8 Archive electronic data to appropriate media when necessary. 17.0 Attachments 17.1 None. 18.0 References 18.1 ETS-9-34.0, Hewlett Packard 1100/MSD Equipment Procedure. 18.2 Hewlett Packard 1100/MSD instruction CD/ROM. 19.0 Affected Documents 19.1 None. 20.0 REVISIONS Revision number Reason for revision Date of Revision ETS-8-181.0 Analysis of Photolysis Samples for Fluorochemicals by HPLC/MS Page 15 of 15 Page 36 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory Method Preparation of Samples for Photolytic Exposure Studies in Aqueous Matrices Method Number: ETS-8-176.0 Adoption Date: Approved By: Laboratoiy Manager Date Exact CopV ------ Date ETS-8-1IS .Q P reparation o f S a m ples f o r P h o to lysis Stu dies in Aqueous M atrices Method Page 1 o f 18 Page 37 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application l.l Purpose. Chemicals dissolved in aqueous solutions are subject to two types of photoreaction. The first type (direct photolysis) occurs when the chemical of interest absorbs sunlight directly and is transformed to products when unstable, excited states of the molecule lead to decomposition. The second type is indirect photolysis, where degradation of the dissolved chemical is the result o f chemical or electronic excitation transfer from light-absorbing species in the water. The simplest reaction involves the absorption of UV energy by hydrogen peroxide (H20 2) to produce 2 hydroxl radicals. These may react with any species in the water, including solvent, buffer, dissolved organic material and target material. Use of water and H20 2is very controlled and predictable. Other sources in other matrices are not as controlled or predicable, but are more environmentally relevant. Natural waters such as lake and sea water can be used for the photolytic reaction matrix because it may contain dissolved organic material that absorbs sunlight and produces reactive intermediates that include singlet oxygen (10 2) which may promote indirect photolysis o f the test substance. Another transient species photochemically produced by the reaction of UV light and dissolved organic materials (humic) is hydrogen peroxide (H20 2) which may react further to form the hydroxyl radical. The addition of H20 2to test solutions may be utilized as a free radicad source to initiate indirect photolytic reactions in controlled test solutions such as MilliQ water or buffers. Further studies involving the use of either naturally occurring metal complexes such as Fe(III) which undergo photoreduction to Fe(II) and free radicals or addition o f T i02as a catalytic surface for indirect photolysis may also be evaluated within this method. 1.2 Compatible analytes. Test substance and degradation products for photolytic exposure include but are not limited to: Compound Acronvm Compound Acronvm Perfluorooctanoic acid PFOA Perfluorobutanoic acid ' PFBA Perfluorooctanesulfonate Perfluorooctanesulfonamide PFOS FOSA Perfluorobutan esulfonate Perfluorabutanesulfonamide PFBS FBSA N-methylperfluorooctanesuIfonamide N-MeFOSA N-methylperfluorobutanesuIfonamide N-MeFBSA N-ethylperfluorooctanesulfonamide N-EtFOSA N-ethylperfluorobutanesulfonamide N-EtFBSA 2-(N-methylperfluoro octanesulfonamido) ethyl alcohol N-MeFOSE-OH 2-(N-methyIperfluorobutanesulfonamido) ethyl N-MeFBSE- alcohol OH 2-(N-ethyIperfluorooctanesulfonamido) N-EtFOSE-OH 2-(N-ethyIperfluorobutanesulfonamido)ethyl ethyl alcohol alcohol N-EtFBSE-OH 1-perfluorooctene -- 1-perfluorobutene -- Perfluorooctanehydride 1H, Cg-hydride Perfluorobutanehydride 1H, C4-hydride ... and other C4 thru Cl0 homologues, and polymeric materials based on the aforementioned compounds. ETS-8-\7 6 .0 P re p a ra tio n o f S a m ples f o r P h o to lysis Studies in A queous M a trices Method Page 2 o f 18 Page 38 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Other possible degradation products include, but are not limited to: C 1H-perfluoroethane (1H-pfC2) 1H-perfluoroethane (lH-pPC2) C3 lH-perfluoropropane (lH-pfC3) 2H-perfluotopropane (2H-pfC3) perfluoro-l -propene (pfC3-lene) C| perfluoro-l-butene (pfC4-lene) Periluoro-2-butene (pfC4-2ene) 2H-perfluon>butane (2H-pfC4) lH-perfluorobutane (lH-pfC4) C5 2H-perfluororpentane (2H-pfC5) pcrfluoro-l-pentene (pfC5-lene) pcrfluoro-2-pentene (pfC5-2ene) C6 perfluoro-2-hexene (pfC6-2ene) 2H-perfluorohexene (2H-pfC6) lH-perfluorohexane (lH-pfC6) perfluoFO-l-hexene (pfC6-lene) IH-pcrfluorohexane (lH-pfC6) C7 2H-perfluorohcptane (2H-pfC5) Perfluoro-l-heptene (pfC7-lene) lH-perfluoroheptan (lH-pfC7) Cg perfluoro-l-octene (pfC8-lene) 2H-perfluorooctane (2H-pfC8) Periluoro-2-ociene (pfC8-2ene) lH-perfluorooctane (lH-piC8) 1.3 Acceptable matrices. Aqueous solution of test substance including but not Limited to the following matrices: pH 7 phosphate buffer, 18.2 M il resistivity water, seawater and metal solutions. 2.0 Summary of Method 2.1 The objective of the photolytic exposure study is to determine whether the test substance undergoes degradation by either direct or indirect photolysis, and to identify and quantify degradation products formed in the test matrix under these conditions. Study samples (5 mL aqueous matrix) are prepared in 40 mL glass VOA vials equipped with screw-top caps with septa. Study sets are prepared in duplicate for separate analysis by LC/MS and dynamic purge and trap GC/MS. When required, the addition of 30% H2O2 solution to initiate radical formation is performed prior to the photolytic exposure and at specified intervals throughout the exposure study. Vials are placed in the photo-reactor and immersed in a water bath controlled at 23-26 C. Samples are exposed to approximately 261 W/m2o f 310-800 nm photo-irradiance for a specified number o f 8-hour periods. An 8-hour period of irradiance is defined as one day's worth of sunlight. Other parameters are acceptable, with the time and settings noted for each study. The number o f days to expose samples is determined by the Team Leader. The amount o f irradiation received by the samples may be monitored in one of the following three ways: 1) calculating the total wattage per length o f exposure 2) use of a radiometer to measure irradiance output, and/or 3) use of a quinine monohydrochloride dihydrate (QMD) actinometer solution exposed along with the samples and monitored for change in UV absorption over time. The use of the radiometer provides an accurate measurement at specified time-points; whereas calculating the total wattage per exposure length and use of the QMD actinometer provide time-averaged total integrated energies. Suntest instruments contain an internal radiometer for maintenance of constant irradiance. A second radiometer may be used as a check for consistency. At the end of the exposure time, samples are removed from the photoreactor and either subsequently analyzed or stored at 1-5 C. Study samples to be analyzed by LC/MS are prepared for analysis by diluting the 5 mL sample volume with 30 mL o f suitable analytical solvent (e.g. methanol) containing internal standard. The GC/MS study samples are stored inverted prior to purge and trap GC/MS analysis. ETS-&-176.(P reparation o f Sam ples f o r P h otolysis Studies in A queous M atrices Method Page 3 o f 18 Page 39 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 2.2 An example of samples to be prepared for each study is shown in the table below. Exact lists may vary, dependent upon the test specifics for each study and will be noted in individual study reports. Typically, there are extra control samples for certain matrices such as Fe2C>3. Description Sample Rep 1 Sample Rep 2 Sample Rep 3 Sample Spike Matrix Blank Matrix Blank Spike Control Sample Control Spike Control Blank Conttol Blank Spike Test Matrix + + + + + 0 0 0 0 Control Matrix "5 0 0 0 0 0 + + + 4* Test Substance + + + + 0 0 4* 4* 0 0 Post Photolysis Target Analyte spike 0 0 4- 40- 0 4- 0 + Sample Type TimeO Time 0 TimeO TimeO TimeO Time 0 TimeO TimeO TimeO Time 0 LC/MS With HjOj Without HjO* GC/MS With h 2o , Without H,Oj X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X X XX X Sample Rep 1 Sample Rep 2 Sample Rep 3 Sample Spike Matrix Blank Matrix Blank Spike Control Sample Control Spike Control Blank Control Blank Spike + + 44- + 4- 0 0 0 0 0 0 0 0 0 0 4- + 44- 44- 4- 4- 0 0 44- 0 0 0 Exposed X XX X 0 Exposed X XX X 0 4- Exposed Exposed X X XX XX X X 0 4- Exposed Exposed X X XX XX X X 40- Exposed Exposed X X XX XX X X 40- Exposed Exposed X X XX XX X X Sample Rep 1 Sample Rep 2 Sample Rep 3 Sample Spike Matrix Blank Matrix Blank Spike Control Sample Control Spike Control Blank Control Blank Spike + + 4- 4- + 4- 0 0 0 0 0 0 0 0 0 0 4- 4- 4* 4- 4- 4440 0 4- 40 0 0 Unexposed X XX X 0 Unexposed X XX X 0 Unexposed X XX X 4* Unexposed X XX X 0 Unexposed X XX X + Unexposed X XX X 0 Unexposed X XX X 4- Unexposed X XX X 0 Unexposed X XX X 4- Unexposed X XX X + - added to test vial; 0 = NOT added to test vial; X = One set W/H2O2, One set w/o H2O2 3.0 Quality Control-Definition/Frequency/Performance Criteria XT Blanks ' 3.1.1 D efinition: M atrix Blank. An analyte-free matrix to which all reagents are added in the same volumes or proportions as used in the sample processing. For photolysis studies, there are multiple matrix blanks to adequately represent the variables within the study in reference to the matrix (e.g. Exposed, Unexposed, Time 0; with peroxide, without peroxide). The matrix blanks are carried through the complete sample preparation, experimental treatment and analytical procedure. The matrix blank is used to document contamination resulting from the experimental treatment and analytical process. Refer to the table below for an example of matrix blank types. The matrix blank is used to document the actual test system without the test substance. The control blank is used to control the test matrix and trace any background levels of target analyte that may be matrix- specific. The table below shows an example of a control blank TZTS-H -nB.QPreparation o f S a m ples f o r P h o to lysis Stu dies in A queous M atrices Method Page 4 of 18 Page 40 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.1.2 Performance C riteria: The frequency o f use and the performance specifications for each of the above defined method blank types shall be as follows: Matrix ID Matrix Blank Control Blank Matrix description Example: 0.01 M Phosphate Buffer, pH 7 Example: ASTM Type II Water Frequency 1 Replicate per light and dark exposure, for each time point and for each analytical methodology. Performance Criteria Any background level of target analyte shall be less than 25% die area counts of the LOQ. 3.2 Lim it of Quantitation (LOQ) 3.2.1 Definition: The lowest concentration that can be reliably measured within specified limits of accuracy during routine laboratory operating conditions. Sample LOQs are highly matrix-dependent. 3.2.2 Q uality Control and Performance C riteria: The LOQ is generally 5 to 10 times the minimum concentration with a 99% confidence limit 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 of the solvent blanks and indicates good accuracy (+ 25%) o f the nominal calibration standard concentration. 3.3 Sample Triplicate 3.3.1 D efinition: Three aliquots prepared as representatives of the same sample source (i.e. test substance) and carried through all steps o f the photolytic study process and analytical procedures in an identical manner. The results from triplicate analyses are used to evaluate variability of the total method, including sample preparation, photolytic process and analysis. 3.3.2 Performance C riteria: The samples in the test matrix will be prepared in triplicate. Each replicate will be prepared for each treatment type: light and dark exposures, with and without hydrogen peroxide, for EACH time-point, and for each analytical methodology (e.g. LC/MS and/or GC/MS). See the following table: Matrix Descrintion Test Matrix containing test analyte(s) Freauencv of Use 3 Replicates per light AND dark exposure, wife AND without H20 2, for each time-point, and for each analytical methodology (i.e. LC/MS and/or GC/MS). Performance Criteria The analyst shall accept %RSD <25%. Precision values not meeting specification must be documented andjustified (if possible). 3.4 Control Sample 3.4.1 D efinition: A known matrix containing the test analyte(s) carried throughout the entire analytical procedure. This is used to document laboratory performance (i.e. precision of sample preparation by comparing spike recoveries from the different matrices and sample types). A control sample consists of a control matrix spiked with test analyte(s). A control sample should be analyzed with each batch of samples processed to verify that the precision and bias of the analytical process ETS-8-176.0Prepora//o o f Sam ples f o r P h otolysis Studies in A queous M atrices Method Page 5 o f 18 Page 41 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.4.2 are within control limits. The results of control sample analyses are compared to control limits established for both precision and bias to determine usability of die data. Performance Criteria, One control sample will be prepared per matrix, per treatment type. See the following table: Matrix Description Control matrix with test analyte(s) added Freauencv of Use 1 Replicate per light AND dark exposure, with AND without H20 2, for each time-point, and for each analytical methodology (i.e. LC/MS and/or GC/MS). Performance Criteria The analyst shall accept recovery values o f 100 25%. Accuracy values not meeting specification must be documented andjustified (if possible). 3.5 Analytical Spike (AS) 3.5.1 D efinition: Prepared by adding a known mass of target analyte(s) to a specified amount o f a diluted and/or aliquoted sample. This assumes that an independent estimate of target analyte concentration is available. Analytical spikes are used to evaluate the recovery efficiency of the analyte and the effect of the matrix on the measurements. 3.5.2 Quality Control and Performance Criteria: One sample spike will be prepared in the actual test matrix sample, and one control spike in the control matrix will be prepared. Each replicate will be prepared per treatment type: for light and dark exposures, with and without hydrogen peroxide, for each time-point, and for each analytical methodology (i.e. LC/MS and/or GC/MS). In addition, one matrix blank spike and one control blank spike will be prepared. See the following table: Matrix Description Test Matrix a n d test substance, spiked with target analyte(s) just prior to analysis Control Matrix a n d test substance, spiked with target analyte(s)just prior to analysis Test Matrix w ithout test substance, spiked with target analyte(s)just prior to analysis Control Matrix w ithout test substance, spiked with target analyte(s)just prior to analysis Freauency of Use 1 Replicate per treatment type. 1 Replicate per treatment type. 1 Replicate per treatment type. 1 Replicate per treatment type. Performance Criteria The analyst shall accept spike recovery values of 100 25% . If spike recoveries are greater than 125% or less than 75%, document that the spike sample is out of the specifications and justify, if possible, the reason. 3.6 Internal Standard/Surrogate 3.6.1 Inte rn a l Standard D efinition (applies to LC/MS and GC/MS samples): A known amount of a compound similar in analytical behavior to the target analyte(s) of interest, added to all samples and standards (post-irradiation), and carried through the entire analytical process. It provides a reference for evaluating and controlling the precision and bias of the applied analytical method. Samples are ETS-8-176.0/Veparai!OH o f Sam ples f o r P hotolysis Studies in Aqueous M atrices Method Page 6 of 18 Page 42 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.6.2 to be quantified using the internal standard providing that the response o f the internal standard is consistent ( 5% relative). Use of external calibration methodology requires written justification by the Team Leader. Surrogate D efinition (applies to LC/MS and GC/MS samples): A known amount of a compound similar in analytical behavior to the target analyte(s) of interest may be added to all samples and standards (pre- or post-irradiation, at the discretion o f the Team Leader) and carried through the remaining sample preparation and analytical process. If added before exposure, it monitors the presence of vial leaks during photolysis, as well as the performance of the purge and trap auto-sampler and concentrator. Surrogate analysis is used to evaluate and control the precision and bias of the analytical method. Surrogates are not used for quantitation. Note: Internal standards are used in all experiments. The use o fsurrogate standards may or may not he used. 3.6.3 Quality Control and Performance Criteria: Matrix Descrintion Freauencv of Use Performance Criteria Sample diluted with 30 mL o f internal standard compound dissolved in a suitable analytical solvent Every LC/MS sample analyzed Sample with surrogate Every GC/MS compound spiked into it. sample analyzed The Coefficient of Variation, or %RSD shall be calculated for the area response o f all appropriate samples per analytical batch. The analyst shall accept %RSD values of <15%. The recovery and precision of the surrogates should be 100 25% and <15%, respectively. Unacceptable values shall be documented and justified, if possible. 3.7 Other Definitions. 3.7.1 Test Analyte/Substance: Any substance (mixture or controlled compound) added or administered to the test system for the purpose of chemical analysis. 3.7.2 Degradation Produces): Secondary analytes of interest produced as a result of chemical reactions during the photolysis and monitored (qualitatively or quantitatively) during the sample analysis procedure. 3.7.3 Target Analyte(s): The analyte(s) singled out in the analytical phase o f the study is tile target analyte. The target analyte may be identical to the test substance used in the experimental phase of the study, a by-product or degradation product that is monitored (qualitatively or quantitatively) during the sample analysis procedure. 3.7.4 Test M atrix: The physical matrix in which the study will be conducted. Also referred to as the test system. 3.7.5 Control M atrix: A known physical matrix to be included with the study for comparison with the test matrix. 3.7.6 Relative Standard Deviation (RSD): A measure o f relative precision for three or more sample replicates; defined as the sample standard deviation divided by the sample average and multiplied by 100. This is expressed as percent (%RSD). 3.7.7 Accuracy: The closeness of agreement between an experimentally determined value and an accepted reference value; defined as the measured value divided by the nominal value and multiplied by 100. ETS-8-176.0 P rep a ra tio n o fS a m p le sf o r P h o to lysis Stu dies in Aqueous M a trices Method Page 7 of 18 Page 43 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 4.0 Health and Safety Warnings 4li Safety ' 4.1.1 Wear the proper lab attire, gloves and eye protection for all parts of these procedures. 4.1.2 Handle all solvents in a hood for all parts o f the described sample preparation procedure. 4.1.3 For potential hazards of each chemical used, refer to material safety data sheets, packing materials, and 3M Environmental Laboratory's Chemical Hazard Review. 4.1.4 No mouth pipetting is allowed. 4.2 Cautions 4.2.1 Glassware in which standards are prepared should be rinsed with solvent to reduce the possibility of accidental contamination. 4.2.2 The photoreactors are equipped with a continuous flow o f cooling water, which poses a threat o f electrocution during the handling o f the photoreactor during irradiation sequences. To avoid possible injury, inspect the units frequently for water leakage and electrical outlets and wiring for wear and tear. Replace any worn parts immediately. 4.2.3 Wear dark protective eyewear when operating the reactor. Do not look directly at the activated lamp. Use caution when handling samples in the reactor; the interior walls o f the reactor and exposed glass vials become extremely hot. 5.0 Interference______________________________________ ________________________ 5.1 Solvents, water and matrix components could interfere with detection thereby decreasing sensitivity in the sample analysis. Care must be taken to prevent all possible contaminants by using fresh reagents, analytical grade solvents and clean glassware during the sample preparation processes. 6.0 Equipment _________ ____________________________________________ 6.1 Analytical balance sensitive to 0.1 mg 6.2 Photoreactor: Suntest CPS+, XLS+, or equivalent, equipped with a xenon arc-lamp and capable o f producing integrated irradiance values from 100-680 W/m2 over the wavelength range of 290-800 nm. Lamp output must be filtered to allow only 290-800 nm wavelengths. A flowing water bath with circulating pump is required. Consult the appropriate 3M SOP for instructions. 6.3 Water recirculating cooler capable of maintaining temperature at 25 C 5 C, from Poly Science, Model 1177-P or equivalent. 6.4 Agilent Technologies UV-visible Spectrophotometer, equipped with tungsten and deuterium lamps, Model 8453, or equivalent. Consult the appropriate 3M SOP for instructions. 6.4.1 Autosampler equipped with eight sample cell holders: Agilent Technologies Model G1120A, Thermostatted Cell Holder: Model 08451-60104, or equivalent. 6.4.1.1 1.0-cm path length quartz spectrophotometer cell from Hewlett Packard, or equivalent. 6.4.2 Long Path-Length Cell Holder, Hewlett Packard (# 89076C) or equivalent. ETS-8-1 IS .Q P reparation o fS a m p les f o r P h o to lysis S tu dies in Aqueous M a trices Method Page 8 o f 18 Page 44 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 6.4.2.1 10-cm path length quartz cell equipped with stopcocks, Hewlett Packard Part # 5061-3392, or equivalent. 6.4.3 Data acquisition and analysis software, HP ChemStation for UV-Visible Spectroscopy, G1116AA, Rev. B.01.02, or later. 6.4.4 PC Computer capable o f running appropriate analysis software to acquire and report data. 6.5 Centrifuge capable of maintaining > 2000 rpm for 10 minutes at ambient temperature. 6.6 Radiometer (optional) capable of monitoring the energy from a xenon source from 290 to 480 nm over time. Model PMA2100, Version 1.16, Solar Light Company, Inc., or equivalent. Consult the appropriate 3M SOP for instructions. 7.0 Supplies and Materials 40 mL amber and clear glass VOA vials with screw caps with septa. 7.2 Crimp cap autovials-1.5 mL, caps, crimper, and decapper. 7.3 Adhesive-backed labels (return address size) for labeling quartz vials and autovials. 7.4 Disposable glass graduated pipettes, 1 mL to 10 mL. 7.5 Disposable glass Pasteur pipettes and rubber bulbs. 7.6 Glass beakers, various sizes. 7.7 Volumetric flasks, from 10 mL to 1000 mL. 7.8 Hamilton Gastight syringes (precision 1% of the total volume), 5 pL to 1000 pL. 7.9 10 mL Bottle-top dispenser, Calibrex, Model # 511, or equivalent. 7.10 Adjustable repeater pipette, Wheaton Step-pette 411, or equivalent, equipped with the appropriate volumetric range pipette tips. 7.11 Ziploc plastic bags, or equivalent. 8.0 Reagents and Standards__________________________________________________ 8ll Methanol (MeOH). HPLC/SPEC/GC and/or purge and trap grade (EM Science, or equivalent. 8.2 Acetonitrile (ACN). HPLC/SPEC/GC and/or purge and trap grad from EM Science, or equivalent. 8.3 Aqueous M atrix, includes but is not limited to the following matrix types: 8.3.1 ASTM Type I water. Milli-Q or equivalent, with a measured resistivity >18.0 Mfi-cm. 8.3.2 0.01 M pH 7.0 Phosphate Buffer. Example: Weigh 1.36 g KH2PO4 into a 2 L volumetric flask and dissolve into 1 L of Type I water. Add 600 mL o f 0.1% NaOH. Adjust to pH 7.0 0.1% with 0.1% NaOH or dilute H2SO4 and dilute to the mark with Type I water for a final cone, of 10 mM. 8.3.3 Lake Surface water. Collected from a known source, with known specifications for Dissolved Organic Carbon (DOC) and Total Organic Carbon (TOC). 8.3.4 Sea water. Collected from a known source, with known DOC and TOC specifications. 8.3.5 Aqueous metal solutions and slurries (e.g. TiOi, Fe2 0 3 ). Example: Dilute 0.015 g of Ti02 (Aldrich Chemical or equivalent) to 500 mL with Milli-Q water. 8.3.6 Aqueous solutions containing soil. Example: Prepare samples containing 0.7g o f characterized soil or sediment in 5 mL of Milli-Q water. K T S -& -n 6 .G P rep a ra tio n o f S a m ples f o r P h otolysis Studies in Aqueous M atrices Method Page 9 of 18 Page 45 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 8.4 Hydrogen Peroxide (H2O2). 30% aqueous solution from EM Science, or equivalent. 8.5 Potassium phosphate. Reagent grade from JT Baker or equivalent. 8.6 Stock Solutions 8.6.1 Stock solutions for the test analyte, target analytes, internal standard are prepared in an organic solvent (e,g. methanol, acetonitrile) at concentrations of approximately 10,000 pg/ml by weighting approximately O.lg o f the appropriate substance into a 10.-mL volumetric flask and diluting to the mark with solvent. This solution is then diluted to make appropriate working solutions. 8.7 Test Analyte Solution: 8.7.1 Example for water soluble analvtefsl: Example: A 1 pg/mL test substance solution in the test matrix (Section 8.3) is prepared by diluting 0.050 mL of stock solution (Section 8.6.1) to 500 mL with test matrix. Aliquots (5 mL) of this solution will transferred to VOA vials for subsequent photolysis. 8.7.2 Example for poor water soluble analvtefsl or those with adsorption difficulties: Prepare a solution of the test substance in acetonitrile (Example: A 500 pg/mL test analyte solution is prepared by diluting 500 pi of stock solution (Section 8.6.1) into a 10 mL volumetric flask and diluting to mark with acetonitrile). Calculate the test analyte concentration such that the organic content in the test vial is no more than 1% of the total sample matrix volume. Example: A 1 pg/mL test analyte in the test matrix (Section 8.3) is prepared by injecting 10 pL of a 500 pg/mL test substance stock (Section 8.6.1) into a VOA vial containing 5 mL of die test matrix. *Acetonitrile is currently the preferred solvent to use when introducing the test substance to the test matrix because it does not interfere. Methanol is a radical scavenger, which canphotooxidize during the exposure and decrease the indirect photolysis o fthe intended test substance. Evidence o fthisphenomenon (approximately 10% decrease in the concentrations o fthefin a l products) has been observed in a study here at 3M (EtFOSE-OHphotolysis in p H 7 buffer, with and without presence ofMeOH). 8.8 Target Analyte(s) Spiking solution: Example: A spike solution of test analyte and target analyte(s) (e.g. projected degradation products) in methanol or acetonitrile is prepared by diluting 500 pL of test analyte stock solution and 100 pL of target analyte(s) stock solution (Section 8.6.1) into 10 mL with MeOH. The final concentration is approximately 500 pg/mL test substance/100 pg/mL target analytes. Addition o f 10 pL of this target analyte spiking solution into the 35-mL diluted sample volume will result in approximately 140 ng/ml and 30 ng/ml concentrations for the test analyte and target analyte(s), respectively.* *Pre-estimation o fthe degradation potential o fthe test analyte and subsequent degradation products is not always possible. I f possible, an analyticalpre screening o frepresentative samples should be perform edfor accurate spiking. General rule o fthumb has been that the test analyte spike amount be ETS-8-176.0P rep a ra tio n o f S a m ples f o r P h o to lysis S tu dies in Aqueous M atrices Method Page 10 of 18 Page 46 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 approximately 25%- 50% o fthe initial concentration. The target analyte(s) spike amount has been 10-100 ng/ml, depending on expected levels under specific conditions. More than one spike solution may be utilized to adequately represent the levels in the samples. Example: A test analyte that undergoes significant degradation during photolysis will require a lower spike concentration in the Exposed sample set due to less test analyte remaining. The Day 0 and Unexposed sample sets, which have not degraded, may require higher test analyte spike concentrations. 8.9 Dilution Solution containing Internal Standard: The diluting solution shall contain internal standard at an area response level equivalent to approximately half the are response o f the test analyte's high standard in the calibration curve. Enough dilution solution shall be prepared for use in all the study samples and in preparation of the calibration curve samples. Example: Internal standard solution is prepared by diluting 100 p,L of stock solution (Section 8.6.1) to 4.0 L with MeOH to a concentration of 250 ng/mL. 8.10 Quinine monohydrochloride dihydrate (QMD). 90% from Aldrich Chemical. 8.11 QMD solution: A 2% (w/v) solution of quinine monohydrochloride dihydrate solution is prepared by weighing approximately 2.0 g into a weigh boat, transferring to a 100 mL . flask and diluting to volume with Milli-Q water. 9.0 Sample Handling 9T Record times of initial preparation, reference numbers of reagents used and the amounts, appropriate dates, times and initials on the photolysis sample preparation worksheet.. Record photolysis reactor used, radiometer ( if applicable), computer for data collection, photolysis start and end on the sample preparation sheet and in the photolysis reactor log books. Record times, dates and initials of sample treatment post-photolysis, reference numbers of reagents used, and storage conditions. 9.2 Upon addition of the test substance solution, invert the 40 mL VOA sample vials (cap side down) to prevent loss of any potential volatile target analytes during the rest o f tide procedure. This is particularly important for the GC/MS samples. GC/MS samples may only be turned upright immediately before being loaded onto the purge and trap autosampler. The LC/MS samples may be turned upright after the photolysis process has been completed. The exception to this being the need to briefly turn the samples upright fo r H2O2 injection through the septa o fthe appropriate VOA sample vials at specified time intervals (See Section 12.6 and Section 12.11.7). 9.3 The completed photolysis samples remain inverted and refrigerated at 1-5 C prior to analysis by LC/MS or sample purge and trap GC/MS. 9.4 Sample preparation prior to LC/MS analysis requires the addition of 30 mL of diluting solvent containing internal standard to the 5-mL photolysis samples. This is to ensure complete recovery of the target analytes from the glass VOA vial surface and to dilute the samples into a working analytical range. Day 0 study samples stored at 1-5 C during the time of photolytic exposure are removed and prepared for analysis at the same time as the exposed and unexposed samples. ETS-8-17 6 .0 P rep a ra tio n o f Sam ples f o r Photolysis S tu dies in A queous M a trices Method Page 11 o f 18 Page 47 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 10.0 Quality Control 10.1 Refer to the definitions section for the quality control specified for each respective sample type. 11.0 Calibration and Standardization___________________ _________________ 11.1 The compounds of interest must be characterized according to laboratory specifications. 11.2 All equipment used, such as the analytical balance, radiometer, etc. should be calibrated prior to use (daily, weekly, etc.) as specified in the standard operating procedure(s). 11.3 All samples analyzed will be run against a standard curve containing varying amounts of target analytes, and a fixed amount of internal standard or surrogate compound. Refer to the appropriate LC/MS and GC/MS methodologies for further analytical information. 12.0 12.1 12.2 12.3 12.4 12.5 Procedure________________________________________ __________________ Obtain the absorbance spectra of the test compound in aqueous solution using a UVVisible Spectrophotometer (ETS-9-46.0). 12.1.1 Using a 10-cm quartz spectrophotometer cell, obtain a blank water absorbance reading over the range 290-800 nm to determine a background or baseline reading. 12.1.2 Aliquot a solution of water containing test substance, at a concentration less than half the solubility limit, into a 10 cm quartz spectrophotometer cell and obtain an absorbance reading over the range 290-800 nm. A positive absorbance may indicate the potential of the analyte to undergo direct photolysis. Non-absorbing analytes would be more likely to undergo indirect photolysis as the potential degradation pathway. Obtain the appropriate number o f clear and amber 40-mL glass vials with caps and cardboard boxes. Label the vial caps using a black permanent pen to distinctly identify samples. Paper labels will be applied post-hydrolysis as they don't stick in water. Prepare appropriate sample preparation worksheets and create labels for each sample to affix to the 40 mL VOA vials and the autovials for analysis after photolysis. The labels should include the study number, sample number, test compound, matrix, exposure type (exposed/ unexposed/ Day 0), date and initials of the analyst. Aliquot 5 mL of the following solutions into clear (for EXPOSED samples) and amber (for UNEXPOSED and DAY 0 samples) 40 mL glass VOA vials: 12.4.1 Matrix with test substance (sample reps 1,2,3, and sample spike). 12.4.2 Matrix without test substance (matrix blank and matrix blank spike). 12.4.3 Control matrix with test substance (control sample and control spike). 12.4.4 Control matrix without test substance (control blank and control blank spike). (When appropriate, test substance may be added after 5mL aliquots o f matrix have been added to the vials. See Section 8.7) All exposed, unexposed, and day 0 samples will contain sample sets with and without peroxide and prepared for LC/MS and GC/MS analyses according to the following table: ETS-8-176.0Pre/)arafion o f Sam ples f o r P hotolysis Stu dies in A queous M atrices Method Page 12 o f 18 Page 48 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Sample Treatment/Type Matrix with test substance Matrix without test substance Control matrix with test substance # of Samples 6 +H20 2(3LC/MS.3GC/MS) 6 -H20 2 (3LC/MS .3GC/MS) 2 +H20 2(1LC/MS.1GC/MS) 2 -H20 2 (11LC/MS, 1GC/MS) 2 +H20 2(1LC/MS.1GC/MS) 2 -H20 2 (11LC/MS.1GC/MS) # of Spikes 2 +H20 2 (1LC/MS, 1GC/MS) 2 -H20 2 (1LC/MS,1GC/MS) 2 +H20 2(1LC/MS.1GC/MS) 2 -H20 2 (1LC/MS, 1GC/MS) 2 +H20 2(1LC/MS, 1GC/MS) 2 -H 20 2 (11LC/MS, 1GC/MS) Control matrix without test substance 2 +H20 2(1LC/MS.1GC/MS) 2 -H20 2 (11LC/MS,1GC/MS) EXPOSED, UNEXPOSED, & DAY 0 24x 3 per exp type = 72 Total Samples per compound per study = 120 (60 for LC/MS, 60 for GC/MS) 2 +H20 2(1LC/MS,1GC/MS) 2 --H20 2 (11LC/MS, 1GC/MS) 16x3=48 12.6 Separate the vials into three boxes labeled "Day 0," "Exposed," and "Unexposed." Initial addition o f peroxide (Section 8.4) is done at this time by removing the cap and injecting the appropriate amount (e.g. 10 - 50 (J.L) into the vial. (Subsequent additions o f peroxide shall be injected through the septa of the VOA vials.) 12.7 F or use of quinine actinometer (Optional): Prepare a batch o f quinine irradiation control samples by aliquoting 5 mL of the 2% aqueous solution (Section 8.8) into the appropriate number of clear and amber 40 mL I-CHEM vials. Prepare one clear and one amber vial per reactor, per day of exposure. Store the vials at 1-5 C and protected from light prior to use. Place one clear vial in the reactor per day, while removing exposed quinine controls. Exposed quinine controls need to be wrapped in foil upon removal to protect from further exposure. Store at 1-5 C prior to measuring the absorbance via UV-Vis Spectrophotometer. The absorbance measurement should be performed as soon as possible, as the absorbance increase rate after light source removal may be 20% of the rate of when light is present. (Reference 18.5). 12.8 Place all the "Day 0" samples immediately in a cooler at 1-5 C or freeze at a continuos temperature of less than 0 C, inverted and protected from light. 12.9 Place "Unexposed" sample vials (amber) into Ziploc bags separated and labeled as "with peroxide" and " without peroxide", respectively. Place the bags in the bottom o f the water bath, under the photoreactor tray that holds the exposed samples. The "unexposed" samples will remain immersed in the 23-26 C water bath under the exposed samples for the duration of the exposure. Include one quinine control sample in an amber vial with the unexposed sample set. 13.0 Photoreactor set up ETS-8-176.0Preparaiion o f S am ples f o r P h otolysis Studies in Aqueous M atrices Method Page 13 o f 18 Page 49 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 13.1 Set the irradiation intensity at the desired output. For most experimental conditions, an intensity of 261 W/m2 is chosen because it yields the equivalent average optimum natural daylight radiation for 300-400 nm at known latitude, (see the table below): Irradiance Source Average Optimum Natural Daylight1 Atlas Photoreactor with integrated irradiance output o f 261 W/nr 300-800 nm using the IR Reflecting and 290 cuton filters `Measured, Miami, Florida (18.3') Aonroximate Inteerated anc Individual Irradiances in W/m2 250-300 nm 300-400 nm 400-800 nm 340 nm 420 nm 0.0 27.8 259.0 0.30 0.67 0.08 27.8 234.36 0.24 0.71 13.2 , For all "Exposed" samples, invert the vials cap-side down and vertical in the photoreactor tray holders to a depth that ensures that half the VOA vial is submersed in the 23-26 C water bath. Include one quinine control sample (Section 12.8) in a clear vial with the exposed sample set. 13.3 Close the door to the photoreactor and turn the door knob completely so the sensor detects that the door is completely shut. 13.4 Turn on the water cooler bath, ensure that it is set between 23-26 C. 13.5 Turn on the photoreactor pump and the power to the reactor. 13.6 Set the irradiation program using the following parameters in the table below: Photoreactor conditions Parameter Setting ' Program #, # of Phases Flowing Water ("FW") 1,1 ON Irradiation intensity Duration of exposure Exam ple: 261 w atts/m 2 Example: 8 hours 13.7 After entering the appropriate parameters within the menu, select program #1 to run and start the irradiation program. Upon lamp ignition, the water bath begins to fill and circulate. Visually check for both lamp and water bath activation as an indication of proper initiation of the photoreactor. When removing samples during the exposure period, select "stop" from the photoreactor keypad. Open the door and carefully remove the sample rack. Caution: The walls o f the chamber are hot and sensitive to scratching. To restart the program, close the door, ensure that the door knob is turned all the way in and press "start" on the reactor module. Visually inspect for proper lamp ignition and water bath circulation. 13.8 At the specified time, spike appropriately labeled sample types with a known volume (e.g. 10-50 pL) of 30% H2O2 solution (Section 8.4) and swirl to ensure adequate mixing. Maintain the inverted position of samples removed for spiking pre- and post- the actual peroxide injection. Return samples to their designated locations (bottom of reactor pan ETS-8-176.OPreparaiion o f Sam ples f o r P hotolysis Studies in Aqueous M atrices Method Page 14 of 18 Page 50 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 for the unexposed samples, reactor tray holder for the exposed samples, or the cooler for the Day 0 samples). Note: Don'tforget to addperoxide to appropriate Day 0 samples! 13.9 Remove the exposed quinine control sample from the reactor tray and visually confirm a color change as an indicator of photoreactor performance. The solution should be a gray/brown color after irradiation. Record the total time exposure of the sample, wrap the sample in foil to protect from light and analyze the quinine sample. Place a new quinine solution vial into the photoreactor tray with the exposed samples. Note: Quinine samples do NO T receiveperoxide. 13.10 Record the chamber temperature daily on the sample prep sheets. 13.11 Upon completion of the photolytic exposure, samples are removed, labeled with adhesive-backed labels and the study sets (Exposed, Unexposed and Day 0) organized for LC/MS or GC/MS analysis. If subsequent analysis can not be performed immediately, store samples in a cooler at 1-5 C. 13.12 Pertinent information regarding start and stop times o f photoreactor exposure study, water bath and chamber temperatures, addition of peroxide, and an explanation of unexpected occurrences shall be documented on the sample preparation worksheets, with appropriate dates, times and initials. 14.0 Sample preparation for analysis.______________________________________ 14.1 LC/MS sample extraction and prep. 14.1.1 Dilute all 5 mL samples by a factor of 1:7 v/v by adding 30 mL of an appropriate analytical solvent containing internal standard (Section 8.6.2) to all vials. 14.1.2 Add spike solution (Section 8.6) containing the target analytes to the appropriate samples. 14.1.3 Ensure the sample vials are inverted several times to ensure adequate mixing. 14.1.4 If samples appear cloudy, and/or the sample matrix appears unclear, it may be necessary to centrifuge the samples, at an appropriate speed and duration (e.g. 2000 rpm for 10 minutes), until no noticeable particulate matter is suspended in the sample. 14.1.5 Aliquot approximately 1 mL into autovials and tightly cap. 14.2 G C/M S sam ple preparation. . 14.2.1 Set up autosampler and concentrator methods. If samples have been kept in cold storage, bring samples to room temperature (approximately 23-26 C). 14.2.2 Spike vials through the septa and place in the autosampler. 15.0 Data Analysis and Calculations________________ ______________________ 15.1 The amount of target analytes in the sample will be quantified against a standard curve regression. 15.2 Means will be calculated by adding the individual entities and dividing the resultant sum by the number o f individual entities. 15.3 Standard deviations will be calculated using either Microsoft Excel or Microsoft Access to calculate standard deviation. The built in function contains the following equation which is based on the individual entities (n) being less than 30: Sx2 -(S * )2 n(n-l) ETS-8-1IS .O P reparation o f Sam ples f o r P h o to lysis Stu dies in Aqueous M atrices Method Page 15 o f 18 Page 51 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 15.4 Sample precision will be reported as % relative standard deviation (% RSD). Sample % RSD will be calculated using the following equation: where: A/B X 100 = Sample % RSD A= standard deviation of averaged samples B= average of samples 16.0 M ethod Performance___________ ' _____________________ 16.1 Refer to the definitions section for the method performance specifications/criteria for each respective sample type. 17.0 Pollution Prevention and Waste Management_________________________ 17.1 Dispose o f sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 18.0 Records__________________________________________ ______ __________ 18.1 Fill out the photolysis sample preparation worksheet documents completely, making sure to include all initials and dates. Store photolysis sample preparation worksheets in the raw data file. 18.2 Enter all standard, stock, solutions, etc. preparation information in the proper preparation logbook(s). Make a photocopy of the logbook pages used, and include the copy in the raw data file. Photocopied logbook pages will be included in the final data packet. 18.3 Archive electronic data to compact disc media. 19.0 Attachments___________________ .___________________________________ 19.1 Attachment A: Example Photolysis Prep sheet. 20.0 References_________________________________________________________ 20.1 Crosby, Helz, and Zepp. Aquatic Surface Photochemistry. t>480 20.2 Inteipersonal conversation with Carrie O'Connor, Optical Systems Engineer, Atlas Electric Devices. ' 20.3 "Suntest CPS/CPS+ Spectral Irradiance Distribution," table distributed by Atlas Electric Devices Company, sent via fax by Richard Sherwin, Sales Representative, 26 Jrdy, 2000. 20.4 "Atlas Xenon Filter Combination and Sunlight Measurements," information generated by Atlas Electric Devices Company sent via fax by Richard Sherwin, Sales Representative, 26 M y, 2000. ETS-8-\1(>.^ P reparation o f S am ples f o r P h o to lysis Studies in Aqueous M atrices Method Page 16 o f 18 Page 52 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 20.5 Bergstrom, David H., Thomas C. Kester, and Shangdong Zhan. "Quinine Chemical Actinometry Studies Under Two Light Sources Specified by die ICH Guideline on Photostability Testing." 21.0 Affected Documents 21.1 None 22.0 Revisions_______ .______________________ Revision Number. Reason For Revision Revision Pate ETS-8- n 6.(Preparation o f S am ples f o r P hotolysis Studies in Aqueous M atrices Method Page 17 of 18 Page 53 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Attachment A --Photolysis Sample Prep Sheet Test Analyte: _____________ Froject/Lab Request Number Exposure Type: Nominal Expos Analysis: _______ LC/MS Sample Matrix: Control Matrix: Fluorocbemical Degradation (Photolysis) Analysis Sample Prep Sheet `Following initial sample prep, all samples will be placed cap-side down and vertical. Followingphotolysis, Day 0 samples will bepulled and extracted with Exposedand Unexposed samples ample Matrix Volume: Date: Time: Test Analyte -- P l r in te rn a l Spike Solution Comments: ----------------- ample No. Description Sample Type Solution .D. Solution 1.0. Solution I.D, Rep i Day 0 Rep 2 DayO Rep 3 DayO Splice DayO Matrix Blank DayO Matrix Blank Spike ____p ? 0 -- ................................ i Control Rep 1 DayO Control Spike DlyO V-.-: :~ ... Control Blank Control Blank Spike -- DayO ...... I BH H M BIBIW W I 1\_______________ ---- .- Rep I Rep 2 Rep 3 Spike Matrix Blank Matrix Blank Spike Control Rep 1 Control Spike Control Blank Control Blank Spike Exposed Exposed Exposed Exposed Exposed Exposed Exposed Exposed Exposed Exposed -1 t ______. ' ` -, .... \ . .. ! . .. Rep 1 Unexposed Rep 2 Unexposed Rep 3 Unexposed Spike Unexposed Matrix Blank Unexposed Matrix Blank Spike Unexposed Control Rep 1 Unexposed Control Spike Unexposed ' !.S 1Control Blank Unexposed / Control Blank Spike Unexposed : Components) and Solution Concentration! i . > I" ' : -T'. ,i - . : ? - -V ' 4 . 1 S o lu tio n I.D . Solution l.D. Comments: mi - *( HHHHHi Vf A : ? ~" ... ;./ / -.r - BBS mm Hi IB Start Date: Time: Freezer/Refriferator Storage I.D>: Initials Start Date: Time: Stop Date: Time: Initials Stop Date: Time: Total Exposure: _________ _ days____________hours Initials Initials Check mark - Indicates the solution added, n/aKnot applicable Comments: E T S -S -l7 6 .0 P re p a ra tio n o f S a m p les f o r P h o to lysis Stu dies in A queous M a tric es Method Page 18 o f 18 Page 54 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory E q uipm en t Procedure O pe r a t io n and M aintenance o f th e Su n lig h t E x posu re Sy stem , I m m er sio n unit, and R ecircu la tin g W a ter C h il l e r Sy stem Procedure Number: ETS-9-44.0 Exact Copy of Original Initial Date Approved by: Laboratory Management Adoption Date: Revision Effective Date: Date Date ETS-9-44.0 Equipment Procedurefo r the A tlas SUNTEST Sunlight Exposure System Page 1 of 8 Page 55 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application_____________________________________________ _________ 1.1 This equipment procedure describes the regular operation and maintenance of the Atlas SUNTEST Sunlight Exposure System equipped with an immersion unit and recirculating water chiller. 2.0 D efinitions__________________________________________ ._______________________ 2.1 Photon energy: U = hv = hc/X where h is Planck's constant, c is the speed of light, and v and Xare the frequency and wavelength of light. Therefore, the energy of a photon, U, is inversely proportionate to the wavelength. 2.2 Irradiance: The energy output ("U" in the above equation for energy of a photon) in Watts/m2 specific to a wavelength or wavelength range. The irradiance output specific to the types of Atlas wavelength filters available (Reference 14.9) should be used as a guide to calculating the global irradiance (in units of W/(m2nm) needed to give a specific energy over a desired wavelength range. 3.0 Description_________________________________________________ ._______________ 3.1 The Atlas SUNTEST Sunlight Exposure System (CPS+ or XLS+) produces visible and ultraviolet light (250-765 W/m2). Light produced is filtered with a filter or combination of filters to allow specific wavelength ranges. Samples are exposed to the light in a reflecting chamber. An immersion unit with water recirculation through a chiller provides a cooled, constant sample temperature. 4.0 Identification_____________________________________________________ _ 4.1 Atlas SUNTEST XLS+, equipped with a xenon arclamp, lamp filter(s) available from Atlas to allow specific irradiance ranges, and immersion unit. 4.2 Atlas SUNTEST CPS+, equipped with a xenon arclamp, lamp filter(s) available from Atlas to allow specific irradiance ranges, and immersion unit. 4.3 Neslab CFT-33 Refrigerated recirculator or equivalent 5.0 W arnings and Cautions______________ ;_______________________________________ 5.1 Health and Safety Warnings: 5.1.1 Wear appropriate laboratory safety personal protective equipment. 5.1.2 The xenon lamp emits ultraviolet light which can cause bums to the skin and permanent damage to the eyes. Never attempt to operate the unit with the test chamber door open. 5.1.3 When filling the sample immersion unit with water, always shut off all power to the SUNTEST device and the immersion unit to prevent electrical shock. 5.2 Cautions: 5.2.1 Handle optical parts carefully, fingerprints on the lamp, filter or quartz dish can result in altered spectral output or early lamp failure. 5.2.2 The reflective coating of the test chamber walls is sensitive to scratches. Do not use any abrasives or harsh cleaning agents that may cause scratches and non uniform illumination of the test chamber. ETS-9-44.0 Equipment Procedurefo r the Atlas SUNTEST Sunlight Exposure System Page 2 of 8 Page 56 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 5.2.3 5.2.4 5.2.5 Keep SUNTEST unit clear o f obstructions that would block vents; overheating may cause blown fuses, shortened lamp life or other damage. After beginning the experiment, always make sure that the sample vials are sufficiently submerged. Excessive heat may affect the results o f the experiment. Manually drain the immersion tank on the XLS+ models after stopping the run; otherwise, the water will overflow. 6.0 Special Instructions_____________________________________________ _ ________ 6.1 None. 7.0 R esponsibility_____________________________________________________ 7.1 The analytical group o f personnel who routinely operates the equipment is collectively responsible for the instrument operation as described in this document. The person responsible for maintenance and calibration (and an alternate) will be identified in the front o f the equipment logbook. 8.0 Supplies and Materials________________________________________________ __ 8.1 Xenon lamp for XLS+, Atlas PN 56077798 8.2 Xenon lamp for CPS+, Atlas PN 56001794 8.3 Hand-tools as required 8.4 Kim-WipesTM 8.5 Optional radiation filter(s) for lamp available from Atlas: Filter/ Atlas P art Number Q uartz Dish w1IR reflective coating, PN 56052388 Q uartz Dish, PN 56052373 UV Special Suprax Filter, PN 56052371 Window Glass Filter, PN 56052372 Window Glass Solar ID 65 Filter, PN 56077769 Solar Standard Filter, PN 56077759 Properties IR reflective coating (supplied standard with unit) Uncoated (to allow higher black standard temperatures) Cut-on at 290 nm, simulates outdoor solar radiation. Cut-on at 310 nm, simulates exposure behind 3 mm (0.118 in.) window glass. Cut-on at 320 nm, simulates exposure behind 6 mm (0.236 in.) window glass. (Must be used with Window Glass Filter above.) Cut-on at 290 nm, simulates outdoor solar radiation at optimal UV intensity. ETS-9-44.0 Equipment Procedurefo r the Atlas SUNTEST Sunlight Exposure System Page 3 of 8 Page 57 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 9.0 Cleaning Procedures________________________________________________________ All routine and non-routine cleaning procedures will be performed by person(s) designated in the front o f the instrument logbook; see Section 12). 9.1 Routine cleaning 9.1.1 Clean the inlet air filters at the back of the SUNTEST unit every 6 months with a mild soap solution. Rinse in clean water. When more severe contamination is present, vacuum the filters or replace them. 9.1.2 Clean the reflector in the test chamber when it is dirty, using a soft cloth and mild soap solution. DO NOT use any abrasive cleaning materials or the reflector may be permanently damaged and irradiance uniformity will be altered. 9.1.3 Clean and/or flush the water tank and water lines on the immersion unit monthly to prevent build up of residue in the circulating water system. 10.0 M aintenance Procedures____________________________________________ ________ 10.1 Routine maintenance will be performed by the person(s) designated in the front of the equipment log (see Section 12): 10.1.1 Replace the xenon lamp after 1500 hours or when the required irradiance level cannot be achieved (e.g. error message reads "E MAX Power reached; CHANGE XENON LAMP") Refer to the SUNTEST instruction manual for details on how to replace the lamp. 10.1.2 If the temperature near the lamp becomes too high, the fuse blows to interrupt power and save the lamp (indicated by the error message "DOOR OPEN or TEMPERATURE FUSE"). Refer to the SUNTEST instmction manual for details on how to replace the fuse. 10.1.3 Record routine maintenance in the equipment log (see Section 12). 10.2 Non-routine maintenance will be performed by the person(s) designated in the front of the equipment log (see Section 12): 10.2.1 If the equipment fails to operate, refer to the equipment manual for further instructions, if necessary. Contact the Team Leader for instructions if the equipment cannot be made operational. 10.2.2 If an abnormal operating situation occurs or if calibration verification fails, contact the responsible individual identified in the equipment log. Label the equipment as "out of service" if it cannot be immediately repaired. 10.2.3 Record non-routine maintenance in the equipment log (see Section 12). 11.0 Instrument Calibration____________________________________________________ 11.1 The photoreactor is set to maintain a specified integrated energy output. The amount of energy output from the lamp may be monitored with the use o f a radiometer. The radiometer system will provide and record instantaneous energy output. Refer to ETS-950.0 Operation and Maintenance o f Radiometer and Detector. 11.2 Calibration o f SUNTEST systems will be performed two times each year by Atlas Electric Devices Company. ETS-9-44.0 Equipment Procedurefo r the A tlas SUNTEST Sunlight Exposure System Page 4 of 8 Page 58 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.0 Operating Procedures _____________________________________________ _ 12.1 For more detailed operating instructions refer to the equipment operating manuals. 12.2 Immersion Unit 12.2.1 To begin operation, fill the tank with water until the level indicator is up to the full mark. 12.2.2 Turn the power on. 12.3 Chiller 12.3.1 Turn the power on. Set water temperature knob to desired set point. Allow temperature to equilibrate before igniting the SUNTEST lamps. 12.4 SUNTEST XLS+ or SUNTEST CPS+ unit set-up 12.4.1 Select the desired wavelength filter from the parts listed under Section 8.4 to achieve the proper irradiation specified in the program, and program the photoreactor with the filter type information: With the photoreactor menu in the "Program" mode, select the appropriate filter combination type: _________________________Optical Filter System Designations A: Coated quartz glass only________________ _______________ B: Coated quartz glass with UV special glass_________________ C: Coated quartz glass with window glass____________________ D: Uncoated quartz glass only ___________________________ E: Uncoated quartz glass with UV special glass_______________ F: Uncoated quartz glass with window glass 12.4.2 Selection/determination of energy output (W/m2) 12.4.2.1 hxadiance control and display is between 250-765 W/m2 (nominally 300 800 nm). The irradiance is determined by the settings of the test program [including type o f filter(s) used]. The selectable range is from 250 W/m2 to 765 W/m2 (page 12, XLS+ Instruction manual). The total (integrated) energy output (300-800 nm) is directly dependent on the type o f lamp filter(s) used. E.g. if the filter has a narrow range such as a cut-on at 400 nm, all irradiance energy coming from wavelengths <400 nm will not reach the samples, and the total integrated irradiance will be less than if the filter's cut-on was at, for example, 290 nm. 12.4.2.2 Once the proper filter(s) is/are designated, the photoreactor will base the energy output on what type o f wavelengths are being allowed to pass through the filter system to reach the samples. To calculate the energy output to program into the system, refer to References 14.7,14.8 and 14.9 as guides to calculate the desired spectral irradiance. Reference 14.7 may be used to calculate the programmed global irradiance necessary to achieve desired irradiances at specific wavelengths or wavelength ranges. Reference 14.8 may be used to reference sunlight measurements and to correlate natural sunlight to the Atlas Suntest photoreactors. Reference 14.9 is a useful reference for determining irradiances at a specific ETS-9-44.0 Equipm ent Procedurefo r the A tlas SUNTEST Sunlight Exposure System Page 5 of 8 Page 59 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 wavelength or a wavelength range using specific filter combinations at specified global irradiances o f 250, 500, and 765 W/(m2nm). 12.5 Photoreactor Analysis set up 12.5.1 Place VOA (volatile organic analysis) vials containing samples (see the appropriate analytical method) into the test chamber. Sample vials to be exposed must be cap side down to allow light to enter the vial. Tighten caps securely to prevent leakage. Secure the vials in the chamber to prevent floating once the water begins to circulate. 12.5.2 Close the chamber door and turn the power on. 12.5.3 From the initial LCD display, use the arrow keys to select Program. Press "Enter". 12.5.4 If programming a new method is necessary, use the arrow key to select Programming. Press "Enter". 12.5.5 Input program number, number of phases, desired irradiance, immersion function, phase time and switch off criteria. Each entry is followed by the "Enter" key. 12.5.6 To start the program, select "Program Start" and press the "Enter" key. 12.5.7 Press "Escape" for the next screens if the filter has not been changed and no printout is desired. Input program number and press "Start" 12.5.8 Program will begin with lamp ignition. Note: Due to the modified sample chamber in the SUNTEST XLS+ models, the water initially present in the immersion tank is not sufficient to fill the sample chamber once a program has started. Refill the immersion unit as the water level drops below the fill line. Once a program has finished, drain the immersion tank so that it does not overflow when water from the sample chamber drains back down into the immersion unit. Failure to do so may result in remote flooding. 12.5.9 To interrupt operation (e.g. to add peroxide reagent)Press "Stop" . If it is necessary to turn the power off (to exchange the lamp, for example) wait until the fan turns off in 1-3 minutes before turning power switch to "O ff' and unplugging the power cord. When ready to continue operation, turn power "On" . 12.5.10 To resume operation, press "Start". The program will continue at the point of interruption. 12.5.11 To read parameters during the program run, scroll through the parameters of the running program by using the arrow keys. This is helpful to see how many more hours are remaining in the running program. 12.5.12 The SUNTEST will shut off automatically when the switch-off criteria are reached. To display the total time and irradiance, press "Enter". Record exposure time in instrument run log. Then turn power "O ff'. 12.5.13 To manually stop the program, press "Stop". Wait until lamp is cooled, then press "Escape". Power can then be turned "O ff'. ETS-9-44,0 Equipm ent Proceduref o r the A tlas SUNTEST Sunlight Exposure System Page 6 of 8 Page 60 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 13.0 Records___________________________________________________________ 13.1 Instrument logbooks 13.1.1 Equipment Log: The person(s) designated at the front o f the equipment log will record all cleaning and maintenance activities in the appropriate log for each SUNTEST system. Records for routine maintenance of equipment must include the dates of the operation, whether the operations followed the SOP, and the initials of the person performing the operation. Records for non-routine repairs performed as a result o f instrument failure or malfunction must include the nature of the defect, how and when the defect was discovered, any remedial action taken in response to the defect, and the date and initials of the person performing the maintenance. Maintenance by outside contractors should include their name and company affiliation. 13.1.2 Run Log: Record each experiment in the appropriate instrument logbook. Enter the operators initials, time and date of exposure, lamp intensity and water temperature as the samples are placed in the chamber. When samples are finished, record the time and date when samples came out, the ending chamber temperature and the actual hours of exposure. All entries made in the run log should be initialed and dated. 13.1.3 SUNTEST Data Output Log: XENOVIEW 2.2 Storage Software will receive and record the measurement data transferred from the SUNTEST system to a computer or printer, while a program is in progress. The measurement data recorded includes: number of phases, phase time, chamber temperature, radiant exposure, irradiance, running time, date and time data is recorded. Refer to XENOVIEW software instruction manual for details on how to operate software. Any printouts of program or other data should be initialed and dated prior to adding to the study file. 13.2 Identification records for each system include equipment ID, manufacturer, model number, and serial number of each individual component. In addition, if components are removed or added, the above information must be written in the logbook including the date the change was made and initials o f the analyst completing the change. 14.0 References________________________________________________________ 14.1 SUNTEST XLS/XLS+ Instruction Manual, Doc. No. 20-8036-00 Rev. 0 12/98 Atlas Electric Devices Company. 14.2 SUNTEST CPS/CPS+ Operating Manual, 6/97 Atlas Company. 14.3 SUNTEST XLS+ Immersion Device Operating Manual, 2/99 Atlas Company. 14.4 SUNTEST CPS+/XLS+ Software Documentation 1.4 Atlas Company. 14.5 XENOVIEW 2.2 Storage Software Operating Instructions. 14.6 ETS-9-50.0, Operation and Maintenance of Radiometer and Detector. 14.7 "SUNTESTIrradiance in W/m2*nm". Tables furnished by Atlas Company. 14.8 "Atlas Xenon Filter Combination". Table furnished by Atlas Company. 14.9 "SUNTEST CPS/CPS+ Spectral Irradiance Distribution". Table furnished by Atlas Company. ETS-9-44.0 Equipm ent Procedure fo r the Atlas SUNTEST Sunlight Exposure System Page 7 of 8 Page 61 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 15.0 A ffected D ocuments 15.1 None. 16.0 Revisions Revision Number. Reason For Revision Revision Date ETS-9-44.0 Equipm ent P roceduref o r the A tlas SUNTEST Sunlight Exposure System Page 8 o f 8 Page 62 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory Analysis o f F luorochem icals by A rch o n P urge and T rap A u to sam pler, T ek m a r P urge and T rap Concentrator and A g ilen t G as C hrom atograph/M ass Spectr o m eter Procedure Num ber: ETS-8-18ZD Exact Copy of Original Initial D a te Approved by: Laboratory Manager Adoption Date: Revision Date: Date Date ^ ETS-8-182.0 ' Analysis ofF C s by Purge & TrapAutosam pler/Concentrator/G C/M S Page 63 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application Til Scope. This method is used for the analysis of selected hydrolysis and photolysis samples for the presence of degradation products such as olefins and hydrides using gas chromatography/mass spectrometry in a full scan mode. An Archon autosampler and Tekmar Purge and Trap concentrator (or an equivalent system) is coupled to a GC for purging analytes from the liquid matrix and concentrating them on the trap column before injecting on to the GC. 1.2 Applicable compounds. Compounds that may be analyzed by this method are listed below. Other fluorochemicals may be detected by monitoring mass spectra and running library comparison. Compounds that are detected but do not have appropriate standards, will be quantified relative to structurally similar standard compounds listed below. 1.2.1 lH-perfluoroethane (lH-pfC2) 1.2.2 Perfluro-2-butene (pfC4-2ene) 1.2.3 lH-perfluropropane (lH-pfC3) 1.2.4 IH-perfluorobutane (lH-pfC4) 1.2.5 Perfluoro-2-heptene (pfC7-2ene) 1.2.6 Perfluoro-l-heptene (pfC7-lene) 1.2.7 lH-perfluorohexane (lH-pfC6) 1.2.8 Perfluoro-2-octene (pfC8-2ene) 1.2.9 IH-perfluoroheptane (lH-pfC7) 1.2.10 2H-perfluorooctane (2H-pfC8) 1.2.11 lH-perfluorooctane (lH-pfC8) 1.3 Instrument Surrogate compounds. Added at the time o f analysis and used to monitor performance of purge and trap autosampler and concentrator. 1.3.1 Dibromofluoromethane 1.3.2 Toluene-d8 1.3.3 4-Bromofiuorobenzene 1.3.4 Pentafluorobenzene 1.3.5 1,4-Difluorobenzene 1.3.6 Chlorobenzene-d5 1.3.7 l,4-Dichlorobenzene-d4 1.4 Sample Surrogate compounds. May be added at the time of sample preparation. 1.4.1 Perfluorocyclohexane 2.0 Summary of Method________________________________ '________________________ 2.1 A dynamic purge and trap system (autosampler and concentrator) is coupled to a temperature programmed GC for analyte separation and subsequent mass spectrometer detection and quanitation. The liquid sample is purged for 20 min. in the sample vial, and file volatile components are swept onto a chemical trap in the concentrator. In the subsequent desorption mode, gas flows in opposite direction and temperature o f the chemical trap increases to 250 C. The trapped analytes are transferred onto the GC column for GC/MS separation, detection, and quanitation. Through this process, a high volume o f sample is injected and most of the non-volatile matrix components stay in the sample vial, allowing low level detection o f fluorochemicals. ETS-8-182.0 Analysis o f FCs b y Purge & Trap Autosampler/Concentrator/GC/M S Page 2 of 12 Page 64 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.0 Definitions_________________________________________________________ 3.1 Calibration Standard. A dilution of various amounts of a stock, intermediate or purchased standard to achieve standard solutions in a concentration range of interest. 3.2 Calibration Curve. The graphical relationship between known values, such as concentration o f a series of calibration standards and their instrumental response. 3.3 External Standard Quantification. Process of establishing the concentration o f a target analyte by plotting the theoretical amount (in units of ng/mL or pg/mL, etc.) versus the response o f the target analyte(s) on column. The resultant curve(s) shall be used to determine unknown concentrations by comparing the area response of target analyte(s) to the area response and corresponding analyte amount on the appropriate analyte's calibration curve. 3.4 Coefficient of Determination (r2). The square of the correlation coefficient. It is the proportion o f the variation in the dependent variable that is accounted for by the independent variable. 3.5 Instrument Surrogate. An organic compound similar to the target analyte(s) in behavior in the analytical process, but is not normally found in the sample(s). A surrogate may be added to sample vial during instrument analysis. 3.6 Sample Surrogate. An organic compound similar to the target analyte(s) in chemical composition and behavior in the analytical process, but is not normally found in the sample(s). A surrogate may be added to sample triplicates and matrix spike samples along with the test analyte (pre-photolysis). 3.7 Continuing 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 Solvent Blank. A sample of analyte-free medium that is not taken through the sample preparation process. This blank is used to evaluate instrument contamination. 3.9 Blank. For photolysis studies, there are multiple blanks to adequately represent the variables o f the study (Exposed, Unexposed and Day 0 samples with/without peroxide addition). The blank is carried through the sample preparation, photolytic and analytical procedures to monitor for contamination during any step. It is also used to establish a chromatographic baseline/background and monitor for analytical interference or suppression o f target analyte(s) from the matrix. 3.9.1 3.9.2 Matrix Blank: A sample of analyte-free matrix (buffered water, lake water, etc.) to which all reagents are added in the same volumes or proportions as used in sample processing. It is used to document the test system without test analyte. Control Blank: A sample of analyte-free matrix (Milli-Q water) to which all reagents are added in the same volumes or proportions as used in sample processing. It is used to control the test matrix and monitor matrix specific ETS-8-182.0 . Analysis o f P C s by Purge & TrapAutosampler/Concentrator/GC/M S Page 3 of 12 Page 65 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 background levels, interferences or suppression o f target analyte(s) from the matrix. 3.10 Lim it of Quantitation (LOQ). The lowest concentration that can be reliably measured within specified limits o f accuracy during routine laboratory operating conditions. The LOQ is generally 5 to 10 times the minimum concentration with a 99% confidence limit 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 matrix blank. 3.11 Sample Triplicates. Three samples taken from and representative of the same sample source. These are prepared separately and carried through all steps o f the exposure, extraction and analytical procedures in an identical manner. There are multiple sets of triplicate samples to adequately represent the photolytic variables o f the study (Exposed, Unexposed and Day 0 with/without peroxide addition). Triplicate samples are used to assess variance of the photolytic method, including sample preparation, photolysis exposure, and analysis. 3.12 Relative Standard Deviation (RSD). A measure of precision defined as the standard deviation of three or more values divided by the average o f the values and multiplied by 100. (Also reported as Coefficient o f Variation (CV)). 3.13 Analytical Spike. Prepared by adding a known mass of target analyte(s) to a specified amount o f a sample or control matrix prior to analysis. This assumes that an independent estimate of target analyte concentration is available. Matrix spikes are used to determine the effect o f the matrix on method recovery efficiency. 3.14 Accuracy. 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 of 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. 3.15 Geometric Mean of the calibration curve: The square root o f the product of the high standard concentration and the low calibration curve standard. When preparing calibration curve standards, the number of calibration standards below the geometric mean shall equal the number of calibration standards above the geometric mean. Having equal distribution o f calibration standards above and below the geometric mean when analyzing and reprocessing data, effectively weights the curve such that both the high and low ends o f the curve are given equivalent significance. 4.0 Warnings and Cautions__________________________________________ 4.1 Health and Safety Warnings: 4.1.1 The operator must be familiar with the purge and trap autosampler/concentrator/GC/MS system and associated hazards, such as high temperature, effluent venting, solvent use, and low-pressure vacuum system. See instrument manuals ETS-8-182.0 Analysis o f FCs by P urge & Trap Autosampler/Concentrator/GC/M S Page 4 o f 12 Page 66 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 4.1.2 All exhaust vents, including the GC oven vent, Tekmar concentrator purge vent, split vent and mass spectrometer pump exhaust must be connected to the laboratory vent system to keep potentially hazardous effluent from mixing with laboratory air. 4.2 Cautions: 4.2.1 It is recommended that a grounded antistatic wrist strap be worn while disconnecting all wires, contacts, or cables which are connected to printed circuit boards within the Archon autosampler, Tekmar concentrator or MS analyzer. 4.2.2 To prevent the breakage o f the Standard Vial on the Archon autosampler, do not use any tool and do not overtighten the thumbnut. 5.0 Interference________________________________________ 5.1 Methanol, water and other co-extracted matrix components could interfere with detection decreasing sensitivity. 6.0 Equipment 6 il System: "Rufus", or equivalent: 6.1.1 Autosampler: Varian, Archon 6.1.2 Concentrator: LSC2000, Tekmar 6.1.3 GC: 6890, Agilent 6.1.4 MS: 5973N, Agilent 6.1.5 Column, GS-GASPRO 60m x 0.23mm, J&W ~ 7.0 Supplies and Materials__________ __________________________________ 7.1 Helium, ultra-high-purity 7.2 40ml VOA vials, e.g. I-Chem, S236-0040 8 .0 R e a g e n t s a n d S t a n d a r d _______________ ________________________________ 8.1 Methanol, Purge and Trap grade or equivalent 8.2 Standards. Typically a minimum of five calibration standards, ranging from 1 ng/ml to 20 ng/ml are prepared. This concentration range should bracket the concentration o f samples and matrix spikes; if the analyte concentration exceeds this range, then the calibration range should be increased. 8.3 Instrum ent Surrogates. Used only to monitor performance of purge and trap autosampler and concentrator and not for quantitation. 8.4 Sample Surrogates. May be used to monitor sample preparation, photolytic exposure and analytical performance. 9.0 Sample Handling_______________________________________________ 9. 1 Store standards and samples in the refrigerator at 4 0 C 3 C until analysis time. ETS-8-182.0 Analysis o f FCs by Purge & Trap Autosampler/Concentrator/GC/M S Page 5 of 12 Page 67 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 9.2 For the analysis, pull samples and standards out o f the freezer and bring them to room temperature. 10.0 Quality Control___________ ________________________ 10.1 Calibration Standards. Calibration standards (Section 11) used to generate a calibration curve. The number of calibration standards and the concentration levels should be sufficient to encompass the expected concentrations of the study samples. In general, a minimum of five calibration standards is required for fit of linear regression. 10.2 Continuing Calibration Verification (CCV). Analyze a mid-range calibration standard after a maximum o f every fifteen samples. 10.3 Solvent blank. Solvent blanks are run before and after every calibration curve, CCV, matrix and control blank (see 3.9.2), and after batches of no more than 30 injections. Acceptable values for the blanks are values below 25% of the limit o f quantitation (LOQ) of the instrument. If analyte carryover is a problem, use back-to-back solvent blanks. 10.4 Sample Triplicates. Prepare and analyze all samples in triplicate to provide a measure o f the precision o f analysis. 10.5 Analytical Spikes. Prepare a matrix spike sample for each sample type as applicable to determine the matrix effect on the recovery efficiency. Concentrations o f the spike should be approximately equal to a mid-range calibration standard. The matrix spike sample should be analyzed periodically to measure the precision associated with the analysis. The analyst shall accept percent spike recoveries of 100 25%. Spike recoveries outside of this range should be noted and used with other criteria to evaluate the condition o f the analytical run. Consult with the Team Leader or designee for direction and final acceptance or rejection of the analytical run. 11.0 Calibration and Standardization 11.1 Analyze standards prior to each set o f samples. The linear regression will be calculated from the plot of all individual calibration points, without including or not forcing through zero, using Target NT Software. A minimum o f five calibration standards is required to generate linear regression for target analyte(s). If the calibration curve residuals are greater than 25% deviation from the theoretical value, quadratic curve fitting and/or dropping low/high curve points may be required if data review shows this to be a consistent and more accurate representation of the instrument response. Document in the raw data the technical justification for any deviation and consult with the team leader or designee for direction and for final acceptance or rejection of the data. 11.2 If the curve does not meet requirements perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze ETS-8-182.0 Analysis o f P C s by Purge & Trap Autosampler/Concentrator/GC/M S Page 6 of 12 Page 68 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.0 Pr o c e d u r e s ________ 12.1 Set Archon autosampler 12.1.1 Archon System Settings US Probe Temp 180 Xfer line Temp 180 US Valve Temp 105 Gripper Open 750 Gripper Closed 999 Standby Pol CLOSED DesDm Pol CLOSED STOP Pol. CLOSED Equilb. Count 0 Equilb. Time 0 12.1.2 Archon System Options Barcode Scanner NO Needle Sparge? YES Ign. Vial Type? YES Ignore No Vial? NO HotWater Rinse? NO Vial Checks? YES Beep on Error? YES 12.1.3 Archon Method Sample Type Soil First Vial 1 Last Vial up to 51 Sample Volume 10 Standard 1 (luL) YES Standard 2 NO S.PreHeat Stir NO Stir Syring Flushes NO 0 PreHeat YES PreHeat Temp 35 PreHeat Time 1.0 Purge Time 20 .0 Desorb Time(m) 0.5 Oper. Mode Remote Cycle Timer 0 .0 Aux. Timer 0 .0 Link to Method 0 .0 Soil Purge Flow 40ml/min Soil Purge Pressure 20psi ETS-8-182.0 Analysis o f FCs by Purge & Trap Autosampler/Concentrator/GC/M S Page 7 o f 12 Page 69 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.2 Set Tekmar options Standby 40C (30C by purge) Purge 2 0 .0 0 min Dry Purge 2 .0 0 min Desorb Preheat 245C Desorb 0.50min at 250C Bake lO.OOmin at 260C BGB OFF BGB Delay Osec Auto Drain ON Valve 180C Line 180C Mount 100C Runs per Sample 1 Purge Flow Purge Pressure Trap 40mL/min 2 0 psi VOCARB 3000 Containing: Carbopack B Carboxen 1000 Carboxen 1001 12.3 Set GC conditions 12.3.1 Oven: Initial temp: 40 C Initial time: 4.00min Ramp at 15.00 C/min to 280 C Final time: 10.00min 12.3.2 Front Inlet: Mode: Split Initial temp: 180C Pressure: 8.50psi (on) Split ratio: 10.7 :1 Split flow: 16.1 ml/min Total flow: 20.6 ml/min 12.4 Set MS conditions 12.4.1 Adjust conditions as needed to optimize system performance and document operating conditions in the instrument run log. Acquisition mode: Scan (from 10 m/z to 650 m/z) MS source temp: 230 C MS quadruple temp: 150C Interface temp: 260 C Multiplier voltage: adjust to give required low standard sensitivity ETS-8-182.0 Analysis ofF C s by P urge & Trap Autosampler/Concentrator/GC/M S Page 8 o f 12 Page 70 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.5 Set up the instrument acquisition method. Name the sequence. The sequence includes a sample list documenting the method used and datafiles created. The sequence should be documented n the run files. 12.6 Sample analysis. 12.6.1 Set up autosampler and concentrator methods. Bring samples to room temperature (~22 C), spike them and place them on autosampler. Generate a mass spectrometer tune report and review. Operating conditions provided above are recommended and may be adjusted to optimize system performance. Analyze all standard, samples, and spiked samples using the same analytical conditions. Document the conditions in the run log. ' 12.6.2 When data acquisition is complete, data files should be transferred to Target version 4.0 for processing. 13.0 Data Analysis and Calculation_______________________________________ 13.1 Each batch o f data should be processed using Target Genie integrator. Integration parameters should be set to minimize the number of manual integrations required yet still result in uniform integration of peaks at all concentration levels. If manual integrations are required, a review code should be assigned to indicate the reason. Review Codes are listed below. Review Code Explanation Peak was not automatically integrated by Target, therefore, integrated M l manually Peak was automatically integrated; was reintegrated manually to M2 improve sample-to-sample integration consistency. Incorrect quantification ion peak was integrated; manual integration M3 was done to select the correct peak. Incorrect monitor ion peak was integrated; manual integration was M4 done to select the correct peak. M5 Others (specify) 13.2 When data processing is complete, summarize the data using an appropriate form. Formula is provided below for some of the calculations that may be required. 13.2.1 Calculate matrix spike percent recoveries using the following equation: (observed concentration - background concentration) x 1 0 0 %Recovery = expected concentration 14.0 Method Performance_______________________________________________ 14.1 Coefficient of Determination (r2). The coefficient of determination (r2) for the initial calibration curves should be 0.990 or greater. The curves should be examined closely for ETS-8-182.0 Analysis o f F C s by Purge & Trap Autosampler/Concentrator/GC/M S Page 9 of 12 Page 71 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 linearity and intercept, particularly for accuracy of quantitation at the low and high ends o f the curve. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. 14.2 Calibration Standards. The acceptance criterion for the calibration standards is that the accuracy of each standard is 75% to 125% ( 25 % difference) of the nominal value. Calibration standards outside this range are to be noted. Document in the raw data the technical justification for deviations. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. 14.3 Instrum ent Surrogate. Review o f the instrument surrogate performance is performed by monitoring instrument surrogate recoveries throughout the run. Inconsistencies in the recoveries may be the result of instrumental changes, or injection error. Consult with the Team Leader or designee for direction and final acceptance or rejection of the analytical nm. 14.4 Sample Surrogate. Sample surrogate performance is evaluated by averaging the area response throughout the analytical run and calculating %RSD. Inconsistencies in the surrogate peak area may indicate instrumental changes, injection error, or changes in the test-system. Consult with the Team Leader or designee for direction and final acceptance or rejection o f the analytical run. 14.5 Continuing Calibration Verification. If the accuracy for the amount of quantified analyte is greater than 25% from the nominal value relative to the initial standard curve, the Team Leader should be consulted. Only those samples analyzed before the last acceptable calibration check standard will be used. Consult with the Team Leader or designee for direction and for final acceptance or rejection for the data. 14.6 Solvent Blanks. Solvent blanks should show no more than a 5% carryover from a high standard or calibration check standard. If so, two sequential solvent blanks may be necessary to rule out instrumental contamination 14.7 M atrix Blanks. Matrix blanks are the basis for determining the LOQ and are monitored at various times in the analytical run. Peaks with greater than 25% o f the peak area o f the designated LOQ value observed in matrix blanks are indicative of either matrix effect, sample contamination or instrument contamination. Use of solvent blanks prior to the matrix blank may be necessary to rule out instrumental contamination or sample contamination. 14.8 Control Blanks. Control blanks are the basis for determining matrix effect (interference or suppression). Peaks with greater than 25% o f the peak area of the designated LOQ value observed in control blanks are indicative of either matrix effect, sample contamination or instrument contamination. 14.9 Limit of Quantitation (LOQ). The LOQ is equal to the lowest acceptable standard (i.e. % accuracy is 5 25 % nominal value) in the calibration curve that is greater than 4 times the level of the matrix blanks. 14.10 Sample Triplicates. The analyst shall accept %RSD values < 25%. %RSD values > 25% should be noted. Data used in the final report that is deemed out of control will be ETS-8-182.0 Analysis ofF C s by Purge & Trap Autosampler/Concentrator/GC/M S Page 10 of 12 Page 72 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 required to have technical justification for why the data is used, documented in the final report and raw data. Consult with the Team Leader or designee for direction, and for final acceptance or rejection of the data. 14.11 Analytical Spikes. The analyst shall accept percent spike recovery values of 100 25%. Spike recoveries outside of this range should be noted. Consult with the Team Leader or designee for direction, and for final acceptance or rejection of 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. 14.12 System Suitability. 14.12.1 Tuning: A mass spectrometer tune report shall be generated before starting each analytical sequence. If the tune parameters do not meet the criteria suggested by the mass spectrometer manual, then the mass spectrometer should be re-tuned. If mass 28 is present in the tune report at >10% relative to mass 69 then an air leak is present in the system. The source o f the leak should be isolated and fixed before the sequence is stated; however if a slight air leak is detected, data can be collected, analyzed, and used as long as the data quality objectives are met. 15.0 Pollution Prevention and Waste Management__________________________ 15.1 Dispose o f sample vials in low BTU and flammable solvent in high BTU containers. Dispose o f glass pipette waste in broken glass containers located in the laboratory. 16.0 Records___________________________________________________________ 16.1 Store chromatograms in the study folder that is labeled with the study number. Include the following information on each chromatogram either in the header or hand written on the chromatogram: injection date, analyst's initial, sample unique number, sample name, preparation date, incubation period, dilution factor (if applicable), and instrument name. Store a copy o f the acquisition conditions with the chromatogram packet. 16.2 Plot the calibration curve by non-weighted linear regression and store in the study/proj ect folder. 16.3 Print the sequence and MS tune report from HP Chemstation. The sequence should be initialed and dated, and stored in the run log binder. The MS tune report should be stored in the tune report file. Copy o f the sequence and MS tune report should be placed in appropriate study/project folder. 16.4 Summarize data using suitable software and store in the study/proj ect folder. 16.5 Back up electronic data to appropriate medium (primarily CD). Record in the study/project folder the filename and location of backup electronic data. 16.6 List the documents and records generated when performing this method and where they are to be archived. ETS-8-182.0 Analysis ofF C s by Purge & TrapAutosampler/Concentrator/GC/M S Page 11 of 12 Page 73 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 17.0 17.1 A t t a c h m e n t ______________________________________________________________________ None. 18.0 References_____________________ ___________________________ 18.1 Archon Purge and Trap Autosampler System Operator's Manual, 1996, Varian. 18.2 Tekmar LSC200 Instruction Manual, 1996, Tekmar. 18.3 Agilent MSD Hardware Manual for 5973N, 1999, Agilent. 18.4 Agilent 6890 Series Gas Chromatograph, volumes 1-3,1999, Agilent 19.0 Affected Documents 19.1 None. 20.0 Revisions Revision Number. Reason For Revision Revision Date ETS-8-182.0 Analysis ofF C s by Purge & Trap Autosampler/Concentrator/GC/M S Page 12 of 12 Page 74 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory Method Indirect Photolysis Screening Test in Synthetic Humic Water M ethod Num ber: ETS-8-177.0 Approved By: Laboratory Management Team Leader Adoption Date: i 7 ^^ Revision Effective Date: Date Date copy 01 Oti9'na' ETS-8-177.0 Indirect Photolysis Screening Tests in Synthetic Humic Water Page 1 o f 16 Page 75 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application_________________________________ 1.1 Purpose. Chemicals dissolved in natural waters are subject to two types of photoreaction. In the first case, the chemical o f interest absorbs sunlight directly and is transformed to products when unstable excited states o f the molecule lead to decomposition. In the second case, reaction o f dissolved chemical is the result of chemical or electronic excitation transfer from light-absorbing species in the water. Synthetic humic water (SHW) is used for the photolytic reaction matrix because it contains dissolved organic material that absorbs sunlight and produces reactive intermediates that include singlet oxygen (10 2) that promotes indirect photolysis o f the test substance. 1.2 The method Is divided into two phases. Phase one includes the preparation o f SHW. Phase Two provides a procedure to calculate solar photolysis rate constants and half lives of test chemicals in pure water (PW) and SHW. This phase also includes parallel solar irradiation o f a radiometer to calculate kI0 (the indirect photolysis rate in the test vessel, e.g. 40 mL glass VOA vial) and k,,E(the near-surface photolysis rate constant in natural water bodies). 1.3 Compatible Analytes. Chemicals that will be subjected to this indirect photolysis screening and testing method include but are not limited to the following compounds: Compound Perfluorooctanoic acid Perfluorooctanesulfonate Perfluorooctanesulfonamide N-methylperfluorooctanesulfonamide N-ethylperfluorooctanesuIfonamide 2 -(N-methylperfluorooctane sulfonamido) ethyl alcohol 2 -(N -eth y ip erfIu o ro o ctan e sulfonamido)ethyl alcohol 1-perfluorooctene Perfluorooctanehydride Acronym PFOA PFOS FOSA N-MeFOSA N-EtFOSA N-MeFOSE-OH N -E tF O S E -O H -- 1H, Cg-hydride Compound Perfluorobutanoic acid Perfluorobutanesuifonate Perfluorobutanesulfonamide N-methylperfluorobutanesulfonamide N-ethylperfluorobutanesulfonamide 2-(N-methylperfluorobutanesulfonamido) ethyl alcohol Acronvm PFBA PFBS FBSA N-MeFBSA N-EtFBSA N-MeFBSEOH 2-{N -elhy! p erflu o ro bu tenesti! fo n am id o )e th y 1 alcohol N -E tF B S E -O H 1-perfluorobutene Perfluorobutanehydride -- 1H, C4-hydride ... and other C4 through C 10 homologues, and polymeric materials based on the above aforementioned compounds. 1.4 Acceptable matrix. Synthetic humic water (SHW), 0.005 M pH 7.0 Phosphate Buffer. ETS-8-177.0 In direct P h o to lysis Screen in g Tests in Synth etic H um ic W ater Page 2 of 16 Page 76 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 2.0 Summary of Method__________________________________________ 2.1 Phase One: A solution of standardized synthetic humic water is prepared by water extraction o f commercial humic material. The SHW is buffered at pH 7 with 0.005 M Phosphate buffer to maintain pH and pre-aged in the photoreactor to produce predictable bleaching behavior. It is then diluted at the time of use to a UV-visible absorbance typical o f most surface fresh waters (approximately 0.5 AU at 370 nm). 2.2 Phase Two: Study samples (5mL aqueous matrix) are prepared in 40 mL glass VOA vials equipped with screw-top caps with septa. Test substance is added to the vials where indicated. (See table below.) Vials are placed in the photoreactor and immersed in a water bath controlled at 25 5 C. Samples to be exposed are photolyzed in the photoreactor at 261 W/m2 (300-800 nm) for designated time intervals. A suggested set of time intervals is listed below. Additional timepoints may be added, if necessary, or as assigned by the Team Leader. Time 0 samples will be refrigerated at 1-5 C. until all timepoints have been completed. Dark controls (unexposed) will also be prepared for each timepoint. Absorbance controls will be used to monitor photo-bleaching o f the SHW. Exposed and unexposed'absorbance controls will be prepared per timepoint. 2.2.1 Samples to be prepared for each timepoint and for each exposure type: Description . . . ... ........................................ | Sample Rep 1 Sample Rep 2 Sample Rep 3 Sample Spike 1 Sample Spike 2 Matrix Blank Matrix Blank Spike Control M atrix(#l) blank Control M atrix(#l) sample Control M atrix(#l) spite Control Matrix(#2) blank C o n tro l Matrix(#2) sample Control Matrix(#2) spike Absorbance Control 1Absorbance Control dup Test Matrix (Buffer /SHW) + + + + + + + 0 0 0 0 0 0 + Control Matrix 1 (Buffer/ PW) 0 0 0 0 0 0 0 + + + 0 0 0 0 0 Control Matrix 2 (PW) 0 0 0 0 0 0 0 0 0 0 + + + 0 0 Test Substance + + + + + 0 0 0 + + 0 -1+ 0 0 Post Photolysis Target Analyte Spike 0 0 0 + r+ 0 + 0 0 + 0 0 + 0 0 LC/MS Analysis A A A A A A A A A A A A .A NA NA G C /M S Analysis UV/Vis Analysis at 370 nm A NA A NA A NA A NA A NA A NA A NA A NA A NA A NA A NA A NA A NA NA A NA A Where "+" = addition o f solution or test substance and "0" = NO addition, A= analysis performed, and NA = no analysis. Time Point 0 8hr 16 hr 32 hr 64 hr 128 hr # of Exposed Samples 0 30 30 30 30 30 # o f Unexposed Samples 30 (Time 0) 30 30 30 30 30 # of samples for LC/MS Analysis ( Exp + Unexp) 13 26 26 26 26 26 # of samples for GC/MS Analysis (Exp + Unexp) 13 26. 26 26 26 26 # o f samples for UV/Vis Analysis (Exp + Unexp) 4 8 8 8 8 8 Total # o f Samples 150 180 143 143 44 ETS-8-177.0 Indirect P hotolysis Screening Tests in Synthetic Hum ic W ater Page 3 of 16 Page 77 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.0 Quality control-pefinition/frequency/Performance criteria____________ 3.1 Blanks 3.1.1 Definitions: M atrix Blank. A sample o f analyte-free matrix (e.g. SHW/buffer) to which all reagents are added in the same volumes or proportions as used in sample processing. For photolysis studies, there are multiple matrix blanks to adequately represent the variables of the study with reference to the matrix (e.g. Exposed, Unexposed and Time 0). The matrix blank is carried through the sample preparation, photolytic and analytical procedures to monitor for contamination during any step. It is also used to establish a chromatographic baseline and monitor for interference or suppression of target analyte(s) from the matrix. Control Blank. A sample of analyte-free control matrix (such as buffer or ASTM Type II water) to which all reagents are added in the same volumes or proportions as used in sample processing. The control matrix serves as a monitor o f the effect of the matrix on the test substance, test analytes and chromatographic behavior. For photolysis studies, there are multiple control blanks to adequately represent the variables of the study with reference to the matrix (e.g. Exposed, Unexposed and Time 0 samples). The control blank is carried through the sample preparation, photolytic and analytical procedures to monitor for contamination during any step. It is also used to establish a chromatographic baseline and monitor for interference or suppression o f target analyte(s) from the control matrix. 3.1.2 Frequency/Performance Criteria: Listed in the following table: Matrix H> Matrix Blank (Buffer/SHW) Control Blank #1 (Buffer/PW ) Control Blank #2 (PW) Matrix descriDtion Freauencv 0.01 M Phosphate Buffer, pH 7: Synthetic Humic Water 0.01 M Phosphate Buffer, pH 7 : ASTM Type n Water ASTM TypeB W ater 1 Replicate p er light and dark exposure, for each time point and for each analytical methodology. Performance Criteria Any background level o f target analyte shall be less than 25% the area counts o f the LOQ. 3.2 Sample Triplicate 3.2.1 Definition: Three aliquots prepared as representatives of the same sample source (e.g. test substance) and carried through all steps of the photolytic study process and analytical procedures in an identical maimer. The results from triplicate analyses are used to evaluate variance of the total method, including sample preparation, photolytic process and analysis. ETS-8-177.0 Indirect Photolysis Screening Tests in Synthetic Humic W ater Page 4 o f 16 Page 78 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.2.2 Frequency/Performance Criteria: Listed in the following table: Matrix Descrintion Freauencv Test Matrix a nd test 3 Replicates per substance treatment type Performance Criteria The analyst shall accept %RSDs <25%. Precision values >25% must be documented andjustified (if possible). 3.3 Analytical Spike (AS) 3.3.1 Definition: A known mass o f target analyte(s) in a specified amount o f a diluted and/or aliquotted sample. This assumes that an independent estimate o f target analyte concentration is available. Analytical spikes are used to evaluate the recovery efficiency of the analyte and the matrix effect. 3.3.2 Freqnency/Performance Criteria: Listed in the following table: Matrix Description Test Matrix a nd test substance, spiked with target analyte(s) just prior to analysis Test Matrix with N O test substance, spiked with target analyte(s)just prior to analysis Control Matrix (#1) and test substance, spiked with target analyte(s) just prior to analysis Control Matrix (#2) a n d test substance, spiked with target analyte(s)just prior to analysis Frequency . 2 spiked samples per treatment type (one in lower half o f the calibration range, and one in the upper half of the calibration range) 1 Replicate per treatment type (mid-range spike concentration) Performance Criteria The analyst shall accept accuracy o f 100 25% . If accuracy is outside o f this range, document andjustify, if possible, the reason for the deviation.. 3.4 Control Sample 3.4.1 D efinition: A known matrix containing the test substance carried through the entire sample preparation, photolytic and analytical procedure. This is used to document laboratory performance by comparing recoveries and matrix effects from the different matrices and sample types. 3.4.2 Frequency/Performance Criteria: Listed in the following table: Matrix Description Control Matrix (#1) and test substance Buffer/PW Control Matrix (#2) and test substance PW Frequency Performance Criteria 1 Replicate per light and dark exposure, for each time point, and for each analytical methodology The analyst shall accept accuracy o f 100 25%. If accuracy is outside o f this range, document and justify, if possible, the reason for the deviation. Indirect Photolysis Screening Tests in Synthetic Humic Water Page 79 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.5 Absorbance Control 3.5.1 Definition: An analyte-free matrix that is carried through the sample processing procedure and analyzed by absorption spectroscopy at 370 nm. It is used to monitor the photo-bleaching rate of the SHW during the testing phase. 3.5.2 Frequency/Performance Criteria: Listed in the table below. Matrix Description Control Matrix (#1) Buffer/SHW only Frequency Performance Criteria 2 Replicates per light and dark exposure, for each time point Absorbance measured at 370 nm is between 0.01AU-0.05 AU (1cm pathlength ceil) 3.6 Internal Standard/Surrogate 3.6.1 Internal Standard Definition (applies to LC/MS samples): A known amount o f a compound similar in analytical behavior to the target analyte(s) of interest (e.g. 3, 3 ,4 ,4 , 5, 5, 6 , 6 ,7 , 7, 8 , 8 , 8 -tridecafluorooctane sulfonic acid (THPFOS) if perfluorooctane sulfonate (PFOS) were to be the target analyte), added to all samples and standards (post-irradiation), and carried through the entire analytical process. It provides a reference for evaluating and controlling the precision and bias of the applied analytical method. Samples are to be quantified using the internal standard. 3.6.2 Surrogate Definition (applies to LC/MS and GC/MS samples): A known amount of a compound similar in analytical behavior to the target analyte(s) that may be added to all samples (pre- or post-irradiation, at the discretion o f the Team Leader), and carried through the remaining sample preparation and/or analytical process. If added before exposure, it monitors the presence o f vial leaks during photolysis, as well as the performance of the purge and trap autosampler and concentrator. Surrogate analysis is used to evaluate the precision and bias of the applied analytical method. Surrogates are not used for quantitation. 3.6.3 Frequency/Performance Criteria: Listed in the following table: Matrix Description Freauencv ofUse Sample diluted with 30 mL o f internal standard compound dissolved in a suitable analytical solvent Every LC/MS sample analyzed Sample with surrogate compound spiked into it. Maybe added to every LC/MS and GC/MS sample analyzed Performance Criteria The %RSD for internal standards shall be calculated for the area response o f all appropriate samples per analytical batch. The analyst shall accept %RSD values o f <15%. %RSD values >15% shall be documented and justified, if possible. The %recovery of internal standards should be 100 25%. .Surrogates are examined for qualitative information only (i.e., area response should be relatively constant). ETS-8-177.0 Indirect Photolysis Screening Tests in Synthetic Humic W ater Page 6 o f 16 Page 80 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3.7 Other Definitions. 3.7.1 Test Substance/Test Analyte: Any substance (mixture or controlled compound) added or administered to the test system for the purpose of chemical analysis. 3.7.2 Degradation Produces): Secondary analytes of interest produced as a result o f chemical reactions during the photolysis and monitored (qualitatively or quantitatively) during the sample analysis procedure. 3.7.3 Target Analyte(s): The analyte(s) singled out in the analytical phase of the study is the target analyte. The target analyte may be identical to the test substance used in the experimental phase o f the study, a by-product or degradation product that is monitored (qualitatively or quantitatively) during the sample analysis procedure. 3.7.4 Test Matrix: The physical matrix in which the study will be conducted. 3.7.5 Relative Percent Difference (RPD): A measure o f precision defined as the absolute value o f the difference o f the two values divided by the average of the two values and multiplied by 1 0 0 . 3.7.6 Relative Standard Deviation (RSD): A measure of relative precision for three or more sample replicates; defined as the sample standard deviation divided by the sample average and multiplied by 100. This is expressed as a percent (%RSD). 3.7.7 Limit of Quantitation (LOQ): The lowest concentration that can be reliably achieved within specified limits of precision and accuracy during routine laboratory operating conditions. The LOQ can be estimated as 10 times the background level in the blank samples. However, it may be nominally chosen within these guidelines to simplify data reporting. For many analytes, the LOQ analyte concentration is selected as the lowest non-zero standard in the calibration curve that is over four times the background level in the blanks. Sample LOQs are highly matrix-dependent. 4.0 Warnings and Cautions____________________ ____________________________ 4.1 Health and Safety Warnings 4.1.1 Wear the proper lab attire for all parts of these procedures. Wear gloves and eye protection at all times. 4.1.2 Handle all solvents in a hood for all parts of the described sample preparation procedure. 4.1.3 For potential hazards of each chemical used, refer to material safety data sheets, packing materials, and 3M Environmental Laboratory's Chemical Hazard Review. 4.1.4 No mouth pipetting is allowed. 4.2 Cautions 4.2.1 The photoreactors are equipped with a continuous flow o f cooling water that poses a threat of electrocution when handling the photoreactor during irradiation sequences. ETS-8-177.0 Indirect P hotolysis Screening Tests in Synthetic Humic Water Page 7 o f 16 Page 81 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 4.2.2 Wear dark protective eyewear when operating the reactor. Do not look directly at the activated lamp. Use caution when handling samples in the reactor; the interior walls of the reactor and exposed glass vials become extremely hot. 5.0 Interference__________________________________ '_______________________ 5.1 Contaminants in solvents, reagents, glassware and other sample processing or analysis hardware may cause interference. To reduce the possibility of interference, glassware in which standards are prepared should be pre-rinsed with methanol and allowed to dry before use. The routine analysis of laboratory method blanks must be used to demonstrate that there is no interference under the conditions of the analysis. 6.0 E quipment_______________________________________________________ ___________ 6.1 Analytical balance sensitive to 0.1 mg 6.2 Photoreactor: Suntest CPS+, XLS+, or equivalent, capable of producing 250-765 Watts/m2, equipped with a xenon arclamp (e.g. 2200 W Xenon Lamp) and the appropriate filters to allow the desired wavelength (e.g. UV Special Suprax with cut-on at 290 nm, and Quartz dish with DR.reflective coating), and a flowing water bath circulating pump or equivalent. 6.3 Water cooler/recirculator capable o f maintaining temperature at 25 C 5 C . 6.4 UV-Visible Spectrophotometer (UV-VIS), equipped with tungsten and deuterium lamps, model 8453, or equivalent 6.4.1 Autosampler: Model G1120A, or 1-cm pathlength cell holder: Model 08451 60104, or equivalent. 6.4.1.1 1-cm pathlength quartz spectrophotometer cell, or equivalent. 6.4.2 Long Path-Length Cell Holder, Hewlett Packard part number 89076C, or equivalent 6.4.2.1 10-cm path length quartz cell equipped with stopcocks, Hewlett Packard Part # 5061-3392, or equivalent. 6.4.3 Data acquisition and analysis software, HP ChemStation for UV-Visible Spectroscopy, G1116AA Rev. B.01.02, or later. 6.5 Data System: A PC capable of controlling the UV-Visible Spectrophotometer system. 6 .6 Centrifuge capable o f maintaining >2000 rpm for 10 minutes at ambient temperature (22 26 C). 6.7 Radiometer, capable o f detecting and recording irradiation output of the photoreactor for the duration of the study. 6 .8 Lab Oven, capable o f maintaining 70-80 C. 7.0 Supplies and Materials___________________________________________________ 7.1 40-mL amber and clear glass vials (VOA) with screw caps. 7.2 Crimp cap autovials: 1.5-mL, caps, crimper, and decapper. 7.3 Adhesive-backed labels (return address size) for labeling quartz vials and autovials. 7.4 Disposable glass graduated pipettes, 1 mL to 10 mL. ETS-8-177.0 In direct P hotolysis Screening Tests in Synthetic H um ic W ater Page 8 o f 16 Page 82 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 7.5 Disposable glass Pasteur pipettes and rubber bulbs. 7.6 Glass beakers, various sizes. 7.7 Volumetric flasks, from 10 mL to 1000 mL. 7.8 Hamilton Gastight syringes (precision 1% of the total volume), 5 pL to 1000 pL. 7.9 10-mL Bottle-top dispenser. 7.10 Teflon filters filter holder apparatus: 0.4 pm pore-size and 0.2 pm pore-size filters (47 mm diameter, GelmanTM or equivalent. 7.11 500-mL glass screw-top containers. 8.0 Reagents and Standards_____________________________________ 8.1 Water/Pure water (PW), ASTM Type II water at a minimum 8.2 Methanol (MeOH), HPLC/SPEC/GC grade from EM Science or equivalent. 8.3 Acetone, HPLC/SPEC/GC grade from EM Science or equivalent. 8.4 Acetonitrile, HPLC/SPEC/GC grade from EM Science or equivalent. 8.5. Humic acid, sodium salt, from Aldrich or equivalent. 8 .6 NaOH, reagent grade from EM ScienceTM or equivalent. 8.7 0.1 % NaOH solution Example: Weigh approximately 1.0 g sodium hydroxide into a weigh boat and transfer quantitatively to a 1 L volumetric flask and dilute to the mark with PW or equivalent. 8.8 Sulfuric Acid (H2S 0 4), reagent grade from Fisher or equivalent. 8.9 Potassium phosphate, reagent grade from JT Baker or equivalent. 8.9.1 0.005 M, pH 7.0 Phosphate Buffer. Example: Weigh 1.36 g KH2P 0 4 into a weigh boat and transfer to a 1 L volumetric flask using PW and dilute to the mark. Transfer the 1 L of solution to a 2 L volumetric flask. Add 600 mL of 0.1% NaOH, adjust the pH to 7.0 0.1 with 0.1% NaOH or dilute H2S 0 4, and dilute to the mark with PW. 8.10 Method Blank Solutions: Method Blank Tvnes Matrix ID Test Matrix B uffer/SH W Control Matrix (#1) B uffer/P W Control Matrix (#2) P W Matrix description Example: 1:10 Solution: Dilute 50 mL Synthetic Humic Water with 0.01 M pH 7.0 Phosphate Buffer solution to 500 mL. Example: 1:10 Solution: Dilute 50 mL Pure Water (ASTM Type H) with 0.01 M pH 7.0 Phosphate Buffer solution to 500 mL. Pure Water (ASTM Type II) 8.11 Stock Solutions. Stock solutions for internal standards and spiking solutions are prepared in MeOH at concentrations of approximately 10,000 pg/mL by weighing approximately 0.1 g of the appropriate substance into a 10-mL volumetric flask and bringing to the mark with MeOH. Dilute to make appropriate working solutions. 8.11.1 Diluting Solution with Internal Standard: The diluting solution shall contain internal standard at an area response level equivalent to approximately half the ETS-8-177.0 Indirect P h otolysis Screening Tests in Synthetic Humic W ater Page 9 o f 16 Page 83 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 area response of the test substance's high standard in the calibration curve. Example: Internal standard solution in MeOH is prepared by diluting 50pL of internal standard stock solution (Section 8.11) to 1 L with MeOH to a nominal concentration of 0.5 AU pg/mL. 8.12 Test Solutions 8.12.1 Test Substance: Prepare a solution of the test substance in acetonitrile. Calculate the concentration so that after the test substance is added to the test vial, no more than 1% of the volume in the test vial will be solvent, (e.g. SOpL added to 5 mL o f matrix = 1% v/v) Then measure the absorbance of the test substance solution diluted with buffer/water matrix to the desired concentration. The maximum absorbance at any wavelength greater than 290nm must be < 0.05, when measured in a standard 1-cm pathlength cell. Example: A 900 pg/mL solution o f test substance in acetonitrile is prepared by weighing 90 mg o f test substance into a 100 mL volumetric flask and diluting to the mark with acetonitrile. 9.0 Sample Handling__________ _________________________________________ 9.1 Record times o f initial preparation and dilution on the fluorochemical degradation (photolysis) analysis sample prep sheet (Attachment A). 9.2 Once the test substance solution has been added, the 40 mL VOA sample vials shall be stored and handled cap-side down to minimize loss of any potential volatile analytes. After the exposure period, the LC/MS samples may be turned upright and stored in a cooler at 1-5 C. After the exposure period, GC/MS samples shall be maintained in an inverted position in a cooler at 1-5 C until they are loaded onto the autosampler. 9.3 Once the 30-mL aliquot o f diluting solvent has been added to the LC/MS photolysis samples, (see Section 12.0), the samples should be analyzed as soon as possible. Alternatively, the samples may be stored at 1-5 C. Day 0 samples are to be stored at 1-5 C during the time of sample exposure, and then diluted along with the exposed and unexposed samples just prior to analysis. 10.0 Quality Control ___________ ____________________________________ 10.1 Quality control parameters (and the frequency of use) are included in Section 3.0. 11.0 Calibration and Standardization_____________________________________ 11.1 The analytes of interest must be standardized according to laboratory specifications. 11.2 All equipment used, such as the analytical balance, photoreactors, etc. should be calibrated prior to use (daily, weekly, etc.) as specified in its standard operating procedure. 11.3 All samples analyzed will be run against a standard curve containing varying amounts of test substance, and a fixed amount of internal standard or surrogate compound. ETS-8-177.0 Indirect P hotolysis Screening Tests in Synthetic Humic Water Page 10 of 16 Page 84 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.0 P r o c e d u r e s_________________________________________________________________________ 12.1 Phase One-Preparation and standardization of synthetic humic water. 12.1.1 Weigh approximately 2.5 g humic acid into a tared 250 mL centrifuge tube. 12.1.2 Add 0.1% NaOH solution to 250 mL. 12.1.3 Screw-cap shut and tape the tube and place horizontally on an orbital shaker. Shake vigorously (e.g. 100-250 rpm) at room temperature for approximately one hour. 12.1.4 Centrifuge the 250 mL of solution at approximately 2000 rpm for 10 minutes or until solution has cleared, and then filter the supernatant through a 0.4pm filter into a clean 500-mL glass screw-top container. 12.1.5 Adjust the pH of the solution to 7.0 with dilute H2S 0 4 or 0.1 % NaOH. 12.1.6 Filter-sterilize the solution through a 0.2pm diameter pore-size filter into a clean 500-mL glass screw-top container. 12.1.7 Seal the container and place cap-side down in the photoreactor chamber. 12.1.8 Expose the SHW 24 hours at 261 W/m2 to pre-age the solution (equivalent to three day's worth o f Miami, Florida sunlight). The EPA's definition o f " 1 Day" o f irradiation is "eight hours." The irradiation intensity o f 261 W/m2 was chosen because it yields the equivalent average optimum natural daylight radiation for 300-400 nm (see the table below): Aunroximate Intearated and Individual Irradiances in W/mz Irradiance Source 250-300 nm 300-400 nm 400-800 nm 340 nm 420 nm Average Optimum Natural Daylight1 0.0 27.8 259.0 0.30 0.67 Atlas Photoreactor with integrated irradiance output o f 261 W/m2300-800 nm using 0.08 27.8 the ER.Reflecting and 290 cut- on filters `M easured --25 "N, M iam i, Florida (See Reference T 8.4) 234.36 0.24 0.71 12.1.9 Aliquot the SHW into a 1 -cm quartz UV-VIS cuvette and analyze the absorbance at 370 nm. 12.1.10 Check the pH o f the solution using pH paper or a pH probe. Adjust the pH if necessary to 7.0 + 0.1 using a dilute H2S 0 4 solution or 0.1% NaOH solution. 12.1.11 Calculate the dilution factor necessary to decrease the absorbance to approximately 0.5 AU (in a 1-cm pathlength cell) in 1 L of SHW: where: 0.5 --A i.m 1L x . AX37Q= th e measured absorbance of the SHW at 370 nm x = the volume of SHW needed to dilute to 1 L with water. 12.1.12 Bring the solution to the exact dilution calculated in 12.1.11 withPW. ETS-8-177.0 Indirect Photolysis Screening Tests in Synthetic Humic W ater Page 11 of 16 Page 85 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.1.13 Verify that the absorbance is approximately 0.5 AU by aliquoting the diluted SHW into a 1-cm UV-VIS cuvette and taking the absorbance reading at 370 nm. 12.1.14 Transfer the SHW stock solution into an amber, or clear foil-wrapped 1 L glass storage bottle, tightly cap and refrigerate. 12.2 Phase Two 12.2.1 Fill out die "Fluorochemical Degradation (Photolysis) Sample Prep Sheet" (Attachment A) as much as possible, assigning sequential unique ID numbers to each sample to be prepared. 12.2.2 Obtain the appropriate number of clear and amber 40-mL glass vials with caps and cardboard boxes. Label the vial caps using a black permanent marker to distinctly identify samples. 12.2.3 Create labels for each sample to be affixed to the 40-mL vials and the autosampler vials after photolysis is completed. The labels should include the sample number, test substance, matrix, exposure type (e.g. exposed/unexposed/Time 0), date and initials of the analyst. 12.2.4 Aliquot 5.0 mL o f Buffer/SHW, Buffer/PW and PW solutions into clear (for exposed samples) and amber (for unexposed and Time 0 samples) 40-mL glas VOA vials. Add test substance to the appropriate vials. See the table below for list of vials, replicates, and sample types. If pre-hydrolysis surrogates are to be used, add them also at this point. 12.2.5 Create one set o f samples (listed below) per time point and for each analytical methodology. (LC/MS and GC/MS): Description Sample Rep 1 Sample Rep 2 Sample Rep 3 Sample Spike 1 Sample Spike 2 Matrix Blank Matrix Blank Spike C ontrol M atrixC #!) b lank Control Matrixftfl) sample Control M atrix(#l) spike Control Matrix(#2) blank Control Matrix(#2) sample Control Matrix(#2) spike Test Matrix (Buffer/SHW) + + + + + + 0 0 0 0 0 0 Control Matrix 1 (Buffer/PW) 0 0 0 0 0 0 0 f + + 0 0 0 Control Matrix 2 (PW) 0 0 0 0 0 0 0 0 0 0 + + -+ Test Substance + + + + + 0 0 0 + + 0 + + Sample Rep 1 Sample Rep 2 Sample Rep 3 Sample Spike 1 Sample Spike 2 Test Matrix Blank Test Matrix Blank Spike Control M atrix(#I) blank Control M atrix(#l) sample Control M atrix(#l) spike Control Matrix(#2) blank Control Matrix(32) sample Control Matrix(32) spike + + + + + + + 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 + + + Where "+" = addition o f solution or test substance and "0" = NO addition + + + + + 0 0 0 + + 0 + ETS-8-177.0 Indirect P hotolysis Screening Tests in Synthetic Humic W ater Vial Type/Exposure Clear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed C lear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed Clear/Exposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amber/Unexposed Amb er/Unexposed Amber/Unexposed Amber/Unexposed Page 12 of 16 Page 86 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Control M atrix(#l)- SHW/Buffer 1 :9 v/v Control Matrix(#2)= PW/Buffer 1:9 v/v Create one set o f samples (listed below) for each time point for UV/VIS analysis: Description Absorbance Control - Light Absorbance Control - Dark Test Matrix (Buffer/SHW) + Control Matrix t (Buffer/PW) 0 0 Control Matrix 2 (PW) 0 0 Test Substance 0 0 Vial Type/Exposure Clear/Exposed Amber/Unexposed 12.2.6 Store the "Time 0" vials in a labeled box at 1-5 C. 12.2.7 Place all the vials that will go into the photoreactor into an oven set to 70-80 C. for 5-10 minutes to acclimate the vials, liquid and headspace to photoreactor conditions. Upon removing the vials from the oven, immediately re-tighten the caps and proceed to load the reactor. 12.2.8 Place the amber "unexposed" vials in plastic bags and arrange on the bottom of the photolysis tray (make sure that they don't float once the tray is filled with water). The "unexposed" vials will remain submerged in the cooling water (25 5 C) during the exposure. (The vials are exposure- and temperature-controlled.) 12.2.9 Place the clear "exposed" vials in the rack in the photoreactor tray cap-side down, and install the rack so that the VOA vial caps will be submerged throughout the duration of exposure. 12.2.10 Prepare the radiometer to read intensity of irradiance over the duration of the exposure. See SOP ETS-9-50.0 for operation o f radiometer. 12.2.11 Expose the samples for the designated time intervals at 261 W/m2. See SOP ETS-9-44.0 for operation of the photoreactor. 12.2.12 Following each exposure interval, remove vials from the photoreactor and store inverted in a cooler at 1-5 C. After all exposures have been completed, remove all sample vials as well as the "Time 0" vials from the cooler and analyze as a single batch for each instrument. 12.2.13 UV-VIS absorbance control analysis 12.2.13.1 Analyze the pH 7.0 SHW/buffer absorbance controls by UV/Visible absorbance spectroscopy at 370 nm by aliquoting the test solution directly into a 1 -cm or greater pathlength quartz cuvette and obtaining the spectra. See SOP ETS-9-46.0 for operation of the UV/VIS instrument. The resultant peak at 370 nm will be analyzed to determine the change in absorbance between the Time 0, exposed and unexposed samples. 12.2.14 LC/MS sample analysis 12.2.15 Dilute the exposed and unexposed samples for all timepoints with 30 mL internal standard solution in methanol (Section 8.11.1). Add spiking solution to the appropriate vials. Invert each vial several times to mix. 12.2.16 Transfer aliquots of LC/MS samples into autovials and then place them in the autosampler for analysis of the parent compound and possible degradation products. Analyze according to ETS-8-181.0. 12.2.17 GC/MS sample analysis ETS-8-177.0 page j3 0f 16 Indirect Photolysis Screening Tests in Synthetic Humic Water Page 87 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 12.2.17.1 Analyze the exposed and unexposed samples for all timepoints as-is by purge and trap GC/MS. Add spiking and surrogate solutions, as required, to the appropriate vials. Analyze according to ETS-8-182.0. 12.2.17.2 Important: Maintain vials in the inverted position until they can be placed in the autosampler. 13.0 Data Analysis and Calculations_____________________________________ 13.1 Not applicable, as this is a sample preparation and analysis method. Consult the appropriate analytical protocol for guidance regarding data analysis and calculations. 14.0 Method Performance_______________________________________________ 14.1 Not applicable. 15.0 Pollution Prevention and Waste Management_________ ___________ ___ 15.1 Dispose of sample waste by placing in high or low BTU (British Thermal Unit) waste containers as appropriate. Use broken glass containers to dispose of glass pipettes. 16.0 Records_________________________________________________ ._________ 16.1 Print out hard copies o f all graphics and data analysis summaries for archiving. 16.2 Sign and date all graphics and label with instrument ID. 16.3 Fill out the sample preparation worksheets) documents completely, making sure to include all initials and dates. 16.4 Archive electronic data to compact disc media. 17.0 Attachments______________________________________________________ 17.1 "Fluorochemical Degradation (Photolysis) Sample Prep Sheet" 18.0 References___________________________________________________________ 18.1 Interpersonal conversation with Carrie O'Connor, Optical Systems Engineer, Atlas Electric Devices. 18.2 "Suntest CPS/CPS+ Spectral Irradiance Distribution," table distributed by Atlas Electric Devices Company, sent via fax by Richard Sherwin, Sales Representative, 26 July, 2000. 18.3 "Atlas Xenon Filter Combination and Sunlight Measurements," information generated by Atlas Electric Devices Company sent via fax by Richard Sherwin, Sales Representative, 26 July, 2000. 18.4 OPPTS 835.5270, Indirect Photolysis Screening Test: Sunlight photolysis in water containing dissolved humic substances. ETS-8-177.0 Indirect P hotolysis Screening Tests in Synthetic Humic Water Page 14 o f 16 Page 88 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 19.0 Affected Documents 19.1 None 20.0 Revisions____________________________________ ._________________________ ___ Revision Number. Reason For Revision . Revision -D--a-t-e- ETS-8-177.0 In direct Photolysis Screening Tests in Synthetic Humic W ater Page 15 of 16 Page 89 of 148 3M Environmental Laboratory Report No. EOO-2192 Attachment A- Photolysis Sample Prep Sheet TEST A N M .Y TE : Lab Raquast Num bar: N o m in a i T im a In ta rva l: R eactor Tam p aratura: FluoracHmical Dagra dation (Photolysis) Sampia Prap Shaat P holoreailoriO : Start Osto: Stop Date: Total Reactor Time: hours ^ Gier Storage ID :_ D ate Aliene ln Date Tlme Out Initials Cooler Storage ID:_ Date &Tim ln^ D ate .Time Out Initials BACK TO MAIN N O TE D ark shaded areas require NO additions NA n o t applicable C o m m e n t e : ______ ETS-8-177.0 In d irect P h o to ly sis S creen in g Tests in S yn th etic H u m ic W ater Page 16 o f 16 Page 90 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory E q uipm en t P rocedure O pera tio n and M aintenance o f th e H ew lett P ackard 8453 U V -V isible Spec tr o ph o to m eter Procedure Num ber: ETS-9-46.0 Exact Copy of Original Approved by: Laboratory Management Adoption Date: I 0 / 7.0J 0 D Revision Effective Date: f Date ETS-9-46.0 Operation and Maintenance o f the HP8453 UV-Vis Spectrophotometer Page 1 of 9 Page 91 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application____________________________________________ 1.1 This equipment procedure describes the operation, cleaning, and maintenance of the Hewlett-Packard 8453 UV-Visible Spectroscopy System. 2.0 Definitions________________________ ________________________________ 2.1 Absorbance: Measure of concentration of material present: expressed as product of molar extinction coefficient (s), pathlength (b), and concentration (c), written as A = s b c (also known as Beer's Law). 2.2 Cuvette or flow cell: Transparent receptacle in which sample solutions are introduced into the light path o f spectrometers. Usually, two sides are equal (e.g. 1 cm x 1 cm) while the third dimension is elongated, possibly as long as 15 cm. For UV work, the material is quartz. Visible work permits the use of glass or plastic cuvettes. 2.3 Pathlength: The distance the light passes through the sample in its holder. In practical terms, the inside dimension of the cuvette (usually 1 cm). 2.4 Slit width; Size o f opening through which light from cuvette emerges. Choice o f slit width depends on wavelength range, separation ability of wavelength selector, and desired isolation of specific wavelength. Slit width is often fixed or automatically programmed. 2.5 Solvent Cutoff: The wavelength at which the solvent absorbs a significant portion of the light, causing a loss of signal. In other words, the solvent becomes opaque to the wavelengths being used. This is common in the ultraviolet, rare in the visible. 2.6 Transm ittance: Ratio of the radiant power transmitted by a sample to the radiant power transmitted by a blank in an equivalent cell or by some other means of compensation for solvent absorption, reflection losses, etc. 2.7 Visible: The portion o f the electromagnetic spectrum, from 400 to 800 nm, detectable by human eyes. 2.8 Ultra-violet (UV): The portion of the invisisble electromagnetic spectrum composed of wavelengths of 10-400 nm. In UV spectrometry we are primarily interested in the nearUV (quartz) region extending from 200 to 380 nm. 2.9 UV Spectrum: a plot o f wavelength (or frequency) o f absorption versus the absorption intensity (absorbance or transmittance). 3.0 Description______________________________ . ' . ... ......... X I The HP 8453 spectrophotometer is a single-beam, microprocessor-controlled, UV-visible spectrophotometer with collimating optics. The ChemStation for UV-Visible spectroscopy software running on a PC with Microsoft NT operating system provides instrument control, data acquisition, and data evaluation. 4.0 I d e n t if ic a t io n ___________ _______________________________________ 4.1 Hewlett Packard G1103A Serial No. CN93500458 4.2 Hewlett Packard 89090A Serial No. DE14300757 5.0 Warnings and Ca u t i o n s ________________________________________ 5.1 Health and Safety Warnings: ETS-9-46.0 Operation and Maintenance o f the HP8453 UV-Vis Spectrophotometer Page 2 of 9 Page 92 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 5.1.1 Eye damage may result from directly viewing light produced by deuterium lamps used in detectors and spectrophotometers. Always turn off the deuterium lamp before opening the lamp door on the instrument. 5.1.2 Some adjustments described in the service manual are made with power supplied to the instrument, and protective covers removed. Electricity and heat available at many points may, if contacted, result in personal injury. 5.1.3 Capacitors inside the instrument may still be charged, even though the instrument has been disconnected from its source of supply. Dangerous voltages, capable of causing serious personal injury, are present in this instrument. Use extreme caution when handling, testing, and adjusting. 5.2 Cautions: 5.2.1 Never touch the quartz envelope of the deuterium lamp with your fingers. Fingerprints absorb UV light and may be burnt in, thus reducing lifetime of the lamp. 5.2.2 Quartz sample cells or sample cells with quartz faceplates are required if you want to use the full 190 to 1100 nm wavelength range o f the spectrophotometer. Good quality glass cells may be used when working above 350 nm. Disposable plastic sample cells are not recommended for use. 5.2.3 For high precision measurements, wait until the spectrophotometer and the lamps have reached thermal equilibrium. The time required is a function of environmental conditions but the instrument should be ready after 45 minutes. To determine if the spectrophotometer is in stable working condition, the HP 8453 Self- test may be performed. (See section 13.1) 5.2.4 Ensure cell windows are free of fingerprints and other contamination. 5.2.5 Avoid the use of alkaline solutions (pH > 9.5) which can attack quartz and thus impair the optical properties of the flow cells. . 5.2.6 Solution in cell should be free of floating particles. 5.2.7 Solution in cell and cell walls should be free of bubbles. 5.2.8 Ensure that solution in cell is homogeneous by thoroughly mixing before measurement. 5.2.9 Blank is measured on the same solvent as sample. 5.2.10 Blank measurement should show a flat baseline. 5.2.11 Cell orientation of blank and sample measurements should be the same. 5.2.12 Ideally, the cell is not removed between sample measurements, which means the cell is filled/rinsed using a pipette or a flow cell is used. 5.2.13 Time between blank and sample measurements should be short. 6.0 Sp e c ia l In st r u c t io n s___________________________________ ___________________________ 6.1 None. 7.0 R e sp o n sib il it y ______________ _______________________________________________________ 7.1 The operator is responsible for routine maintenance and cleaning. ETS-9-46.0 Operation and Maintenance of the HP8453 UV-Vis Spectrophotometer Page 3 of 9 Page 93 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 7.2 The person responsible for the equipment (and an alternate) will be identified in the front of the equipment logbook, and are responsible for all routine and non-routine maintenance and associated documentation. 8.0 S u p p l ie s a n d M a t e r ia l s_____________________________________________________________ 8.1 Pozidriv screwdriver 8.2 Isopropanol, reagent grade 8.3 Canister of compressed oil-free air. 8.4 Surgical cotton swabs, lint-free. 8.5 Deuterium lamp assembly, Agilent 8453 (Part No. 2140-0605). 8.6 Lamp, Tungsten G1315A, Agilent 8453A (Part No. G1103-60001). 9.0 In st r u m e n t C l e a n in g P r o c e d u r es_________________________________________________ 9.1 Cleaning the Stray Light Filter. (Recommended at one-yearly intervals or more frequently when operating the spectrophotometer in a particularly dirty environment.) 9.1.1 Turn off the instrument and disconnect the power cord. Take the plastic and sheet metal rear covers off, see "Removing and Replacing Covers" on page 109 of the HP 8453 Service Manual. 9.1.2 Remove any accessory board or MIO board that may be plugged in from the rear side of the instrument. 9.1.3 Remove the upper rear foam block. 9.1.4 Disconnect the shutter cable from the SPM board. Open the screw that fixes the shutter assembly to the optical unit and remove the shutter assembly. 9.1.5 Dampen a lint-free, surgical cotton swab with reagent grade isopropanol and gently swab the surface of the stray light filter. Repeat several times with clean swabs and alcohol each time. 9.1.6 Use a canister of compressed oil-free air to further clean the stray light filter. 9.1.7 Position the shutter assembly above the source lens and fix the screw that holds it at the optical unit, see Figure 39 on page 124 o f the Service manual. Connect the shutter cable to the SPM board. 9.1.8 Replace the upper rear and upper front foam blocks. 9.1.9 If available, replace any accessory board or MIO board (plugged in from the rear side of the instrument). 9.1.10 Replace the plastic and sheet metal rear covers. Push the plastic rear cover down so that it locates on both sides, see "Removing and Replacing Covers" on page 109 o f the HP 8453 Service Manual. 9.1.11 Reconnect the line power and turn on the instrument. Check that the spectrophotometer passes the self-test, this means that the green light on the front panel comes on and that you can do a blank measurement from the software. 9.2 Cleaning the Source Lens from the Sample Compartment Side. (Recommended at oneyearly intervals or more frequently when operating the spectrophotometer in a particularly dirty environment.) 9.2.1 Turn off the instrument and disconnect the power cord. 9.2.2 Remove any cuvette holder from the sample compartment. ETS-9-46.0 Operation and Maintenance of the HP8453 UV-Vis Spectrophotometer Page 4 of 9 Page 94 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 9.2.3 To have better access you may want to take the plastic and sheet metal rear covers off, see "Removing and Replacing Covers" on page 109 o f the HP 8453 Service Manual. 9.2.4 Dampen a lint-free, surgical cotton swab with reagent grade isopropanol and gently swab the surface of the source lens. Repeat several times with clean swabs and alcohol each time. 9.2.5 Use a canister of compressed oil-free air to further clean the source lens. 9.2.6 If you have taken the covers off, replace them. 9.2.7 Replace the cuvette holder: Reconnect line power and turn on the instrument. Check that the spectrophotometer passes the self-test, this means that the green light on the front panel comes on and that you can do a blank measurement from the software. 9.3 Cleaning the Spectrograph Lens. (Recommended at one-yearly intervals or more frequently when operating the spectrophotometer in a particularly dirty environment.) 9.3.1 Turn off the instrument and disconnect the power cord. 9.3.2 Remove any cuvette holder from the sample compartment. 9.3.3 To have better access you may want to take the plastic and sheet metal rear covers off, see "Removing and Replacing Covers" on page 109 of the HP 8453 Service Manual. 9.3.4 Dampen a lint-free, surgical cotton swab with reagent grade isopropanol and gently swab the surface of the spectrograph lens. Repeat several times with clean swabs and alcohol each time. 9.3.5 Use a canister o f compressed oil-free air to further clean the spectrograph lens. 9.3.6 If you have taken the covers off, replace them. Replace the cell holder in the sample compartment. 9.3.7 Reconnect line power and turn on the instrument. Check that the spectrophotometer passes the self-test, this means that the green light on the front panel comes on and that you can do a blank measurement from the software. 10.0 M a in t e n a n c e P r o c e d u r es_________________________________________________________ 10.1 Routine maintenance. 10.1.1 Cleaning the stray light filter. Indicators for a dirty stray light filter include: 10.1.1.1 After exchanging the lamps, the intensity test executed by the ChemStation software still falls below the specified level. 10.1.1.2 One of the stray light tests fails. 10.1.1.3 The photometric accuracy test fails. 10.1.2 Cleaning the lenses that are accessible from the sample compartment side. An indication for dirty lenses is when, after exchanging the lamps, the intensity test executed by the ChemStation software still falls below the specified level. 10.2 Non-routine: Document any non-routine maintenance. 10.2.1 Exchange the deuterium or the tungsten lamp when the intensity test, which is executed through the ChemStation software, falls below the specified level or when one of the lamps no longer ignites. See HP 8453 Service Manual for lamp replacement procedure (p.96). ETS-9-46.0 Operation and Maintenance of the HP8453 UV-Vis Spectrophotometer Page 5 of 9 Page 95 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 11.0 11.1 11.2 11.3 11.4 11.5 O pe r a t in g P r o c ed u r es_____________________________________________________________ Powering Up the HP 8453 UV-Visible Spectrophotometer and PC controller. 11.1.1 Switch on the PC and boot the PC operating system. 11.1.2 Switch on the spectrophotometer and wait until the spectrophotometer's indicator light turns green. This process includes the spectrophotometer's self test and takes about one minute. 11.1.3 Launch a measurement session by pressing the operating system's "Start" button and select "Programs", "HP UV-Visible ChemStations", "spectrometer 1 online" 11.1.4 The system is ready to use if the blue "busy " status display on the system's bottom message line turns off. Note: For high precision measurements wait until the spectrophotometer and the lamps have reached thermal equilibrium. The time required is a function of environmental conditions but should be ready after 45 minutes. 11.1.5 The first measurement to perform is a reference measurement. After this alignment you are ready to measure absorbance data and spectra Inserting a Cell. 11.2.1 The HP 8453 is shipped with the standard single-cell holder which accommodates standard cells or flow cells. 11.2.2 Move the locking lever to its up position. 11.2.3 Insert the sample cell, making sure you orient it correctly. The frosted (non-clear) sides of the sample cell should not be in the path o f the light beam. 11.2.4 Lock the sample cell in place by pushing the locking lever back down. 11.2.5 Small volume flow cells and particularly any cells with less than a 2 mm aperture may require use of the optional adjustable cell holder. This device helps you ensure the cells are properly centered in the light path. Entering a Cell's Path Length. 11.3.1 Click "Setup" on the Instrument Panel. 11.3.2 Type the path length in cm in the "Setup Manual" dialog box. 11.3.3 Click "OK" to set the specified path length. Starting a Measurement Session. 11.4.1 Start a measurement session by selecting Instrument 1 online from the menu. 11.4.2 Perform a reference measurement. Typically the cell containing the solvent used with your samples is put in the measurement position and a blank measurement performed. To start this measurement, click the "Blank" button on the Instrument Panel or press the spectrophotometer's "Blank" button. 11.4.3 Perform a sample measurement. To get the most precise results, use the same cell in the same orientation to the measurement beam. Flush the cell about three times with the sample solution and start the measurement by clicking the Instrument Panel's "Sample" button or by pressing the spectrophotometer's "Sample" button. Setting up a Method for Single Component Analysis. 11.5.1 Choose the "Quantification" task from the data analysis panel. 11.5.2 Enter the used wavelength in the input fields. ETS-9-46.0 Operation and Maintenance o f the HP8453 UV-Vis Spectrophotometer Page 6 of 9 Page 96 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 11.5.3 If background correction is desired, select "Single Reference Wavelength", "Subtract Average Over a Range" or "Three-point Drop Line" from the background correction combo box. 11.5.3.1 "Single Reference Wavelength" requires the input of one wavelength in the adjacent wavelength edit field on the right side of the combo box. 11.5.3.2 For "Subtract Average Over a Range" or "Three-point Drop Line" you must define the range/baseline by entering the start and end wavelengths in the two adjacent wavelength edit field on the right side o f the combo box. 11.5.4 Specify a name for the analyte. 11.5.5 Choose "Concentration" to enter the analyte concentration directly or "Weight & Volume" to have the ChemStation calculate the concentration. Enter the units for the concentration or the weight and volume. 11.5.6 If you want to be prompted for the concentration of the standards during measurement, select "Prompt for Standard Information". In the combo box you can select whether the prompt asks you for the concentration, or calculates the concentration based on volume, weight and purity. 11.5.7 If you have diluted samples and want to correct for dilution, select "Prompt for Dilution Factor" of sample. 11.5.8 Choose the desired calibration curve type. 11.5.9 Select the desired data type and display range of the spectra in the graphical window. 11.5.10 Choose "OK" to close the dialog box. 11.5.11 Perform the following steps if you want to calibrate the method. 11.5.11.1 Measure a blank on the solvent if necessary using fire "Blank" button in the instrument panel. 11.5.11.2 Measure the standards using the "Standard" button in the instrument panel. If you have selected one of the prompts, the appropriate values will be requested in a dialog box. The spectrum is displayed automatically in the "Standard Spectra" window as they are measured. Note: There is no fixed limit to the number of calibration standards that can be incorporated into a calibration. However, each of the calibration curve types requires a m in im u m number of standards of different concentrations, which can be found in Table 7, page 45 of the HP Manual: Understanding Your UVVisible Spectroscopy System. The ChemStation software calibrates automatically when at least the minimum number of standards has been measured. A table with the used standards and values as well as calibration curve is displayed. 11.5.11.3 If the calibration is successful, the calibration curve icon of the data analysis panel changes from red to green. 11.6 Measuring and Displaying an Absorbance Spectrum 11.6.1 Select the "Clear" icon using the Toolbar, to delete any spectral data that you do not wish to keep. 11.6.2 Select the "Spectrum/Peaks" task from the Method menu or data analysis panel. 11.6.3 Select the boxes for "Peak/Valley Find" if necessary. ETS-9-46.0 Operation and Maintenance of the HP8453 UV-Vis Spectrophotometer Page 7 of 9 Page 97 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 11.6.4 Select "Absorbance" as data type. 11.6.5 Set the display range you want to see in the graphical window. 11.6.6 Choose "OK" to close the "Spectrum/Peaks Parameters" dialog box. 11.6.7 Measure a Blank on the solvent if necessary using the "Blank" button in the instrument panel. 11.6.8 Measure all the samples using the "Sample" button in the instrument panel. The spectra are displayed automatically in the "Sample Spectra" window as they are measured and depending on the selected parameters you will get a table o f results. 11.7 Loading Spectra: 11.7.1 Choose "Load" from the `Tile" menu, then choose the type of spectra that you want to load (Samples or Standards) from the submenu to display the "Load Spectra" dialog box. 11.7.2 If the spectra you wish to load are not in the "File Name" list, select a different directory from the "Directories" list. 11.7.3 Select the spectra you wish to load from the File Name list and choose OK to close the dialog box. 11.8 Saving Spectra: 11.8.1 Choose "Save" from die "File" menu, then choose the type of spectra that you want to save (Samples, Standards or Selected Spectra) from the submenu to display the "Save Spectra As" dialog box. 11.8.2 If you wish to save the spectra in a directory other than the current one, select the new directory from the "Directories" list. 11.8.3 Type the name you wish to save the spectra as in the "File Name" field and choose "OK" to close the dialog box. 11.8.3.1 A valid file name consist o f eight alphanumeric characters and the file extension .sd or .std. Usually, the extension .std is used for standards only. 11.8.4 You can also save spectra using the ToolBar. 12.0 R eco rds_________________________________________________________________ 12.1 Document all cleaning and maintenance performed on the instrument in the maintenance or run/maintenance logbook. Include a description of the procedure(s) performed, name o f person who performed procedure(s), any unusual observations, parts replaced or needing replacement, and whether the procedure was routine or non-routine. Logbooks are archived when complete. 13.0 Testing, C alibration and/or Standardization P rocedures 13.1 HP 8453 Self-Test: 13.1.1 Make sure that you are in the "Verification and Diagnostics" mode. The mode is indicated on the tool bar of the HP ChemStation session. 13.1.2 Select the "Self-Test" task in the analysis panel's selection box. 13.1.3 Choose "Self-Test". Start from the "Task" menu or click "Start" to start the self test. 13.1.4 The self-test results will be displayed in a window with pass/fail criteria. 13.2 Calibrating for a single component analysis. ETS-9-46.0 Operation and Maintenance of the HP8453 UV-Vis Spectrophotometer Page 8 of 9 Page 98 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 13.2.1 Calibration for single component analysis is based on the measurement of standard samples with known concentrations. During the calibration process, the software calculates the calibration coefficients, which are then used for the quantification o f unknown samples. 13.2.2 The status for calibration can be seen in the data analysis panel: 13.2.2.1 Uncalibrated: RED dashed calibration curve icon. 13.2.2.2 Calibrated: GREEN continuous calibration curve icon. 13.2.3 To calibrate for single component analysis: 13.2.3.1 Load or set up a method for the "Quantification" task. 13.2.3.2 If the "Standard" spectra window is not displayed, choose "Show" standards in the data analysis panel. 13.2.3.3 You can either load the standards from file or measure them. 13.2.3.4 To measure standards: 13.2.3.4.1 Measure a bank on the solvent if necessary using the "Blank" button in the instrument panel. 13.2.3.4.2 Measure the standards using the "Standard" button in the instrument panel. If you have selected one o f the prompts in the method, the appropriate values will be requested in a dialog box. 13.2.4 The spectra are displayed automatically in the "Standard Spectra" window as the standards are measured. A minimum number of standards is required, depending on the selected calibration curve. The ChemStation software calibrates automatically when at least the minimum number of standards have been measured. A table with the used standards and values as well as a calibration curve is displayed. 13.2.5 If the calibration is successful, the calibration curve icon o f the data analysis panel changes from red to green. 14.0 References__________________________________________________________________ 14.1 Definitions obtained from www.spectroscopymag.com. 14.2 HP Manual: Understanding Your UV-Visible Spectroscopy System, Hewlett-Packard: Wilmington, DE, 1997. Part No. G l 115-90005. 14.3 HP 8453 UV-Visible Spectrophotometer Operator's Manual, Hewlett-Packard: Wilmington, DE, 2000. Part No. G l 115-90012. 14.4 HP 8453 UV-Visible Spectrophotometer Service Manual, Hewlett-Packard: Wilmington, DE, 1998. Part No. Gl 115-90003. 15.0 Affected D ocuments 15.1 None. 16.0 R e v isio n s Revision Number. Reason For Revision Revision Date ETS-9-46.0 Operation and Maintenance of the HP8453 UV-Vis Spectrophotometer Page 9 of 9 Page 99 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 3M Environmental Laboratory E quipm ent P rocedure R o u tin e M aintenance o f Arch o n Pu rg e and T ra p Au to sa m pler , T ek m a r P urge and T rap C oncentrator and A g ilen t G as Chrom atograph/M ass Spectro m eter Procedure Number: ETS-9-49.D Approved by: Laboratory Manager Adoption Date: Revision Date: Date Date ETS-9-49.0 Routine Maintenance o f the Purge & Trap Autosampier/Concentrator/GC/M S Page 100 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.0 Scope and Application (Use Numbered Tier I)____________________________ 1.1 This equipment procedure describes the maintenance required for optimal operation of the Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent gas chromatograph / mass spectrometer (GC/MS) system. Specific items requiring routine maintenance include occasional tightening vial escalator's nuts and refilling the Standard Vial and water bottle in the Archon autosampler and periodic cleaning o f the mass spectrometer ion source. 2.0 Definitions_______________________________________________________________ 2.1 None. 3.0 Description______________________________________________________________ 3.1 Archon purge and trap autosampler equipped with Tekmar purge and trap concentrator and Agilent gas chromatograph and mass spectrometer. 4.0 I d e n t if ic a t io n _____________________________________________________________________ 4.1 System : "Rufus". (An equivalent system may be used). 4.1.1 Autosampler: serial number 13006, Varian, Archon 4.1.2 Concentrator: serial number 90297002, LSC2000, Tekmar 4.1.3 GC: serial number US00034972,6890 G1530A, Agilent 4.1.4 MS: serial number US01180105,5973N G2589A, Agilent 4.1.5 PC: serial number US94850812, D6720T, HP Kayak XA 5.0 W a r n in g s a n d C a u tio n s___________________________________________________________ 5.1 Health and Safety Warnings: 5.1.1 Cooling the Tekmar before removing the side cover for maintenance prevents contact bums. 5.1.2 Turning off power source for Tekmar before removing the side cover will prevent electric shock. 5.2 Cautions: 5.2.1 It is recommended that a grounded antistatic wrist strap be worn while disconnecting all wires, contacts, or cables which are connected to printed circuit boards within Archon autosampler, Tekmar concentrator or MS analyzer. 5.2.2 To prevent the breakage o f the Standard Vial on Archon autosampler, during the refill, do not use any tool and do not overtighten the thumbnut. 5.2.3 Never add oil while the foreline pump is on. 6.0 Special Instructions 6.1 Not applicable. ETS-9-49.0 Routine M aintenance o f the Purge & Trap Autosampler/Concentrator/GC/M S Page 2 of 5 Page 101 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 7.0 Responsibility______________________________________________________________ 7.1 Routine maintenance procedures may be performed by a primary custodian, and by any analyst who has been trained to perform these maintenance procedures by a primary custodian. 8.0 Supplies and Materials__________________________________________________ 8.1 Graphite Ferrules, 0.4 mm I.D. for 0.25 mm columns 8.2 Abrasive paper, Hewlett-Packard, part no. 5061-5896 8.3 Alumina abrasive powder, Hewlett-Packard, part no. 8660-0791 8.4 Acetone, reagent grade 8.5 Dichloromethane, reagent grade 8 .6 Methanol, purge and trap grade 8.7 Gloves (clean, lint-free, cotton), Hewlett-Packard, part no. 8650-0030 (large); 8650-0029 (small) 8 .8 Cotton swabs 8.9 Chem-wipes 8.10 Glass beakers 8.11 Sonicator 8.12 Base deactivated 2 mm ID gooseneck splitless injection port liners, Restek Corporation, part #20796-210.5, or equivalent 8.13 11 mm diameter Thermogreen LB2 septa, Supelco, part #23163, or equivalent 8.14 Viton injection port o-rings, Restek Corporation, part #20377, or equivalent 8.15 Septum wrench, Hewlett-Packard, part #19251-00100 8.16 Tweezers 9.0 C l e a n in g P r o c e d u r e s N/A 10.0 M aintenance Procedures ___________________________________ 10.1 Routine; Tighten the elevator's assembly nuts when Archon autosampler displays error message " Elevator not homed position ". 10.1.1 Stop autosampler run by pressing STOP button on the front display twice. 10.1.2 Open the back cover of autosampler 10.1.3 Tighten top and bottom nuts on the elevator's assembly, do not over tighten them. 10.1.4 Close the back cover of autosampler . 10.2 Routine; Fill the water bottle, empty the waste bottle. 10.3 Routine: If internal standards or surrogates are to be used, be certain the Standard Vial is filled with the required internal standard or surrogate. 10.3.1 Turn the helium gas "OFF" with the toggle switch. 10.3.2 Push "System" key, choose Maintenance, Standard Control, Front Park. 10.3.3 Grasp the vial and loosen the black thumbnut. Slide the vial down. ETS-9-49.0 Routine M aintenance o f the Purge & Trap Autosampler/Concentrator/GC/M S Page 3 of 5 Page 102 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 10.3.4 Clean the vial with methanol, dry it thoroughly and fill the vial with approximately 5 ml of standard or surrogate 10.3.5 Slide the vial back up into standard mount. Finger-tighten thumbnut until it is snug. Do not use any tool and do not overthighten. 10.3.6 Turn the helium gas into ON position. Prime Standard Loop. 10.4 Routine: GC/MS foreline pump maintenance. 10.4.1 Examine the oil level window daily. If the oil level is near or below the lower line then add foreline pump oil. Never add oil while the foreline pump is on. 10.4.1.1 Vent the MSD according to the MSD Hardware Manual. 10.4.1.2 Remove the fill cap. 10.4.1.3 Add pump fluid until the oil level in the window is near, but not above, the upper line. 10.4.1.4 Reinstall the fill cap. 1 0.4.1.5 Pump down the MSD according to the MSD Hardware Manual 10.4.2 Change foreline pump oil eveiy 6-12 months according to the MSD Hardware manual. 10.5 Nonroutine: Document any nonroutine maintenance in the instrument's maintenance logbook. 11.0 O perating Procedures ___________________________________________________ 11.1 For operating procedures, refer to an appropriate analytical method, or to the Archon Purge and Trap Autosampler System Operator's Manual, Tekmar LSC200 Instruction Manual and the Hewlett-Packard MSD Hardware Manual for HP 5973N & HP 6890 Series Mass Selective Detectors. 12.0 Recorps____________________ ____________ _____________ .______ 12.1 Document any maintenance performed on the instrument in the maintenance or run/maintenance logbook. Include a description o f the procedure(s) performed, any unusual observations, parts replaced or needing replacement, and whether the procedure was routine or non-routine. Be sure to date and initial the entry. Logbooks are archived when complete. 13.0 Testing, Calibration and/or Standardization Procedures__________________ 13.1 After cleaning the source and allowing sufficient time for the vacuum to pump down and the mass spectrometer to equilibrate to operation temperature, run an autotune; check for improved performance and for the presence or absence of air leaks. A tune report can also be used to check for leaks after performing injection port maintenance. 14.0 References 14.1 Archon Purge and Trap Autosampler System Operator's Manual 14.2 Tekmar LSC200 Instruction Manual ETS-9-49.0 Routine M aintenance o f the P urge & Trap Autosampler/Concentrator/GC/M S Page 4 of 5 Page 103 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 14.3 Hewlett-Packard MSD Hardware Manual for HP 5973N & HP 6890 Series Mass Selective Detectors. 15.0 A ffected Documents____________________________________________ ___ 15.1 None. 16.0 Revisions Revision Number. Reason For Revision Revision Date ETS-9-49.0 Routine M aintenance o f the Purge & Trap Autosampler/Concentrator/GC/M S Page 5 of 5 Page 104 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix B: Chemical Characterization This appendix includes chemical characterization information for both reference substances and control substances. Page 105 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Chemical Characterization S u b sta n c e IUPAC Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity S u b sta n c e IUPAC Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity PFOA Perfluorooctanoic acid, ammonium salt c 8f 15o 2n h 4 3825-26-1* 3M Specialty Chemicals 2002 Frozen 332 White wax or powder 95% C 7 Perfluoroheptenes (90/10 m ix) 10% 2-Perfluoroheptene, 90% Perfluoroheptene c 4f 9c f = c 2f 5, c 5f ,, c f = c f 2 355-63-5* Lancaster Synthesis 2005 Frozen 90004250, TNA 3025 Clear ambient liquid 100% PFHpA Perfluoroheptanoic acid c 7f ,, o 2h 375-85-9* Aldrich 2005 Frozen TCR-99131-25 Clear Crystals 98%< C7 Hydride 1, 1,2 ,2 ,3,3,4,4,5,5,6 ,6 ,7,7,7 pentadecafluoroheptane C 7F 15H 27213-61-2* Lancaster Synthesis 2005 Frozen 900591 l,TN A -3026 Clear ambient liquid 97% PFPA Perfluoropentanoic acid c 5f 9o 2h 2706-90-3* Aldrich 2002 Frozen SD-043 Clear Liquid 95%< C 6 Hydride 1, 1,2 ,2 ,3,3,4,4,5,5,6 ,6 ,6 tridecafluorohexane Q F 15H 355-37-3* Lancaster Synthesis 2005 Frozen 90005941, TNA-3027 Clear ambient liquid 98% Page 106 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Reference Substances (continued) Reference Substance IUPAC Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity Reference Substance IUPAC Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity C8Hydride/Olefin Mix 1 , 1, 1,2 ,2 ,3,3,4,4,5,5,6 ,6 ,7,8 ,8 ,8 pentadecafluorooctane, 2 perfluorooctene C6F 13C FH C F3'C6F 13C F= C F2 TCR-99030-18 3M Specialty Chemicals 6/6/99 Frozen 1 Clear ambient liquid 85% C8 Hydride, 15% C8 Olefin C4Interior Olefin 2-Perfluorobutene c 4f 8 360-89-4* Lancaster Synthesis 2002 Frozen G00195, TNA-4298 Clear ambient liquid 97% C8Terminal Hydride 1, 1,2 ,2 ,3,3,4,4,5,5,6 ,6 ,7,7,8 ,8 ,8 heptadecafluorooctane C 8F I7H ' 335-65-9* Aldrich Chemical 2002 Frozen 04307PN, TNA-2983 Clear ambient liquid 99% C3 Terminal Hydride 1, 1,2 ,2 ,3,3,3 heptafluoropropane C3F 7H 2252-84-4* Lancaster Synthesis 2010 Flammable G0062B, TNA-4294 Gas 97% ' C4 Terminal Hydride 1,1,1,2,3,3,4,4,4 nonafluorobutane C4F 9H 375-17-7* Crescent Chemical 2002 Frozen 6A-46, TNA-3997 Clear ambient liquid 99% C2 Terminal Hydride 1, 1,2 ,2 ,2 pentafluoroethane c 2f 5h 354-33-6* Lancaster Synthesis 2010 Flammable G00492, TNA-3021 Gas 99% Page 107 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Chemical Characterization (Control Substances)* Control Substances Structure IUPAC Name Use Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity Control Substances PFCH C sF .2 Perfluorocyclo-hexane Surrogate Standard For GC/MS analysis Aldrich Chemical 2005 Frozen 01911AU Colorless M oist Solid 97% Chlorobenzene-ds PFBS Perfluorobutane-sulfonate, potassium salt C4F 9SO2K Internal Standard for LC/MS analysis 3M Specialty Chemicals 2002 Frozen TCR-99030-028 White Powder 97%< Toluene-dg Penta fluorobenzene C 6H F 5 Pentafluorobenzene Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 6 /2 0 0 2 Frozen A013256 M ethanol solution 99% (2500 pg/mL 0.2% ) Dibromofluoromethane Structure IUPAC Name Use Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity Control Substances Structure IUPAC Name Use Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity *CAS Number QCID 5 Chlorobenzene-d5 Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 6 /2 0 0 2 Frozen A 013256 M ethanol solution 99% (2500 pg/mL + 0.2%) 1,4 Difluorobenzene Q H 4F 2 1,4 Difluorobenzene Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 6 /2 0 0 2 Frozen A 013256 M ethanol solution 99% (2500 pg/mL + 0.2%) C7 D 8 Toluene-dg ' Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 2 /2 0 0 2 Frozen A012973 Methanol solution 99% (2500 pg/mL 0.2%) 4-Bromofluoro-benzene C6H 4BrF 4-B ro m o flu o ro -b en zen e Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 2 /2 0 0 2 Frozen A 012973 . M ethanol solution 99% (2500 pg/mL + 0.2%) C H B r2F D ibrom ofluorom ethane Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 2 /2 0 0 2 Frozen A012973 Methanol solution 99% (2500 pg/mL 0.2%) 1,4-Dichlorobenzene-di C 6C12D 4 1,4-D ichlorobenzene-d4 Instrumental Surrogate Standard For GC/MS analysis Restek Corp. 6/2 0 0 2 Frozen A013256 M ethanol solution 99% (2500 pg/mL 0.2%) Page 108 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix C: Kinetics Model and Kinetics Calculations This appendix presents the mathematical description of the kinetics model employed in this study and the application of this model in the determination of the estimated half-lives presented. Page 109 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Kinetics Model C1. Reaction Components and Rates The arguments below are based on the following idealized set of reactions representing the photodegradation of a parent compound P and its products A m, which number N. The actual reactions that occur are subsumed in these equations, and are assumed to proceed with pseudo-first order rates k Pm(forthe parent) and k Am(forthe products). P + p h o to n --> n m A m+ Yml (m= ltoN ) (C1) A m + p h o to n -> Ym2 (m = 1 to N) (C2) where "photon may either represent a photon of light or it may represent some other species in solution that reacted with a photon to produce a new reactive species and the general symbols Y ml and Y m2 represent all the other hydrolysis products. C2. Parent Compound Concentrations Equation C1 indicates that the pseudo-first order differential change in the parent concentration P at a constant flux of light or a constant concentration of radicals is given by (C3) which is equivalent to the separable differential equation (C4) Equation C4 may be directly integrated to obtain the general solution With the initial condition P(t = 0) = P0, the specific solution to Equation C4 is using the additional definition of the total parent photolysis rate (C5) (C6) Page 110 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 kp "Pm (C7) Equation C6 can be re-written in a form that allows a least-squares estimate of the total parent hydrolysis rate: kpt = -In (C8) 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 photolysis rate. Equation C6 indicates that over a period of time T 1^ (the parent half-life) the parent concentration P is reduced through hydrolysis by a factor of two, where (C9) A least squares estimate T ^ of the parent photolysis half-life is therefore available from (C10) C3. Product Compound Concentrations The pseudo-first order differential changes in the product concentrations An (using Equations C2 and C6) re d A m= ( n mk PmP - k AmA m) dt = ( n mkpmP0 e"kp ` - k AmA m) dt (C 11) and the (first order, non-separable) differential equation governing the product concentrations is dAm dt + k AmA m - n mkpmP0 e -kpt (C12) The "standard form" of Equation C12 is Page 111 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 A m+ S ( t ) A m = Q ( t ) where the "function" S (t) is actually a constant: S(t) = k Am and Q(t) = n mk PmPo e-kpt The general solution A mto Equation C12 is contained in (C13) (C14) (C15) A eJS(,)dt = | q (.) eJS(t')dt' dt + C (C16) where eJs(t)<u = = e kAm Jd t _ (C17) and JjQ (t)e ^ s(t')dt' dt + C = n mk PmP0 1 ekto` e 'kptdt + C (C18) There are two cases of Equation C18 to consider. In the circumstance that k Am = k p , which occurs only when the rate of the mth product is identical to the total parent photolysis rate, the general solution to Equation B18 is (for k Am= k p) A m e kpt = n mk PmP0 t + C (C19) and, using the initial condition A m(t = 0 ) = A ^ , the specific solution to Equation18 is (for k Ajn = k p ) A m = (n m k PmP 0 (C20) Page 112 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 We note that when = k p = 0 (that is, when both the parent and potential product are photolytically stable), Equation C7 requires (also) that k Pm= 0, so Equation C20 becomes (C21) 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 photolysis rate k pand the product photolysis rate k ^ approach each other. In the more probable case, for which k Am # k p (i.e. that the rate of the mth product is different from the total parent rate), the general solution to Equation C18 is (C22) and the specific solution to Equation C18 with the initial condition A m(t = 0) = A m0 is (C23) Of greatest interest here is the case in which the product compounds are known to be photolytically stable, that is, when k ^ = 0 for all m. In this case, Equation C23 becomes (for stable products) (C24) C4, Relationships Between the Parent and Compound Concentrations Equations C7 and C24 can be combined to obtain (for stable products) (C25) Page 113 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 so that or N k p y (A m - A ra0) kP= X m=l n "> k p m = ` J - e ' kp` ) f (for stable products) c-kPt j _ ^ (Am- A m0) I Po (for stable products) (C26) k pt = - I n l jj? (Am- A m0) =, Po (C27) If the changes in the product concentrations are all small compared to the original parent concentration, that is, if ". i A m - Am_______ mm nl) A P0 1 we may use the expression (valid for -1 < X < 1 ) (C28) ln(l + X ) = X - - X 2 + - X 3 - - X 4 +. 234 and Equation B23 becomes (for stable products and ^ ^ A m A m0 < < P 0) ( ^N AA m- A* m0 \ k pt s - m=l or (C29) (C30) Page 114 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 (for stable products and <<P0) (C31) C5. Parent Half-Life Estimates Based on Limits of Quantification of the Products In every experimental determination of k p, there is some set of values A'm0Q (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= Aj;0Q. With these assumptions, the experimental data indicate that the reaction rate k pis less than some maximum value (kP)raax as follows: (for photolytically stable products at concentrations below the limits of quantitation) k p i ( k , L = 0 m-1 (C 3 2 ) Under the same circumstances and assumptions, the experimental data indicate that the parent half-life T'/p (see Equation C9) is greater than the value (t ^p) as follows: (for photolytically stable products at concentrations below the limits of quantitation) -i-l > (t ^ ) = 7 M i L = A t P, H 2 ) P ' PAnin (V i 1 A l 0Q V P /max . m=l (C33) The reader should note that Equations C32 and C33 are valid only when both 1) the products are stable and 2) the concentrations of all the potential products are measured. Otherwise, the quantity (k p)maxin Equation C32 may not actually represent the maximum possible value of the rate constant k p, and the related result in Equation C33 for (t ^ ) is also questionable. Page 115 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 C6. Parent Half-Life Estimates Based on Limits of Quantification and Experimental Precision of Product Concentrations In certain experiments, some 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 o 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+ iimat 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 m. This reasoning suggests that the following estimate of the maximum parent photolysis rate is appropriate: (for stable products at either 1) constant measured concentrations with standard deviation amor 2) concentrations below the limits of quantitation) LOQ BelowLOQ Cons tan t (C34) Under these circumstances and assumptions, the experimental data indicate that the parent half-life T 1^ is greater than the value ( T ^ ) minas follows: (for stable products at either 1) constant measured concentrations with standard deviation omor 2) concentrations below the limits of quantitation) t* 2 (Tl? L = 7vr^Pf/m-ax= At p ta<2) Below LOQ Cons tan t (C35) The reader should note that Equations C34 and C35 are valid only when both 1) the products are photolytically stable and 2) the concentrations of all the potential products are measured. C7. Parent Half-Life Estimates Based on the Experimental Precision of Parent Concentrations In certain experiments, the concentration of the parent remains essentially constant 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 } i pand standard deviation o p, the data do not exclude the possibility that the product concentration increased Page 116 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 from the initial value p p - o P to the value p p + g p at time t = A t . This reasoning suggests that the following estimate of the maximum parent photolysis rate is appropriate: (for essentially constant parent concentrations with mean value ( i Pand standard deviation o P) k p" < (Vk Pr /)m ax 2op Up A t ' (C36) Under these circumstances and assumptions, the experimental data indicate that the parent half-life T '/p is greater than the value (t ^ ) ^ as follows: (for essentially constant parent concentrations with mean value p p and standard deviation o P) T 1/2 b (2 ) _ p.p A tln (2 ) p (\ k pr )/max 2 o p" (C37) References to Appendix C: C1 I. N Levine, "Physical Chemistry," McGraw-Hill (New York), pp. 498-501 (1978). 02 F. Daniels, et al., "Experimental Physical Chemistry" , McGrawHill (New York), p.131 (1962). Kinetic Calculations - Indirect Photolysis Only two values of the parent concentration were recorded (P0 and P ) which represent the values before and after the exposure period of length At. In this case, no least squares regression is possible to determine the rate k p In Equation C8: . ^P^ k pt -In (C38) The measured values of for P0 and P are 1427 ng/ml and 1432 ng/ml, respectively. Equation C39 is appropriate: Page 117 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 k P * ( k p)m ax = V-2ta Ap t (C39) The observed rate using At = 69.5 hours, |Ip = 1478ng/ml, and O p = 3 3.2 ng/ml is k p = 6.46 x 10`4 hours'1. The rate of photolysis in the reactor k P is related to the actinic rate of photolysis k ACT by (I N ACT k ACT ~ k P I (C40) where I ACT = 261 w/m2 is the actinic solar intensity (at 45 south latitude) and the measured reactor intensity is I R = 680 w/m2. This gives ^ 261 w / m 2 ^ k ACT = 6 . 46 x l O -4 h r -1 : 2 .48x l O -4 hr' 1 680 w / m 2 (C41) For samples under constant illumination, the reaction rate and half-life are related by Equation C9: T 1/2: K 2 ) 'ACT (C42) However, the actinic half-life is three times larger, according the standard eight-hour exposure day. This leads to (indirect photolysis) T 31n( 2) = 349 days. k ACT (C43) Page 118 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix D: Representative Chromatograms C h ro m a to g ra m s fro m th e p re s e n t stu d y a re in clud ed in this a p p e n d ix . Page 119 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 2 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0002.D Sample Name: MeOH blank ===ss=t===s= Injection Date : 5/18/00 3:07:45 PM Sample Name : MeOH blank Acq. Operator : kej Acq. Instrument : Rush Seq. Line : 2 Vial : 99 Inj : 1 Acq. Method : C :\HPCHEM\1\METHODS\PF0A516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518A X .M Last changed : 7/10/00 9:43:20 AM by kej (modified after loading) (Results are from a previously saved SIM Analysis (ES- ) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:43:20 AM kej Page 1 of 2 Page 120 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 3 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0003.D Sample Name: 1:7 MilliQ/MeOH Injection Date 5/18/00 3:26:02 PM Seq. Line : 3 Sample Name 1:7 MilliQ/MeOH Vial : 98 Acq. Operator kej Inj : 1 Acq. Instrument Rush Inj Volume : 5 Pi Acq. Method C:\HPCHEM\l\METHODS\PFOA516 .M Last changed 5/18/00 11:35:03 AM by kej Analysis Method C :\HPCHEM\1\METHODS\R0518A X .M Last changed 7/10/00 9:43:24 AM by kej (modified after loading) SIM Analysis (ES--) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column 4x35mm. Instrument 1 7/10/00 9:43:25 AM kej Page 1 of 2 Page 121 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 4 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0004.D Sample Name: 00028-37-00 Injection Date : 5/18/00 3:44:20 PM Seq. Line : 4 Sample Name : 00028-37-00 Vial : 1 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Acq. Method : C :\HPCHEMX1\METHODS\PFOA516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518AX.M Last changed : 7/10/00 9:43:28 AM by kej (modified after loading) (Results are from a previously saved SIM Analysis (ES- for PFOS/PFBS/PFOA using Dionex lonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:43:28 AM kej Page 1 of 2 Page 122 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 5 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0005.D Sample Name: 00028-37-01 Injection Date ; 5/18/00 4:02:35 PM Seq. Line : 5 Sample Name : 00028-37-01 Vial : 2 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Inj Volume : 5 pi Acq. Method C :\HPCHEM\l\METHODS\PF0A516.M Last changed 5/18/00 11:35:03 AM by kej Analysis Method C :\HPCHEM\1\METHODS\R0518AX.M Last changed 7/10/00 9:43:32 AM by kej (modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:43:32 AM kej Page 1 of 2 Page 123 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 6 of 62 Data File C:\HFCHEM\l\DATA\R051800B\rush0006.D Sample Name: 00028-37-02 Injection Date 5/18/00 4:20:50 PM Seq. Line : 6 Sample Name 00028-37-02 Vial : 3 Acq. Operator kej Inj : 1 Acq. Instrument Rush Inj Volume : 5 pi Acq. Method C :\HPCHEM\l\METHODS\PFOA516.M last changed 5/18/00 11:35:03 AM by kej Analysis Method C:\HPCHEM\1\METHODS\R0518AX.M last changed 7/10/00 9:43:36 AM by kej (modified after loading) SIM Analysis (ES-) for PF0S/PFBS/PF0A using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:43:36 AM kej Page 1 of 2 Page 124 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 13 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0013.D Sample Name: 00028-37-09 Injection Date : 5/18/00 6:28:34 PM Seq. Line : 13 Sample Name : 00028-37-09 Vial : 10 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Inj Volume : 5 pi Acq. Method : C:\HPCHEM\l\METHODS\PFOA516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518AX.M Last changed : 7/10/00 9:44:04 AM by kej (modified after loading) SIM Analysis (ES- ) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. MSD1 263, E10=262.7:263.7 (R051800B\RUSK0013.D) API-ES Negative 1000000 800000 Calibration Std 9 | 600000 400000-j 200000J o -i--.--.-.-.------- -_------ .------j------ ------ <------ .---- -,--.-.-.-..-..--.-..-.-.-..-..>-.-..-.-.-..-..--.-..-.-.-..-..1-.-..-.-.-..-..-..-.-.-..-.-..--..-.-D--- " ---- j------ 2468 MSD1 363, EIC*362.7363.7 (R051800BVRUSH0013.D) API-ES Negative 1200000 1000000 800000 600000 400000 200000 0- ------ ,------ ------- ,------ ,------ ------- ------- *-- n -- -------- *------'------1------ ------ '------t_ 32 46 MSD1 299, EIC=298.7:299.7 (R05ia00B\RUSH0013.D) API-ES Negative 8i 150000 125000 100000 75000 - 50000 25000 0- -- ...... - ... .. ............. - 2 '4 6 MSD1 413, EIC*412.7:413.7 (R051800B\RUSH0013.D) API-ES Negative ............. ... L '8 1i0 '..... ' min i 1 ... 10 min _ 1_ 1I0 1 1 min 1500000 1000000 500000- o - rTM-------- ----- --- r -- 02 ---- -------- ------r------------4 ------ ------- 1-----6 ------- ------- r_----8rr----r- 11 ' ....... 10 min Instrument 1 7/10/00 9:44:04 AM kej Page 1 of 2 Page 125 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 29 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0029.D Sample Name: 0515-PFOAfe-ll Injection Date : 5/18/00 11:20:33 PM Seq. Line : 29 Sample Name : OS15-PFOAfe-ll Vial : 21 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Inj Volume : 5 pi Acq. Method : C:\HPCHEM\l\METHODS\PFOA516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C :\HPCHEM\l\METHODS\R0 51BAX.M Last changed : 7/10/00 9:45:05 AM by kej (modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFQA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:45:05 AM kej Page 1 of 2 Page 126 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 30 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0030.D Sample Name: 0515-PFOAfe-12 Injection Date : 5/18/00 11:38:48 PM Seq. Line : 30 Sample Name : 0515-PFOAfe-12 Vial : 22 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush inj Volume : 5 jil Acq. Method : C:\HPCHEM\1\METHODS\PFOA516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518AX.M Last changed : 7/10/00 9:45:09 AM by kej (modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column. 4x35mm. MSD1 263, EIC=282.7:263.7"(R051800B\RUSH0030 D) API-ES Negativo Exposed - 72hrs Blatik Spiked Fe203 NoPeroxidc 5 Q ' ------------------- J iliil ^ t .. i .. i .. i . . i ., 11. i . i .. -- > T'*---- '---- --~T ..............--------- ---------1-------- 1-------- --------- *--------1-------- --------- 1-------- *-------1-------- '-------- '------- * 2 4 .6 8 10 mir MSD1 363, EIC=362.7:363.7 (R051800B\RUSH0030.D) API-ES Negativo 125000 \ 100000 - 75000 4 50000 -I 25000 -j 0' * 1 i ....... '-------- 1-- ...'-------- 1-------- '--------1-------- 1--------'--------'--------r------- -------- -------- -------- 1-------- '-------- -------- 2 4 8 8 10 mlr M S b l 239, EiC=298.V:299.7 (R0518008\RUSH0030.D) API-ES Negative 175000- 150000*5 125000 4 100000 75000 ~ 5ooo4 25000-^ 04 > i r 1 t 'r ' 1 1 i ... . ..... _2 4 ..... 6 MSD1 413, EIC412.7:413.7 (R051800B\RUSH0030.D) API-ES Negative 350000 - 300000 250000 200000 - 150000 100000 50000 0' ' ' 1 ' l~~ 2 J '1 ' ' J J 46 l T -r-- i j 8 1>. , 8 i 1 . 1 i-- ---- -------- --------10 mir <........|---------1---- ----1-------10 mir Instrument 1 7/10/00 9:45:10 AM kej Page 1 of 2 Page 127 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 31 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0031.D Sample Name: 0515-PFOAfe~13 Injection Date : 5/18/00 11:57:01 PM Seq. Line : 31 Sample Name : 0515"PFOAfe-13 Vial : 23 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Inj Volume : 5 pi Acq. Method : C :\HPCHEM\l\METHODS\PFOA516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518AX.M Last changed : 7/10/00 9:45:13 AM by kej (modified after loading) . SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. MSD1 263, EIC*262.7:263.7(R051800B\RUSH0031.0) API-ES Negative Exposed - 72hrs Sample F e203 N o Peroxide 6000 5000 -i MSD1 363, E!C=362.7:363.7 (R051800B\RUSH0031.D) API-ES Negative 12000 10000 8000 -i 6000 4000 2000 MSD1 299, E!C=29Q.7:299.7(R051800B\RUSH0031.D) APf-ES Negative 150000125000 100000i 75000J 50000 2 5 0 0 0 - 0^ 6 MSD1 413, IC 4 1 2.7:413.7 (R 051800B\RUSH0031.D ) A PI-E S Negative 1 6 0 0 0 0 0 -i 1250000 -i 1000000 750000 500000 -j 250000H 0- 10 -- 10 10 mir Instrument 1 7/10/00 9:45:14 AM kej Page 1 of 2 Page 128 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 37 of 62 Data File C:\HPCHEM\l\DATA\R051800B\rush0037.D Sample Name: 0515-PFOAfe-19 Injection Date : 5/19/00 1:46:30 AM Seq. Line : 37 Sample Name : 0515-PFOAfe-19 Vial : 29 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Inj Volume : 5 pi Acq. Method : C:\HPCHEM\1\METH0DS\PF0A516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518AX.M Last changed : 7/10/00 9:45:36 AM by kej (modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. MSD1 263, EtC=262.7:263.7 (R051800B\RUSH0037.D) API-ES Negative 7000 - Exposed - 72hrs Control M illiQ No Peroxide 600 0- sooo-i 4000 3000- 246 MSD1 363, E10=362.7:363.7 (R0518008\RUSH0037.D) API-ES Negative 14000 " 12000- 10000 8000 6000 4000 _ 2000 ^ .............. ................................................................................ 2 46 MSD1 299, 10=298.7:299.7 (R051800B1RUSH0037.D) API-ES Negative > _/ ---------,-------- --------- --------8 10 min i 1, i -v 8 10 mir 150000-t 125000 -i 100000 75000 50000 25000i 1 0 ^ 1' ........' ----- T------- 1-------- '- " T -- --------------------- -------" -------- ' 1........ ' '----------- ' *" 2468 MS01 413, EIC=412.7:413.7 (R051800B\RUSH0037.D) API-ES Negative 1500000 1250000 1000000 750000 500000 250000 I0 - __________________________________________________________________________________________ ... , . I- ` i 1 > i l -i- i a2 468 10 mir .... I......... ......... ........... 10 mir Instrument 1 7/10/00 9:45:36 AM kej Page 1 of 2 Page 129 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 40 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0091.D ' Sample Name: 0515-PFOAfe-51 Injection Date : 5/19/00 6:13:39 PM Seq. Line : 91 Sample Name :0515-PFOAfe-51 Vial : 61 Acq. Operator :kej Inj : 1 Acq. Instrument :Rush Acq. Method :C:\HPCHEM\l\METHODS\PFOA516.M Last changed :5/18/00 11:35:03 AM by kej Analysis Method :C:\HFCHEM\1\METHODS\R0518BX.M Last changed :7/10/00 9:16:40 AM by kej (modified after loading) (Results are from a previously saved SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. MSD1 263, E10=262.7:263.7 (R0518008\RUSH0091.D) API-ES Negative 7000 -j Day 0 Blank Fe203 No Peroxide 6000 5000 -, J 4000 -i 3000- 1 * i ' 1' ' ' i ' - ' 1 1 >246 6 MSD1 363, EIC=362.7:3Q3.7 (RO510OOB\RUSHOO91.D) APl-ES Negative i 10 M mi n liiilil i i ' ' i ' ' ` 2468 MSD1 299,HC=298.7:299.7(R0S1800B\RUSH0091.O) APl-ES Negative 150000 125000 100000 75000 50000 25000- 1 ......... ......... ,-------- ,--------,---------------- --------- ,----- - ] ----------------*----------------!------------- ------------*----246 8 MSD1 413, EIC=412.7:413.7 (R051800B\RUSH0091.D) API-ES Negative i 10 i 10 min min Instrument 1 7/10/00 9:16:40 AM kej Page 1 of 2 Page 130 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 41 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0092.D Sample Name: 0515-PFOAfe-52 3=HI35= =2= = = Injection Date 5/19/00 6:31:53 PM Seq. Line : 92 Sample Name 0515-PFOAfe-52 Vial : 62 Acq. Operator kej Inj : 1 Acq. Instrument Rush Inj Volume : 5 pi Acq. Method C:\HPCHEM\l\METHODS\PFOA516.M Last changed 5/18/00 11:35:03 AM by kej Analysis Method C:\HPCHEM\1\METHODS\R0518BX.M Last changed 7/10/00 9:16:44 AM by kej (modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. MSD1 263, EIC=262.7:263.7 (R051800B\RUSH0092D) API-ES Negative 120000 100000 -j 8 0 0 0 0 -i DayO Dlank Spiked Fe203 No Peroxide 60000 40000 2 0 0 0 0 - 0 -i ... . _ -------- ,-------- ,-------- --- 1--- ................... ........................ 1 i-----------r-- --- -- -- ---1-- ------ ---- \-- --- --,-------- ,-------- ,--- ---- .--- 2 4 6 8 10 mir MSD1 363,EIC=362.7;363.7{R051800B'RUSH0092.D) API-ES Negative 125000 i 100000-^ 7 5 0 0 0 - 50000 -j 25000 V i > i 1 - i 1 ---1--- '-- . >246 8 MSD1 299, EIC=298.7:299.7 (R051800BRDSH0092.D) API-ES Negative i- 10 mf n 150000 A 125000 100000- 75000i 50000 128000 o 4-------- ,-------- .--- ---- 1--- ---- ....................................................................... ---- *---1--- --- 1--- 1--- 1--- --- 1-- 7 >246 MSD1 413, 10412.7:413.7 (R051800B\RUSH0092.D) API-ES Negative , 8 T ----- 1---1-------- 10 mi n 300000 250000 200000 150000 100000 50000- o--------- ,--- --- '--- I--- --- '--- ----1 32 4 1 ---*---1-------- 6 --- !--- '-a ` i1 _ - .... 10 .. 1 .. mfn Instrument 1 7/10/00 9:16:44 AM kej Page 1 of 2 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 42 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0093.D Sample Name: 0515-PFOAfe-53 Injection Date Sample Name Acq. Operator Acq. Instrument Acq. Method 5/19/00 6:50:09 PM Seq. Line 0515-PFOAfe-53 Vial kej Inj Rush Inj Volume C:\HPCHEM\1\METHODS\PFOA516.M : 93 : 63 :1 : 5 pi Last changed 5/18/00 11:35:03 AM by kej Analysis Method C:\HPCHEM\1\METHODS\R0518BX.M Last changed 7/10/00 9:16:48 AM by kej {modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:16:48 AM kej Page 1 of 2 Page 132 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 47 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0098.D Sample Name: 0515-PFOAfe-58 Injection Date 5/19/00 8:21:32 PM Seq. Line 98 Sample Name Acq. Operator 0515-PFOAfe-58 kej Vial Inj 68 1 Acq. Instrument Rush Inj Volume 5 (ll Acq. Method C :\HPCHEM\1\METHODS\PFOA516.M Last changed 5/18/00 11:35:03 AM by kej Analysis Method C:\HPCHEM\1\METHODS\R0518BX.M Last changed 7/10/00 9:16:48 AM by kej (modified after loading) SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:17:10 AM kej Page 1 of 2 Page 133 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 14 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0065.D Sample Name: 0515-PFOAfe-31 :SS= ssSS=S== =S38 = SS: Injection Date 5/19/00 10:17:53 AM Seq. Line 65 Sample Name 0515-PFOAfe-31 Vial 41 Acq. Operator kej Acq. Instrument Rush nj 1 Inj Volume 5 pi Acq. Method C :\HPCHEMM \METHODS \PF0A516.M Last changed 5/18/00 11:35:03 AM by kej Analysis Method C :\HPCHEM\1\METHODS\R0518BX.M Last changed 7/10/00 9:15:03 AM by kej (modified after loading) (Results are from a previously saved SIM Analysis (ES-) for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:15:03 AM kej Page 1 of 2 Page 134 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 15 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0066.D Sample Name: 0515-PFOAfe-32 Injection Date Sample Name Acq. Operator Acq. Instrument Acq. Method Last changed 5/19/00 10:36:09 AM Seq. Line : 0515-PFOAfe-32 kej Rush Vial : 42 Inj : 1 Inj Volume : 5 pi C:\HPCHEM\1\METH0DS\PF0A516.M 5/18/00 11:35:03 AM by kej Analysis Method C :\HPCHEMU \METHODS \R0518BX.M Last changed 7/10/00 9:15:07 AM by kej (modified after loading) SIM Analysis (ES-) for PF0S/PFBS/PF0A using Dionex IonPac NG1 column, 4x35mm. MSD1 263, EIC*262.7:2B3.7 (R051800B\RUSH0066.D) API-ES Negative 120000 100000 80000 60000 40000 20000 0- Uncxposed - 72hrs Blank Spiked Fe203 No Peroxide ................ - ............ 24 6 M5D1 363, c 1C'*362.7:363.7 (R051800BVRUSH0066.D) API-ES Negative I 8 r M in M p M o lllil .t....r....f....i __i .... i .... i . 24B MSD1 299, EIC=298.7:299.7 (RO518OOB\RUSHOO60.D) APFES Negative ' ' 150000 125000 4 100000-i 7 5 0 0 0 -j 50000 25000 0-i - ............... - -- - , In * -------- -------- 1-------- -------- -------- 2 4 '6 ' ` ' w a rn 413, tlL J *4 l 2.7:413.7 (R051800B\RUSH0066.D) API-ES Negative 350000 i 300Q0Q A 250000 200000 150000i 100000 5 0 0 0 0 -i j0 -i _________________________________________________________________________________________ 2 4 fl ' ' -- -----------------------------J 0 _____ _ mir Instrument 1 7/10/00 9:15:07 AM kej Page 1 of 2 Page 135 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. EOO-2192 Batch Run # 16 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0067.D Sample Name: 0515-PFOAfe-33 Injection Date : 5/19/00 10:54:22 AM Seq. Line : 67 Sample Name : 0515-PFOAfe-33 Vial : 43 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Acq. Method : C:\HPCHEM\l\METHODS\PFOA516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518BX.M Last changed : 7/10/00 9:15:10 AM by kej (modified after loading) (Results are from a previously saved SIM Analysis (ES-) for PF0S/PFBS/PFQA using Dionex IonPac NGl column, 4x35mm. Instrument 1 7/10/00 9:15:11 AM kej Page 1 of 2 Page 136 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Batch Run # 21 of 61 Data File C:\HPCHEM\l\DATA\R051800B\rush0072.D Sample Name: 0515-PFOAe-38 Injection Date : 5/19/00 12:26:11 PM Seq. Line : 72 Sample Name : 0515-PFOAfe-38 Vial : 48 Acq. Operator : kej Inj : 1 Acq. Instrument : Rush Acq. Method : C:\HPCHEM\1\METH0DS\PF0A516.M Last changed : 5/18/00 11:35:03 AM by kej Analysis Method : C:\HPCHEM\1\METHODS\R0518BX.M Last changed : 7/10/00 9:15:29 AM by kej (modified after loading) (Results are from a previously saved SIM Analysis (ES- for PFOS/PFBS/PFOA using Dionex IonPac NG1 column, 4x35mm. Instrument 1 7/10/00 9:15:29 AM kej Page 1 of 2 Page 137 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix E: Soil Types and Characterizations This appendix presents the physical descriptions and chemical characterizations of the three soils used in the present investigation Page 138 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 STANDARD LABORATORY SOILS PARAMETER ST CROIX CO., Vfl MORGAN CO* ALA METALS Ag, mg/Kg dried basis <3 <3 Al. mg/Kg dried basis As. mg/Kg dried basis Ba, mg/Kg dried basis _ Be, mg/Kg dried b'asts ' Ca, mg/Kg dried basis Cd. mg/Kg dried basis 22000 <10 210 '0.6 3000 <0.5 40000 <10 9.7 < b .s 1000 <0.5 Co, mg/Kg dried basis 8 5 . Cf, mg/Kg dried basis 36 41 Cu, mg/Kg dried basis . 12 8.5 Fe, mg/Kg driedbasis 20000 22000 . Hg. mg/Kg dried basis 0.04 ' 0.11 - Mg, mg/Kg dried basis 2800 1500 Mn, mg/Kg dried basis 600 120 Mo. mg/Kg dried basis 52 84 Na, mg/Kg dried basis 120 73 Ni, mg/Kg dried basis IS 14 P, mg/Kg dried basis 620 150 Pb, mg/Kg dried basis Se, mg/Kg dried basis <30 <200 <30 <200 Zn, mg/Kg dried basis 49 47 EXCHANGE ACTIVITY Extractable Ca, meq/IOOg 8.14 3.50 Extractable P, meq/100g 0.68 0.10 Extractable Mg, meq/tOOg 2.97 0.43 Extractable Na. meq/lOOg 0.03 0.02 Base Saturation % CEC, me^lOOg 42 28.3 18 22.0 ESP, % 0.11 0.09 SAR, units 0.04 0.05 NUTRIENTS - Available P. mg/L 61 5 NH3-N, mg/L 2.00 .- 0.25 N03-N, mg/L . 84 14 KJeldahl N, mg/Kg 1470 280 ORGANIC MATTER ' TOC, % O M ,% 1.79 3.09 0.277 0 .4 7 8 PHYSIOCHEMICAL Field Capacity, % 19.1 22.3 pHw, units 5.7 4.6 pHs, units 5.7 4.5 Ume-req, Tons/Acre Soluble Salts, mmhos/cm 1.8 0.98 1.8 0.32 CLASSIFICATION % Clay 22 26 %SM 44 36 %Sand 34 38 Soil Type LOAM CLAY LOAM BASE NEUTRAL PESTICIDES BDL BDL ACID PESTICIOES BOL BDL BDL a Below Detection Limits EPA'SSM <3 46000 <10 to o 0.6 66000 <0.5 6 25 13 18000 0.02 30000 710 2 220 13 ' 710 <30 <200 47 26.1 0.16 3.58 0.06 100 21.3 0.28 0.05 ' IS 1.80 6.0 250 0 .4 6 7 0.80S 16.0 7.7 7.5 NONE 0.66 22 26 52 SANDY CLAY LOAM BDL BDL . ________ ;------------------------------ -- ---------- Page 139 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix F: Light Intensity Measurements at 45 South Latitude (Miami FL) Page 140 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 with THA ?mJ S<ki L?ni h Ontor f jui;r, ms Miami uiiliijht. 25 S'ailli foposiiro -s u n iiy n t rhJA '_'oc3(.'* v '-v n.vtl > v Uv ,,u' 0,1 ot W:'us Coniliinations 10.S3 W/m' -^34a<im> ts Miami Sunliijht. ?fi South Exposure Atlas Xenon Filter Combination Filter Combination* Inner Outer Filter Gin* Filter GUu* Test Conditions type*? BmalHeate ty p e -tr- Boroiffleala ry^>**8 ',. ^ ^ V (w h ja -L____ Bem ineete . **"i V ',*.4/`C*rM' ori'/c.oW,m*1MmjOtti.t**i*:etnoimg'tb*]t#lf*ntfasVit.on;K . SodaLime . nohtfaetnd**testevwtndw Q tiirtz , VjEtonjaH tag, ORA ' 8oroII!o*t* ` trradtanc* Ranges* WAn* Wattage 260-300 300-400 400400 CIRA ' Sode U rn e ' Type*".'- BoroaHicate Sodi Unie Sunlight Measurements -- 4$^fiejr$itt' Peak Natural Daylight ! ' Peak Natural . Daylight Standard ' *Small variations are possible, depending on condition o f lamp andfilters. -/? Page 141 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Appendix G: Characteristics of the Spectral Output of the Suntest Instruments This appendix contains an Excel spreadsheet of the characteristics of the spectral output of the Suntest Photoreactors used in the present investigation. Page 142 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 Suntest Irradiance in W /m A2*nm Assuming the use of 300-800nm Global Sensor Only Filters Used Wavelength nm 250 252 254 256 258 260 262 264 266 268 270 272 274 276 278 280 282 284 286 288 290 292 294 296 298 300 302 304 306 308 310 312 314 316 318 320 322 324 326 328 IR Q/Suprax (UV) 0.001 0 0 0 0.001 0 0 0 0 0 0 0 0 0 0 0 0 0 0.003 0.005 0.009 0.014 0.021 0.028 0.045 0.054 0.07 0.085 0.116 0.138 0.151 0.175 0.21 0.24 0.263 0.279 0.329 0.352 0.369 0.4 Irradiances factored to yield 680 W/mA2 in 300-800nm band IR Q/Suprax (UV) 0.001119 0 0 0 0.001119 0 0 0 0 0 0 0 0 0 0 0 0 0 0.003357 0.005595 0.010071 0.015666 0.023499 0.031332 0.050355 0.060426 0.07833 0.095115 0.129804 0.154422 0.168969 0.195825 0.23499 0.26856 0.294297 0.312201 0.368151 0.393888 0.412911 0.4476 Page 143 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 330 0.431 0.482289 332 0.449 0.502431 334 0.475 0.531525 336 0.495 0.553905 338 0.525 0.587475 340 0.549 0.614331 342 0.565 0.632235 344 0.566 0.633354 346 0.587 0.656853 348 0.614 0.687066 350 0.61 0.68259 352 0.635 0.710565 354 0.656 0.734064 356 0.685 0.766515 358 0.662 0.740778 360 0.675 0.755325 362 0.719 0.804561 364 0.714 0.798966 366 0.73 0.81687 368 0.813 0.909747 370 0.858 0.960102 372 0.767 0.858273 374 0.8 0.8952 376 0.827 0.925413 378 0.864 0.966816 380 0.962 1.076478 382 0.992 1.110048 384 0.974 1.089906 386 0.996 1.114524 388 1.028 1.150332 390 1.111 1.243209 392 1.126 1.259994 394 1.227 1.373013 396 1.642 1.837398 398 1.552 1.736688 400 1.243 1.390917 402 1.228 1.374132 404 1.241 1.388679 406 1.284 1.436796 408 1.473 1.648287 410 1.395 1.561005 412 1.551 1.735569 414 1.416 1.584504 416 1.369 1.531911 418 1.426 1.595694 420 1.644 1.839636 422 1.453 1.625907 424 1.472 1.647168 426 1.462 1.635978 428 1.466 1.640454 Page 144 of 148 430 432 434 436 438 440 442 444 446 448 450 452 454 456 458 460 462 464 466 468 470 472 474 476 478 480 482 484 486 488 490 492 494 496 498 500 502 504 506 508 510 512 514 516 518 520 522 524 526 528 1.466 1.487 1.5 1.566 1.714 1.616 1.616 1.57 1.563 1.573 2.267 2.031 1.796 1.811 2.127 1.835 3.267 2.476 2.541 5.277 2.487 1.922 2.924 1.837 1.813 2.289 2.516 2.651 1.952 1.842 1.898 3.012 2.089 1.871 1.888 1.898 1.973 2.005 1.94 1.927 1.934 1.963 2.013 2.031 2.021 1.995 1.971 1.98 1.966 1.978 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.640454 1.663953 1.6785 1.752354 1.917966 1.808304 1.808304 1.75683 1.748997 1.760187 2.536773 2.272689 2.009724 2.026509 2.380113 2.053365 3.655773 2.770644 2.843379 5.904963 2.782953 2.150718 3.271956 2.055603 2.028747 2.561391 2.815404 2.966469 2.184288 2.061198 2.123862 3.370428 2.337591 2.093649 2.112672 2.123862 2.207787 2.243595 2.17086 2.156313 2.164146 2.196597 2.252547 2.272689 2.261499 2.232405 2.205549 2.21562 2.199954 2.213382 Page 145 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 530 1.954 2.186526 532 1.932 2.161908 534 1.942 2.173098 536 2.008 2.246952 538 2.014 2.253666 540 2.113 2.364447 542 1.996 2.233524 544 1.977 2.212263 546 1.962 2.195478 548 1.921 2.149599 550 1.876 2.099244 552 1.84 2.05896 554 1.938 2.168622 556 2.17 2.42823 558 2.106 2.356614 560 1.936 2.166384 562 1.912 2.139528 564 1.749 1.957131 566 1.706 1.909014 568 1.77 1.98063 570 1.997 2.234643 572 2.032 2.273808 574 1.802 2.016438 576 1.624 1.817256 578 1.572 1.759068 580 1.654 1.850826 582 2.215 2.478585 584 2.034 2.276046 586 1.738 1.944822 588 2.118 2.370042 590 2.159 2.415921 592 1.996 2.233524 594 2.185 2.445015 596 1.607 1.798233 598 1.492 1.669548 600 1.471 1.646049 602 1.37 1.53303 604 1.263 1.413297 606 1.207 1.350633 608 1.184 1.324896 610 1.292 1.445748 612 1.494 1.671786 614 1.31 1.46589 616 1.631 1.825089 618 2.672 2.989968 620 2.172 2.430468 622 1.416 1.584504 624 1.181 1.321539 626 1.256 1.405464 628 1.251 1.399869 Page 146 of 148 630 632 634 636 638 640 642 644 646 648 650 652 654 656 658 660 662 664 666 668 670 672 674 676 678 680 682 684 686 688 690 692 694 696 698 700 702 704 706 708 710 712 714 716 718 720 722 724 726 728 1.51 2.325 1.246 0.959 0.927 0.834 0.856 0.876 1.013 1.571 1.431 1.12 1.071 0.86 0.801 1.103 0.763 0.762 0.855 1.037 0.575 0.682 0.912 0.526 0.567 0.514 0.738 1.065 1.214 2.331 1.16 0.73 0.603 0.432 0.688 0.347 0.327 0.298 0.31 0.275 0.424 2.069 0.594 0.302 0.289 0.254 0.297 0.384 0.592 0.817 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 1.68969 2.601675 1.394274 1.073121 1.037313 0.933246 0.957864 0.980244 1.133547 1.757949 1.601289 1.25328 1.198449 0.96234 0.896319 1.234257 0.853797 0.852678 0.956745 1.160403 0.643425 0.763158 1.020528 0.588594 0.634473 0.575166 0.825822 1.191735 1.358466 2.608389 1.29804 0.81687 0.674757 0.483408 0.769872 0.388293 0.365913 0.333462 0.34689 0.307725 0.474456 2.315211 0.664686 0.337938 0.323391 0.284226 0.332343 0.429696 0.662448 0.914223 Page 147 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00-2192 730 0.593 732 0.634 734 0.472 736 0.316 738 0.375 740 1.097 742 0.368 744 0.272 746 0.264 748 0.439 750 0.356 752 0.231 754 0.273 756 0.406 758 0.823 760 1.344 762 0.554 764 1.555 766 1.114 768 0.388 770 0.194 772 0.152 774 0.188 776 0 .2 1 778 0.247 780 0.388 782 0.327 784 0.225 786 0.171 788 0.335 790 0.693 792 0.137 794 0.18 796 0.326 798 0.632 800 0.233 Total Integrated Irradiance in 300-800nm Wavelength Band 607.6 W/mA2 0.663567 0.709446 0.528168 0.353604 0.419625 1.227543 0.411792 0.304368 0.295416 0.491241 0.398364 0.258489 0.305487 0.454314 0.920937 1.503936 0.619926 1.740045 1.246566 0.434172 0.217086 0.170088 0.210372 0.23499 0.276393 0.434172 0.365913 0.251775 0.191349 0.374865 0.775467 0.153303 0.20142 0.364794 0.707208 0.260727 680.0 W/mA2 Page 148 of 148