a 3M Environmental Laboratory Report No. E00 2192 Study Tit/ e 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:

Study Identification Number: E00 2192 Screening Studies on the Aqueous Photolytic Degradation of Perfluorooctonic Acid (PFOA)

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.

Sponsor Representative Date Page 3 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 03/ 30/ 01 03/ 3 I /O 1 04/ 18/ 0 1 04/ 19/ 0 1 Quality Assurance Statement Data 03/ 3 1 /O 1 03/ 3 1 /O 1 Draft Report 0411 910 1 04/ 19/ 01 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 Date Reported to Management I Study Director Phase Quality Assurance Unit 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 Study Personnel and Contributors ......................................................................................... 7 Table of Contents ................................................................................................................... 5 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 Signatures .............................................................................................................................. 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 FL) ....................... 140 Appendix G: Characteristics of the Spectral Output of the Suntest Instruments .................. 142 Page 5 of 148 BACK TO MAIN 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 lntertec 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 I O 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. lS2 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 (FenOs) 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 phot~ lysis? '~ 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 H202 was added as a well characterized source of .OH radicals?* 6 This was used to test for the propensity of PFOA to undergo indirect photolytic decomposition. The second matrix contained Fez03 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. 7g8 The third matrix contained a standard humic material, diluted to levels that have been shown to approximate an environmental environment." '

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  perfluoro hexanoic acid by LC/ MS. Structures of the pertinent compounds are illustrated in Figure 1.

Figure I. Structures of the Compounds Targeted by LClMS Analysis PFOA F F F F F F 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 pemuoronated olefins (e. g. c8F16) and 1 or 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 (kp) 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 p,, and a standard deviation of q,.)

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.

UVNisible analysis were performed following 3M Environmental Laboratory Method ETS 9 46.0 "Operation and Maintenance of the Hewlett Packard 8453 UV Visible Spectrophotometer" An aqueous saturated solution of PFOA was prepared and an initial UVNlS spectrum recorded. No absorbance above baseline over the range 190 1 I00 nm was detected.

Sample preparation for this analysis followed 3M Environmental Laboratory Method ETS 8 177.0 "Indirect Photolysis Screening Test in Synthetic Humic Water" or ETS 8476.0 "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 HZOZ at 1: l 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 4S" C. 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 f 3° C. The temperature of the chamber itself was allowed to drift to 70 t I 0° C. 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 Description 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.

Test Matrix Control Test Post Photolysis Sample Type (e. g. Fe203 Matrix Substance Target Analyte (Conditions) in water) (e. g. (PFOA) spike Table 1. Typical Sample Preparation Scheme Used in the Present Investigation I I water) I Ind. Photo. Sample Rep 1 + 0 + 0 Initial Ind. Photo. Sample Rep 2 + 0 + 0 Initial Ind. Photo. Sample Rep 3 + 0 + 0 Initial Ind. Photo. Sample Spike + 0 + + Initial Matrix Blank + 0 0 0 . Initial Matrix Blank Spike + 0 0 + Initial Direct Photo. Sample 0 + + 0 Initial Direct Photo. Spike 0 + + + Initial Control Blank 0 + 0 0 Initial Control Blank Spike 0 + 0 + Initial Ind. Photo. Sample Rep 1 + 0 + 0 Light Ind. Photo. Sample Rep 2 + 0 + 0 Light. Ind. Photo. Sample Rep 3 + 0 + 0 Light Ind. Photo. Sample Spike + 0 + + Light Mabix Blank + 0 0 0 Light Matrix Blank Spike + 0 0 + Light Direct Photo. Sample 0 + + 0 Light Direct Photo. Spike 0 + + + Light Control Blank 0 + 0 0 Light Control Blank Spike 0 + 0 + Light LC/ MS* With W/ out Y 4 YO2 X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X Ind. Photo. Sample Rep 1 + 0 + 0 NoLight 11 X X Ind. Photo. Sample Rep 2 + 0 + 0 No Light Ind. Photo. Sample Rep 3 + 0 + 0 No Light Ind. Photo. Sample Spike + 0 + + No Light Matrix Blank + 0 0 0 No Light Matrix Blank Spike + 0 0 + No Light Direct Photo. Sample 0 + + 0 No Light Direct Photo. Spike 0 + + + No Light Control Blank 0 + 0 0 No Light Control Blank Spike 0 + 0 + No Light + = added to test vial; 0 = NOT added totest vial; *Duplicate sets, one with H202, one wit1 material test where H202 was not added) X X X X X X X X X X X X X X X X X X u t (excludes GC/ MS* With W/ out Y 4 w 2 X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X e Humic 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 21 9 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 I. 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 051 5 PFOAfe 21,3Il45,55 showed low recovery. The GC/ MS CCV R0602046 failed to pass criteria. The samples numbered 0515 PFOAshw 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 051 5 PFOA 08, 50, 14 and 48 failed to meet spike recovery criteria in the pH 7 buffer portion of this investigation. Control sample 051 5 PFOA 48 showed low PFOA recovery while four sample triplicate sets for PFPA analysis had high RSD. The GC/ MS samples numbered 051500 PFOAfe 067,077 and 090 failed to meet spike recovery criteria in the iron oxide portion of the investigation. Two samples, 051500 PFOAshw 12 and 051 500 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 k 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 Q5% 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 =5% 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 GUMS 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 H202 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 H202 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 Dir. Photo. Sample' Initial Time Point ND Nc Ind. Photo. Sample' Exp. to Light ND Nc Dir. Photo. Sample` Exp. to Light ND Nc 1 Ind. Photo. SamDle' Not Exuosed ND Nc 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 H202 ~

Dir. Photo. Sample' Not Exposed I M )I Nc Detection Limits I 0.292 I r4 0.178 Sample Conditions I PFPA I PFHxA 164 hour Exposure Initial Time Point Ind. Photo. Sample' nanomoles nanomols ND Nc Sample Conditions Fe203 W/ yO; Initial Time Point Fez03 WO/@ O,` Initial Time Point F e z 0 3 W/ yO: Exp. to Light F e z 0 3 w/ yO: Not Exposed Detection Limits 69.5 hour exposure Fez03 WO/ yO, ' Exp. to Light Fe203 WO/ yO, ' Not Exposed Mass PFPA PFHxA PFHpA PFOA Volatiles Balance (percent) ND ' Nc 0.846 122 ND 106% ND Nc 0.808 121 ND 105% ND NC 1.27 105 ND 91.8% ND Nc 0.808 116 ND 101% ND Nc 0.769 121 ND 105% ND NC 0.789 118 . ND 102% nanomoles nanomoles nanomoles nanomoles nanomoles 0.729 f 0.348 0.500 2.03 0.0095` 0.394 99.3% 0.385 97.9% 0.404 55.1 95.8% 0.385 56.8 98.8% 0.356 56.4 ND 97.9%

Sample Conditions PFPA 164 hour exposure nanomoles Humic Water` Initial Time Point ND Water* Initial Time Point ND Humic Water` Exp. to Light ND Humic Water` Not Exposed ND water' Exp. to Light NA 0.356 I 55.7 1 96.8% 0.2019 I 4.06 I 0.0095* I PFHxA PFHpA PFOA Mass Balance nanomoles nanomoles nanomoles (percent) ~

Nc 0.433 55.7 96.8% Nc 0.469 60.2 105% Nc 0.404 52.0 90.5% NA N4 NA NA Nc 0.394 59.3 103% Vials initially contained 57.9 nMoles PFOA. 1. Indirect Photloysis Sample: These samples had H202 added as a radical source, results are from triplicate samples. 2. Direct Photolysis Sample: These samples did not have H 2 4 added, 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.

2b. Matrix: Fe203 in Water, With and Without H202 WaterZ NotExposed I ND Nc I 0.391 I 55.0 I 95.6% Vials initially contained 116 nMoles PFOA. 1. Samples had H202 and Fe2O3 added as a radical generating species, results are from triplicate samples. 2. Samples contained just Fe203 as 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.

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 Dashed lines indicate the l a limits. Solid line indicates the maximum slope. 0 0 The Fez03 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 H202, was pooled to determine the experimental error of the analysis. This data, shown graphically in Figure 2, yielded an estimated environmental half life of 2 349 days. The exact mathematical solution is shown in Appendix C.

Figure 2. Pooled concentration data from the iron oxide rich matrix.

I 0 0 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 k 10% under all experimental conditions. A minimum half life for PFOA based upon these preliminary studies was calculated to be 2 349 days.

Page 18 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 References 1. Fafe, Transporf and Transformation Test Guidelines, OPPTS 835.5270 lndirecf Phofolysis Screening Test; EPA712 C 98 099; United States Environmental Protection Agency, U. S. Government Printing Office: Washington, DC, 1998, pp. 1 22.

2. Of CD Guideline for Tesfing of Chemicals, Phofofransformation of Chemicals in Wafer Direct and lndirecf Phofoysis, (Draft Document); OECD, 2000, pp. 1 59.

3. Scrano, L.; Bufo, S. A.; Perucci, P.; Meallier, P.; Mansour, M. Photolysis and Hydrolysis of Rimsulfuron. fesfic. Sci. 1999, Vol. 55, pp. 955 961.

4. Nubbe, M. E.; Adams, V. D.; Moore, W. M. The Direct and Sensitized Photeoxidation of Hexachlorocyclopentadiene. Waf. 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. Phofochem. 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. 429 435.

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:

09/ 2A/ Management Date 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 ETS 9 44.0 ET S 9 49 .O ETS 8 182.0 ETS 8 177.0 ETS 8 176.0 ETS 8 181 .O Operation and Maintenance of the Hewlett Packard 8453 UV Visible Spectrophotometer Operation and Maintenance of the Sunlight Exposure System, Immersion Unit, and Recirculating Water Chiller System Routine Maintenance of Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent Gas Chromatograph/ Mass Spectrometer Analysis of Fluorochemicals by Archon Purge and Trap Autosampler, Tekmar Purge and Trap Concentrator and Agilent Gas Chromatograph/ Mass Spectrometer Indirect Photolysis Screening Tests in Synthetic Humic Water Preparation of Samples for Photolytic Exposure Studies in Aqueous Matrices 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 METEOD ANAlLYSIS OF PHOTOLYSIS SAMPLES FOR FLUOROCHEMICALS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH MASS SPECTROMETRY DETECTION Method Number: ETS 8 181.0 Adoption Date: jd/ L 4 1 Effective Revision Date:

Approved by:

uc/ aJ ~t )

Date &/& Laboratory Management Page 22 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Comuound Perfluorooctanoic acid Pefluorooctanesul fonate Perfluorooctanesul fonamide N methylpefluorooctanesulfonamide N ethylperfluorooctan~~ ulfonamide 2( NmefhylperfluoIooctanesulfonamido) ethyl alcohol ~( N e~ yl~~ uo~ terfluorooctanesulfonamido)

ethyl alcohol Acronvm COfnOOUIId Acronvm PFOA Pduorobutanoic acid PFBA PFOS Perfluorobutanesutfonate PFBS FBSA FOSA Pdluorabucanesu! fanamide N MeFOSA N methylperfluorobutanesulfonamide N MeFBSA N EtFOSA N eth ylperfluorobutanesul fonamide N EtFBSA N MeFOSE OH 2( N methylperfluorobutanesulfonamido) N MeFBSE OH ethyl alcohol N EtFOSE OH 2( N ethylperfluorobutanesulfonamido) N EtFBSE OH ethyl alcohol I I 1.3 Compatible matrices for analysis. Aqueous (Millipore ASTM Type I water), buffered water, lake water, sea water and metal slurries (TiO2, Fez03, 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 detectiodquantification. The analysis is performed by separating target analytes on an HPLC analytical column such as a Dionex NGl (35x 4.6mm, lOpmpar! kle), Betasil C18 column (50x2 mm, 5 pm particle) or equivalent using an ammonium acetateh4eOH 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 (ie. 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 Sohare.

ETS 8 1 8 1 .O Page 2 of 15 AnaIysis of Photolysis Samples for Fluorochemicals by HpLC/ MS Page 23 of 14% BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 3.0 3.1

3.2 3.3 3.4 3.5 3.6 3.7 3.8 ,3.9 DEFINITIONS Calibration Standard. A dilution of various amounts of a stock, intermediate or purchased standard to achieve standard solutions in a concentration range of interest.

CaIibration Curve. The graphical relationship between known values, such as concentration of a series of calibration standards and their instrumental response.

Internal Standard Quantification. Process of establishing a relationship betw; een 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. External Standard Quantification. Process of establishing the concentration of a target mdyte by plotting the theoretical amount (in units of ppb or ppm, etc.) versus the response of the target andyte( 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 masdvolume analyzed, if noted, must be compensated for by a factor applied to the value. 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 of r may range .from 1 to +l. A value of +1 denotes perfect direct functional relationship between two variables. A value of 1 also denotes a perfect inverse relationship. When r = 0, there is no effect of one variable upon the other variable. 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. Internal standard. A horn amount of a compound or element similar in analytical behavior to the compound( s) or element@) 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 of the applied analytical method. Samples are to be quantified using the internal standard. 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@). A surrogate may be added to samples along with the test analyte &re 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. Continuing Calibration Verification (CCV). Standards analyzed during an analytical run to verify the continued accuracy of 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 18 1.0 Page 3 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLClMS Page 24 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 3.10 Solvent Blank. A sample of analyte fiee medium (for example, methanol, 1: 7 diluted buffer: methanol) that is not taken through the sample preparation process. This blank is used to evaluate instrument contamination.

Blank. For photolysis studies, there are multiple blanks to adequately represent the variables of the study exposed, Unexposed and Day 0 samples WitWwithout 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 baselinebackground and monitor for analytical interference or suppression of target analyte( s) from the matrix. 3.11.1 Matrix 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 II 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. 3.1 1 3.12 Limit 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 simplie 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. Sample Triplicates. Three samples taken from and representative of the same sampIe some and separately carried through aI1 steps of the extraction, photolysis and analytical procedures in an identical manner. There are multiple sets of triplicate samples to adequately represent the photolytic variables of the study (Exposed, Unexposed and Day 0 WiWwithout peroxide addition). Triplicate samples are used to assess variance of the photolytic method, including sample preparation, photolysis, and analysis.

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 of triplicate samples to adequately represent the photolytic variables of the study (Exposed, Unexposed and Day 0 witWwithout peroxide addition). This is used to document method performance and matrix effects by comparing recoveries from the different matrices and sample types.

Relative Standard Deviation (RSD). A measure of precision defined as the standard deviation of three or more values divided by the average of the values and multiplied by 100. (Also reported as Coefficient of Variation (CV)). Analytical Spike (AS). Prepared by adding a known mass of target analyte( s) to a specified amount of 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 of the matrix on recovery efficiency. There are multiple 3.13 3.14 3.15 3.16 ETS 8 18 1 .O Page 4 of 15 Analysis of Photolysis Samples for Fluorochemicals by HpLClMS Page 25 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 3.17 3.18 3.19 3.20 3.21 3.22 4.0 types of spiked samples to adequately represent the photolytic variables of the study Gxposed, Unexposed and Day 0 ; withlwithout peroxide addition.) 3.16.1 Matrix 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 II water) sample containing the test analyte or blank to which a known mass of target analyte( s) is added prior to analysis.

Accuracy. The closeness of 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 of the study, the acceptance criterion is 75% to 125% of the nominal value. Dilution. A step in the sample preparation procedure in which a solvent (Le. methanol, acetone) is added to the test analyte/ sample matrix (i. e. water, buffer, etc.) to prepare it for instrumental analysis. Atmospheric Pressure Ionization (MI): The Agilent Technologies HPLC 1100MSD 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. Electrospray Ionization @S, 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 of a strong electrical field. Mass Spectrometry, Mass Spectrometer (MS), Mass Spectrometer Detector (MSD): The API HP1100 MSD system equipped with a quadrupole mass selective detector, Ions are selectively discriminated by mass to charge ratio (dz) and subsequently detected, Geometric Mean of the calibration curve: The square root of 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 of 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.

. 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 .O Page 5 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLC/ MS Page 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 runs.

5.0 INTERFERENCE 5.1 Contaminants in solvents, reagents, glassware, and other sample processing or analysis hardware may cause interference. Use the routine analysis of laboratory method blanks to demonstrate that there is no such interference. Contamination fiom columns, HPLC tubing, and detector components may cause interference at low detection levels. The routine analysis of solvent blanks must be used to demonstrate that there is no such interference. 5.2 6.0 EQUIPMENT 6.1 6.2 Analytical balance sensitive to 0.1 mg. Hewlett Packard (HP) 1100 HPLC System, or equivalent.

6.2.1 Pump, binary, Model G1312; Quaternary, Model G1311 A, or equivalent.

6.2.2 Solvent degasser, Model G1322A or equivalent.

6.2.3 Autosampler, ALS Model G1313A, variable injection volume or equivalent.

6.2.4 Column heater, Model G1316A, or equivalent.

Betad* C18,50 x 2 mrn; Dionex IonPac@ NG1 Guard column, 4 x 35 mm; or equivalent. Mass spectrometer. Hewlett Packard MSD Model G1946A, or equivalent.

Refrigerator capable of maintaining 4 5 3 "C. Data system. A personal computer capable of controlling the HPLC system as well as recording and processing signals from the detector. 6.3 6.4 6.5 6.6 ETS 8 181 .O Page 6 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLCMS Page 27 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 6.7 System control/ data analysis software: Hewlett Packard ChemStationQ 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 7.2 7.3 Labels 7.4 7.5 Pasteur pipets, gIass, disposable 7.6 7.7 Volumetric flasks, various sizes 7.8 Beakers, glass, various sizes 7.9 Vials, 40 mL, VOA (I Chem or equivalent) Crimp cap autovials, 1.8 mL Graduated pipets, glass, disposable, 1 mL to IO mL Hamilton Gastight@ syringes (precision f 1% of total volume), 10 pL 1000 pL Automatic pipettor, capable of dispensing 10 5000 pL 8.0 REAGENTS AND STANDARDS 8.1 8.2 8.3 8.4 Methanol (MeOH). HPLC/ SPEC/ GC grade from EM Science, or equivalent Acetone. HPLC/ SPEC/ GC grade from EM Science, or equivalent ASTM Type I1 Water. Water with lower resistance must not be used.

Ammonium acetate, 2 mM in water. This solution is chromatographic solvent A (see ammonium acetate crystals to a l L volumetric flask containing about 500 mL water, adding 10 mL of methanol, diluting to the mark with 18.0 MQ water and mixing.) 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:

85.1 Fluorochemical or target malyte prepared in acetonitrile (or suitable analytical solvent). (Example: A stock solution is prepared at a concentration of approximately 30,000 p g /d 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 of 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, Section 12.2.1). (Example: An acceptable eluent solution is made by adding 0.15 g 8.5 ETS 8 1 81.0 Page 7 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLCMS 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 OC f 3 "C or room temperature, until analysis can be performed. Document storage conditions on sample prep worksheet with date and initials.

10.0 QUALXTY CONTROL 10.1 Calibration Standards. Calibration standards (Section 11) used to .generate a calibration 10.2 10.3 10.4 10.5 curve should be prepared in the same type of solvent or matrix as in the study samples. 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. Broad calibration ranges (greater than three orders of magnitude between low and high standards), may require use of a quadratic fit of the data and requires more points to adequately represent the calibration range. Continuing Calibration Verification (CCV). Analyze a mid range calibration standard after a maximum of every fifteen samples. 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 of quantitation (LOQ) of the instrument. If analyte carryover is a problem, use back to back solvent blanks. Sample Triplicates. Analyze all sets of triplicate samples to provide a measure of the precision of 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 of each study sample batch. AX1 samples (matrix and control samples, blanks and spikes) from a specified exposure type or time may be analyzed within the same analytical batch.

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 k 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 fmal 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 .O Page 8 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLCMS 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 of five calibration standards is required to generate linear regression for target analyte( s). If the calibration curve residuals are greater than 25% deviation fiom the theoretical value, quadratic curve fitting and/ or dropping lowhigh 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. 11.1.1 Use the following documentatiodfootnotes maybe used to justify dropping highflow curve points.

1) "Higb/ 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 &om the curve to disqualify a data range that may have been significantly affected by background levels of the analyte." 3) "Hiwow calibration point( s) (list points) were excluded as they were not within the 4 2 5 % 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 SootWare window, turn the system "on" to: turn on the 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. MSD set up. Turn the MSD "on" in the software to equilibrate the system. 12.2.1 Check the level of nitrogen in the tank and ensure there is enough to complete the 12.2 impending run.

12.2.2 Clean the MSD according to the Equipment Procedure ETS 9 34.0 Operation and Maintenance of HP LCMS System.

ETS 8 181.0 Page 9 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLCMS Page 30 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Polarity Acquisition mode Gain Fragmentor Dwell time Carillaw voltaae 12.2.3 Perform a Check Tune or Autotune to ensure system operational qualification and performance verification of the MSD. Log the Tune results and keep a copy with the analytical raw data ~~

NegaUve (or Positive)

SIM (or SCAN)

1 .O (up to 7.0)

70 (may be set to one voltage, or ramped for each ion)

183 msec (time is a function of the amount of ions).

3500. or eauivalent 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': I MSD: 1 Ionization mode I API ES lor API APCI)

Drying gas Nebulizer pressure Nitrogen, or equivalent 30 pig, or equivalent Drying gas flow Drying gas temp ~ ~.~

8 Umin, or equivalent 300' C, or equivalent 12.3 LCCheck TIME (MIN)

0.0 1 .o 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 LDjtemperature, 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:

*% A %0 FLOW RATE 60 40 0.3 mUmin 60 40 0.3 mUmin 4.0 11.0 5 95 0.3 mumin 5 95 0.3 mUmin Page 31 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 ~

AUTO SAMPLER PROORAM:

INJECTION VOLUME: 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:

None 5.0 vL, or equivalent I AUTO SAMPLER: I ALS Model GI 31 3A I I 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.

<IP_ADDRESS> 123.5.2 <IP_ADDRESS> <IP_ADDRESS> Verify that all samples and standards are positioned correctly. Enter the identification code for each standard and samples. For solvent blanks, identie the solvent and the traceability number. Use one injection per sample. 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 HP1100iMSD 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 injectiodrun number (e. g. for samples acquired on "Hillary7', data files shall be "HILLOOOl". . . . "`~ LLOO##" '). 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 of samples can be set up in the sequence table with each set bracketed by calibration standards. Analyze a single continuing calibration standard (CCV) after a maximum of 15 injections. Solvent blanks shall be analyzed before and after the CCV and before method and control blanks, if Page 11 of 15 ETS 8 18 1 .O Analysis of Photolysis Samples for FIuorochemicals by HPLCNS 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 cany over from the highest standard concentration. Solvent blanks may also be used to separate groups of samples and evaluate for carry over problems fiom 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 of 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.

Integration Codes. The following integration codes may be utilized to document what type of manual integration was performed. 13.2 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 Matrix spikes. Calculate the percent recovery for each of the matrix spikes. Calculate the matrix spike percent recoveries using the following equation:

% Recovery = (observed stiked samtde result  observed samule result) x 100 Using the observed matrix spike recoveries, calculate the average spike recovery. Nominal amount spiked '

13.4 Accuracy, Calculate the accuracy of each calculated calibration standard and CCV samples using the following equation. Accuracy = (Measured Conc.) x 100 Nominal Conc.

Page 12 of 15 ETS 8 18 1 .O Analysis of PhotoIysis Samples for Fluorochemicals by HPLCMS 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 Samde Set x 100 Average of Sample Set .

14.0 METHOD PERFORMANCE 14.1 14.2 14.3 14.4 14.5 14.6 Coefficient of Determination (9). The coefficient of determination (r') for the calibration curves should be 0.990 or greater. The curves should be examined closely for linearity and intercept, particularly for accuracy of quantitation at the low and hi& ends of the curve. The accuracy of 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. Calibration Standards. The acceptance criterion for the calibration standards is that the accuracy of each standard is 75% to 125% (k 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. 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 of the analytical run.

Continuing Calibration Verification. If the accuracy for the amount of 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. Solvent Blanks. Solvent blanks should show no more than a 5% canyover 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 of the designated LOQ value are observed in sequential solvent blanks, this is indicative of instrument contamination. The instrument shall be serviced by thoroughly cleaning the electrospray source, and replacingkleaning c o l m s , 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. Matrix 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 of the designated LOQ value observed in matrix blanks are indicative of matrix effect,

ETS 8 181 .O Page 13 of 15 Analysis of Photolysis Samples for Fluomchemicals by IPLCIMS Page 34 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 .

14.7 14.8 14.9 sample contamination or instrument contamination. Evaluation of the solvent and control bIanks 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. Control 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. Limit of Quantitation (LOQ). The LOQ is equal to the lowest acceptable standard (i. e. % accuracy is S 25 % nominal value) in the calibration curve that is greater than 4 times the level of the matrix blanks. 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 required to have technical justification for why the data is used, documented in the final report and raw data. Consult with the Te, m Leader or designee for direction, and for find 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% of 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 rt 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 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 SuitabWty. Without performing a method validation, system suitability can be demonstrated by acceptable instnunental checks (e. g. abbreviated &z check tune, 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 15.2 15.3 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containers to dispose of glass pipettes. 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. Use smaller bore columns when possible to minimize waste generation.

ETS 8 181 .O Page 14 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLCMS Page 35 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 16.0 RECORDS 16.1 16.2 16.3 16.4 16.5 16.6 16.7 16.8 Print hard copies of all graphics and data analysis summaries for archiving.

Sign and date all graphics and label with instnunent ID. Fill out appropriate preparation worksheets completely, making sure to include all initids and dates, along with the study number and sample identification. 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. Print chromatograms, reprocessing sequence and batch reports for all analyses. Print calibration tables and curve information and store in the raw data file. Enter all standard preparation infomation in the standards preparation logbook. Make a photocopy of the logbook page and include the copy in the raw data file. Archive electronic data to appropriate media when necessary.

17.0 ATTACHMENTS 17.1 None.

18.0 REFF, RENCES 18.1 18.2 ETS 9 34.0, Hewlett Packard 1 1OOiMSD Equipment Procedure. 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 18 1.0 Page 15 of 15 Analysis of Photolysis Samples for Fluorochemicals by HPLClMS 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:

Laboratory Manager fl Date ETS 8 176. OPreparation of Samples for Photolysis Studies in Aqueous Matrices Method Page 1 of 18 Page 37 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Perfiuomoctanoic acid Perfluorooctanesulfonate PcrfYuor ooctanesulfonamide 1.0 SCOPE AND APPLICATION PFOA Peffluorobutanoic acid PFBA PFOS Perfluorobutanesulfonate PFBS FOSA Perfluorobutanesul fonamide FBSA 1.1 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 of chemical or electronic excitation transfer fkom light absorbing species in the water. The simplest reaction involves the absorption of W energy by hydrogen peroxide (H202) to produce 2 hydroxl radicals. These may react with any species in the water, including solvent, b a e r , dissolved organic material and target material. Use of water and H202 is 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 (' 0 2 ) which may promote indirect photolysis of the test substance. Another transient species photochemically produced by the reaction of W light and dissolved organic materials (humic) is hydrogen peroxide (H202) which may react further to form the hydroxyl radical. The addition of HzO2 to test solutions may be utilized as a free radical 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( II1) which undergo photoreduction to Fe( I1) and free radicals or addition of Ti02 as a catalytic surface for indirect photolysis may also be evaluated within this method.

CompatibIe analytes. Test substance and degradation products for photolytic exposure include but are not limited to: 1.2 Z( N methylperfluoro octanesulfonamido) ethyl alcohol Z( N ethylperfluorooctanesuIfonamido) ethyl alcohol 1 oeduorooctene 1 Comaound I Acronym I Compound I Acronym I N MeFOSE OH Z( N methylperfluorobutancsulfonamido) ethyl N EtFOSE OH 2( N ethy$ e~ uorobutanesuIfonamido) ethyl alcohol alcohol  1 ueffluombutene N methylperfluorooctanesulfonamide 1 N MeFOSA N methylperfluorobutanesulfonamide I N MeFBSA ~~

N ethvluemuomoctanesulfonamide I N EtFOSA I N ethvl~ erfiuombutanesulfonamide 1 N EtFBSA I N MeFBSE OH N EtFBSE OH 4 ~  ~~

r . . , and other C., thru Cla homologues. and oolymeric materials based on the aforementioned comKunds. I ETS 8 176. OPreparation of Samples for Photolysis Studies in Aqueous Matrices Method Page 2 of 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:

C2 lMpcrfluomthanc( lH pE2) IH perfluoroethane (IH pfCz) 1 C, 1 H perfluompropane (IH pE3) LH perfluobpmpane (ZH plC3) pcffluom 1 propene (pE3 lenc)

C, perfluom l buttoe (pR; Q lene) Pcrtluom 2 butenc (pE4 2ene) 2H pcfflwmbutane (2H prOr) 1H perfluombutane (1H pE4)

C, 2H perfluororpcnme OH PES) perfluom 1 pen~ e @fCS lcne) pcrfluorPZ pentcac @CS Zenc)

Cs perUuom 2lmene (pfC6 2cnc) lfi perfluorohcxaoe (1H pE6) perfluom 1 hcxene (pE6 lenc) IH perfluomhcxme (IHpE6)

C, 2H pcrfluorohcptane (2H pE5) Perfluom l hcptene (ptC7 lene) IH ptrfluoroheptane (IH ptC'?)

Ce perfluom lsctene (pfC8 lene) 2H perfluomoctane (2H pE8) Perfluor0 2 octene (pfC8 2ene) IH perfluomme (IH pE8) 2H pcrfluorohexene (2H pE6)

1.3 Acceptable matrices. Aqueous solution of test substance including but not limited to the following matrices: pH 7 phosphate b a e r , 18.2 MR 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 quanti@ 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 LCMS and dynamic purge and trap GC/ MS. When required, the addition of 30% H20z 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 OC. Samples are exposed to approximately 261 W/ m2 of 3 10 800 nm photo irradiance for a specified number of %hour periods. An %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 of days to expose samples is determined by the Team Leader. The amount of irradiation received by the samples may be monitored in one of the following three ways: 1) calculating the total wattage per length of exposure 2) use of a radiometer to measure irradiance output, a d o r 3) use of a quinine monohydrochloride dihydrate (QMD) actinometer solution exposed along with the samples and monitored for change 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 LCMS are prepared for analysis by diluting the 5 mL sample volume with 30 mL of suitable analytical solvent (e. g. methanol) containing internal standard. The GCMS study samples are stored inverted prior to purge and trap GCMS anaIysis.

ETS 8 176, OPreparation of Samples for Photolysis Studies in Aqueous Matrices Method Page 3 of I8 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 Fe203.

Sample Rep 2 + 0 + 0 Time 0 Sample Rep 3 + 0 + 0 Time 0 Sample Spike + 0 + + Time 0 MatrixBlank + 0 0 0 Time 0 MatrixBlankSpike + 0 0 + Time 0 Control Sample 0 f + 0 Time 0 Contml Spike 0 + + + Time 0 Control Blank 0 + 0 0 Time 0 Control Blank Spike 0 + 0 + Time 0 kc+ p.,.

7 0 + 0 Exposed Sample Rep 2 + 0 + 0 Exposed Sample Rep 3 + 0 + 0 fipo= d Sample Spike + 0 + + Exposed Matrix Blank + 0 0 0 Exposed Matrix Blank Spike + 0 0 + Expo= d Cantml Sample 0 + + 0 Exposed Conhol Spike 0 + + + Exposed Control Blank Spike 0 + 0 + Exposed Control Blank 0 + 0 0 Exposed _I___

7 Ssmp e Rep 1 + 0 U m X p i Z Sample Rep 2 Sample Rep 3 Sample Spike Matrix Blank Control Sample I Matrix Blank Spike , CMltrol Blank Unexposed Unexposed Unexposed Unexposed Unexposed Unexposed Unexposed Unexwsed LCMS 7?%= mEz WA Hi01 X X X X X X X X X X X X X X X X ..

X X X X  GCMS 77E YEz HzOr H20,

X X X X X X X X X X X X ._ x X X X X X X X w/ H24, One set w/ o H202 3.0 QUALITY CONTROL DEFINITION/ FREQUENCY/ PERFORMANCE CRITERIA 3.1 Blanks 3.1.1 Definition: Matrix 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 ETS 8 176. OPreparation of Samples for Photolysis Studies in Aqueous Matrices Method Page 4 of 18 Page 40 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Matrix ID Matrix Blank Conbpi Blank Matrix descrintian Freauency Performance Criteria Example: 1 Replicate per light and backWJmd level of 0.0 1 M Phosphate Buffer, pH 7 Example: ASTM Type I1 Water dark exposure, for each h e point and for each analytical methodology. *e LOQ. target analfie shall be less than 25% the area Counts Of Matrix Descridion I I I I J Frequencv of Use Performance Criteria 3.2 Limit 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 Qual@ Control and Performance Criteria: 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 blaaks and indicates good accuracy (2 25%) of the nominal calibration standard concentration.

3.3.1 Definition: Three aliquots prepared as representatives of the same sample source (i. e. test substance) and carried through all steps of the photolytic study process and analytical procedures in an identical manner. The results fiom triplicate analyses are used to evaluate variability of the total method, including sample preparation, photolytic process and analysis. 3.3.2 Petformcmce Criteria: The samples in the test matrix will be prepared in triplicate. Each replicate wilI 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 andor GCMS). See the following table: 3.3 Sample Triplicate Test Matrix containing test analyte( s)

3.4 Control Sample 3.4.1 Definition: A known matrix containing the test analyte( s) carried throughout the entire analytical procedure. This is used to document laboratory performance (Le. 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 8 176.0Preparation of Samples for Photolysis Studies in Aqueous Matrices Method Page 5 of 18 Page 41 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 with target analyte( s) just prior to analysis 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 the data. 3.4.2 Performance Criteria. One control sample will be prepared per matrix, per treatment type. See the following table:

The analyst shall accept Il) latrixDescription Control Matrix rmdtest substance spiked with target analyte( s) just prior to analysis Test Matrix without test substance, spiked with target analyte( s) just prior to anaiysis Control Matrix without test substance, spiked with target analyte( s) just prior to analysis Control matrix with test anal@(@ added 1 Replicate per treatment type. of 100 2 25%. Ifspike recoveries are greater than 125% or less than 7% document that the spike sample is out of the specifications and justie, if possible, the reason, 1 Replicate per treatment type.

~

1 Replicate per treatment type. Freauencv of Use I Performance Criteria 1 3.5 Analytical Spike (AS) 3.5.1 Deftition: Prepared by adding a known mass of target analyte( s) to a specified amount of a diluted andor 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. LCMS andor GCMS). In addition, one matrix blank spike and one control blank spike will be prepared. See the following table:

I Performance Criteria 1 I Matrix Descriotion I Freauency of Use E s t Ma& andtest substance, spiked I 1 Replicate per treatment type. I I ETS 8 176. OPreparafion of Samples for Photolysis Studies in Aqueous Matrices Method Page 6 of 18 Page 42 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Matrix Description Sample diluted with 30 mL of internal standard compound dissolved in a suitable analytical solvent to be quantified using the internal standard providing that the response of the internal standard is consistent (k 5% relative). Use of external calibration methodology requires written justification by the Team Leader. 3.6.2 Surrogaie DeJinition (applies io L W S and GWSsampies): A known amount of a compound similar in analytical behavior to the target analyt~( s) of interest may be added to all samples and standards (pre or post irradiation, at the discretion of the Team Leader) and carried through the nzmaining 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.

Freauencv of Use Performance Criteria Every LclMS sample analyzed The Coefficient of Variation, or %RSD shall be calculated for the area response of all appropriate samples per analytical batch. The analyst shall accept %RSD values of 45%. The recovery and precision of the surroEates Note: Internal standardr are used in all experiments. The use of surrogate standards may or may not be used.

with surrogate compound spiked into it.  should be 100 %? S% and 4 5 %, respectively. Unacceptable values shall be documented and Every sample analyzed justified, if poss,. le.

3.7 Other Definitions. 3.7.1 3.7.2 3.7.3 3.7.4 3.7.5 3.7.6 3.7.7 Test AnalyteISubstance: Any substance (mixture or controlled compound) added or administered to the test system for the purpose of chemical analysis. Degradation Product(@: Secondary d y t e s of interest produced as a result of chemical reactions during the photolysis and monitored (qualitatively or quantitatively) during the sample analysis procedure. 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 of the study, a by product or degradation product that is monitored (qualitatively or quantitatively) during the sample analysis procedure. Test Matrix: The physical matrix in which the study will be conducted. Also referred to as the test system. Control Matrix: A known physical matrix to be included with the study for comparison with the test matrix. 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 percent (% RSD). 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. OPreparation of Samples for Photolysis Studies in Aqueous Matrices 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 4.1 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 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.

4.2.1 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 of cooling water, which poses a threat of electrocution during the handling of 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 of the reactor and exposed glass vials become extremely hot. 4.2 Cautions 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 a11 possible contaminants by using fresh reagents, analytical grade solvents and clean glassware during the sample preparation processes.

6.0 EQUIPMENT 6.1 6.2 Analytical balance sensitive to 0.1 m g Photoreactor: Suntest CPS+, =Si, or equivalent, equipped with a xenon arc lamp and capable of producing integrated inadiance 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. Water recirculating cooler capable of maintaining temperature at 25 O C f 5 OC, from Poly Science, Model 1 177 P or equivalent. Agilent Technologies W 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 GI 1 ZOA, Thermostatted Cell Holder: Model 0845 1 601 04, or equivalent. 1.0 cm path length quartz spectrophotometer cell fiom Hewlett Packard, or equivalent. 6.3 6.4 <IP_ADDRESS> 6.4.2 Long Path Length Cell Holder, Hewlett Packard (# 89076C) or equivalent.

ETS 8 176. OPreparation of Samples for Photolysis Studies in Aqueous Matrices Method Page 8 of 18 Page 44 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 <IP_ADDRESS> 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 W Visible Spectroscopy, GI 1 16AA, Rev. B. 01.02, or later. 6.4.4 PC Computer capable of d n g appropriate analysis software to acquire and report data. Centrifige capable of maintaining > 2000 rpm for 10 minutes at ambient temperature. Radiometer (optionaI) capable of monitoring the energy from a xenon source from 290 to 480 ntn over time. Model PMA2100, Version 1.16, Solar Light Company, Inc., or equivalent. Consult the appropriate 3M SOP for instructions. 6.5 6.6 7.0 7.1 7.2 7.3 7.4 7.5 7.6 7.7 7.8 7.9 7.10

7.1 1 SUPPLIES AND MATERIALS 40 mL amber and clear glass VOA vials with screw caps with septa. Crimp cap autovials 1.5 mL, caps, crimper, and decapper. Adhesive backed labels (return address size) for labeling quartz vials and autovials. Disposable glass graduated pipettes, 1 mL to 10 mL. Disposable glass Pasteur pipettes and rubber bulbs. Glass beakers, various sizes. Volumetric flasks, from 10 mL to 1000 mL. Hamilton Gastight@ syringes (precision f 1% of the total volume), 5 pL to 1000 pL. 10 mL Bottle top dispenser, Calibrex, Model # 5 1 1, or equivalent. Adjustable repeater pipette, Wheaton Step pette 4 1 1, or equivalent, equipped with the appropriate volumetric range pipette tips. Ziploc@ plastic bags, or equivalent.

8.0 REAGENTS AND STANDARDS 8.1 8.2 8.3 Methanol (MeOB). HPLC/ SPEC/ GC andor purge and trap grade (EM Science, or equivalent. Acetonitrile (ACN). HPLC/ SPEC/ GC and/ or purge and trap grad fiom EM Science, or equivalent. Aqueous Matrix, includes but is not i i t e d to the following matrix types: 8.3.1 ASTM Type I water. Milli Q@ or equivalent, with a measured resistivity >I 8.0 MR cm. 8.3.2 0.01 M pH 7.0 Phosphate Buffer. Example: Weigh 1.36 g mzP04 into a 2 L volumetric flask and dissolve into 1 L of Type I water. Add 600 mL of 0.1 % NaOH. Adjust to pH 7.0 k 0.1% with 0.1% NaOH or dilute Hzs04 and dilute to the mark with Type I water for a fmal conc. 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. TiOz, Fe203). 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 of characterized soil or sediment in 5 mL of MilIi Q@ water.

ETS 8 176. OPrepuration ofSumples for Photolysis Studies in Aqueous Matrices Method Page 9 of 18 Page 45 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 8.4 8.5 8.6 8.7 8.8 Hydrogen Peroxide (Hz02). 30% aqueous solution fiom EM Science, or equivalent. Potassium phosphate. Reagent grade fiom JT Baker or equivalent. 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/ d by weighting approximately 0. I g of the appropriate substance into a lO. mL volumetric flask and diluting to the mark with solvent. This solution is then diluted to make appropriate working solutions. Test Analyte Solution: 8.7.1 Example for water soluble analvtefs): 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 analvtefs) 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 pl 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 the 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 can photooxidize during the exposure and decrease the indirect photolysis of the intended test substance. Evidence of this phenomenon (approximately 10% decrease in the concentrations of the final products) has been observed in a study here at 3M (EtFOSE OHphotolysis in pH 7 buffer, with and without presence of Meow).

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 p g /d test substance/ 100 pg/ mL target analytes. Addition of 10 pL of this target analyte spiking solution into the 35 mL diluted sample volume will result in approximately 140 ng/ ml and 30 ndnd concentrations for the test analyte and targ6t analyte( s), respectively.

We estimation of the degradation potential of the test analyte and subsequent degradation products is not always possible. Ifpossible, an analyticalpre screening of representative samples should be performed for accurate spiking. General rule of thumb has been that the test analjte spike amount be ETS 8 176. QPreporation of Samples for Photolysis Studies in Aqueous Matrices Method Page 10 of 18 Page 46 of 148 BACK TO MAIN 8.9 8.10 8.1 1 9.0 9.1

9.2 9.3 9.4 3M Environmental Laboratory Report No. E00 2192 approximately 25% 50% of the initial concentratioR The target analyte( s) spike amount has been 10 I 00 ng/ ml, depending on expected levels under specifc conditions, More than one spike solution may be utilized to adequately represent the levels in the samples. &ample: 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 amlyte spike concentrations.

Dilution Solution containing Internal Standard: The diluting solution shall contain internal standard at an area response level equivalent to approximately half the are response of 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 pL of stock solution (Section 8.6.1) to 4.0 L with MeOH to a concentration of 250 ng/ mL. Quinine monohydrochloride dihydrate (QMD). 90% from Aldrich Chemical. QMD solution: A 2% (wh) 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.

SAMPLE HANDLING 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. 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 of the procedure. This is particularly important for the GCMS samples. GCMS samples may only be turned upright immediately before being loaded onto the purge and trap autosampler. The LCMS 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 for H202 injection through the septa of the appropriate VOA sample vials at specified time intervals (See Section 12.6 and Section 12. I l , 7). The completed photolysis samples remain inverted and refrigerated at 1 5 'C prior to analysis by LC/ MS or sample purge and trap GCMS. Sample preparation prior to LCMS 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 fkom the glass VOA vial surface and to dilute the samples into a working analytical range. Day 0 study samples stored at 1 5 OC during the time of photolytic exposure. are removed and prepared for analysis at the same time as the exposed and unexposed samples.

ETS 8 176. OP~ eparatjon of Samplesfor Photolysis Studies in Aqueous Matrices Method Page 11 of 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 11.2 11.3 The compounds of interest must be characterized according to laboratory specifications. 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). All samples analyzed will be run against a standard curve containing varying amounts of target andytes, and a fixed amount of internal standard or surrogate compound. Refer to the appropriate LC/ MS and GCIMS methodologies for further analytical information.

12.0 PXOCEDURE 12.1 Obtain the absorbance spectra of the test compound in aqueous solution using a W Visible 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 d y t e s would be more likely to undergo indirect photolysis as the potential degradation pathway. 12.2 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 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, fest substance may be added after 5mL aliquots of 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 LCMS and GCMS analyses according to the following table: 12.3 12.4 12.5 ETS 8 176. OPreparation of Sumpies for Photolysis Studies in Aqueous Matrices Method Page 12 of 18 Page 48 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Sample Treatmenmype Matrix with test substance Matrix without test substance # of Samples # of Spikes 6 +HZ02 (3LC/ MS, 3GC/ MS)

6 HzOz (3LCMS ,3GC/ MS) 2 i& O2 (1LCIMS, lGCIMS)

2 HZ02 (1 ILCIMS, IGCMS) 2 +HzO2 (lLcIMS, lGCIMS)

2 HzOz (lLC/ MS, lGC/ MS)

2 +HZ02 (lLCIMS, lGCIMS)

2 H24 (lLC/ MS, 1GCIMS)

Control matrix with test substance Control matrix without test substance EXPOSED, UNEXPOSED, & DAY 0 12.6 12.7 12.8 12.9 13.0 2 +HzO2 (lLC/ MS, IGCiMS)

2 HzOz (IlLCIMS, lGC/ MS)

2 +HzOz (lLC/ MS, lGC/ MS)

2 HzOz (I lLCIMS, IGCIMS) 2 +HZ02 (lLC/ MS, lGC/ MS)

2 H24 (1 lLC/ MS, lGC/ MS)

2 +HzOz (ILC/ MS, lGCIMS)

2 Hz02 (1 lLC/ MS, lGCMS)

24x 3perexptypes72 16x3= 48 Separate the vials into three boxes labeled "Day 0," "Exposed," and "Unexposed." Initial addition of peroxide (Section 8.4) is done at this time by removing the cap and injecting the appropriate amount (e. g. 10  50 pL) into the vial. (Subsequent additions of peroxide shall be injected through the septa of the VOA vials.) For use of quinine actinometer (Optional): Prepare a batch of 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 ofexposure. Store the vials at 1 5 OC 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 W 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). Place all the "Day 0" samples immediately in a cooler at 1 5 OC or freeze at a continuos temperature of less than 0 "C, inverted and protected from light. Place "Unexposed" sample vials (amber) into Ziploc@ bags separated and labeled as "with peroxide" and " without peroxide", respectively. Place the bags in the bottom of 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.

1 PHOTOREACTOR SET UP ETS 8 176.0Preparation ofSamp/ es for Photolysis Studies in Aqueous Matrices Method Page 13 of 18 Page 49 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Irradiance Source Average Optimum Natural Daylight' 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 d i t i o n for 300 400 nm at known latitude. (see the table below):

Approximate Inteerated and Individual Irradiances in W/ mz 250 300 nm 300 400 nm 400 800 nm 340 nm 420 nm 0.0 27.8 259.0 0.30 0.67 Atlas Photoreactor with intepted irradiance output of 261 W/ m 300 800 nm using the IR Reflecting and 290 cut on filters 0.08 ~

27.8 234.36 0.24 0.71 Parameter ETS 8 176. OPrepurration of Samples for Photolysis Studies in Aqueous Matrices Method Page 14 of 18 Setting Page 50 of 148 Flowing Water CIFW")

Irradiation intensity Duration of exposure ON Example: 261 watts/ rn*

Example: Bhours 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't forget to add peroxide to appropriate Day 0 samples! Remove the exposed quinine control sample from the reactor tray and visually confm a color change as an indicator of photoreactor performance. The solution should be a grayhrown 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 NOT receive peroxide. 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 GCMS analysis. If subsequent analysis can not be performed immediately, store samples in a cooler at 1 5 OC.

13.12 Pertinent information regarding start and stop times of 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, 13.9 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 centrifige 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.

14.2.1 Set up autosampler and concentrator methods. If samples'have been kept in cold 14.23 Spike vials through the septa and place in the autosampler. 14.2 GCMS sample preparation.

storage, bring samples to room temperature (approximately 23 26 "C).

15.0 DATA ANALYSIS AND CALCULATIONS 15.1 15.2 15.3 The amount of target analytes in the sample will be quantified against a standard curve regression. Means will be calculated by adding the individual entities and dividing the resultant sum by the number of individual entities. Standard deviations will be calcdated 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:

nCx2  (Cn) 2 J n( n 1)

ETS 8 176.0Prepurution of Samples for Photolysis Studies in Aqueous Mairices Method Page 15 of 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:

A B X 100 =Sample % RSD where: A= standard deviation of averaged samples B= average of samples 16.0 METHOD PERFORMANCE 16.1 Refer to the definitions section for the method performance specificationshiteria for each respective sample type.

17.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 17.1 Dispose of 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, makiig sure to include all initials and dates. Store photolysis sample preparation worksheets in the raw data file. 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. Archive electronic data to compact disc media. 18.2 18.3 19.0 ATTACHMENTS ~ ~

19.1 Attachment A: Example Photolysis Prep sheet.

20.0 REFERENCES 20.1 20.2 20.3 20.4 Crosby, Helz, and Zepp. Aauatic Surface Photochemistrv. p 480 Interpersonal conversation with Carrie O'Connor, Optical Systems Engineer, Atlas Electric Devices. "Suntest CPS/ CPS+ Spectral Irradiance Distribution," table distributed by Atlas Electric Devices Company, sent via fax by Richard Sherwin, Sales Representative, 26 July, 2000. "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.

ETS 8 176. OPreparation of Samplesfor Photolysis Studies in Aqueous Matrices Method Page 16 of 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 the ICH Guideline on Photostability Testing." 21.0 AFFECTED DOCUMENTS 21.1 None 22.0 REVISIONS Revision Number.   ___ ~ ~

Reason For Revision ETS 8 176. OPreparation of Samplesfor Photolysis Studies in Aqueous Mutrices Method Page 17 of 18 Revision  Date Page 53 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Attachment A  Photolysis Sample Prep Sheet Fluorocbemical Degradation (Photolysis) Analysis Sample Prep Sheet Tu1 Andyte P r o j a t h h Requat Number ExpamreType: Nomind Eipor Annlylir: *FoNowlng lnltld sample ptvp all sumplea 1~ 111 ba placed caprl& Lwn mrdverrlcd. Fd/ owit@ oto/ pk Lky 0 smnplrc wfll kpulled mrdeXUUWd with Erparrdd Wnuparedsumplcs Control Matrix:

I I I I I ETS 8 176. OPreparation of Samples for Photolysis Studies in Aqueous Mowices Method Page 18 of 18 Page 54 of 148 BACK TO MAIN 3M Environ mental Laboratory Report No. E00 2192 3M ENVIRONMENTAL LABORATORY EQUIPMENT PROCEDURE OPERATION AND MAINTENANCE OF THE SUNLIGHT EXPOSURE SYSTEM, IMMERSION U", AND RECIRCULATING WATER CHILLER SYSTEM Procedure Number: ETS 9 44.0 Exact Copy of Original @? ,3 zy 4J Initial Date Revision Effective Date:

Approved by:

ETS 944.0 Page 1 of8 Equipment Procedure for the Atlm S W E S T Sunlight Exposure System 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 DEFINITIONS 2.1 Photon energy: U = hv = hc/ h where h is Planck's constant, c is the speed of light, and v and h are the frequency and wavelength of light. Therefore, the energy of a photon, U, is inversely proportionate to the wavelength. Irradiance: The energy output ('TJ" in the above equation for energy of a photon) in Wattsfm2 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. 2.2 3.0 DESCRIPTION 3.1 The Atlas SUNTEST" Sunliht Exposure System CPS+ or XLS+) produces visible and ultraviolet light (250 765 W/ lm2). Light proiuced is filtered with a filter or combination of filters to aIIow 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 4.2 4.3 Atlas SUNTEST@ XLS+, equipped with a xenon arclamp, lamp filter( s) available from Atlas to allow specific irradiance ranges, and immersion unit. Atlas SUNTEST@ a s +, equipped with a xenon arclamp, lamp filter(@ available from Atlas to allow specific irradiance ranges, and immersion unit. Neslab CFT 33 Refrigerated recirculator or equivalent 5.0 WARNINGS AND CAUTIONS 5.1 Health and Safety Warnings: 5.1.1 5.1.2 Wear appropriate laboratory safety personal protective equipment. The xenon lamp emits ultraviolet light which can cause burns to the skin and permanent damage to the eyes. Never attempt to operate the unit with the test chamber door open. 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.

Handle optical parts carefully; fingerprints on the lamp, filter or quartz dish can result in altered spectral output or early lamp failure. 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. 5.1.3 5.2 Cautions: 5.2.1 53.2 Page 2 of 8 ETS 9 44.0 Equipment Procedure for the Atlus SUNTEST Sunlight Exposure System Page 56 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 Filter1 Atlas Part Number Quartz Dish w/ IR reflective coating, PN 56052388 Quartz Dish, PN 56052373 W Special SupraxQ Filter, PN 56052371 5.2.3 5.2.4 5.2.5 Keep SLJNTES'P unit clear of 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 of the experiment. Manually drain the immersion tank on the XLS+ models after stopping the run; otherwise, the water will overflow.

ProDerties IEt reflective coating (supplied standard with unit) Uncoated (to allow higher black standard temperatures) Cut on at 290 nm, simulates outdoor solar 6.0 SPECIAL INSTRUCTIONS 6.1 None.

Window Glass Solar ID 65 Filter, PN 7.0 RESPONSIBILITY 7.1 The analytical group of 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 h n t of the equipment logbook.

mm (0.118 in.) window glass. Cut on at 320 nm, simulates exposure behind 6 8.0 SUPPLIES AND MATERIALS 8.1 Xenon lamp for XLS+, Atlas PN 56077798 56077769 Solar Standard Filter, PN 56077759 mm (0.236 in.) window glass. (Must be used with Window Glass Filter above.) Cut on at 290 nm, simulates outdoor solar i radiation at optimal UV intensity. 8.2 8.3 Hand tools as required 8.4  WipesTM 8.5 Xenon lami for CPS+, Atlas PN 56001794 Optional radiation filter( s) for lamp available h m Atlas:

1 radiation. I Cut on at 3 10 nm, simulates exposure behind 3 Window Glass Filter, PN 56052372 ETS 9 44.0 Page 3 of 8 Equipment Procedure for the Atlas SUNTEST Sunlight Exposure Systein 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 of 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. 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. 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. 9.1.2 9.1.3 10.0 MAINTENANCE 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 L W ") Refer to the SUNTESP instruction manual for details on how to replace the lamp. If the temperature near the lamp becomes too high, the h e blows to interrupt power and save the lamp (indicated by the error message "DOOR OPEN or TEMPERATURE FUSE"). Refer to the SUNTEST" instruction manual for details on how to replace the fuse. Record routine maintenance in the equipment log (see Section 12). 10.1.2 10.1.3 Non routine maintenance will be performed by the person@) designated in the ftont of the equipment log (see Section 12): 10.2.1 10.2 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. 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. Record non routine maintenance in the equipment log (see Section 12). 10.2.2 10.2.3 11.0 INSTRUMENT CALIBRATION 11.1 The photoreactor is set to maintain a specified integrated energy output. The amount of energy output fkom the lamp may be monitored with the use of a radiometer. The radiometer system will provide and record instantaneous energy output. Refer to ETS 9 50.0 Operation and Maintenance of Radiometer and Detector. Calibration of SUNTEST" systems will be performed two times each year by Atlas Electric Devices Company. 11.2 ETS 9 44.0 Page 4 of 8 Equipment Procedure for the Atlas SUNTEST Sunlight EXpOSltre System Page 58 of 148 BACK TO MAIN 3M Environmental Laboratory Report No. E00 2192 12.0 OPERATING PROCEDURES 12.1 12.2 Immersion Unit For more detailed operating instructions refer to the equipment operating manuals.

12.2.1 To begin operation, fill the tank with water until the level indicator is up to the full mark. 12.2.2 T u the power on.

12.3.1 SUNTEST" XLW or SUNTEST@ CPS+ unit set up 12.4.1 12.3 Chiller Turn the power on. Set water temperature knob to desired set point. Allow temperature to equilibrate before igniting the SUNTEST" lamps.

Select the desired wavelength filter b m 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: 12.4 With the photoreactor menu in the ''Program" mode, select the appropriate filter combination type:

ODtical filter System Designations A: Coated quartz glass only B: Coated quartz glass with W special glass C: Coated quartz glass with window glass D: Uncoated quartz glass only E: Uncoated quartz glass with W special glass F: Uncoated quartz glass with window glass I I 12.4.2 Selectioddetemination of energy output (W/ m') <IP_ADDRESS> Irradiance 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 of filter@) used]. The selectable range is from 250 W/ m2 to 765 W/ m2 (page 12, XLS t Instruction manual). The total (integrated) energy output (300 800 nm) is directly dependent on the type of lamp filter@) used. E. g. if the filter has a narrow range such as a cut on at 400 nm, all irradiance energy coming from wavelengths c400 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. Once the proper filter( s) idare designated, the photoreactor will base the energy output on what type of 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 Suntestm photoreactors. Reference 14.9 is a useful reference for determining irradiances at a specific <IP_ADDRESS> Page 5 of 8 ETS 9 44.0 Equipment Procedure for the Atlas SUNTESTStmlight Exposure System 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 of 250,500, and 765 W/( m2nm). 12.5 Photoreactor M y s i s set up 12.5.1 12.5.2 12.5.3 12.5.4 12.5.5 12.5.6 12.5.7 12.5.8 12.59 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. Close the chamber door and turn the power on. From the initial LCD display, use the arrow keys to select Program. Press "Enter". If programming a new method is necessary, use the arrow key to select Programming. Press "Enter". Input program number, number of phases, desired irradiance, immersion hction, phase time and switch off criteria. Each entry is followed by the "Enter" key. To start the program, select "Program Start" and press the "Enter" key. Press "Escape" for the next screens if the filter has not been changed and no printout is desired. Input program number and press "Start" 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 immekion unit. Failure to do so may result in remote flooding. 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 "Off' and unplugging the power cod. 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.