Document jyX2NL93Ld8voEM5nkZO27q2

r <77 19 U U 6 -J t6 9 7 DuPont-13202 TRADE SECRET Study Title [ ]: Hydrolytic Stability o f [ ] as a Function o f pH Test Guidelines: OECD Guideline 111 Author: Bogdan Szostek, Ph.D. Study Completed on: November 24,2003 Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Laboratory Project ID : DuPont-13202 W ork Request Number: 14643 Service Code Number: 392 Page 1of34 C om pany Sanitized. D o es not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT This study was conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, which are consistent with the OECD Principles o f Good Laboratory Practice (as revised in 1997) published in ENV/MC/CHEM(98)17 and MAFF Japan Good Laboratory Practice Standards (59 NohSan Number 3850. The test substance was characterized by the sponsor prior to the initiation o f this study, except the percent content o f 8-2 TBA. The analysis o f percent content o f 8-2 TBA in the test substance was completed after this study completion. Although the characterization was not performed under Good Laboratory Practice Standards, the characterization was done by an ISO 9000 certified laboratory, and the accuracy o f the data is considered sufficient for the purposes o f this study. Applicant / Sponsor ELLdu Pont de Nemours and Company Wilmington, Delaware 19898 U.S .A. Study Director Bogdan Szostek, PhD. Senior Research Chemist Date Applicant / Sponsor:______________________________________ _________ DuPont Representative Date -2 Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability of [ ] as a Function o f pH QUALITY ASSURANCE STATEMENT DuPont-13202 Haskell Sample Number(s): 25707 Dates o f Inspections: Protocol: July 25,2003 Conduct: August 1, 2003 Records, Reports: September 30, 2003; October 1-3,6, 2003 Dates Findings Reported to: Study Director: October 15, 2003; November 20, 2003 Management: October 15, 2003; November 20, 2003 Reported by: WondaK.lQdly Quality Assurance Auditor Date -3 Company Sanitized. Does not contain TSCA CBI. AR-226 f ]: Hydrolytic Stability of [ 40** ] as a Function o f pH DuPont-13202 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study. Approved by: S. Mark KennedyA 'h.D. Manager, Analytical Chemistry 2 y - /OpV*-.03 Date Issued by Study Director: Bogdan Szostek, FkD . Senior Research Chemist Date -4 Company Sanitized. Does not contain TSCA CBI. AR-226 Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 TABLE OF CONTENTS Page GOOD LABORATORY PRACTICE COMPLIANCE STA TEM EN T.......................................2 QUALITY ASSURANCE ST A T E M E N T .............................................................................................3 CERTIFICATIO N....................................................................................................................................... 4 LIST OF T A B L E S........................................................................................................................................6 LIST OF FIG U R E S..................................................................................................................................... 6 STUDY INFO R M A TIO N .......................................................................................................................... 7 STUDY PERSONNEL.................................................................................................................................8 SU M M A R Y ....................................................................................................................................................9 INTRODUCTION...................................................................................................................................... 10 STUDY D ESIG N .........................................................................................................................................10 M ATERIALS AND M E T H O D S............................................................................................................10 A. M aterials........................................................................................................................................... 10 1. Test Substance............................................................................................................................................................... 10 2. Preparation o f buffer solutions..................................................................................................................................11 3. Equipment......................................................................................................................................................................12 B. Methods............................................................................................................................................. 12 1. Preliminary te st.............................................................................................................................................................12 2. pH measurement and temperature............................................................................................................................ 12 3. Analysis o f test substance.......................................................................................................................................... 13 4. Analysis o f 8-2 T B A ................................................................................................................................................... 13 5. Calculations....................................................................................................................................................................14 RESULTS AND D ISC USSIO N ..............................................................................................................16 C O N C L U SIO N S.........................................................................................................................................17 RECORDS AND SAMPLE ST O R A G E ..............................................................................................17 REFERENCES............................................................................................................................................ 17 T A BLE S........................................................................................................................................................ 18 FIG U R ES......................................................................................................................................................29 -5 Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 LIST OF TABLES Page 1. Analytical results for aqueous stability o f [ ], pH 1 .2 ,37C, Days 0 (10-Sep-2003) and 5 (15-Sep-2003).................................................................................................................................................................. 19 2. Analytical results for aqueous stability o f [ ], pH 4.0, 50C, Days 0 (28-Aug-2003) and 5 (02-Sep-2003).................................................................................................................................................................. 20 3. Analytical results for aqueous stability o f [ ], pH 7.0, 50C, Days 0 (28-Aug-2003) and 5 (02-Sep-2003)....................................................................................... 21 4. Analytical results for aqueous stability o f [ ], pH 9.0, 50"C, Days 0 (03-Sep-2003) and 5 (08-Sep-2003).................................................................................................................................................................. 22 5. Measured concentrations o f 8-2 TBA for pH 1.2, 37C, Days 0 (10-Sep-2003) and 5 (15-Sep-2003)...................23 6. Measured concentrations o f 8-2 TBA for pH 4.0, 50C, Days 0 (28-A ug-2003) and 5 (02-Sep-2003)..................24 7. Measured concentrations o f 8-2 TBA for pH 7.0, 50C, Days 0 (28-A ug-2003) and 5 (02-Sep-2003)..................25 8. Measured concentrations o f 8-2 TBA for pH 9.0, 50C, Days 0 (03-Sep-2003) and 5 (08-Sep-2003)...................26 9. Daily temperature readings o f the shaking water bath........................................................................................................ 27 10. pH measurements for Day 0 and Day 5.................................................................................................................................. 28 LIST OF FIGURES Page 1. LC chromatograms o f four Day 5, pH 1.2 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards o f [ ].............................................................................................................. 30 2. LC chromatograms o f four Day 5, pH 4.0 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards o f f ].............................................................................................................. 30 3. LC chromatograms o f four Day 5, pH 7.0 replicate samples (A), 7500 m g/L (B), and 15000 mg/L (C) calibration standards o f f ].............................................................................................................. 31 4. LC chromatograms o f four Day 5, pH 9.0 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards o f f ] ...............................................................................................................31 5. Representative LC chromatograms o f blank samplesfor Day 5, pH 1 .2 ,4 .0 ,7 .0 , and 9 .0......................................... 32 6. Representative calibration curve for 8-2 TBA .......................................................................................................................33 7. Representative chromatograms o f 25.5 pg/L 8-2 TBA calibration standard and 520 pg/L D-8-2 TBA internal standard.................................................................................................................................................................... 33 8. Representative chromatograms for Day 0, pH 4.0 hydrolytic stability sample and 520 pg/L D-8-2 TBA internal standard............................................................................................................................................................................ 34 9. Representative chromatograms for Day 0, pH 4.0 blank sample...................................................................................... 34 -6 Company Sanitized. Does not contain TSCA CBI. AR-226 f ]: Hydrolytic Stability o f [ jas a Function o f pH DuPont-13202 STUDY INFORMATION Substance Tested: [ ] Svnonvms/Codes: H-25707 Submitter's Notebook Numberts): E105186-11 Haskell Number: 25707 Purity: 100% Fluorinated polymer Known Impurities: 0.016% 8-2 Telomer B Alcohol (1H,1H,2H,2Hperfluorodecan-l-ol, CAS# 678-39-7) Physical Characteristics: Off-white solid Stability: The test substance appeared to be stable under the conditions o f the study; no evidence o f instability was observed. Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: June 6, 2003 / (see report cover page) Experimental Start/Completion: June 9, 2003 / September 15,2003 -7 Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability of [ ] as a Function o f pH STUDY PERSONNEL Study Director: Bogdan Szostek, Ph.D. Primary Technician: Keith B. Prickett, B.S. Management: S. Mark Kennedy, Ph.D. Analytical Associate: Keith B. Prickett, B.S. Analytical Chemist: Bogdan Szostek, Ph.D. Management: S. Mark Kennedy, Ph.D Toxicology Report Preparation: Lisa G. Burchfield, A.A. Management: Nancy S. Selzer, M.S. DuPont-13202 -8 Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 SUMMARY The hydrolytic stability o f [ ], a fluorotelomer-based acrylate copolymer, was tested in buffered aqueous solutions at pH 4.0, 7.0, 9.0 at 50C and pH 1.2 at 37C for five days. The [ ] was demonstrated to be hydrolytically stable under these test conditions. The stability was demonstrated by the comparison o f the percent recoveries o f [ ] for Day 0 and Day 5 samples and analysis o f chromatographic peak shape for Day 5 samples. The average percent recoveries for the Day 0 and Day 5, pH 1.2 replicate samples are 101 1.2% and 101 1.0%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 4.0 replicate samples are 100 0.5% and 97 2.1%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 7.0 replicate samples are 101 1.3% and 100 0.8%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 9.0 replicate samples are 100 1.0% and 104 2.5%, respectively. The difference in percent recoveries for Day 0 and Day 5 samples did not reach the critical level o f 10% that would indicate the hydrolysis o f the test substance for any o f the investigated test conditions. The observed increase o f 8-2 TBA concentration is approximately two orders o f magnitude lower than the concentration o f 8-2 TBA expected if 10% o f the test substance hydrolyzed. In addition, the measured concentrations o f 8-2 Telomer B Alcohol (8-2 TBA, C8F I7CH2CH2OH) indicated an increase o f the 8-2 TBA concentration between Day 0 and Day 5. The increase o f the 8-2 TBA concentration can be accounted for by the presence o f the residual 8-2 TBA raw material present in the test substance. -9Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 INTRODUCTION The purpose o f the study was to investigate the hydrolytic stability o f [ ], a fluorotelomer-based acrylate copolymer, in pH 1.2, 4.0, 7.0, and 9.0 aqueous buffer solutions. The study was conducted according to the protocol: "Hydrolytic stability o f [ ] as a function o f pH", which was patterned after the OECD Guideline 111 .(1^ Additionally, the concentration o f the 8-2 Telomer B Alcohol (8-2 TBA) was monitored to investigate if 8-2 TBA is released from the test substance during the test. STUDY DESIGN A preliminary test was performed on the test substance at 50 0.1 C at each o f the pH 4.0, 7.0, 9.0 and at 37 0.1 C for pH 1.2. If less than 10% o f the test substance hydrolysis is observed after 5 days, the test substance was considered hydrolytically stable and no additional testing was performed. In addition to the test substance determination during the preliminary test, the samples were analyzed for 8-2 TBA. The study design complies with the following test guidelines: OECD Guideline 111(1) with the exception that the OECD Guideline 111 requires that the amount o f the test substance used is the smaller o f 0.01 M or half the saturation concentration in water. The test substance ([ ]) is insoluble in water and the analytical method does not allow detection o f the test substance at trace levels. Therefore, the study was conducted with an excess o f undissolved test substance. MATERIALS AND METHODS A. Materials 1. Test Substance Name: Composition: CAS Number: Test Substance Chemistry: Haskell number: [] 100% Fluorinated Polymer [] [ [ [ 25707 1 1 1 The test substance was characterized to determine the 8-2 TBA content, % fluorine, and polymer characterization parameters: Mn (number average), Mw (weight average), Mz (z average), and PDI (polydispersity index) with the following results: - 10Company Sanitized. Does not contain TSCA CBI. AR-226 [ - Hydrolytic Stability of [ ] as a Function o f pH % 8-2 TBA (w/w): % Fluorine (w/w): Mn: Mw: Mz: PDI: 0.016% [ [ [ [ [ ] ] ] ] ] 2. Preparation o f buffer solutions DuPont-13202 The following buffer solutions were prepared as described in the Annex to OECD guideline 111(1). a. pH 1.2: The 0.2 M potassium chloride (KC1) was prepared by dissolving 3.73 g o f KC1 (purity 99.0%, Sigma) in 250 mL o f water. The 0.2 M hydrochloric acid (HC1) was prepared by diluting 1.83 g o f hydrochloric acid (36.5-38%, Sigma) in 250 mL water. The pH 1.2 buffer solution was prepared by placing 125 mL o f 0.2 M KC1 and 161.3 mL o f a 0.2 M HC1 solution into a 500-mL flask and bringing the flask to volume with water. The pH o f the buffer was adjusted to 1.2 with hydrochloric acid. b. pH 4: The 0.1 M solution o f potassium hydrogen phthalate was prepared by dissolving 10.21 g o f potassium hydrogen phthalate (purity 99+%, Sigma) in 500 mL o f water. The pH 4 buffer solution was prepared by placing 2 mL o f a 0.1 M sodium hydroxide solution and 250 mL o f a 0.1 M potassium hydrogen phthalate solution into a 500-mL flask and bringing the flask to volume with water. The pH o f the buffer was adjusted to 4.0 with hydrochloric acid. c. pH 7: The 0.1 M solution o f potassium phosphate was prepared by dissolving 6.81 g o f potassium phosphate, monobasic (KH2PO4, purity 99.5%, Sigma) in 500 mL o f water. The pH 7 buffer solution was prepared by placing 148 mL o f a 0.1 M sodium hydroxide solution and 250 mL o f the 0.1 M potassium phosphate solution into a 500-mL flask and bringing the flask to volume with water. The pH o f the buffer was adjusted to 7.0 with sodium hydroxide. d. pH 9: The 0.1 M boric acid (H3BO3) solution in 0.1 M potassium chloride (KC1) was prepared by dissolving 3.73 g o f KC1 (purity 99.0%, Sigma) and 3.09 g o f H3BO3 (purity 99.5%, Sigma) in 500 mL o f water. The pH 9 buffer solution was prepared by placing 250 mL o f a 0.1 M H3BO3 solution in 0.1 M KC1 and 106.5 mL o f a 0.1 M sodium hydroxide solution into a 500-mL flask and bringing the flask to volume with water. The pH o f the buffer was adjusted to 9.0 with sodium hydroxide. Water used for buffer preparation was obtained from a Bamstead NANOpure Diamond water purification system. The water resistivity is >18.2 megaohmcm . All buffer solutions were -11Company Sanitized. Does not contain TSCA CBI. AR-226 [ I: Hydrolytic Stability o f [ ] as a Function of pH DuPont-13202 sterilized prior to use by filtering through Coming Sterilization Filter systems with 0.22 pm cellulose acetate filters. The buffer solutions were stored at room temperature. 3. Equipment pH Meter: Water Bath: Autoclave: Automatic Pipettes: Balance: Sonicator: Centrifuge: Vortexer: Glass Vials: Orion Model 250 A Water bath: Lab-Line Force Reciprocating Water Bath Shaker, Model 4682 AMSCO Model 3023 Research Pro 100 ,1 0 0 0 , 5000; Eppendorff Mettler AE-100 Branson Model 1210 or Branson Model 5200 Sorvall RT7 Multi-tube vortexer, VWR 10 mL borosilicate crimp top glass vials (Supelco) with 20 mm T/butyl septa and aluminum foil. The vials, foil, and septa were autoclaved at 121 C for 30 min before use. B. Methods 1. Preliminary test The preliminary test was performed on [ ] at 50C at each o f the pH 4.0, 7.0, 9.0 and at 37C for pH 1.2. The same sample preparation procedure and study design was followed for each o f the pH levels tested. Approximately 100 mg o f the [ ] was weighed into each o f sixteen 10-mL autoclaved glass vials and 3 mL o f appropriate buffer solution was added. The vials were crimp capped with the bottom o f the septum covered with autoclaved aluminum foil. Eight o f the vials were processed for analysis as the Day 0 samples. The remaining eight samples were wrapped with the aluminum foil, submerged in the water bath, and shaken at 100 rpm in the water bath for five days at the appropriate temperature. Eight vials containing only 3 mL o f appropriate buffer were prepared to serve as the blanks. Four o f them were used as the Day 0 blanks and the other four were wrapped in aluminum foil and placed in the water bath to be the Day 5 blanks. Additionally, six vials containing 3 mL o f appropriate buffer were spiked with 8-2 TBA stock solution (4.5 pL o f 1020 mg/L 8-2 TBA). The vials were crimp capped with the bottom o f the septum covered with autoclaved aluminum foil. Three o f the vials were used as the quality control (QC) samples for 8-2 TBA analysis on Day 0. The other three vials were wrapped in ahmunum foil and placed in the water bath together with eight o f the test substance vials. These served a role o f QC samples for 8-2 TBA analysis on Day 5. Four o f the Day 0 or four o f the Day 5 vials and two blank vials were processed for the pH measurement and the [ ] determination. The other four o f the Day 0 or Day 5 vials, two blank vials, and 8-2 TBA QC vials were processed for the 8-2 TBA determination. 2. pH measurement and temperature. The pH was measured on Day 0 and Day 5 in the vials containing the buffer and the test substance that were designated for the [ ] analysis. The vials were uncapped, the pH electrode was introduced to the vials and the pH reading was recorded. These vials were further processed for [ ] analysis. The temperature o f the water - 12Company Sanitized. Does not contain TSCA CBI. AR-226 Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 bath was monitored daily. The water bath temperature readings from the automatic temperature control were checked and recorded in the study records daily. 3. Analysis o f test substance After the pH measurement, 0.3 mL o f diluted [ ] fluorosurfactant (H-24678) was added to each vial, the vials were recapped, vortexed for ten minutes, and centrifuged for fifteen minutes at 3200 rpm. The addition o f the [ ] fluorosurfactant is necessary for wetting o f the [ ] and facilitates the pelleting o f the material at the bottom o f the vial after centrifugation. After centrifugation the vials were uncapped and the supernatant was discarded using a manual pipetter. The remaining solid material was dissolved by adding 10 mL o f tetrahydrofuran (THF) and sonication for 30 minutes. Approximately, 1 mL o f the THF solution was placed in a glass LC vial and subjected to LC-GPC analysis. The blank samples were subjected to the same procedure except the pH measurement. The stock solution o f the [ ] was made by dissolving 0.15 g o f [ ] in 10 mL o f THF. Thirty minutes o f sonication were required to completely dissolve the test substance. Calibration standards for the [ ] analysis were made by appropriate dilution o f the stock. The calibration standards ranged from 3000 to 15000 mg/L [ ]. Fresh calibration standards were prepared for each instance o f analysis. Linear regression o f the peak height vs. standard concentration was used to establish the calibration curves. Instrumentation and conditions: LC System: Column: LC Solvent: Flow Rate: Column Temperature: RI Detector Temperature: Data Collection: Injection Volume: Agilent HP 1100 with Agilent Refractive Index Detector (RID) Styragel HR 4 ,7 .8 mm x 300 mm, Waters Tetrahydrofuran (THF) 1 mL/min 40 C 35 C 0-15 min 50 pL 4. Analysis o f 8-2 TBA Samples designated for 8-2 TBA analysis were processed by injecting 6 mL acetonitrile through the vial septum using a glass syringe. Neither Day 0 nor Day 5 samples were uncapped before acetonitrile injection to avoid any losses o f 8-2 TBA. The vials containing injected acetonitrile were vortexed for 15 minutes on a multitube vortexer. Subsequently, the vials were uncapped and 1 mL o f the extract was transferred to a glass LC vial, spiked with internal standard, crimp capped, and subjected to LC/MS analysis for 8-2 TBA. 1H, 1H, 2H, 2H-perfluorodecan-l-ol (97.6%, Oakwood Products, West Columbia, SC) was used as the analytical standard o f 8-2 TBA. ID, ID, 2D, 2D, 3 -13C-heptadecafluoro decanol (M+5) (abbreviated as D-8-2 TBA) was used as the internal standard. Stock solutions (1000 mg/L) o f the analytical standard and the internal standard were prepared in methanol and were stored refrigerated. The calibration standards were prepared fresh for each calibration in 50/50% - 13Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 water/acetonitrile. The calibration standards were made in the range from 25 to 2550 pg/L 8-2 TBA. A constant concentration o f internal standard was used: 520 pg/L D-C8-2A. The calibration curves were contracted using the ratio o f the peak area for ions m/z monitored for 8-2 TBA and peak area for ions monitored for D -8-TBA and the concentrations o f 8-2 TBA. Instrumentation and conditions: HPLC Instrument: M S Instrument: M odel 2795, W aters ZQ, Micromass LC Parameters: Column: Mobile Phase: Column Temperature: Injection Volume: Column Switch: Luna 3u C18 (2), 20 mm x 2mm A - water B - methanol Gradient: Time (min %A 0 30 1 30 20 60 6.1 30 8 30 30C 20 pL 2 and 6 min %B Flow (mL/min) 70 0.25 70 0.25 100 0.25 100 0.25 70 0.25 70 0.15 MS Parameters: Ionization mode: Capillary voltage: Cone voltage: Source Temperature: Desolvation Temperature: Data Acquisition Function: Electrospray, negative ions 2.5 kV 10 V 120C 300C 8-2 TBA: SIR* o f 463, 509 ,5 2 3 ; 0-6 min D- 8-2 TBA: SIR o f 468, 514, 528 m/z; 0-6 min * These ions represent the deprotonated molecular ion o f 8-2 TBA (m/z 463), formate adduct o f 8-2 TBA (m/z 509), and acetate adduct o f 8-2 TBA (m/z 523). The formate and acetate adduct are formed because o f residual presence o f acetate and/or formate in the system originating from other methods that are run on the system and utilize these components in the mobile phase. 5. Calculations The following calculations were used to determine the nominal concentrations o f [ ] (Tables 1-4). For example, if 0.1009 g o f [ ] was weighed into the vial and subsequently dissolved in 10 mL o f THF during sample processing, this would represent the nominal concentration o f 100.9 mg/0.01 L=10,090 mg/L. The percent recovery is calculated by dividing the measured concentration by the nominal concentration, times 100%. The expected (nominal) concentration o f 8-2 TBA in the spiked samples was calculated as follows: the 4.5 pL o f 1020 mg/L 8-2 TBA spike represents 4.5 pL x 1020 ng/pL= 4590 ng 8-2 TBA. Total volume o f extract is 9 mL: 3 mL o f buffer and 6 mL o f acetonitrile. The expected - 14Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 nominal concentration o f 8-2 TBA in the extract is: 4590 ng/9 mL=510 ng/mL. The percent recovery is calculated by dividing the measured concentration by the nominal concentration, times 100%. The expected concentration o f 8-2 TBA in the acetonitrile extract if 10% o f the [ ] hydrolyzed can be estimated in the following way. Ten percent o f 100 mg o f [ ] represents 10 mg. Taking into account that the test substance contain [ ] fluorine and assuming that all fluorine is present in the form o f bound 8-2 TBA, the hydrolysis o f the 10 mg o f the test substance would release: [ ] [ ]5.56 mg 8-2 TBA. This amount o f 8-2 TBA would be contained in 9 mLs o f extract. Therefore, expected concentration o f 8-2 TBA would be: 5.56 mg/0.009 L= 618 mg/L. The [ ] contains residual raw material, 8-2 TBA. The expected, maximum concentration o f 8-2 TBA derived from the 8-2 TBA present in the extract, assuming complete extraction, is: 100 mg o f [ ] x (0.016%/100)/0.009 L= 1.78 mg/L (1780 pg/L). -15C om pany S an itized . D o e s n o t con tain TSCA CBI. [ ]: Hydrolytic Stability o f [__________________ ] as a Function o f pH______________________________ DuPont-13202 RESULTS AND DISCUSSION Tables 1-4 present the results for measured concentrations o f [ ] and percent recoveries obtained for Day 0 and Day 5 replicate samples for pH 1.2,4.0, 7.0, 9.0, respectively. The average percent recoveries for the Day 0 and Day 5, pH 1.2 replicate samples are 101 1.2% and 101 1.0%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 4.0 replicate samples are 100 0.5% and 97 2.1%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 7.0 replicate samples are 101 1.3% and 100 0.8%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 9.0 replicate samples are 100 1.0% and 104 2.5%, respectively. The relative difference between the Day 5 and Day 0 concentrations o f the test substance was less than 10% for any o f the investigated pHs. This indicates that the [ ] is hydrolytically stable at investigated pHs. Figures 1 to 4 compare the LC chromatograms o f four replicates o f Day 5 samples with the chromatograms o f two calibration standards for pH 1.2, 4.0, 7.0, 9.0, respectively. The shape o f the chromatographic peak, eluting from 7 to 11 minutes, is indicative o f the molecular weight distribution o f the [ ]. The hydrolysis o f [ ] would result in change o f the observed peak shape. Comparison o f the peak shape o f the freshly made calibration standards o f [ ] and the Day 5 samples indicates that no evident change o f the peak shape for Day 5 samples was observed for any o f the investigated pHs. This further supports the conclusion o f [ ] hydrolytic stability. Figure 5 presents the LC chromatograms o f the Day 5 blank samples for pH 1.2, 4.0, 7.0, and 9.0. Additional evidence supporting the hydrolytic stability o f [ ] can be derived from the measured concentration o f 8-2 TBA. The hydrolysis o f [ ] would result in the release o f 8-2 TBA and the increase o f 8-2 TBA concentration in the test system. Based on the charcterization data, the test substance contains 0.016% o f residual 8-2 TBA. Assuming that the buffer extracts all the 8-2 TBA during the time o f hydrolytic stability experiment, the expected concentration o f 8-2 TBA in the processed extract (3 mL buffer and 6mL acetonitrile) would be 1.78 mg/L (see B.5 Calculations). Similarly, the expected concentration o f 8-2 TBA in the extract, if 10% o f [ ] was hydrolyzed, is 618 mg/L (see B.5 Calculations). Tables 5-8 present the measured concentrations o f 8-2 TBA in the extracts for Day 0 and Day 5 replicate samples for pH 1.2,4.0, 7 .0,9.0, respectively. The data shows a significant increase o f 8-2 TBA concentration in the Day 5 samples, when the average concentration o f 8-2 TBA for Day 0 and Day 5 samples are compared. However, the concentrations reach only some o f the expected level o f fully extracted 8-2 TBA (1780 pg/L) and these are approximately two orders o f magnitude lower than the concentrations expected when 10% o f the test substance would hydrolyze. The increase o f the 8-2 TBA concentration between the Day 0 and Day 5 samples is simply a result o f extraction o f the 8-2 TBA from [ ] during the hydrolytic stability experiment at elevated temperature (50C or 37C). The percent recovery results for 8-2 TBA (Tables 5-8) support the validity o f the 8-2 TBA measurement in the Day 0 and Day 5 samples for all investigated pHs. The Day 5 sample percent recovery results demonstrate that no significant loss o f 8-2 TBA occurred from the test system during the hydrolytic stability experiment. Figure 6 shows representative calibration curve obtained for 8-2 - 16Company Sanitized. Does not contain TSCA CBI. AR-226 Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 TBA. Figures 7-9 show chromatograms for 8-2 TBA calibration standard, Day 0, pH 4.0 test sample, and Day 0, pH 4.0 blank sample, respectively. Table 9 summarizes the water bath temperature data for pH 1.2, 4.0, 7.0, and 9.0. In all cases, the water bath temperature was maintained at the specified level with deviation not exceeding 0 .1 C . Table 10 summarizes results o f the pH measurements for Day 0 and Day 5 samples for pH 1.2, 4.0, 7.0, and 9.0. The measured pH for Day 0 and Day 5 replicate samples for all o f the investigated pHs was at the specified nominal level with the deviations not exceeding 0.1. CONCLUSIONS The hydrolytic stability o f [ ], a fluorotelomer-based acrylate copolymer, was tested in buffered aqueous solutions at pH 4.0, 7.0, 9.0 at 50C and pH 1.2 at 37C for five days. The [ ] was demonstrated to be hydrolytically stable under these test conditions. The stability o f the [ ] was demonstrated by the comparison o f the percent recoveries o f [ ] for Day 0 and Day 5 samples and analysis o f chromatographic peak shape for Day 5 samples. The difference in percent recoveries for Day 0 and Day 5 samples did not reach the critical level o f 10% that would indicate the hydrolysis o f the test substance for any o f the investigated test conditions. In addition, the measured concentrations o f 8-2 TBA indicated an increase o f the 8-2 TBA concentration between Day 0 and Day 5. The increase o f the 8-2 TBA concentration can be accounted for by the presence of the residual 8-2 TBA in the test substance. The increase o f 8-2 TBA concentration is approximately two orders o f magnitude lower than the concentration o f 8-2 TBA expected if 10% o f the test substance hydrolyzed. RECORDS AND SAMPLE STORAGE Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. REFERENCES 1. Organisation for Economic Cooperation and Development (OECD). May 12,1981. OECD Guidelines for Testing o f Chemicals, Hydrolysis as a Function o f pH, OECD Guideline No. 111. - 17Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [__________________ ] as a Function of pH DuPont-13202 TABLES - 18Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]= Hydrolytic Stability o f [ ] as a Function o f pH Table 1: Analytical results for aqueous stability o f f Days 0 (10-Sep-2003) and 5 (15-Sep-2003). DuPont-13202 ], pH 1.2, 37C, 1 11 1 pH Day Rep Nominal Concentration (mg/L) Measured Concentration (mg/L) 1.2 0 1 1.2 0 2 1.2 0 3 1.2 0 4 10100 10100 10000 10200 10100 10300 10200 10200 Average Standard Deviation 1.2 5 1 1.2 5 2 1.2 5 3 1.2 5 4 10200 10000 10200 10300 10100 10100 10300 10400 Average Standard Deviation % Recovery 100 102 102 100 101 1.2 99 101 101 101 101 1.0 - 19Company Sanitized. Does not contain TSCA CBI, AR-226 I ] Hydrolytic Stability o f [ ] as a Function o f pH Table 2: Analytical results for aqueous stability o f [ Days 0 (28-Aug-2003) and 5 (02-Sep-2003). DuPont-13202 ], pH 4.0, 50C, l )t 1 pH Day Rep Nominal Concentration (mg/L) Measured Concentration (mg/L) 4.0 0 1 4.0 0 2 4.0 0 3 4.0 0 4 10100 10100 10000 10200 9960 10100 10000 10200 Average Standard Deviation 4.0 5 1 4.0 5 2 4.0 5 3 4.0 5 4 10200 10100 10100 10100 9660 9810 10100 9810 Average Standard Deviation % Recovery 99 100 100 100 100 0.5 95 97 100 97 97 2.1 -20Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH Table 3: Analytical results for aqueous stability o f f Days 0 (28-Aug-2003) and 5 (02-Sep-2003). DuPont-13202 ], pH 7.0, 50C, l 11 1 PH Day Rep Nominal Concentration (mg/L) Measured Concentration (mg/L) 7.0 0 1 7.0 0 2 7.0 0 3 7.0 0 4 10200 10100 10100 10000 10100 10200 10300 10100 Average Standard Deviation 7.0 5 1 7.0 5 2 7.0 5 3 7.0 5 4 10200 10200 10100 10000 10200 10200 10200 9910 Average Standard Deviation % Recovery 99 101 102 101 101 1.3 100 100 101 99 100 0.8 -21Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function of pH Table 4: Analytical results for aqueous stability o f [ Days 0 (03-Sep-2003) and 5 (08-Sep-2003). DuPont-13202 ], pH 9.0, 50C, l 1[ 1 PH Day Rep Nominal Concentration (mg/L) Measured Concentration (mg/L) 9.0 0 1 9.0 0 2 9.0 0 3 9.0 0 4 10100 10000 10000 10100 9950 10100 10000 10000 Average Standard Deviation 9.0 5 1 9.0 5 2 9.0 5 3 9.0 5 4 10000 10100 10000 10300 10200 10300 10500 11000 Average Standard Deviation % Recovery 99 101 100 99 100 1.0 102 102 105 107 104 2.5 - 22Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 Table 5: Measured concentrations o f 8-2 TBA for pH 1.2, 37C, Days 0 (10-Sep-2003) and 5 (15-Sep-2003). Nominal 8-2 TBA pH Day Rep* Concentration (pg/L) Measured 8-2 TBA Concentration (pg/L) b % Recovery 1.2 0 1 1.2 0 2 1.2 0 3 1.2 0 4 - 727 774 762 895 - Average Standard Deviation 790 73 1.2 0 QCl 1.2 0 QC2 1.2 0 QC3 510 510 510 595 117 594 116 633 124 Average Standard Deviation 119 4.4 1.2 5 1 1.2 5 2 1.2 5 3 1.2 5 4 - 493 688 658 490 - Average Standard Deviation 582 110 1.2 5 QCl 1.2 5 QC2 1.2 5 QC3 510 510 510 440 86 471 92 476 93 Average Standard Deviation ` Q C l, QC2 and QC3 represent three 8-2 TBA recovery samples for each test day. b Concentrations of 8-2 TBA in the extract (3 mL buffer and 6 mL acetonitrile). 90 3.8 - 23Company Sanitized. Does not contain TSCA CBI. AR-226 t ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 Table 6: Measured concentrations of 8-2 TBA for pH 4.0, 50C, Days 0 (28-Aug-2003) and 5 (02-Sep-2003). Nominal 8-2 TBA pH Day Rep* Concentration (pg/L) Measured 8-2 TBA Concentration (pg/L) b % Recovery 4.0 0 1 4.0 0 2 4.0 0 3 4.0 0 4 - 778 699 795 740 - Average Standard Deviation 753 43 4.0 0 QCl 4.0 0 QC2 4.0 0 QC3 510 510 510 495 97 508 100 487 95 Average Standard Deviation 97 2.5 4.0 5 1 4.0 5 2 4.0 5 3 4.0 5 4 - 1560 1680 1680 1430 - Average Standard Deviation 1590 120 4.0 5 QCl 4.0 5 QC2 4.0 5 QC3 510 510 510 464 91 471 92 447 88 Average Standard Deviation * Q C l, QC2 and QC3 represent three 8-2 TBA recovery samples for each test day. b Concentrations of 8-2 TBA in the extract (3 mL buffer and 6 mL acetonitrile). 90 2.1 -24Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function of pH DuPont-13202 Table 7: Measured concentrations o f 8-2 TBA for pH 7 .0 ,50C, Days 0 (28-Aug-2003) and 5 (02-Sep-2003). Nominal 8-2 TBA pH Day Rep * Concentration (pg/L) 7.0 0 1 7.0 0 2 7.0 0 3 7.0 0 4 - Measured 8-2 TBA Concentration (pg/L) b % Recovery 796 875 841 801 - Average Standard Deviation 828 37 7.0 0 QCl 7.0 0 QC2 7.0 0 QC3 510 510 510 519 102 494 97 499 98 Average Standard Deviation 99 2.7 7.0 5 1 7.0 5 2 7.0 5 3 7.0 5 4 - 1670 1300 1800 1380 - Average Standard Deviation 1540 240 7.0 S QCl 7.0 5 QC2 7.0 5 QC3 510 510 510 565 I l l 411 81 462 91 Average Standard Deviation * Q Cl, QC2 and QC3 represent three 8-2 TBA recovery samples for each test day. b Concentrations of 8-2 TBA in the extract (3 mL buffer and 6 mL acetonitrile). 94 15 - 25Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 Table 8: Measured concentrations o f 8-2 TBA for pH 9.0, 50C, Days 0 (03-Sep-2003) and 5 (08-Sep-2003). Nominal 8-2 TBA pH Day R e p ' Concentration (pg/L) 9.0 0 1 9.0 0 2 9.0 0 3 9.0 0 4 - M easured 8-2 TBA Concentration (pg/L) b % Recovery 796 519 864 704 - Average Standard Deviation 721 150 9.0 0 QC1 9.0 0 QC2 9.0 0 QC3 510 510 510 483 95 492 96 -- Average Standard Deviation 96 0.7 9.0 5 1 9.0 5 2 9.0 5 3 9.0 5 4 - 800 757 868 1010 - Average Standard Deviation 859 110 9.0 5 QC1 9.0 5 QC2 9.0 5 QC3 510 510 510 437 86 472 93 417 82 Average Standard Deviation " QC1, QC2 and QC3 represent three 8-2 TBA recovery samples for each test day. b Concentrations of 8-2 TBA in the extract (3 mL buffer and 6 mL acetonitrile). 87 5.6 -26Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH Table 9: Daily temperature readings o f the shaking water bath. DuPont-13202 Time (Day) 0 1 2 3 4 5 0 1 2 3 4 5 0 1 2 3 4 5 0 1 2 3 4 5 T em perature pH Date (C) 1.2 10-Sep-2003 37.1 1.2 11-Sep-2003 37.0 1.2 12-Sep-2003 37.0 1.2 13-Sep-2003 37.0 1.2 14-Sep-2003 37.1 1.2 15-Sep-2003 37.0 4.0 28-Aug-2003 50.0 4.0 29-Aug-2003 50.1 4.0 30-Aug-2003 50.0 4.0 31-Aug-2003 49.9 4.0 01-Sep-2003 50.0 4.0 02-Sep-2003 50.0 7.0 28-Aug-2003 50.0 7.0 29-Aug-2003 50.1 7.0 30-Aug-2003 50.0 7.0 31-Aug-2003 49.9 7.0 01-Sep-2003 50.0 7.0 02-Sep-2003 50.0 9.0 03-Sep-2003 50.0 9.0 04-Sep-2003 50.0 9.0 05-Sep-2003 49.9 9.0 06-Sep-2003 50.0 9.0 07-Sep-2003 50.0 9.0 08-Sep-2003 50.0 -27Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [__________________ ] as a Function o f pH Table 10: pH measurements for Day 0 and Day 5. DuPont-13202 Day Rep Nominal pH Measured pH 0 1 1.2 1.25 0 2 1.2 1.25 0 3 1.2 1.26 0 4 1.2 1.25 5 1 1.2 1.29 5 2 1.2 1.30 5 3 1.2 1.28 5 4 1.2 1.29 0 1 4.0 4.10 0 2 4.0 4.10 0 3 4.0 4.10 0 4 4.0 4.10 5 1 4.0 4.10 5 2 4.0 4.10 5 3 4.0 4.10 5 4 4.0 4.10 0 1 7.0 7.05 0 2 7.0 7.06 0 3 7.0 7.05 0 4 7.0 7.05 5 1 7.0 7.10 5 2 7.0 7.10 5 3 7.0 7.10 5 4 7.0 7.10 0 1 9.0 9.05 0 2 9.0 9.06 0 3 9.0 9.05 0 4 9.0 9.05 5 1 9.0 9.10 5 2 9.0 9.08 5 3 9.0 9.09 5 4 9.0 9.08 -28Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 FIGURES -29Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ as a Function o f pH DuPont-13202 Figure 1: LC chromatograms o f four Day 5, pH 1.2 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards o ff ]. Figure 2: LC chromatograms o f four Day 5, pH 4.0 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards o ff ]. -30Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 Figure 3: LC chromatograms of four Day 5, pH 7.0 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards of [ ]. Figure 4: LC chromatograms of four Day 5, pH 9.0 replicate samples (A), 7500 mg/L (B), and 15000 mg/L (C) calibration standards of [ ]. -31Company Sanitized. Does not contain TSCA CBI. AR-226 [ ]: Hydrolytic Stability o f [ ] as a Function o f pH DuPont-13202 Figure 5: Representative LC chromatograms of blank samples for Day 5, pH 1.2,4.0, 7.0, and 9.0. - 32Company Sanitized. Does not contain TSCA CBI, AR-226 f ] Hydrolytic Stability o f [ ] as a Function o f pH Figure 6: Representative calibration curve for 8-2 TEA. DuPont-13202 Figure 7: Representative chromatograms of 25.5 pg/L 8-2 TBA calibration standard and 520 pg/L D-8-2 TBA internal standard. - 33Company Sanitized. Does not contain TSCA CBI. AR-226 r 1: Hydrolytic Stability of [ ] as a Function o f pH DuPont-13202 Figure 8: Representative chromatograms for Day 0, pH 4.0 hydrolytic stability sample and 520 pg/L D-8-2 TBA internal standard. Figure 9: Representative chromatograms for Day 0, pH 4.0 blank sample. -34Company Sanitized. Does not contain TSCA CBI. AR-226