Document a1X1xL8w5LGLNwMknVXr9Qnkb

P-1 Sanitized Version for the AR-226 Public Printed Copy File fCBI removed: copy restricted materials included as read only) Index DOCUMENT DESCRIPTION NO. OF PAGES CBI OR OTHER RESTRICTION? 1 March 7, 2007 letter from DuPont 1 No restriction to EPA 2 DuPont Investment in 1 CBI - removed Fluorotelomer Environmental Fate Science 2004-2007 3 Appendix I Divider Sheet 1 No restriction 4 Cost Summary - Telomers 1 CBI - removed 5 Testing Invoices - Set 1 1 CBI - removed 6 Testing Invoices - Set 2 1 CBI - removed 7 Protocol H-26634 15 No restriction 8 Protocol H-26665 15 No restriction 9 Appendix II Divider Sheet 1 No restriction 10 Testing Invoices 1 CBI - removed 11 Appendix III Divider Sheet 1 No restriction 12 DuPont Investment in 1 CBI - removed Fluorotelomer Environmental Fate Science 13 Appendix IV Divider Sheet 1 No restriction 14 Summary of EPA Submissions 7 No restriction (dated 07/11/2002) 15 Biodegradation studies of 2 Subject to copyright - document is read only and fluorotelomer-based polymers in may not be copied without author permission; soils Contact information to request author permission: Dr. Robert Buck (302-892-8935 robert.c .buck@usa.dupont.com) 16 Biodegradation studies of 27 Not for further distribution without author approval - fluorotelomer-based polymers in document is read only and may not be copied without activated sludge, soil, and author permission; sediments Contact information to request author permission: Dr. Robert Buck (302-892-8935 or Robert.C.Buck@usa.dupont.com) Com pany Sanitized P-2 DuPont Chemical Solutions Enterprise P. 0. Box 80023 Wilmington, DE 19880-0023 March 7, 2007 Dear Jim, Thank you for your time in meeting with us February 20th, as well as for the good discussion that ensued. Enclosed please find the additional information you requested on: 1) The comprehensive, peer reviewed study commissioned by DuPont and conducted by Environ on the global fate and exposure potential from DuPont Fluorotelomer products and 2) Information on the studies that DuPont has conducted and commissioned on the biodegradation o f fluorotelomer intermediates and products. Specifically, you will find the Wildlife International invoices in Appendix I (outlined in the attached document pages 2 and 3), invoices from ENVIRON in Appendix II (outlined in the attached document page 4), and Academic Studies we have supported in Appendix III pages 9, 10, and 19-41 (outlined in the attached document page 5). As well, I have included a document in Appendix IV that reviews the information we have shared with the EPA and at various scientific meetings. Included in this Appendix is the most recent information shared at the SETAC meeting in November. If you would like any o f these documents please don't hesitate to ask. As a follow up I would like to arrange a time to discuss these items and their significance for future work with you at your earliest convenience. With best regards, Henry E. Bryndza Technology Directory DuPont Chemical Solutions Enterprise DuPont Central Research & Development P-3 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED Document Number in Index: 2 Document Description: DuPont Investment in Fluorotelomer Environmental Fate Science 2004-2007 S P-4 APPENDIX I cbdc.3 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED Document Number in Index: 4 Document Description: Cost Summary - Telomers 7 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED Document Number in Index: 5 Document Description: Testing Invoices - Set 1 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED Document Number in Index: 6 Document Description: Testing Invoices - Set 2 p.8 PROTOCOL H-26634: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS This protocol complies with the Organisation for Economic Cooperation and Development OECD Guideline 307 Adopted April 2002 Submitted to E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 USA Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 February 17, 2005 boo.7 P-9 Wildlife International, Ltd. 2- - H-26634: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS SPONSOR: E.I. du Pont de Nemours and Company SPONSOR'S REPRESENTATIVE: William Berti TESTING FACILITY: Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 STUDY DIRECTOR: Edward C. Schaefer Wildlife International, Ltd. LABORATORY MANAGEMENT : Henry O. Krueger, Ph.D. Director of Aquatic Toxicology and Non-Target Plants Proposed Dates: FOR LABORATORY USE ONLY Experimental Start Date: I W lch 2 uc $' Experimental Termination Date: /S' 00 c Project No.: H Z -tO t Test Concentrations: 00 ^ Test Substance No.: ^ ^& ;__ Z<jQ , |0 i v i * , z i , e>2`/yt ty? Reference Substance No. (if applicable)-M *i, cyy5 t y y r PROTOCOL APPROVAI. fX/ p STUDY DIRECTOR cl / j y ------ ------------------------------- -LABORATORY MANAGEMENT 0 DATE to e s ' SPONSOR'S REPRESENTATIVE DATE PROTOCOL NO.: 112/021705/SED-TRANSa/SUBl 12 // Wildlife International, Ltd. -3 - INTRODUCTION Soil may be exposed to chemicals by direct application, spray drift, run-off, drainage, waste disposal, industrial or agricultural effluent and atmospheric deposition. This protocol describes a laboratory test method to assess the transformation o f the test substance in aerobic and anaerobic soil systems. No bias is expected in this type o f study. OBJECTIVE The objective of the study is to assess the potential for transformation of the test substance in aerobic and anaerobic soils. EXPERIMENTAL DESIGN The test will be conducted with four test soils under both aerobic and anaerobic condition. Four groups will be established for each soil type and condition (aerobic or anaerobic): (1) untreated (control) live soil; (2) 200 mg test substance/kg dw treated live soil; (3) 200 mg test substance/kg dw treated sterile soil; (4) 250 |Xg 8-2 telomer B alcohol/kg dw treated sterile soil; 10 pg 8-2 telomer B acid /kg dw; 10 pg 8-2 unsaturated telomer B acid/kg; and 10 pg perfluorooctanoic acid/kg dw, C9, CIO, and Cl 1 at 10 pg/kg dw all added to a sterile soil. The test systems will be incubated in sealed containers at approximately 20C for up to 1 year. Two incubation chambers from each of the groups will be removed at appropriate time intervals and the soil samples will be extracted and analyzed for potential metabolites. In addition to the day zero samples, a minimum o f eight additional samplings will be performed. Sufficient incubation chambers to allow for 3 additional sampling intervals will be prepared for the untreated control (1) and 200 mg test substance/kg dw treated live soil (2) groups. These chambers will be sampled as needed at the request of the study monitor. Additional untreated and test substance treated soil chambers will be prepared for use as matrix fortification samples, biomass determinations, viability controls and to monitor aerobic conditions as necessary. The test groups will be prepared using 25 grams (dry weight equivalent) of soil. While this amount of soil is less than the 50 to 200 grams specified in the OECD 307 guideline, it was chosen to allow for 1) the extraction o f the entire test chamber contents and internal surfaces; 2) sufficient PROTOCOL NO. : 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. -4 headspace to minimize the need for active aeration which could result in a loss of potential transformation products. METHODS AND MATERIALS The test system and study conditions are based on the OECD Guideline for Testing of Chemicals, Guideline 307, Aerobic and Anaerobic Transformation in Soil (1). As this guideline is designed for low molecular weight substances, but not for polymers, adaptations o f the method may be necessary for feasibility reasons. Nevertheless, the study will be carried out in such a way that the recommendations given in the guideline will be followed as closely as possible. Test Substances Information on the characterization of test, control or reference substances is required by Good Laboratory Practice (GLP) Standards and Principles. The Sponsor is responsible for providing Wildlife International, Ltd. verification that the test substance has been characterized prior to its use in the study. If verification of GLP test substance characterization is not provided to Wildlife International, Ltd., it will be noted in the compliance statement of the final report. The Sponsor is responsible for all information related to the test substance and agrees to accept any unused test substance and/or test substance containers remaining at the end o f the study. The test substance is not radiolabeled and has no single molecular formula. It is a mixture containing perfluoroalkyl groups o f varying carbon numbers as polymer side chains. Test Soils Four freshly collected US soils will be used to evaluate the test substances under aerobic and anaerobic conditions. The soils will be collected from each of the following soil orders: PROTOCOL NO.: 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. -5- Soil O rder Percent of US Land* Alfisol 13 Mollisol 25 Inceptisol 16 Ultisol 13 Percent of total land in the US, from Foth, H.D. 1990. Fundamentals of Soil Science 8 ed. John Wiley & Sons, New York) The soils will be processed as soon as possible after sampling. Vegetation, larger soil fauna and stones will be removed prior to passing the soil through a 2 mm sieve. The soil used in the sterile groups will be sterilized by cobalt irradiation. The soils may be stored in the dark at 4 2 C, if necessary. However, storage and pre-incubation time together will not exceed 3 months. Characterization o f the soils will be conducted by Agvise Laboratories (Northwood, North Dakota, USA). The following is a list o f the minimum physicochemical properties o f the soil to be determined. Texture (i.e., percentage o f sand, silt and clay) pH Organic carbon Bulk density Field moisture capacity Cation exchange capacity The percent moisture and water holding capacity o f each o f the soil types will be determined. In addition, the microbial biomass o f a live sample of each soil type will be determined. The soil microbial biomass will be determined at the beginning o f the test prior to adding the test substance and in test and control soil samples at four months, at six months, and at 12 months using the fumigation-extraction method. Test Apparatus and Conditions The test chambers will be glass serum bottles with foil lined closures and will be identified by project number, test substance ID, test concentration, and unique identifier. The chambers will be incubated statically at approximately 20 2C. PROTOCOL NO. : 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. 6- - Soil Pre-Incubation The soil moisture of each test soil content will be adjusted to 40 to 60% of water holding capacity (which is equivalent to a pF o f between 2.0 and 2.5, or from 0.1 to 0.33 bar). The test soil then will be allowed to warm to the test temperature for a least 2 days. A pre-incubation period of at least 7 days for the aerobic soils and 14 days for the anaerobic soils will be performed under the appropriate conditions (i.e. temperature, soil moisture content, aerobic or anaerobic). Approximately 25 grams dry weight equivalent of test soil will be added to the test chambers. Sterile soil treatments will be dosed at approximately 200 mg/kg (dw) with both chloramphenicol and cycloheximide to inhibit microbial growth. The chambers to be incubated under aerobic conditions will be sealed with septa and incubated. The chambers to be incubated under anaerobic conditions will be flushed with anaerobic mixed gas, sealed with septa and incubated. The soil moisture content should be maintained in the optimal microbial growth range of 40 to 60% water holding capacity (WHC). The soil moisture content should be checked and adjusted, if necessary, at least once during the pre-incubation period by weighing the test vessels. Sterile-filtered demineralized water (degassed sterile-filtered demineralized water for the anaerobic soils) will be added as necessary to compensate for water losses. Preparation of the Test Chambers The soil moisture content will be checked and adjusted, if necessary, prior to the addition of the test substance. Sterile-filtered demineralized water (degassed sterile-filtered demineralized water for the anaerobic soils) will be added as necessary to compensate for water. Anaerobic treatments and controls will be prepared within an anaerobic environment. The test substance will be applied to the soil within the test chambers as described below. Background Blank Control: Soils with no test substance added will be tested in duplicate to check for background concentrations of analytes (see Sampling and Measurements). The entire bottle will be extracted after a sample o f the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Test Substance: The test substance dosed treatments will be prepared by dosing approximately 25 grams (dry weight equivalent) o f soil with sufficient test substance to deliver 200 mg/kg dry weight. The test substance will be administered by direct weight addition. The test chambers will be sealed with septum lined with aluminum foil, mixed and incubated. The entire bottle PROTOCOL NO. : 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. -7- will be extracted after a sample of the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Abiotic Controls: The abiotic controls will be prepared by dosing approximately 25 grams (dry weight equivalent) o f Co-sterilized soil with sufficient test substance to deliver 200 mg/kg dry weight. The test substance will be administered by direct weight addition. The entire bottle will be extracted after a sample of the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Spike Recovery Controls: The spike recovery controls will be prepared by dosing approximately 25 grams (dry weight equivalent) of Co-sterilized soil with appropriate volumes of 8-2 TBA stock (250 pg/mL in ethanol) and fluorinated acid stocks (10 pg/mL in water). The stocks will be injected directly into the soil using a glass microsyringe. The test chambers will be immediately sealed with septum lined with aluminum foil and the content of the chambers will be mixed. The entire bottle will be extracted after a sample o f the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Viability Controls 1untreated): The metabolic activity of untreated test soil will be assessed on a monthly basis. Duplicate incubation chambers for each test soil and condition (aerobic and anaerobic) will be dosed at approximately 100 mg/kg dw with a combination of radiolabeled and non-labelled glucose. The evolved l4C 02 (aerobic and anaerobic conditions) and l4CH4 (anaerobic conditions) will be measured and the percent biodegradation calculated. The methods and procedures used will be documented in the study records and in the final report Aerobic Controls (treated!: The aerobic controls will be prepared by dosing approximately 25 grams (dry weight equivalent) of soil with sufficient test substance to deliver 200 mg/kg dry weight. The test substance will be administered by direct weight addition. The test chambers will be sealed with septum lined with aluminum foil, mixed and incubated. The oxygen content of a sacrificial chamber will be measured at monthly intervals. The methods and procedures used will be documented in the study records and in the final report. Maintenance of Test Chambers The soil moisture content should be maintained in the optimal microbial growth range of 40 to 60% water holding capacity (WHC; a pF o f between 2.0 and 2.5, or from 0.1 to 0.33 bar). The soil moisture content should be checked and adjusted, if necessary, at regular 1 to 2 week intervals by weighing the test vessels. Adding sterile-filtered demineralized water (degassed sterile-filtered demineralized water for the anaerobic soils) will compensate water losses. PROTOCOL NO.: 112/021705/SED-TRANSa/SUBl 12 W ildlife International, Ltd. 8- - The oxygen content in the headspace o f two aerobic control chambers will be assessed on a monthly basis to ensure aerobic conditions. If the mean measured oxygen content within the control chambers is less than 19%, aerobic test chambers will be aerated with filter-sterilized, oil-free air. If test chambers have to be opened to add water or for monitoring purposes, a sample of the head space will be taken first using a C l8 cartridge as described below. The chamber will then be recapped as soon as possible. The septum and cap previously removed will be stored at -10C or lower. The C18 will subsequently extracted and analyzed. Sample extracts will be stored at -10C or lower if analysis is delayed more than 24 hours. Sampling and Measurements Duplicate chambers from each of the control, treated and spike recovery soils will be extracted (e.g., acetonitrile or other suitable solvent) and analyzed. Proposed sampling intervals will be at 0, 1 and 2 weeks and 1, 2, 4, 6, 9, and 12 months (9 sampling times), however the actual sampling intervals will be documented in the study records and in the final report. More or less frequent intervals may be conducted at the discretion of the Study Director. Potential volatile transformation products in the headspace o f the soils chambers will be collected using an appropriate trap (i.e. C 18 cartridge). At each sampling time prior to opening the test vessel, the septum will be pierced using a needle connected to a Cl 8 cartridge and syringe. A volume of headspace gas from within the chamber will be pulled through the C l8 cartridge using the attached syringe. The C18 will be subsequently extracted and analyzed. Sample extracts will be stored at --10C or lower if analysis is delayed more than 24 hours. Extracts will be analyzed for: 1. CF3(CF2)7CH2CH2OH (perfluorooctyl ethanol, 8-2 TBA, CAS# 678-39-7) 2. CF3(CF2)7CH2COOH (2-perfluorooctyl ethanoic acid, 8-2 Saturated A cid, CAS#27854-31- 5) 3. CF3(CF2)6CF=CH2COOH (2-H-hexadecafluoro-2-decenoic acid, 8-2 Unsaturated Acid, CAS# 70887-84-2) 4. CF3(CF2)6COOH (Octanoic acid, pentadecafluoro-; PFOA; CAS# 335-67-1 ) 5. Heptadecafluorononaoic Acid; CAS#375-95-l (C9) 6. Nonadecafluorodecanoic Acid; CAS#335-76-2 (C10) PROTOCOL NO. : 112/021705/SED-TRANSa/SUB 112 /7 p. 16 W ildlife International, Ltd. -9 7. Perfluoroundecanoic Acid; CAS#4234-23-5 (C 11) Analytical Methods Methods of analysis will be verified prior to the start o f the study. Analytical reference standards that are used to aid the identification o f the test substance and its potential degradation products will be documented in the study records. All chemicals and solvents will be reagent grade or purer. Certificates of analysis (COA) will be provided for all test substances, reference standards, chemicals and solvents, when available. Sources will be documented in the study records. Demineralized water will be used in the study. Quality Criteria Recovery for a given sampling time point should range between 70 to 120% for the analysis of the 8-2 TBA, 8-2 Saturated Acid, 8-2 Unsaturated Acid, C9, CIO, C l 1, and PFOA fiom the spike recovery control samples. These ranges should be interpreted as targets and should not be used as criteria for acceptance o f the test. If recoveries and analysis do not meet the criteria of between 70 to 120%, then the study director will consult with the Sponsor's Representatives. 8-2 TBA Liquid Chromatography/ Mass Spectrometry (LC/MS), Gas Chromatography/Mass Spectrometry (GC/MS), or other suitable method can be used to analyze 8-2 TBA in the extracts of soil and headspace samples. Sample collection, preparation, and analytical methods used will be documented in the study records and the final report. 8-2 S aturated Acid, 8-2 U nsaturated Acid, C9, CIO, C l l , and PFOA Liquid Chromatography with Tandem Mass Spectrometry (LC/MS/MS) or other suitable method, will be used to analyze 8-2 Saturated Acid, 8-2 Unsaturated Acid, C9, CIO, C l 1 and PFOA in the extracts of soil samples. Sample collection, preparation, and analytical methods used will be documented in the study records and the final report. EVALUATION OF THE RESULTS Statistical methods including means, standard deviations, and regression lines, will be used as appropriate. The concentration of the transformation products in the soil and headspace will be given as PROTOCOL NO.: 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. - 10 mg kg'1(dry weight) and as mole kg'1(dry weight) for each sampling interval. Transformation products listed in soil and headspace samples will be plotted against time. HEALTH AND SAFETY It is advisable to read the MSDS for the reference chemical and test substance when available. If an MSDS is not available (e.g., the substance is a product o f research and development) it is important to review what is known about the substance with a person knowledgeable about the safe handling o f the substance, such as the person supplying the substance for testing. It is advisable to follow guidelines on Storage and Handling o f Chemicals; Personal Protective Equipment, Waste Disposal Guide. TEST SUBSTANCE DISPOSAL After the issuance of the final report, the remaining test substance will be stored at the testing lab until its expiration date and then destroyed, unless other arrangements are made between the Sponsor and the testing lab. RECORDS TO BE MAINTAINED Records to be maintained will include, but not limited to, the following: 1. A copy o f the signed protocol. 2. Identification and characterization o f the test substance as provided by Sponsor. 3. Study initiation and termination dates. 4. Experimental initiation and termination dates. 5. Test and reference substance preparation and dosing calculations. 6. Soil source and pretreatment data. 7. Results o f analytical methods performed. 8. Temperature range recorded during test period. 9. Copy of final report. PROTOCOL NO.: 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. -il - FINAL REPORT A final report o f the results of the study will be prepared by Wildlife International, Ltd. and submitted to the Study Monitor no later than one month after the conclusion of the definitive study. The report will include, but not be limited to the following, when applicable: 1. Name and address o f facility performing the study. 2. Dates on which the study was initiated and completed. 3. Objectives and procedures stated in the approved protocol, including any changes in the original protocol. 4. Identification and characterization of the test substance as provided by Sponsor. 5. A summary and analysis of the data . 6. A description of the transformations and calculations performed on the data. 7. A description o f the methods used and reference to any standard method employed. 8. A description o f the test system. 9. A description o f the preparation o f the test solutions, the testing concentrations, and the duration of the test. 10. A description of all circumstances that may affect the quality or integrity o f the data. 11. The name o f the study director, the names o f other scientists or professionals, and the names of all supervisory personnel, involved in the study. 12. The signed and dated reports o f each o f the individual scientists or other professionals involved in the study, if applicable. 13. The location where the raw data and final report are to be stored. CHANGES TO THE FINAL REPORT If it is necessary to make corrections or additions to the final report after it has been accepted, such changes shall be made in the form of an amendment issued by the Study Director. The amendment shall clearly identify the part of the study that is being amended and the reasons for the alteration. Amendments shall be signed and dated by the Study Director and Laboratory QA. CHANGES TO PROTOCOL Planned changes to the protocol will be in the form o f written amendments signed by the Study Director and approved by the Sponsor's Representative. Amendments will be considered as part o f the protocol and will be attached to the final protocol. Any other changes will be in the form PROTOCOL NO.: 112/021705/SED-TRANSa/SUBl 12 W ildlife International, Ltd. - 12of written deviations signed by the Study Director and filed with the raw data. All changes to the protocol will be indicated in the final report. GOOD LABORATORY PRACTICES This study will be conducted in accordance with Good Laboratory Practice Standards for EPA; will be consistent with OECD Principles o f Good Laboratory Practice. Each study conducted by Wildlife International, Ltd. is routinely examined by the Wildlife International, Ltd. Quality Assurance Unit for compliance with Good Laboratory Practices, Standard Operating Procedures and the specified protocol. A statement of compliance with Good Laboratory Practices will be prepared for all portions of the study conducted by Wildlife International, Ltd. The Sponsor will be responsible for compliance with Good Laboratory Practices for procedures performed by other laboratories (e.g., residue analyses or pathology). When the final report is completed, original copies of the study data and magnetically encoded records generated by Wildlife International, Ltd. for this study will be sent to the Sponsor. A certified copy will be retained in the archives of Wildlife International, Ltd. PROTOCOL NO.: 112/021705/SED-TRANSa/SUB 112 W ildlife International, Ltd. -13 REFERENCES 1 Organisation for Economic Cooperation and Development. April 2002. Aerobic and Anaerobic Transformation in Soil. OECD Guideline 307. PROTOCOL NO. : 112/021705/SED-TRANSa/SUB 112 Wildlife International, Ltd. Project No.: 112E-108 Page 1 of 1 AMENDMENT TO STUDY PROTOCOL STUDY TITLE: H-26634: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS PROTOCOL NO: 112/021705/SED-TRANSa/SUBl 12 AMENDMENT NO.: 1 SPONSOR: E.I du Pont de Nemours and Company P R O JE C T N O .: 112E-108 EFFECTIVE DATE: 06 May 2005 AM ENDM ENT: Maintenance o f Test Chambers, Page -7- CHANGE: The soil moisture content should be checked and adjusted, if necessary, at regular 1to 2 week intervals by weighing the test vessels. The soil moisture content should be checked and adjusted, if necessary, at monthly intervals by weighing the test vessels. REASON: The frequency at which the soil moisture content was checked was reduced based on observed losses. S' STUDY DIRECTOR (p DATE oof' 5 c .t 0 DATE L (0-4- Wildlife International Ltd. p. 22 PKO.fi;('T NO : ; 121.-10$ iOyi 1 of 1 STUDY TITLE: PROTOCOL NO: SPONSOR: DEVIATION TO STUDY PROTOCOL H-26634: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS 112/021705/SED-TRANSa/SUBl 12 E.I du Pont de Nemours and Company DEVIATION NO.: 1 P R O JE C T NO.: 112E-108 DEVIATION: Preparation o f Test Chambers; Viability Controls (untreated), Page -7- The protocol indicated that duplicate incubation chambers for each test soil and condition (aerobic and anaerobic) would be dosed at approximately 100 mg/kg dw with a combination of radiolabelled and non-labelled glucose. The test chambers were actually dosed at 3000 mg/kg dw with a combination o f radiolabelled and non-labelled glucose. REASON: Oversight by study personnel. IMPACT: In the best judgm ent o f the Study Director, this deviation did not impact the integrity o f study. A concentration o f 2000 to 4000 mg glucose per kg dry weight soil is a common for the determination o f glucose-induced respiration rates. STUDY DIRECTOR P f S e p / fl-ooi. DATE T O c t 0(,, DATE p. 23 PROTOCOL H-26665: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS This protocol complies with the Organisation for Economic Cooperation and Development OECD Guideline 307 Adopted April 2002 Submitted to E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 USA Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 February 17, 2005 W ildlife International, Ltd. 2- - H-26665: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS SPONSOR: E.I. du Pont de Nemours and Company SPONSOR'S REPRESENTATIVE: William Berti TESTING FACILITY- Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 STUDY DIRECTOR- Edward C. Schaefer Wildlife International, Ltd. LABORATORY MANAGEMENT: Hemy O. Krueger, Ph.D. Director o f Aquatic Toxicology and Non-Target Plants Proposed Dates: FOR LABORATORY USE ONLY Experimental Start Date: Project No.: / f WVXti 2oe c /iz e ./o q Experimental Termination Date: i l oot Test Concentrations: ?t'i) ^ / K cj , ill ^ / < `i ^ >0^ /{j Test Substance No.: f / f j>U, U`-24, J1, m i Reference Substance No. (if applicable): if ys' PROTOCOL APPROVAI 10 DATE 2.QC C / DATE {JO U T " PROTOCOL NO. : 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. -3 - INTRODUCTION Soil may be exposed to chemicals by direct application, spray drift, run-off, drainage, waste disposal, industrial or agricultural effluent and atmospheric deposition. This protocol describes a laboratory test method to assess the transformation o f the test substance in aerobic and anaerobic soil systems. No bias is expected in this type o f study. OBJECTIVE The objective o f the study is to assess the potential for transformation o f the test substance in aerobic and anaerobic soils. EXPERIMENTAL DESIGN The test will be conducted with four test soils under both aerobic and anaerobic condition. Four groups will be established for each soil type and condition (aerobic or anaerobic): (1) untreated (control) live soil; (2) 200 mg test substance/kg dw treated live soil; (3) 200 mg test substance/kg dw treated sterile soil; (4) 250 pg 8-2 telomer B alcohol/kg dw treated sterile soil; 10 pg 8-2 telomer B acid /kg dw; 10 pg 8-2 unsaturated telomer B acid/kg; and 10 pg perfluorooctanoic acid/kg dw, C9, CIO, and Cl 1 at 10 pg/kg dw all added to a sterile soil. The test systems will be incubated in sealed containers at approximately 20C for up to 1 year. Two incubation chambers from each of the groups will be removed at appropriate time intervals and the soil samples will be extracted and analyzed for potential metabolites. In addition to the day zero samples, a minimum o f eight additional samplings will be performed. Sufficient incubation chambers to allow for 3 additional sampling intervals will be prepared for the untreated control (1) and 200 mg test substance/kg dw treated live soil (2) groups. These chambers will be sampled as needed at the request o f the study monitor. Additional untreated and test substance treated soil chambers will be prepared for use as matrix fortification samples, biomass determinations, viability controls and to monitor aerobic conditions as necessary. The test groups will be prepared using 25 grams (dry weight equivalent) of soil. While this amount o f soil is less than the 50 to 200 grams specified in the OECD 307 guideline, it was chosen to allow for 1) the extraction o f the entire test chamber contents and internal surfaces; 2) sufficient PROTOCOL NO.: 112/021705/SED-TRANSb/SUBl 12 W ildlife International, Ltd. -4 headspace to minimize the need for active aeration which could result in a loss of potential transformation products. METHODS AND MATERIALS The test system and study conditions are based on the OECD Guideline for Testing of Chemicals, Guideline 307, Aerobic and Anaerobic Transformation in Soil (1). As this guideline is designed for low molecular weight substances, but not for polymers, adaptations o f the method may be necessary for feasibility reasons. Nevertheless, the study will be carried out in such a way that the recommendations given in the guideline will be followed as closely as possible. Test Substances Information on the characterization of test, control or reference substances is required by Good Laboratory Practice (GLP) Standards and Principles. The Sponsor is responsible for providing Wildlife International, Ltd. verification that the test substance has been characterized prior to its use in the study. If verification o f GLP test substance characterization is not provided to Wildlife International, Ltd., it will be noted in the compliance statement of the final report. The Sponsor is responsible for all information related to the test substance and agrees to accept any unused test substance and/or test substance containers remaining at the end of the study. The test substance is not radiolabeled and has no single molecular formula. It is a mixture containing perfluoroalkyl groups of varying carbon numbers as polymer side chains. Test Soils Four freshly collected US soils will be used to evaluate the test substances under aerobic and anaerobic conditions. The soils will be collected from each o f the following soil orders: PROTOCOL NO. : 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. -5- Soil O rder Percent of US Land* Alfisol 13 Mollisol 25 Inceptisol 16 Ultisol 13 Percent of total land in the US, from Foth, H.D. 1990. Fundamentals of Soil Science 8 ed. John Wiley & Sons, New York) The soils will be processed as soon as possible after sampling. Vegetation, larger soil fauna and stones will be removed prior to passing the soil through a 2 mm sieve. The soil used in the sterile groups will be sterilized by cobalt irradiation. The soils may be stored in the dark at 4 2 C, if necessary. However, storage and pre-incubation time together will not exceed 3 months. Characterization of the soils will be conducted by Agvise Laboratories (Northwood, North Dakota, USA). The following is a list o f the minimum physicochemical properties of the soil to be determined. Texture (i.e., percentage of sand, silt and clay) pH Organic carbon Bulk density Field moisture capacity Cation exchange capacity The percent moisture and water holding capacity of each o f the soil types will be determined. In addition, the microbial biomass of a live sample of each soil type will be determined. The soil microbial biomass will be determined at the beginning of the test prior to adding the test substance and in test and control soil samples at four months, at six months, and at 12 months using the fumigation-extraction method. Test Apparatus and Conditions The test chambers will be glass serum bottles with foil lined closures and will be identified by project number, test substance ID, test concentration, and unique identifier. The chambers will be incubated statically at approximately 20 2C. PROTOCOL NO.: 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. 6- - Soil Pre-Incubation The soil moisture o f each test soil content will be adjusted to 40 to 60% o f water holding capacity (which is equivalent to a pF o f between 2.0 and 2.5, or from 0.1 to 0.33 bar). The test soil then will be allowed to warm to the test temperature for a least 2 days. A pre-incubation period of at least 7 days for the aerobic soils and 14 days for the anaerobic soils will be performed under the appropriate conditions (i.e. temperature, soil moisture content, aerobic or anaerobic). Approximately 25 grams dry weight equivalent o f test soil will be added to the test chambers. Sterile soil treatments will be dosed at approximately 200 mg/kg (dw) with both chloramphenicol and cycloheximide to inhibit microbial growth. The chambers to be incubated under aerobic conditions will be sealed with septa and incubated. The chambers to be incubated under anaerobic conditions will be flushed with anaerobic mixed gas, sealed with septa and incubated. The soil moisture content should be maintained in the optimal microbial growth range of 40 to 60% water holding capacity (WHC). The soil moisture content should be checked and adjusted, if necessary, at least once during the pre-incubation period by weighing the test vessels. Sterile-filtered demineralized water (degassed sterile-filtered demineralized water for the anaerobic soils) will be added as necessary to compensate for water losses. Preparation of the Test Chambers The soil moisture content will be checked and adjusted, if necessary, prior to the addition of the test substance. Sterile-filtered demineralized water (degassed sterile-filtered demineralized water for the anaerobic soils) will be added as necessary to compensate for water. Anaerobic treatments and controls will be prepared within an anaerobic environment. The test substance will be applied to the soil within the test chambers as described below. Background Blank Control: Soils with no test substance added will be tested in duplicate to check for background concentrations o f analytes (see Sampling and Measurements). The entire bottle will be extracted after a sample o f the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Test Substance; The test substance dosed treatments will be prepared by dosing approximately 25 grams (dry weight equivalent) o f soil with sufficient test substance to deliver 200 mg/kg dry weight. The test substance will be administered by direct weight addition. The test chambers will be sealed with septum lined with aluminum foil, mixed and incubated. The entire bottle PROTOCOL NO.: 112/021705/SED-TRANSb/SUB 112 Wildlife International, Ltd. -7 - will be extracted after a sample o f the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Abiotic Controls: The abiotic controls will be prepared by dosing approximately 25 grams (dry weight equivalent) o f Co-sterilized soil with sufficient test substance to deliver 200 mg/kg dry weight. The test substance will be administered by direct weight addition. The entire bottle will be extracted after a sample o f the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Spike Recovery Controls: The spike recovery controls will be prepared by dosing approximately 25 grams (dry weight equivalent) of ^Co-sterilized soil with appropriate volumes o f 8-2 TBA stock (250 pg/mL in ethanol) and fluorinated acid stocks (10 |ig/m i. in water). The stocks will be injected directly into the soil using a glass microsyringe. The test chambers will be immediately sealed with septum lined with aluminum foil and the content of the chambers will be mixed. The entire bottle will be extracted after a sample of the headspace has been taken. The methods and procedures used will be documented in the study records and in the final report. Viability Controls (untreated): The metabolic activity of untreated test soil will be assessed on a monthly basis. Duplicate incubation chambers for each test soil and condition (aerobic and anaerobic) will be dosed at approximately 100 mg/kg dw with a combination of radiolabeled and non-labelled glucose. The evolved l4C 02 (aerobic and anaerobic conditions) and l4CfL, (anaerobic conditions) will be measured and the percent biodegradation calculated. The methods and procedures used will be documented in the study records and in the final report. Aerobic Controls (treated): The aerobic controls will be prepared by dosing approximately 25 grams (dry weight equivalent) of soil with sufficient test substance to deliver 200 mg/kg dry weight. The test substance will be administered by direct weight addition. The test chambers will be sealed with septum lined with aluminum foil, mixed and incubated. The oxygen content o f a sacrificial chamber will be measured at monthly intervals. The methods and procedures used will be documented in the study records and in the final report. Maintenance of Test Chambers The soil moisture content should be maintained in the optimal microbial growth range of 40 to 60% water holding capacity (WHC; a pF o f between 2.0 and 2.5, or from 0.1 to 0.33 bar). The soil moisture content should be checked and adjusted, if necessary, at regular 1 to 2 week intervals by weighing the test vessels. Adding sterile-filtered demineralized water (degassed sterile-filtered demineralized water for the anaerobic soils) will compensate water losses. PROTOCOL NO. : 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. 8- - The oxygen content in the headspace of two aerobic control chambers will be assessed on a monthly basis to ensure aerobic conditions. If the mean measured oxygen content within the control chambers is less than 19%, aerobic test chambers will be aerated with filter-sterilized, oil-free air. If test chambers have to be opened to add water or for monitoring purposes, a sample of the head space will be taken first using a C l8 cartridge as described below. The chamber will then be recapped as soon as possible. The septum and cap previously removed will be stored at -10C or lower. The C18 will subsequently extracted and analyzed. Sample extracts will be stored at -10C or lower if analysis is delayed more than 24 hours. Sampling and Measurements Duplicate chambers from each of the control, treated and spike recovery soils will be extracted (e.g., acetonitrile or other suitable solvent) and analyzed. Proposed sampling intervals will be at 0, 1 and 2 weeks and 1, 2, 4, 6, 9, and 12 months (9 sampling times), however the actual sampling intervals will be documented in the study records and in the final report. More or less frequent intervals may be conducted at the discretion o f the Study Director. Potential volatile transformation products in the headspace o f the soils chambers will be collected using an appropriate trap (i.e. C l 8 cartridge). At each sampling time prior to opening the test vessel, the septum will be pierced using a needle connected to a C18 cartridge and syringe. A volume of headspace gas from within the chamber will be pulled through the C l8 cartridge using the attached syringe. The C l8 will be subsequently extracted and analyzed. Sample extracts will be stored at -10C or lower if analysis is delayed more than 24 hours. Extracts will be analyzed for: 1. CF3(CF2)7CH2CH2OH (perfluorooctyl ethanol, 8-2 TBA, CAS# 678-39-7) 2. CF3(CF2)7CH2COOH (2-perfluorooctyl ethanoic acid, 8-2 Saturated A cid, CAS#27854-315) 3. CF3(CF2)6CF=CH2COOH (2-H-hexadecafluoro-2-decenoic acid, 8-2 Unsaturated Acid CAS# 70887-84-2) ' 4. CF3(CF2)6COOH (Octanoic acid, pentadecafluoro-; PFOA; CAS# 335-67-1) 5. Heptadecafluorononaoic Acid; CAS#375-95-l (C9) 6. Nonadecafluorodecanoic Acid; CAS#335-76-2 (CIO) PROTOCOL NO.: 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. -9 7. Perfluoroundecanoic Acid; CAS#4234-23-5 (C11) Analytical Methods Methods o f analysis will be verified prior to the start of the study. Analytical reference standards that are used to aid the identification of the test substance and its potential degradation products will be documented in the study records. All chemicals and solvents will be reagent grade or purer. Certificates o f analysis (COA) will be provided for all test substances, reference standards, chemicals and solvents, when available. Sources will be documented in the study records. Demineralized water will be used in the study. Quality Criteria Recovery for a given sampling time point should range between 70 to 120% for the analysis of the 8-2 TBA, 8-2 Saturated Acid, 8-2 Unsaturated Acid, C9, CIO, C l 1, and PFOA from the spike recovery control samples. These ranges should be interpreted as targets and should not be used as criteria for acceptance of the test. If recoveries and analysis do not meet the criteria o f between 70 to 120%, then the study director will consult with the Sponsor's Representatives. 8-2 TBA Liquid Chromatography/ Mass Spectrometry (LC/MS), Gas Chromatography/Mass Spectrometry (GC/MS), or other suitable method can be used to analyze 8-2 TBA in the extracts of soil and headspace samples. Sample collection, preparation, and analytical methods used will be documented in the study records and the final report. 8-2 Saturated Acid, 8-2 Unsaturated Acid, C9, CIO, C ll, and PFOA Liquid Chromatography with Tandem Mass Spectrometry (LC/MS/MS) or other suitable method, will be used to analyze 8-2 Saturated Acid, 8-2 Unsaturated Acid, C9, CIO, C l 1 and PFOA in the extracts o f soil samples. Sample collection, preparation, and analytical methods used will be documented in the study records and the final report. EVALUATION OF THE RESULTS Statistical methods including means, standard deviations, and regression lines, will be used as appropriate. The concentration o f the transformation products in the soil and headspace will be given as PROTOCOL NO.: 112/021705/SED-TRANSb/SUBl 12 33 W ildlife International, Ltd. - 10 mg kg*1(dry weight) and as mole kg*1(dry weight) for each sampling interval. Transformation products listed in soil and headspace samples will be plotted against time. HEALTH AND SAFETY It is advisable to read the MSDS for the reference chemical and test substance when available. If an MSDS is not available (e.g., the substance is a product o f research and development) it is important to review what is known about the substance with a person knowledgeable about the safe handling o f the substance, such as the person supplying the substance for testing. It is advisable to follow guidelines on Storage and Handling o f Chemicals; Personal Protective Equipment, Waste Disposal Guide. TEST SUBSTANCE DISPOSAL After the issuance o f the final report, the remaining test substance will be stored at the testing lab until its expiration date and then destroyed, unless other arrangements are made between the Sponsor and the testing lab. RECORDS TO BE MAINTAINED Records to be maintained will include, but not limited to, the following: 1. A copy of the signed protocol. 2. Identification and characterization o f the test substance as provided by Sponsor. 3. Study initiation and termination dates. 4. Experimental initiation and termination dates. 5. Test and reference substance preparation and dosing calculations. 6. Soil source and pretreatment data. 7. Results o f analytical methods performed. 8. Temperature range recorded during test period. 9. Copy o f final report. PROTOCOL NO.: 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. -h - FINAL REPORT A final report o f the results of the study will be prepared by Wildlife International, Ltd. and submitted to the Study Monitor no later than one month after the conclusion o f the definitive study. The report will include, but not be limited to the following, when applicable: 1. Name and address o f facility performing the study. 2. Dates on which the study was initiated and completed. 3. Objectives and procedures stated in the approved protocol, including any changes in the original protocol. 4. Identification and characterization of the test substance as provided by Sponsor. 5. A summary and analysis of the data . 6. A description of the transformations and calculations performed on the data. 7. A description o f the methods used and reference to any standard method employed. 8. A description o f the test system. 9. A description o f the preparation o f the test solutions, the testing concentrations, and the duration o f the test. 10. A description o f all circumstances that may affect the quality or integrity o f the data. 11. The name o f the study director, the names of other scientists or professionals, and the names of all supervisory personnel, involved in the study. 12. The signed and dated reports o f each o f the individual scientists or other professionals involved in the study, if applicable. 13. The location where the raw data and final report are to be stored. CHANGES TO THE FINAL REPORT If it is necessary to make corrections or additions to the final report after it has been accepted, such changes shall be made in the form o f an amendment issued by the Study Director. The amendment shall clearly identify the part of the study that is being amended and the reasons for the alteration. Amendments shall be signed and dated by the Study Director and Laboratory QA. CHANGES TO PROTOCOL Planned changes to the protocol will be in the form of written amendments signed by the Study Director and approved by the Sponsor's Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. Any other changes will be in the form PROTOCOL NO.: 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. - 12of written deviations signed by the Study Director and filed with the raw data. All changes to the protocol will be indicated in the final report. GOOD LABORATORY PRACTICES This study will be conducted in accordance with Good Laboratory Practice Standards for EPA; will be consistent with OECD Principles of Good Laboratory Practice. Each study conducted by Wildlife International, Ltd. is routinely examined by the Wildlife International, Ltd. Quality Assurance Unit for compliance with Good Laboratory Practices, Standard Operating Procedures and the specified protocol. A statement o f compliance with Good Laboratory Practices will be prepared for all portions o f the study conducted by Wildlife International, Ltd. The Sponsor will be responsible for compliance with Good Laboratory Practices for procedures performed by other laboratories (e.g., residue analyses or pathology). When the final report is completed, original copies of the study data and magnetically encoded records generated by Wildlife International, Ltd. for this study will be sent to the Sponsor. A certified copy will be retained in the archives o f Wildlife International, Ltd. PROTOCOL NO.: 112/021705/SED-TRANSb/SUB 112 W ildlife International, Ltd. -13 REFERENCES 1 Organisation for Economic Cooperation and Development. April 2002. Aerobic and Anaerobic Transformation in Soil. OECD Guideline 307. PROTOCOL NO.: 112/021705/SED-TRANSb/SUBl 12 Wildljfe International, Ltd. Project No.: 112E-109 Page 1 of 1 STUDY TITLE: AMENDMENT TO STUDY PROTOCOL H-26665: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS PR O T O C O L NO: 112/021705/SED-TRANSb/SUB 112 SPONSOR: E.I du Pont de Nemours and Company EFFECTIVE DATE: 06 May 2005 AMENDMENT NO.: 1 P R O JE C T NO. : 112E-109 AM ENDM ENT: Maintenance o f Test Chambers, Page -7- CHANGE: The soil moisture content should be checked and adjusted, if necessary, at regular 1 to 2 week intervals by weighing the test vessels. TO : The soil moisture content should be checked and adjusted, if necessary, at monthly intervals by weighing the test vessels. REASON: The frequency at which the soil moisture content was checked was reduced based on observed losses. STUDY DIRECTOR LABORATORY MANAGEMENT J C S t {? ' J o t e DATE S & i /u, DATE Wildlife International Ltd. PR( 1of 1 STUDY TITLE: PROTOCOL NO: SPONSOR: DEVIATION TO STUDY PROTOCOL H-26665: TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS 112/021705/SED-TRANSb/SUB 112 DEVIATION NO.: 1 E.I du Pont de Nemours and Company P R O JE C T NO.: 112E-109 DEVIATION: Preparation o f Test Chambers; Viability Controls (untreated), Page -7- The protocol indicated that duplicate incubation chambers for each test soil and condition (aerobic and anaerobic) would be dosed at approximately 100 mg/kg dw with a combination of radiolabelled and non-labelled glucose. The test chambers were actually dosed at 3000 mg/kg dw with a combination of radiolabelled and non-labelled glucose. REASON: Oversight by study personnel. IM PA C T: In the best judgm ent ofthe Study Director, this deviation did not impact the integrity of study. A concentration o f 2000 to 4000 mg glucose per kg dry weight soil is a common for the determination o f glucose-induced respiration rates. STUDY DIRECTOR LABORATORY MANAGEMENT J ''*<? /___ (V(: DATE 5 <9d Q(, DATE APPENDIX II p. 39 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED D ocum ent N um ber in Index: 10 Document Description: Testing Invoices p. 40 APPENDIX m p. 41 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED D ocum ent N um ber in Index: 12 Document Description: DuPont Investment in Fluorotelomer Environmental Fate Science & S0& / 2 - p. 42 APPENDIX IV he. B p. 43 U.S. EPA - DuPont Created By: Robert C Buck on 07/11/2002 at 02:50 PM Workgroup. Category: Sub Category: Product Stewardship General General Security U.S. ENVIRONMENTAL PROTECTION A G E N C Y 4 March 2007 : f ) DuPont Investment in Fluorotelomer Environmental Fate Science: sM- DuPont Investmert ir Fluorotelomer nvuimri;)l Fntr Science 20DMBfdt tS- Presentation: 2007 ?**<(, u .fpasi, ^ RCBndf (DuPont Confidential Information) 2) Wildlife Inti a) Invoices: 1- . V#? 3007_Feb_2005-06 wticarfeh i invi c. p.cjt Qgote for 307/311 ?ons WLF_Coststo date andEst Futire Studies Costs pdf (DuPont Confidential Information) S3f- 3) ENVIRON Invoices: Environiwwces, DEMStudy n i r-n (D uPont Confidential Information) 13?.. .^ Historical Archive Document of EPA Presentations : SirtaceProtections Solutions -.pur Soil Siodegradtion Study Protocols: a. Urethane Polymer Protocol: i:i iorwncol_CBl.pdf Acrylate Polymer Protocol: i i 2E-iDsurut^,H . iil|..n Atote. The studies started with a suite o f 7 analytes. Two additionalanalytes w s re a d c M a i 15 m onths. ta- SETAC Presentati o ns : -E1Ad rj. 1'lyrrufiK'dca.pdl 2006_SETACNA_PolymerBiodegPoster_RC8_DRAFT_OnePage.pdf(not confidential) 3 November 2006 : Preliminary Aerobic Soil Studies Information sent as DuPont Confidential Information. Contains T5CA CBI. Subject to Copyright. Contents o f CD 1 o f 3 iga 'd> Cover Letter: 2006_3 Nov_Cover Letter Sol Studies_USEPA_CBI.pdf m* . Draft Study Reports: 200B_02Nqv_H2666S_draft report_rpti09_CBi.pdf .~ 2006_02 Nov_H26634_DRAFTREP0RT_RPT10B_(H.pdf Raw Data Index: 20Q6_02 Nov_DRAFT_5oi Skudes Records IndexjCa^df iiiM Analytical Data Summary: 2G06_02 nov_DRAFT 12 MonthAnalytical Summary_CBt.pdf SS Test Substance Certificates of Analysis: saaaf'. ^k 3_2006_110ct_DRAFI_H-26fi^_.;OA_CBlp<ll it 4_-'uuG_nucr_UKAr Contents of CD 2 of 3 A_k.brpar ~ Contents of CD 3 & 3 DuPont Soil Biodegradation Study Update. October 2006, Washington. DC CBI Documents Provided in Advance: tSp Jjte 1_1 r |. ettei- i<vrg ( r>cn ?n<~t?nnp_rpi prjf Test Substance COAs: p. 45 1_K26634_COAJ7Oct2005.pdf 2_H26665_COAI_17Od2005.pdf <iZa jzm ---- 3_2006_1 1 0ct_DRAFT_H-26634_COA.pdf 4_2006_11Oct_DRAFT_H-26665_C0A.pdi iE * SEM- Study Protocols: S_H26634_Soil Study_n 2 E-1 O&ptotocoi.pai ._hjuua.. -_..*i_.ti*t* 11 :t 1 ofl| .i[4ccrtpdf -st Analytical Metehods: 7- So'1Study Analytical Methodi t.ui i <gaf. a i:iMir.-ji M<-th,.d Added analytec.pdf "J Data: 10_Mollisol Raw Data AnayUcal Summary.It IM Interpretation: 8_2006_noctjnterpetaiiL<-.AA^tihm aj '.-.iinjiitw pdf C l '} 9_2006J0Oct_Merpretation AgorithmVvorl&ooli.dL Modeling the Study Results: ii-Mudcimn ni i tnddonier-hnsed Polymeric Products_Soi! Biodeg Studies_DRAFT.p<tf Non-CBI Documents Provided in Advance: B33P". A A_12 October 2006 Email to DLynch EPA NOTC8l.pdf tfa 'A C_6-2 FTOK Biodegradation P^hwaysJ40T c a p d f iM- B_2006J1 Oct_Pub#c*on &Report Kciein usa?" .Jk D_20O6_XlroniAJ 11u_pp II- I:a F.di r+.T .; nt.ffl DuPont Presentation, 15 September, 2006 2UCfcj5Sept_L)U Review with ustPA.ppt DuPont Presentation, 16 August 2006 PMN Meeting ' P Premanufacture Ntrtce slides 081606 final Company Sanitized.ppt EFA Meeting 0616UC m,d ,m ^ 13 ! cka rweangnouo rinai company ianmzeajopt epa meeting imrouucnori utn ;,ui. m ,,i.. i( |14 77 EPAMeeting Introduction081506 final CompanyCammed ppt PremanuiLik'turrrmii ,. imimF Inal cpi ppl DuPont Presentation, 8 September 2005 : epa Mg 9-b-u-z-Ci^ DuPont Presentation, 8 April 2005: r-upplv Chain Review 08 April'05 EPA.pi* 9 i EPA Meeting 06 AprTOS.pp* GDEM April 2005 SHK.ppt Paper PS Summary Apr! 2005 SHK.ppt DuPont Presentation, 31 January 2005 : DuPont Global Strategy Presentation : 3lJen2005 DuPont Pre-ierJahuMu U'-lka ai .vi. n w p rif DuPont Supply Chain Review : Supply uri Review ji j ^im:. i n.>i : f Fft DuPont Presentation, 15 December 2004 ; DuPont Biodegradation Studies -- j Um^iqu 15U6C2UU4_L'Uf'urit-f.F`A M-v'tirm I'i> . ml.itinn prf 15Pec2004_DuPont-EPA Meefrig Agenda .pdf DuPont Presentation, 10 November 2004 : J II i I mfiint Ongoing Research Summary_9Nov2004_NOCBi.ppt "Cl n Mnv.-vwM FPa_0uDont Ongoing Research Summary_CBI-RWRppt p. 47 DuPont Presentation. 17 June 2004 : DuPont Consumer Article Exposure & Risk Characterization (FINAL VERSION) *2 Purnrut Risk Assessmert - Article Presentatioryj-iEf^.j .Mua-i'irx F-lf DuPont Risk Assessmert - Article Presentation Tenfv. li-.iunemns ppt DuPont Presentation, 30 April 2004 (FINAL CBI VERSION) DnPonl PFDA RHnrtion R DuPont PFDA Rprtnrtinn Rpryvt I NON-CSI VERSION DnPrinl PFDA Rerlnrtinn Rnnnrt fM ^W I fuP n n t Ronnrt rnAnri]?nrU n. U S . EPA-OPPTS DuPont Presentation, 16 March 2004 (FINAL: CBI VERSION) Rranri PresenT*TM i jk f p a 1RMs U S. EPA-OPPTS DuPont Presentation, 25 November 2002 (FINAL: CBI VERSION) fcRIUrarimfW tIRFPA Di.Pnnt PR I Jnrfciln (FINAL: NON-CSi VERSION) ?rimnu?nn? il s f p a nuPnm p.q Unrlsi* F Copy of the Non-CBI Document in the Public Record (AR-226) PSKInuSDf]? D uPnnt P im e n ta tin n tn I IS FF U.S. EPA-OPPTS DgPont Presentation, 17 Decem ber2001 p. 48 17 DEC 2001 DUPONT i7 r w .? n m n i iPnnt f p a p r i {CBI VERSION : 17 Dec 2001 RC 3) 17 DEC 2001 DUPONT i7rVn?nm rtnPnnt f p a NTMr:R(NON-CBI VERSION : 17 Dec 2001 RCB) 17Dec2001 EPA Presentation with Post Meeting Corrections/Updates fluPnntPPA Prpcontqfinfl Hnov w P U.S. EPA-OPPTS DuPont Presentation, 17 December 2001, RESUBMITTED 1 2 F E B , 20 0 2 i7 n r^ ? n m nnPnnt f p a wnnCRi i7 n e r ? n m nu P nm f p a c r i 1?Fnh SWV IJ^FPA l Inrlatorl Prociirvtatinn fViyor I Ptfi U.S. EPA-OPPTS TRP Presentation (20 JUN 2000) 20 June 2000 Mtg TRP rnlg_w EPA Chart; U.S. EPA-OPPTS TRP Presentation (23 OCT 2000) EPA Review Presentation OCT23_2000_tina U S-EPArOPPTS Presentations DuPont & TRP (21 Feb 2001) (DO NOT MAKE ANY CHANGES) FPA Revinw n i IPONT Prr^r^nt^tian FFH?1 FPA Fil F TRP PreRfinlalion FFR: (CHARTS GIVEN TO THE EPA-OPPTS on 21 FEB 2001 non-CBI Versions o f the Above) "iTvf".V Di rPH N T Pj fiRi'DJiri;' "t *** r 3; ; rFfi7 p. 49 U.S. EPA-ORD, Duiuth, MN, 7 June 2001 (R. C, Buck) 7Ju;i2n01 EPA_ORD Discussion Miscellaneous Presentations to and by U.S. EPA August 3. 2000 EPA Staff Presentation on PFOS 226-0619.pdf SPI FMG Presentation to U S EPA 23 April 2002 Final No CBI April 23 EPA Presentation Edit History: Rey. 70. Editor Robert C Buck m . Robert c Buck 68. Robert C' Buck 67. Roiwrl C Buck 66. Rotieri C Buck *Only p a st five edits arc shonn Edit Date 03/08/2007 03:48:37 pm 03/07/200704:32:26 PM 03/07/200/ 07:53-07 AM 03/07/2007 07:48:16 AM 03/07/2007 07:47:59 AM p. 50 COPY RESTRICTED DOCUMENT DOCUMENT IS READ ONLY NO COPYING WITHOUT AUTHOR PERMISSION D ocum ent N um ber in Index: 15 Document Description: Biodegradation studies o f fluorotelomer-based polymers in soils C o m m en t: The printed copy o f the document may be viewed, but may not be copied without author permission. DuPont contact information for requesting author permission to copy: Dr. Robert Buck 302-892-8935 robert.c.buck@ usa.dupont.com -^ 2 /s mm Biodegradation studies of fluorotelomer-based polymers in soils. The mincies o f science Subject to Copyright. Do not cite or quote without author's written permission. Abstract _ Buck1, W. R. Berti1, M. H. Russell1, B. Szostek1, N. Wang1, E. Schaefer2, R. Van Hoven2 1 E. I. duPont de Nemours &Co., Inc. Wilmington, Delaware U.S.A. ______ 2 Wildlife International, Ltd., Easton, Maryland, U.S.A Study Methodology W 'ihlliji hitmiritituiiil. / t,l Poster presented at SETAC North American Meeting November 2006 Montreal, Canada Kinetic Modeling Biodegradation o f fluorotelomer-based polymers in the environment is o f interest to determine if they may be a potential source of perfluorocarboxyltc acids (PFCAs) We have conducted studies on a fluorotelomer-based acrylate polymenc product in four soils to determine whether the fluorotelomer moieties covalently bonded in these polymers can be liberated and transformed to PFCAs Treatments included an untreated control, live (non-sterile) treatm ents containing the test sub sta ice stenle treatments with the test substance, and sterile treatments spiked with several analytes to assess recoveries The test vessels were incubated statically for one year. Separate headspace and soil samples were collected and analyzed The polymenc test substance contained residual fluorotelomer raw m aten^s that are known to degrade to the target analytes The experimental data after 15 months were analyzed using a kinetic degradation model PFCAs quantified in the test systems ongmated exclusively from transformation o f the residual fluorotelomer raw m atenals There was no indication that polymer degradation contributed to PFCAs observed The estimated fluoroacrylate polymer degradation half-life was > 10,000 years Introduction Perfluoro carboxylates (PFCAs) have been identified as widely present in th e e nvironm ent1 Recently a complete picture o f PFCA sources has been review ed2 Direct emissions o f PFCAs are the largest source o f g lo b * histone PFCA emissions Indirect PFCA sources, or "precursors' , included perfluoroalkyl sulfonyl- and fluorotelomer-based substances For these indirect sources, residual unreacted raw matenals and impunties present in sales products were identified as a potential source of PFCAs A key uncertainty in the assessment of indirect PFCA sources was whether fluorotelomer-based products, largely composed o f acrylic polymers, or perfluoroalkylsulfonyl-based products degrade in th e environment to form PFCAs In order for this to occur! the OECD 307 as a guideline 4 soils, aerobic and anaerobic conditions initially. Measurements at 0 ,1 , 2 weeks; 1 ,2 ,4 ,6 ,9 ,1 2 ,1 5 ,1 8 , 24 months Analyzed 12 and 15 month samples for 7-3 A d d and 7-2 sFTOH. Completing 18 months sampling and analysis. Final samples to be taken at 24 months. 1. Alfisol: Howard County, Missouri Sandy loam (11% d ay ), pH 5.8, Organic carbon 1.4% 2. Inceptlsol: Deer Park (Spokane Co.), Washington Loam (23% clay), pH 6.6, Organic carbon 2.0% 3. Molllsol: Grand Forks County, North Dakota Sandy d a y loam (23% d ay), pH 6.9, Organic carbon 2.1% 4. U ltis d : New Castle Co, Delaware Sandy loam (14% clay), pH 4.8, Organic carbon 2.9% General Experimental Design Background Blank C ontrd: no test Item added Test Item; testitem concentration o f2 0 0 mg active Ingredient kg-1 dry w t soli Abiotic Control: sterilized test medium with added test Item concentration o f 200 mg a.I. kg-1 DW soli. Spike Recovery C ontrds: sterilized medium spiked with following analytes: 8-2 FTOH, 8-2 FTA, 8-2 FTUA, PFO, PFN, PFD, PFU, 7-2 sFTOH, 7-3 FTA Test vessels are glass serum bottles with aluminum foil-lined d osures Incubated statically a t 20*C. Soli m dsture content assessed regularly by weighing each bottle and adding water as needed to maintain a level o f 40 to 60% w ater-hddlng capacity. A headspace sample from each vessel Is passed through a C1B cartridge. The entire contents o f the test vessel are then extracted first using acetonitrile and second with an aliquot o f a 200 mM NaOH solution. Polymer Degradation Kinetic Model kl degradami ralaof polymer k2* dgradt* ralecf alcohol/ n Conceptual Polymer Degradation Pathway Equationa Polymer. dPfdl - (kl ' a * P) Alcohol: dAfdt * (k1` c ` PI-lk2a( 'A l Acid. 4Aeidr> (k2 p A) ' awoiv<a u ty Optimize ONLY degradation of Residual Materials to form PFCA Optimize ONLY degradation of Polymer to form PFCA polymer backbone and/or the covalent bond or "linkage* between the fluorotelomer o r perfluoroalkyl sulfonyl functionality and the polymer backbone would have to be severed AE Mass Balance Interpretation Biodegradation is a transformation route that will determine polymer environmental fate A limited number of experimental biodegradation studies have been conducted on fluorotelomer alcohol (FTOH), a residual raw matenal, which have identified transformation products that can be used to probe polymer degradation 3 * The present study was designed to determine whether and to what extent a fluoroacrylate polymer derived from fluorotelomer alcohol degraded in aerobic soils to form perfluorooctanoate (PFO) Test Substance & Degradation Pathways Alcohol Equivalents (AE) enfimikm AE = Moie* of 8-2 FTOH equivalents or "Alcohol Equivalents' derived from 8-2 eubstencee (e g 8-2 fluorotelomer acrylate) present at the Test Substance (TS) Residuals = The sumof raw matenals and impurities idenhfied and present n Polymei AE ornai Af mgTestScintane* , 172 nmolAE mg TS AEInRolymtrperrager Test Substance ol AEInResiduilapermgorTestSubstwice AE Mass Balance Approach Fluorotelomer-based Acrylate Polymeric Product Test Substance OCH,CHjC,F,, Fkjorotlom*i acrybk monomer the Test Substance Input Values The tim e course o f residual fluorotelomer alcohol degradation provides a much more reasonable fit to th e expenmental At aqueous dispersion of fluoroacrylate polymer m water - Fkiofotelomar ecrytot* monomer emufeton potymerced w*h other m Produc* by process Composition - Fluoroecrylale polymer Hydrocarbon surfactants JC i ( / V X /\ Polymer Degradation F(cfj,ichchjoh F(CT7),,CH.CH,OC(0)0*CH; Cleave Ester Linkage (hydrolysis) J Polymer Backbone i Degradationto I Olnomert CHl=CHC(=0)-i-OCH,CH,C,FII Residual Materials Degradation Oeave Ester Linkage (hydrolysis) C,F,,Ch,CH?OH B-2Fluorotelomer Alcohol * X1 = weghl percent (wt%) Fluorine ai the Test Substance (TS) X2 - weight percent (wt%) Fluorine r the Fluorotelomer Alcohol (FTOH) RawMatenal used to make the Test Substance X3 = weight percent(wt%) 8-2 Fluorotelomer Alcohol (8-2 FTOH) n the Fluorotelomer Alcohol Raw Matenal AeiyuPdymr Incapttcl V ( S3 FTOH Acryl*Permei si UHnd FTUA -* PFO FTOH 72FTOH< 00 \ ............. ............. .... data than the corresponding bme course o f polymer degradation alfisol inceptisol molfisol ultisol P o ly n w r hatete > 10,000 > 10,000 > 10.000 > 10.000 Residual M aterials h a te ilte 33 38 45 16 Kinetic Modeling Summary Primary source of PFCA fits with a model for degradation of Residua! Materials alone. Calculated degradation half-life of Residual Materials ranges between 16 and 45 days in the four soils. Calculated Acrylate Polymer degradation half-life is greater than F(CFJ)71*CFjCHjCHO 10,000 years. S3 FTAL FJCF^-CFjCHjOOjH S-3PTA FlCF^COjH PFNA fF(Cf)7uCF=CHi J[ 8-1 Olefin FlCF^MCFaCHCOjH |F(CF)714CHOHCH;COOHI S-2FTUA f(CFj)7CH=CHj'1 T F(CF2))'*CH=CHCOjH I IIon 397 [ 7-2 Olefin J ' RCf^-CHfOHJCH, 7-J uAeid 1 _ FtCFJ.COjH - " CO, PFHA 7-2 FTOH FlCFA^CHjCHiCOjH 7-8 AsM - .PFHpA F(CF,)yuCOjH I J-Oxidatic 1 k PFOA ** ------- - j Monitored Study Analytes Aova* Pdpn r Affo 1------b . ------ -----------a_____ a___ <00 500 T: Summary & Conclusions Degradation analytes determined in the test system can be exclusively attributed to degradation of residual fluorotelomer raw materials and impurities in the test substance. The results obtained from kinetic modeling the degradation kinetics of the fluoroacrylate polymer further support the conclusion that PFO formation is dominantly due to the transformation of the residual materials. 'P The degradation of the polymeric structure appears to be negligible with calculated degradation half-lives that are > 10,000 years. 8-2 Fluorotelomer AWotid (FTCH) F(CF,CH,CHflH 8-2Fluorotelomer Aod (FTA) F(CF,)1CH,COOH 8-2 Fluorotelomer UnsaturetedAcid (FTUA) F(CF,),CF=CI-lCOOH 7-3Aad 7-2 sFTOH F(CFp,CH2CH2COOH F(CFj)tCH(OH)CHj PFO. PFN. PFO PFU(PFG8) FfCF^COOH, n =7, 8, 9 ' PFN, PFD and PFU levels observed are consistent with expectations based on test substance composition and known degradation pathways (e g P F N -1 % o f PFO) PFN, PFD and PFU levels observed are consistent with expectations based on test substance composition and known degradation pathways, (e.g PFN -1% of PFO) Demonstrated analytical methods & study operation for over 15 months. ' f & v nSS S l s k f f t f e 40(115, 3463 2 p,cveaourosetal Sa Tf^nol 40(1) 32 3 W arnet Environ So Tschod 3 p. 52 COPY RESTRICTED DOCUMENT DOCUMENT IS READ ONLY NO COPYING WITHOUT AUTHOR PERMISSION D ocum ent N um ber in Index : 16 Docum ent Description: Biodegradation studies o f fluorotelom er-based polym ers in activated sludge, soil, and sediments C om m ent: The printed copy o f the document may be viewed, but may not be copied without author perm ission. DuPont contact information for requesting author permission to copy: Dr. Robert Buck 302-892-8935 robert.c .buck@ usa.dupont.com 0 Biodegradation studies of fluorotelomer-based polymers in activated sludge, soil, and sediments. W.R. Berti. B.Szostek, R.C. Buck, N. Wang, and J.T. Gannon E. I. duPont de Nemours & Co., Inc. E. Schaefer and R.L. Van Hoven Wildlife International The miracles o f science p. 54 Outline Do not cite or quote. Introduction Biodegradation studies Biodegradation studies Biodegradation studies Biodegradation studies Summary in activated sludge in soils in sediments in anaerobic digester sludge p. 55 Do not cite or quote. Not for further distribution without author approval. Summary Study results to date Soil and Sludge methodologies and analytical methods have been demonstrated No evidence of polymer transformation observed - Inherent Biodegradation in Sludge - 21 Days - Biodegradation in four Soils - 180 Days Ongoing studies Soil Studies continuing to one-year Sediment studies commenced continuing to one year Anaerobic sludge studies to commence shortly Do not cite or quote. Not for further distribution without author approval. Polymeric Product Dispersion Aqueous Dispersion of.... Polymeric Particles 100-200 nm Hydrocarbon Surfactant(s) "Typical" Polymeric Product 20% Polymer wt.% Active Ingredient (~ 20-30 %) wt.% Fluorine (~ 6-10%) ~ 500 ppm Residual Raw Materials - Telomer B Alcohol - Telomer B Olefin - Telomer Acrylate Monomer Polymer M w > 10,000 Do not cite or quote. Not for further distribution without author approval. Product Routes to the Environment Soil (e.g. landfill) - Municipal and Industrial Waste Landfill Waste-Water -S lud ge & Sediment Incineration -M unicipal and Industrial Waste - No PFOA formed under typical municipal waste incinerator conditions Yamada et al. Chemosphere 2005, 61, 974-984. Do not cite or quote. Not for further distribution without author approval. p. 58 Study Objectives : Sludge, Soils, and Sediments Preliminary studies: - Develop, demonstrate and optimize experimental methods and analytical procedures. Main studies -A ssess degradation potential of fluorotelomer-based polymers by monitoring formation of potential products of degradation 8-2 Fluorotelomer Alcohol (FTOH): F(CF2)8CH2CH2OH 8-2 Fluorotelomer Acid (FTA): F(CF2)8CH2COOH 8-2 Fluorotelomer Unsaturated Acid (FTUA): F(CF2)7CF=CFICOOFI Perflurorooctanoic Acid (PFOA): F(CF2)7COOH ^ p. 59 Do not cite or quote. Not for further distribution without author approval. Aerobic Activated Sludge Studies Used experimental set-up for a Modified ZahnWellens Study (OECD 302B) 300 mg DOC as test substance/L 1 g sludge (D.W .)/L Up to 28 days Measured Dissolved Organic Carbon, F-, and specific metabolites - Trapped gasses in C 18 resin column - Initial experim ents to examine polymer degradation in sludge Do not cite or quote. Not for further distribution without author approval. O<0 Q. Inherent Biodegradability : Acrylate Polymer No evidence of polymer transformation observed after 21 days 60 50 </) ^o4 0 E =L w 30 (D I < 10 0 8-2 FTOH in solution 8-2 FTA PFOA 8-2 FTOH in off-gas 8-2 FTUA 45 |jmoles 8-2 FTOH measured in product Day 7 Day 21 p. 61 Do not cite or quote. Not for further distribution without author approval. General Experimental Design : Soil Studies Background Blank Control: no test item added Test Item: test item concentration of 200 mg active ingredient kg-1 dry wt. Abiotic Control: sterilized test medium with added test item concentration of 200 mg a.i. kg-1 DW soil. Spike Recovery Controls: sterilized medium spiked with following analytes: - 8-2 FTOH, 8-2 FTA, 8-2 FTUA, PFOA p. 62 Do not cite or quote. Not for further distribution without author approval. Soil Studies : OECD 307 as a guideline Preliminary Study -T w o test items and one soil under aerobic conditions for 28 days Sandy clay loam from North Dakota; pH 7.9; 1.2 % organic carbon -Demonstrated experimental and analytical capabilities, recoveries of analytical spikes. -N o evidence of polymer transformation observed Do not cite or quote. Not for further distribution without author approval. Main soil studies : In-progress Four soils - Alfisol: Howard County, Missouri Sandy loam (11% clay), pH 5.8, Organic carbon 1.4% - Inceptisol: Deer Park (Spokane Co.), Washington Loam (23% clay), pH 6.6, Organic carbon 2.0% - Mollisol: Grand Forks County, North Dakota Sandy clay loam (23% clay), pH 6.9, Organic carbon 2.1% - Ultisol: New Castle Co, Delaware Sandy loam (14% clay), pH 4.8, Organic carbon 2.9% p. 64 Do not cite or quote. Not for further distribution without author approval. Soils Studies Test vessels are glass serum bottles with aluminum foil-lined closures incubated statically at 20C. Soil moisture content assessed regularly by weighing each bottle and adding water as needed to maintain a level of 40 to 60% water holding capacity. A headspace sample from each vessel is passed through a C18 cartridge. The entire contents of the test vessel are then extracted first using acetonitrile and second with an aliquot of a 200 mM NaOH solution. Do not cite or quote. Not for further distribution without author approval. Soils Studies : Main Studies in-progress p. 65 Measurements at 0, 1, and 2 weeks, and 1,2,4, and 6 months -A ll analytes extracted from control (untreated), treated live, treated sterile, and spiked (with all analytes, no test substance) sterile soils, each replicated twice - 8 - 2 FTOH measurements of headspace samples -M oisture content, viability assessments, 0 2 content of treatments, biomass (months 0, 4 and 6) K \$ Status -T e s t systems working well; fast mineralization of 14C-glucose; moisture content constant (-5 0 % W HC) -C hanging to LC/MS to determine 8-2 FTOH in soil extracts - N o analytes in controls; 8-2 FTOH not measured in headspace samples p. 66 Do not cite or quote. Not for further distribution without author approval. Recoveries from four soils 120 Includes all four soils incubated under sterile 100 conditions Spikes sO I - 8-2 FTOH: 250 ug/Kg f 01 -Acids: 10 ug/Kg Average recoveries up to 6 months - 8-2 FTOH up to 60 days - -7 0 % and greater. 8-2 FTOH 8-2 FTA 8-2 FTUA PFOA p. 67 Do not cite or quote. Not for further distribution without author approval. Recoveries from Soils up to 180 days Four soils, sterile conditions Recoveries of PFOA and 8-2 FTUA > 80% 8-2 FTOH and 8-2 FTA recoveries decreasing with time. - Not recovered - bound residue Days 120 \5 Do not cite or quote. Not for further distribution without author approval. Recoveries from Soils up to 180 days Average recoveries up to 6 months under sterile conditions - 60 days for 8-2 FTOH. Recoveries of 8-2 FTOH variable Differences by soil type Alfisol Inceptisol Mollisol Ultisol p. 69 Do not cite or quote. Not for further distribution without author approval. S o il: Acrylate Polymer Product 8-2 FTOH concentration Four soils combined decreases indicating biodegradation After 180 days, PFOA is less than the 8-2 FTOH added to soil as an residual Month 6 samples taken. Continue to one year No evidence of polymer transformation observed 2.1 pmoles 8-2 FTOHKg1 soil added as residual 130 pmole equiv.8-2 FTO H *K g1 soil within polymer LOQ of 8-2 FTA and 8-2 FTUA < -0 .0 0 4 pmoles-Kg1 8-2 FTOH not measured at 120 and 180 days PFOA |jmole/Kg soil Acrylate Polymer Product oval. 2.5 * PFOA formed appears to be leveling off at the concentration of residual 8-2 FTOH. PFOA formation is only from the degradation of residual 8-2 FTOH and not from the polymer. No evidence of polymer transformation observed p. 71 Do not cite or quote. Not for further distribution without author approval. Summary - Soil Studies No evidence of polymer degradation observed thus far Demonstrated analytical methods & study operation Sterile spike recoveries - Good overall recoveries, especially for PFOA and 8-2 FTUA - Validating alternative method and will reanalyze samples for 8-2 FTOH 8-2 FTOH was not found in the head space of any samples, - including sterile soils to which it had been fortified at 0.54 pmole kg-1 (250 pg kg-1, LOQ < 0.0125 pg m L 1 headspace). Acrylic Polymer - 8-2 FTOH, 8-2 FTA, 8-2 FTUA, and PFOA extracted from soils can be accounted for by transformation of fluorinated residuals alone. - Similar results for other polymer products under study p. 72 Do not cite or quote. Not for further distribution without author approval. Sediment Studies Treatments and methods for the sediment similar to soil studies. Sediment and water added to each test vessel - watensediment volume ratio between 3:1 and 4:1 ; the minimum sediment layer is ~ 2 cm. Headspace gases continuously purged a low air flow rate; passed through a C18 cartridge. Anaerobic test vessel conditions are imposed as in the soil test. In addition to headspace and sediment extract samples, separate aqueous samples are collected and analyzed. p. 73 Do not cite or quote. Not for further distribution without author approval. Sediment System Studies : OECD 308 Guideline Preliminary Studies : complete - In-life completed (28 days) - Anaerobic sediment from Turkey Creek (Talbot County, MD USA) High organic carbon content (6.4% O C), pH 5.6, and fine texture (loam) - Changing to LC-MS (from GC-MS) for 8-2 FTOH to improve analytical after validating method Main Study - Review preliminary study results for recoveries once 8-2 FTOH analysis complete and revise protocol - 2 sediments Aerobic sediment from Choptank River (Denton, MD USA) Low organic carbon content (0.5% OC) and a course texture (sand). S am e source of anaerobic sediment as that used in preliminary study - 12 month duration; may extend to 18 months depending on data from soil studies Do not cite or quote. Not for further distribution without author approval. Preliminary studies in anaerobic aquatic sediments: Spike Recoveries PFOA 140 One sediment, sterile conditions Good recoveries of PFOA and 8-2 FTUA Sediment Water DayO Day28 Do not cite or quote. Not for further distribution without author approval. Preliminary studies in anaerobic aquatic sediments: Spike Recoveries 8-2 FTOH One sediment, sterile conditions Low recoveries of 8-2 FTOH Recoveries of 8-2 FTA inconsistent Day 0 Day 28 p. 76 Do not cite or quote. Not for further distribution without author approval. Summary - Preliminary sediment studies No evidence of polymer degradation observed Low recoveries of 8-2 FTOH from spike recovery control systems -Validating LC-MS method (from GC-MS) for 8-2 FTOH and reanalyze samples ^ ^ Revising and finalizing protocol for main study -A ssess ability of test vessels and systems to contain analytes of interest, especially 8-2 FTOH p. 77 Do not cite or quote. Not for further distribution without author approval. Anaerobic Biodegradability in Digester Sludge Study OECD 311 Guideline Preliminary Study -Develop and validate analytical methods in digester sludge -Protocol under review 1 anaerobic digester sludge Incubate at 35 degrees C Sample on days 0, 14, and 28 ^ p. 78 Do not cite or quote. Not for further distribution without author approval. Summary Study results to date Soil and Sludge methodologies and analytical methods have been demonstrated No evidence of polymer transformation observed g u? | - Inherent Biodegradation in Sludge - 28 Days - Biodegradation in four Soils - 180 Days 1 -C g Ongoing studies Soil Studies continuing to one-year Sediment studies commenced continuing to one year Anaerobic sludge studies to commence shortly Cfc