Document bXq8Qoa7MvDkJbov6myzQVnk

~J<7 .~ ~ - 3175~ Sanitized Version for th e AR-226 Public Electronic Docket (CBI and copy restricted documents removed) Index DOCUMENT DESCRIPTION NO. CBI OR OTHER RESTRICTION? OF PAGE S 1 March 7, 2007 le tter from DuPont to EPA 1 No restriction 2 DuPont Investment in Fluorotelomer 1 CBI - removed Environmental Fate Science 2004-200 7 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 restri ction 8 Protocol H-26665 15 No restriction 9 Appendix II Divider Sheet 1 No restri ction 10 Testing Invoices 1 CBI - removed 11 Appendix III Divider Sheet 1 No restriction 12 DuPont Investment in Fluorotelomer 1 CBI - removed Environmental Fate Scienc e 13 Appendix IV Divider Sheet 1 No rest ri ction 14 Summary of EPA Submissions (dated 7 No restriction 07/11/2002) 15 Biodegradation studies of fluorotelomer- 1 Subject to copyright - removed ; based polymers in soils Contact information to request authorization to copy : Dr . Robert Buck (302-892-8935 robert .c .buck@usa.dupont .com) 16 Biodegradation studies of fluorotelomer- 1 Not for further distribution withou t based polymers in activated sludge, soil, author approval - removed ; and sediments Contact information to request authorization to copy : Dr . Robert Buck (302-892-8935 or Robert .C .Buck usa .du ont.com) Company SWifted p. 2 DuPont Chemical Solutions Enterprise P . 0 . Box 80023 Wilmington, DE 19880-0023 March 7, 200 7 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 an d 2) Information on the studies that DuPont has conducted and commissioned on the biodegradation of 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 of 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 8 p. 3 CONFIDENTIAL BUSINESS INFORMATIO N DOCUMENT REMOVED Document Number in Index : 2 Document Description : DuPont Investment in Fluorotelomer Environmental Fate Science 2004-200 7 C ..C/ ~- `- ' C~- p. 4 APPENDIX I ~~r~C~ .3 p. 5 CONFIDENTIAL BUSINESS INFORMATIO N DOCUMENT REMOVED Document Number in Index : 4 Document Descri ption : Cost Summary - Telomers ~ ~_~~~ p. 6 CONFIDENTIAL BUSINESS INFORMATIO N DOCUMENT REMOVED Document Number in Index : 5 Document Description : Testing Invoices - Set 1 ~ bo e p. 7 CONFIDENTIAL BUSINESS INFORMATIO N DOCUMENT REMOVED Document Number in Index : 6 Document Description : Testing Invoices - Set 2 p. 8 PROTOCO L 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 D rive Easton, Mary land 21601 (410) 822-860 0 February 17, 2005 ~ c~~~~-~l ~ 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 D ri ve Easton, Maryland 2160 1 STUDY DIRECTOR: Edward C . Schaefer Wildlife International, Ltd. LABORATORY MANAGEMENT : Henry O . Krueger, Ph .D . Director of Aquatic Toxicology and Non-Target Plants FOR LABORATORY USE ONL Y Proposed Dates : Experi mental Experimental Start Date : A'4 cc ~ 2 uCs Termination Date : !S n2.rrJ 1()C16 Project No . : l~Z G' ~ f~ p Test Concentrations : Test Substance No . : <0 0 "-J%K~ ; zSo 5 /Ka 415 2 " , r 6 t zL, 66 Ly GEY9 Reference Substance No . ( if applicable) :E y*9, C993 . 4 9 y5- PROTOCOL APPROVAL C STUDY DIRECTOR DATE r ~C LABO OR M AGEMENT DA TE SPONSOR'S REPRESENTATIVE DATE PROTOCOL NO . : 112/021 705/SED-TRANSa/SUB 112 ~~ P . 10 Wildlife International, Ltd. -3 - INTRODUCTION Soil may be exposed to chemicals by direct application, spray drift, run-off, drainage, wast e disposal, indus trial or agricultural effluent and atmosphe ri c deposition . This protocol desc ribes a laboratory test method to assess the tran sformation of the test substance in aerobic and anaerobic soil systems . No bias is expected in this type of study . OBJECTIVE The objective of the study is to assess the potential for tr ansformation of the test substance in aerobic and anaerobic soils . EXPERIMENTAL DESIGN The test will be conducted with four test soils under both aerobic an d an aerobic condition. Four groups will be established for each soil type and condition (aerobic or anaerobic) : (1) un treated (con trol) live soil ; ( 2) 200 mg test subst an ce/kg dw treated live soil ; ( 3) 200 mg test substance/kg dw treated ste rile soil ; (4) 250 g 8-2 telomer B alcohol/kg dw treated sterile soil ; 10 g 8-2 telomer B acid /kg dw ; 10 g 8-2 unsaturated telomer B acid/kg; an d 10 g perfl uorooctanoic acid/kg dw, C9, C 10, and C 11 at 10 g/kg dw all added to a steri le 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 appropri ate time intervals and the soil samples will be ex tracted and analyzed for potential metabolites . In addition to the day zero samples, a minimum of eight additional samplings will be performed . Sufficient incubation chambers to allow for 3 additional sampling inte rvals will be prepared for the un tr eated control ( 1) and 200 mg test subst ance/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 ma trix fo rtification samples, biomass determinations, viabi lity con trols and to monitor aerobic conditions as necessary . The test groups will be prepared using 25 grams ( d ry 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 extracti on of the entire test chamber contents and internal surfaces ; 2) sufficient PROTOCOL NO . : 112/021705/SED-TRANSa/SUB 112 - P . 11 Wildlife International, Ltd. -4headspace 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 of 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 characterizati on 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 substan ce has been characterized p ri or to its use in the study . If verifi cation of GLP test subst ance characteri zation is not provided to Wildlife Inte rnational, Ltd ., it will be noted in the complian ce 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 re maining at the end of the study . The test substance is not radiolabeled an d has no single molecular formula . It is a mixture containing perfl uoroalkyl groups of varying carbon numbers as polymer side chains . Test Soils Four freshly collected US soils will be used to evaluate the test substan ces under aerobic and anaerobic conditions . The soils will be collected from each of the following soil orders : PROTOCOL NO . : I 12/021705/SED-TRANSa/SUB 112 p . 12 Wildlife International, Ltd. -5- Soil Order Percent of US Land* Alfisol 13 Mollisol 25 Inceptisol 16 Ultisol 1 3 *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 2C, 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 of 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 capacit y Cation exchange capacity The percent moisture and water .holding capacity of each of 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 . : 112/021 705/SED-TRANSa/SUB 112 p . 13 Wildlife 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 of 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 Chamber s 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 of 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) 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 entire bottle PROTOCOL NO . : 112/021705/SED-TRANSa/SUB 112 119,3 p . 14 Wildlife International, Ltd. 7- will be extracted aft er a sample of the headspace has been taken . The methods and procedures used will be documented in the study records an d in the final report. Abiotic Controls : The abiotic controls will be prepared by dosing approximately 25 grams (dry weight equivalent) of 6OCo-ste ri lized soil with sufficient test substance to deliver 200 mg/kg d ry 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 an d procedures used will be documented in the study records and in the fi nal report. Spike Recovery Controls : The spike recovery controls will be prepared by dosing approximately 25 grams ( dry weight equivalent) of 60Co-sterilized soil with approp ri ate volumes of 8-2 TBA stock (250 g/mL in ethan ol) an d fluorinated acid stocks (10 g/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 bo ttle will be extracted after a sample of the headspace has been taken. The methods an d procedures used will be documented in the study records and in the final repo rt. Viability Controls (untreated) : The metabolic activity of untreated test soil will be assessed on a monthly bas is . 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 '4COZ (aerobic and an aerobic conditions) and 14CHa ( anaerobic conditions) wi ll be measured and the percent biodegradation calculated_ The methods and procedures used wi ll be documented in the study records an d in the final repo rt. Aerobic Controls (treated) : The aerobic controls will be prepared by dosing approximately 25 grams (dry weight equivalent) of soil with sufficient test subst an ce to deliver 200 mg/kg dry weight. The test substance wi ll be administered by direct weight addition . The test chambers will be sealed with septum lined with aluminum foil, m ixed an d incubated . The oxygen content of a sacrificial chamber will be measured at monthly intervals . The methods an d procedures used will be documented in the study records and in the final report . Maintenance of Test Chamber s The soil moisture content should be maintained in the optimal microbial growth r an ge of 40 to 60% water holding capacity (WHC ; a pF of 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 interv als by weighing the test vessels . Adding steri le-filtered demineralized water ( degassed ste rile-fi ltered demineralized water for the anaerobic soils) will compensate water losses . PROTOCOL NO . : 1 1 2/02 1 705/SED-TRANSaISUB 112 p . 15 Wildlife 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 C18 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 . Samp ling 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 of 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 C18 cartridge and syringe . A volume of headspace gas from within the chamber will be pulled through the C18 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 Acid, 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#375-95-1 (C9) 6 . Nonadecafluorodecanoic Acid ; CAS#335-76-2 (C10) PROTOCOL NO . : 112!021705/SED-TRANSa/SUB 112 /J p .16 Wildlife International, Ltd. -97 . Perfluoroundecanoic Acid; CAS#4234-23-5 (C11) Analytical Methods Methods of 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 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 Criteri a 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, C10, C11, 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 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 Saturated Acid, 8-2 Unsaturated Acid, C9, C10, C11, 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, C 10, C 11 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 RESULT S 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 9~ p . 17 Wildlife International, Ltd . -10- mg kg- ' (dry weight) and as mole kg" ( dry weight) for each sampling interv al. Transformation products listed in soil and headspace samples will be plo tted against time. HEALTH AND SAFET Y It is advisable to read the MSDS for the reference chemical and test substan ce when available . If an MSDS is not available ( e .g., the substance is a product of research and development) it is important to review what is known about the substance with a person knowledgeable about the safe handling of the substan ce, such as the person supplying the substance for testing . It is advisable to follow guidelines on Storage and H andling of Chemicals ; Personal Protective Equipment, Waste Disposal Guide . TEST SUBSTANCE DISPOSA L A fter the issuance of the final repo rt, the remaining test substance will be stored at the testing lab until its expiration date and then destroyed, unless other arr an gements are made between the Sponsor and the testing lab . RECORDS TO BE MAINTAINE D Records to be maintained wi ll include, but not limited to, the following : 1 . A copy of the signed protocol . 2 . Identification and characte ri zation of the test substance as provided by Sponsor . 3 . Study initiation an d termination dates . 4 . Experimental initiation and termination dates . 5 . Test and reference substan ce preparation an d dosing calculations . 6 . Soil source an d pretreatment data . 7. Results of an alytical methods performed . 8 . Temperature range recorded duri ng test period. 9 . Copy of final report . PROTOCOL NO . : 112/021705/SED-TRANSa/SUB112 9~ p . 18 Wildlife International, Ltd. -11- FINAL REPORT A final report of 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 repo rt will include, but not be limited to the following, when applicable : 1 . Name and address of facility performing the study . 2 . Dates on which the study was initiated and completed . 3 . Objectives an d procedures stated in the approved protocol, including any changes in the original protocol. 4 . Identification an d characterization of the test substance as provided by Sponsor . 5 . A summary and analysis of the data . 6 . A desc ription of the transformations and calculations performed on the data . 7 . A desc ri ption of the methods used and reference to any stan dard method employed. 8 . A description of the test system. 9 . A description of the preparation of the test solutions, the testing concentrations, and the duration of the test . 10 . A description of all circumstan ces that may affect the quality or integri ty of the data . 11 . The name of 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 report s of each of the individual scientists or other professionals involved in the study, if app li cable . 13 . The location where the raw data an d final repo rt are to be stored . CHANGES TO THE FINAL REPORT If it is necessa ry to make corrections or additions to the final report after it has been accepted, such ch anges 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 PROTOCO L Planned changes to the protocol will be in the form of w ritt en amendments signed by the Study Director an d approved by the Sponsor's Representative . Amendments will be considered as part of the protocol and will be a ttached to the final protocol . Any other changes will be in the form PROTOCOL NO . : 1 1 2/02 1 705/SED-TRANSa/SUB 112 ~g P . 19 Wildlife International, Ltd. -12of written deviations signed by the Study Director an d fi led with the raw data . A ll changes to the protocol will be indicated in the final report. GOOD LABORATORY PRACTICE S This study will be conducted in accordance with Good Laborato ry Practice St andards for EPA; will be consistent with OECD Principles of Good Laborato ry Practice . Each study conducted by Wildlife International, Ltd . is routinely examined by the Wildlife Inte rn ational, Ltd . Quality Assurance Unit for compliance with Good Laborato ry Practices, Standard Operating Procedures and the specified protocol . A statement of complian ce with Good Laboratory Practices will be prepared for all portions of the study conducted by Wildlife Inte rn ational, Ltd . The Sponsor will be responsible for compliance with Good Laborato ry Practices for procedures performed by other laboratories ( e .g., residue analyses or pathology) . When the final repo rt 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 Intern ational, Ltd . PROTOCOL NO . : 112/021705/SED-TRANSa/SUB 112 9~ p . 20 Wildlife International, Ltd . 13 REFERENCES 1 Organisation for Economic Coopera ti on and Development. April 2002 . Aerobic and Anaerobic Transformation in Soil. OECD Guideline 307 . PROTOCOL NO . : 1 12/02 1 705/SED-TRANSa/SUB 112 10 p . 21 Wildlife International, Ltd . Project No . : 112E-10 8 Page 1 of 1 AMENDMENT TO STUDY PROTOCOL STUDY TITLE : H-26634 : TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOIL S PROTOCOL NO : I 1 2/02 1 705/SED-TRANSa1SUB112 AMENDME NT NO. : 1 SPONSOR: E .1 du Pont de Nemours and Comp any PROJECT NO . : 112E-108 EFFECTIVE DATE : 06 May 200 5 AMENDMENT : Mainten ance of Test Chambers, Page -7CHANGE : The soil moisture content should be checked and adjusted, if necessary, at regular 1 to 2 week inte rvals by weighing the test vessels . TO : The soil moisture content should be checked an d adjusted, if necessary, at monthly interv als by weighing the test vessels . REASON : The frequency at which the soil moisture content was checked was reduced based on obse rv ed losses . ~_(^ z STUDY DIRECT /R DATE LABO T RY MA AGEMENT DATE to-q106 /~>/ p . 22 ~ r`,~ ~ ~ ~ ~ ion at, U~.~ tid. ~ 1 of 1 DEVIATION TO STUDY PROTOCO L STUDY TITLE : H-26634 : TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOIL S PROTOCOL NO : 11 2/02 1 705/SED-TRANSa/SUB 112 DEVIATION NO . : 1 SPONSOR: E .1 du Pont de Nemours and Company PROJECT NO . : 112E-108 DEVIATION : Preparation of Test Chambers; Viability Controls (untreated), Page -7- The protocol indicated that duplicate incubation chambers for each test soil and condition ( aerobic an d anaerobic) would be dosed at approximately 100 mg/kg dw wi th 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. IMPACT : In the best judgment of the Study Director, th is deviation did not impact the integrity of study . A concentration of 2000 to 4000 mg glucose per kg d ry weight soil is a common for the determination of glucose-induced respiration rates . STUDY DIRECTOR DATE LABORA RY A AGEMENT DATE ~0,2- p . 23 PROTOCO L H-26665 : TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS This protocol complies with th e Organisation for Economic Cooperation and Development OECD Guideline 307 Adopted April 2002 Submitt ed to E .I. du Pont de Nemours and Compan y Wilmington, Delaware 19898 USA Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryl and 21601 (410) 822-860 0 Februa ry 17, 200 5 1 2)3 c~~ ~ p . 24 Wildlife International, Ltd. -2H-26665 : TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOIL S SPONSOR : E .I. du Pont de Nemours and Company SPONSOR'S REPRESENTATIVE : William Be rti TESTING FACILITY : Wildlife International, Ltd . 8598 Commerce D ri ve Easton, Marylan d 2160 1 STUDY DIRECTOR: Edward C . Schaefer Wildlife International, Ltd. LABORATORY MANAGEMENT : Henry O . Krueger, Ph.D . Director of Aquatic Toxicology an d Non-Target Plants FOR LABORATORY USE ONLY Proposed Dates : Experimental Experi mental Start Date : Termination Date : I S^~ V~~ 1u n 6 Project No . : E-_ l0 Test Concentrations : Wo L 1 Sy ~~ny~ lcJ~ ~Ky b Ex ; 6~E~ 6 d14, 6F ~ 9 Test Substance No . : Reference Substance No . (if applicable) : ~QYy 6f Y3 G4vt' PROTOCOL APPROVAL 1 0 ~"4~Crf Z00 STUDY DIRECTOR DATE LABO Y AGEMENT DATE SPONSOR'S REPRESENTATIVE DATE PROTOCOL NO . : 112/021705/SED-TRANSb/SUB 112 /O / p . 25 Wildlife International., Ltd. -3INTRODUCTION Soil may be exposed to chemicals by direct application, spray dri ft , run-off, drainage, waste disposal, industrial or agricultural effluent and atmospheri c deposition. This protocol describes a laborato ry test method to assess the transformation of the test subst an ce in aerobic and anaerobic soil systems . No bias is expected in this type of study . OBJECTIVE The objective of the study is to as sess 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 subst ance/kg dw treated live soil ; ( 3) 200 mg test substance/kg dw treated sterile soil; (4) 250 g 8-2 telomer B alcohol/kg dw treated sterile soil; 10 g 8-2 telomer B acid /kg dw ; 10 g 8-2 unsaturated telomer B acid/kg; and 10 p.g perfl uorooctanoic acid/kg dw, C9, C10, and C 11 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 appropri ate time interv als and the soil samples will be ex tracted and analyzed for potential metabolites . In addition to .the day zero samples, a minimum of eight additional samplings will be performed . Sufficient incubation chambers to allow for 3 additional sampling interv als will be prepared for the untr eated contr ol ( 1) and 200 mg test substan ce/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 substan ce treated soil chambers will be prepared for use as matrix fortification samples, biomass determinations, viability con trols and to monitor aerobic conditions as necessary_ The test groups wi ll 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 of the entire test chamber contents and internal surfaces ; 2) sufficient PROTOCOL NO . : 112/021705/SED-TRANSb/SUB 112 M~ p . 26 Wildlife International, Ltd. 4headspace to minimize the need for active aeration which could result in a loss of potential transformation products . METHODS AND MATERIAL S 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 subst ances, but not for polymers, adaptations of the method may be necessary for feasibility reasons . Nevertheless, the study wi ll be carried out in such a way that the recommendations given in the guide line will be followed as closely as possible . Test Substance s 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. veri fication that the test substance has been characte ri zed p ri or to its use in the study . If verification of GLP test substance characteri zation is not provided to Wildlife International, Ltd ., it will be noted in the compliance statement of the final report . The Sponsor is responsible for al l information related to the test substance an d agrees to accept any unused test substance an d/or test substance containers remaining at the end of the study . The test substan ce 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 subst an ces under aerobic and anaerobic conditions . The soils will be collected from each of the fo ll owing soil orders : PROTOCOL NO . : 112/021705/SED-TRANSb/SUB 112 ~~~ p . 27 Wildlife International, Ltd. -s- Soil Order 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 aft er sampling . Vegetation, larger soil fauna an d stones will be removed pri or to passing the soil through a 2 mm sieve . The soil used in the ste ri le groups will be steri lized by cobalt irradiation . The soils may be stored in the dark at 4 2C, if necessary. However, storage and pre-incubation time together will not exceed 3 months. Characterization of the soils will be conducted by Agvise Laborato ries (Northwood, No rth Dakota, USA) . The following is a list of 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 of 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 p ri or to adding the test substance an d in test and control soil samples at four months, at six months, and at 12 months using the fumigation-extraction method . Test Apparatus and Condition s The test chambers will be glass serum bottles with foil lined closures and will be identified by project number, test subst ance ID , test concentration, an d unique identifier . The chambers will be incubated statically at approximately 20 2C . PROTOCOL NO . : 112/021 705/SEll=TiLANSb/SUB 112 ~a~ p . 28 Wildlife International, Ltd. -6- Soil Pre-Incubatio n 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 of 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 . Preparati on of the Test Chamber s 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 of 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) 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 entire bottle PROTOCOL NO . : 1 1 2/02 1 705/SED-TRANSb/SUB 112 /v~ p . 29 Wildlife International, Ltd. -7wi ll 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) of 6OCo-sterilized soil with sufficient test substan ce to deliver 200 mg/kg dry weight. The test substan ce will be administered by direct weight addition . The entire bo ttle 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 repo rt . Spike Recovery Controls : The spike recovery controls will be prepared by dosing approximatelyy 25 grams (dry weight equivalent) of C60o-sterilized soil with appropriate volumes of 8-2 TBA stock (250 g/mL in eth anol) and fluorinated acid stocks (10 g/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 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 metabo li c activity of untreated test soil will be assessed on a monthly basis . Dup licate incubation chambers for each test soil and condition (aerobic and an aerobic) will be dosed at approximately 100 mg/kg dw with a combination of radiolabeled an d non-labe lled glucose . The evolved 14CO2 (aerobic and anaerobic conditions) and 14CH4 (an aerobic 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 repo rt. 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 administe red by direct weight addition . The test chambers will be sealed with septum lined with aluminum foil, mixed and incubated. The oxygen content of a sac ri ficial chamber wi ll be measured at monthly inte rvals. The methods and procedures used will be documented in the study records and in the final repo rt . Maintenance of Test Chamber s The soil moisture content should be maintained in the optimal microbial growth range of 40 to 60% water holding capaci ty (WHC ; a pF of between 2 .0 and 2 .5, or from 0 .1 to 0.33 bar) . The soil moisture content should be checked an d adjusted, if necessary, at regular I to 2 week interv als by weighing the test vessels. Adding ste ri le-filtered demineralized water (degassed ste rile-filtered demineralized water for the anaerobic soils) will compensate water losses . PROTOCOL NO . : 112/021 705/SED-TRANSbrS UB112 ~a~ p . 30 Wildlife International, Ltd. -8- The oxygen content in the headspace of two aerobic control chambers wi ll 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-steri lized, oil- fr ee air. If test chambers have to be opened to add water or for monito ring purposes, a sample of the head space will be taken first using a C18 cart ri dge as described below . The chamber will then be recapped as soon as possible . The septum an d cap previously removed will be stored at -10C or lower. The C18 will subsequently extracted an d analyzed . Sample extracts will be stored at -10C or lower if analysis is delayed more than 24 hours . Samp li ng and Measurements Duplicate chambers from each of the control, treated and spike re cove ry soils will be extracted (e .g., acetonitri le or other suitable solvent) an d analyzed. Proposed sampling intervals wi ll 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 repo rt . More or less frequent intervals may be conducted at the discretion of the Study Director . Potential volatile transformation products in the headspace of 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 wi ll be pierced using a needle connected to a C 18 cartridge and syringe . A volume of headspace gas from within the chamber will be pulled through the C18 cartridge using the attached syringe . The C 18 will be subsequently extracted and analyzed. Sample extracts will be stored at - 10C or lower if an alysis is delayed more th an 24 hours . Extracts will be an alyzed for : 1 . CF3(CF2)7CH2CH2OH (perfluorooctyl eth anol, 8-2 TBA, CAS# 678-39-7 ) 2 . CF3(CF2)7CH2COOH (2-perfluorooctyl ethan oic acid, 8-2 Saturated Acid, 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, pentadecafl uoro- ; PFOA; CAS# 335-67-1) 5 . Heptadecafluorononaoic Acid; CAS#375-95-1 (C9) 6 . Nonadecafluorodecanoic Acid; CAS#335-76-2 (C10 ) PROTOCOL NO : 112/021 705/SED-TRANSb/SUB 112 - ~~/) p . 31 Wildlife International, Ltd. 97 . Perfluoroundecanoic Acid ; CAS#4234-23-5 (C11) Analytical Method s Methods of 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 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 Criteri a 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, C10, C11, 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 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 Saturated Acid, 8-2 Unsaturated Acid, C9, C10, C11, 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, C10, C11 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 RESULT S 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-TRANSb/SUB112 /~/ p . 32 Wildlife International, Ltd. -10- mg kg-' (dry weight) and as mole kg-' (dry weight) for each sampling interval. Transformation products listed in soil an d headspace samples will be plotted against time . HEALTH AND SAFETY It is advisable to read the MSDS for the reference chemical and test subst ance when available . If an MSDS is not available (e .g ., the substance is a product of research and development) it is important to review what is known about the substance with a person knowledgeable about the safe handling of the substance, such as the person supplying the substance for testing . It is advisable to follow guidelines on Storage and Han dling of Chemicals ; Personal Protective Equipment, Waste Disposal Guide . TEST SUBSTANCE DISPOSA L After the issuance of the final report , the remaining test substance will be stored at the testing lab until its expiration date an d then destroyed, unless other arrangements are made between the Sponsor and the testing lab . RECORDS TO BE MAINTAINE D Records to be maintained will include, but not limited to, the following : 1 . A copy of the signed protocol. 2 . Identification and characterization of the test substance as provided by Sponsor . 3 . Study initiation and termination dates . 4 . Expe rimental initiation and termination dates . 5 . Test and reference subst ance preparation and dosing calculations . 6 . Soil source and pretreatment data. 7 . Results of analytical methods performed . 8. Temperature range recorded during test period . 9 . Copy of final report . PROTOCOL NO . : 1 12/02 1 705/SED-TRANSb/SUB 112 ~~~~ p . 33 Wildlife International, Ltd. -11- FINAL REPORT A final repo rt of the results of th e study wi ll be prepared by Wildlife International, Ltd. and submi tted to the Study Monitor no later than one month after th e conclusion of th e definitive study . The report will include, but not be limited to the following, when app licable : 1 . Name and address of facility performing the study . 2 . Dates on which the study was initiated an d completed . 3 . Objectives and procedures stated in the approved protocol, including any changes in the original protocol . 4 . Identification and characterization of the test substan ce as provided by Sponsor. 5 . A summary and analysis of the data . 6 . A description of th e tr ansformations and calculations performed on the data . 7 . A desc ription of the methods used an d reference to any stan dard meth od employed . 8_ A descri ption of the test system. 9 . A desc ription of the preparation of the test solutions, the testing concentrations, and the duration of th e test . 10 . A desc ription of all circumstan ces that may affect th e quali ty or integrity of the data . 11 . The name of the study director, the names of other scientists or professionals, and the names of all supe rvisory personnel, involved in the study . 12_ The si gned and dated repo rts of each of th e 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 chan ges shall be made in the form of an amendment issued by th e 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 PROTOCO L Planned ch anges to the protocol will be in the form of wri tt en amendments si gn ed by the Study Director an d 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 chan ges will be in th e form PROTOCOL NO . : 1 1 2/02 1 705/SED-TRANSb/SUB 112 /~~ p . 34 Wildlife International, Ltd. -12- of written deviations signed by the Study Director and filed with the raw data . All changes to the protocol will be indicated in the final repo rt. GOOD LABORATORY PRACTICES This study will be conducted in accordance with Good Laborato ry Practice Standards for EPA ; will be consistent with OECD P ri nciples of Good Laboratory Practice . Each study conducted by Wildlife International, Ltd . is routinely examined by the Wildlife International, Ltd. Quality Assurance Unit for complian ce with Good Laboratory Practices, Standard Operating Procedures and the specified protocol . A statement of compliance with Good Laborato ry 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 laborato ries (e .g ., residue analyses or pathology) . When the final report is completed, original copies of the study data and magnetically encoded records generated by Wild li fe International, Ltd . for this study wi ll be sent to the Sponsor. A cert i fi ed copy will be retained in the archives of Wildlife Intern ational, Ltd . PROTOCOL NO . : 112/021705/SED-TRANSb/SUB 112 //~z p . 35 Wildlife International, Ltd. -13REFERENCES 1 Organisation for Economic Cooperation and Development . Apri l 2002 . Aerobic an d Anaerobic Transformation in Soil. OECD Guideline 307 . PROTOCOL NO . : 1 1 2/02 1 705/SED-TRANSb/SUB 112 I/v p. 36 Wildlife International, Ltd . Project No. : 112E-10 9 Page 1 of 1 AMENDMENT TO STUDY PROTOCO L STUDY TITLE : H-26665 : TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOILS PROTOCOL NO : 1 1 2/02 1 705/SED-TRANSb/SUB 112 AMENDMENT NO. : 1 SPONSOR : E .1 du Pont de Nemours and Company PROJECT NO . : 112E-109 EFFECTIVE DATE : 06 May 200 5 AME NDMENT : Maintenance of Test Chambers, Page -7- CHANGE : The soil moisture content should be checked and adjusted, if necessa ry, at regular I to 2 week intervals by weighing the test vessels . TO : The soil moisture content should be checked and adjusted, if necessa ry, at monthly intervals by weighing the test vessels . REASON : The frequency at which the soil moisture content was checked was reduced based on obse rved losses . ~ ~-- -2 C STUDY DIRECTOR DATE LABORATORY MANAGEMENT DATE I~~ ~.e. cew~-~ ~~ Q~ lU-9-06 p . 37 ^ _ ?` w . . ~ ~ ff 4..... .~~~su~,~~. + s...~' I 1of1 ~,y DEVIATION TO STUDY PROTOCO L STUDY TITLE : H-26665 : TRANSFORMATION POTENTIAL IN AEROBIC AND ANAEROBIC SOIL S PROTOCOL NO : 112/021705/SED-TRANSb/SUB 112 DEVIATION NO. : I SPONSOR : E .I du Pont de Nemours and Company PROJECT NO . : 112E-109 DEVIATION: Preparation of 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 . IMPACT : In the best judgment of the Study Director, this deviation did not impact the integrity of study. A concentration of 2000 to 4000 mg glucose per kg dry weight soil is a common for the determination of glucose-induced respiration rates . STUDY DIRECTOR DATE LABORATORY MANAGEMENT DAT E i~~ p . 38 -A...P._P.~ END- IX II ~~~ ~ p . 39 CONFIDENTIAL BUSINESS INFORMATION DOCUMENT REMOVED Document Number in Index : 1 0 Document Desc ription : Testing Invoices ~/ 9 ~ p . 40 APPENDIX III ~zo p . 41 CONFIDENTIAL BUSINESS INFORMATIO N DOCUMENT REMOVED Document Number in Index : 1 2 Document Description : DuPont Investment in Fluorotelomer Environmental Fate Science 121 ~~ / ~-) p . 42 APPENDIX IV 122, 13 p . 43 U.S . EPA - DuPont Created By : Robert C Buck on 0-17 .11 .1200-2 at 02 :50 Pfvl Workgroup : Product Stewardshi p Category : General Sub Category : Genera l Security U .S . ENVIRONMENTAL PROTECTION AGENCY 4 March 2007 : 1) DuPont Investment in Fluorotelomer Environmental Fate Science : ~ DuPont Investment in Fluorotelomer Environmental Fate Science 2004-06 .pdf Presentation : 2007_7March USEPA Slides RCB .pdf (DuPont Confidential Information) mail 2) Wildlife Intl a) Invoices : 2007_Feb_2005-061MIdlife Intl Invoices .pdf b) Quote for 307/311 2006 VvU Costs to date and Est Future Studies Costs .pd f (DuPont Confidential Information ) 3) ENVIRON Invoices : Environ invoices, DEM Study 2005-2006_ALL .pdf (DuPont Confidential Information) EW Historical Archive Document of EPA Presentations : Surface Protections Solutions - .pd f Soil Biodegradtion Study Protocols : Urethane Polymer Protocol : 112E-106protocol_CBl .pdf Acrylate Polymer Protocol : 11 2E-1 ospratocol_CBi .pdf Note: The studies started with a suite of 7ana/ytes. Two additiona/ana/ytes were added at 15 months. t,,m;e SETAC Presentations : SETAC NA 2005-642-Polymer Biodeg .pdf /22.3 ~~'~ .~ p . 44 ~- . 2006 SETAC NA Polymer Biodeg Poster RCB_DRAFT One Page .pdf ( not confidential) 3 November 2006 : Prelimina ry Aerobic Soil Studies Informatio n sent as DuPont Confidential Information . Contains TSCA CBI . Subject to Copyright . Contents of CD 1 of 3 mat-. Cover Letter : 2006_3 Nov Cover Letter Soil Studies_USEPA_CBl .pd f Draft Study Reports : 2006_02 Nov_H26665_DRAFT REPORT_RPT109_CBl .pdf 2006_02 Nov-H26634-DRAFT REPORT RPT108_CBl .pd f Raw Data Index : 2006 02 Nov_DRAFT_Soil Studies Records Index CBl .pdf 4i& Analytical Data Summa ry : 2006 02 Nov DRAFT 12 Month Analytical Summary_CBl .pdf Test Substance Ce rtificates of Analysis : IT 3200611 OctDRAFT_H-26634_COA_CBl .pdf Contents of CD 2 of 3 _2006 11Oct_DRAFT_H-26665_COA_CBl .pdf Contents of C.0 3 & ' DuPont Soil Biodegradation Study Update, October 2006, Washington, DC C81 Documents Provideid in Advance : ~Az 1_1 Cover Letter for CBI Documents_120ct2006_CBI .pdf Test Substance COAs : /~ ~ p . 45 ~ ~ ~ 1_H26634 COA17 Oct2005 .pdf 2_H26665 COAI_170ct2005 .pd f 3200611 OctDRAFT_H-26634_COA .pdf 4_2006_11Oct_DRAFTH-26665_COA .pd f Study Protocols : 5_H26634 Soil Study_112E-108protocol .pdf 6 H26665_Soil Study 112E-109protocol .pdf Analytical Metehods : 7-Soil Study Analytical Methods .pdf 12-Analytical Method-Added analytes .pd f ~et Data : 10_Mollisol Raw Data Anayltical Summary .xls Interpretation : 8_2006_1 1 Oct_Interpretation Algortihm 8 AE Calculations .pdf 9 2006 10Oct_Irrterpretation Agorithm Workbook .xls Modeling the Study Results : 11 Modeling of Fluorotelomer-based Polymeric Products_Soil Biodeg Studies DRAFT .pdf Non-CBI Documents Provided in Advance : law ,, 69 A-1 2 October 2006 Email to D Lynch EPA_NOT CBl .pdf B20061 1 Oct_ Publication & Report References-NOT CBl .pdf C-8-2 FTOH Biodegradation Pathways-NOT CBl .pdf D_2006 JChromA_1110,pp117-124 .pdf DuPont Presentation, 15 September, 2006 2006 15Sept_DD Review with USEPA .ppt DuPont Presentation, 16 August 2006 PMN Meetin g 0 Premanufacture Notice slides 081606 final Company Sanitized .ppt EPA Meeting 081606 final CBl .ppt Ele EPA Meeting 081606 final Company Sanitized .ppt EPA Meeting Introduction 081506 final CBi .ppt / ~~ p . 46 EPA Meeting Introduction 081506 final Company Sandized .ppt Premanufacture Notice slides 081606 final CBl .ppt ~ ,r' DuPont Presentation, 8 September 2005 : EPA Mtg 9-8-05-CeI .ppt ~~ DuPont Presentation, 8 April 2005 : Supply Chain Review 08 April'05 EPA .ppt EPA Meeting 08 April'05 .ppt GDEM April 2005 SHK .ppt Paper PS Summary April 2005 SHK .ppt DuPont Presentation, 31 Janua ry 2005 : ~ DuPont Global Strategy Presentation : 31 Jan2005 DuPont Presentation to USEPA_AR226-1914 .pdf ~ DuPont Supply Chain Review : Supply Chain Review 31 Jan'05 Final_CBl .ppt DuPont Presentation, 15 December 2004 : DuPont Biodegradation Studies : C -- ,di a 15Dec2004_DuPont-EPA Meeting Presentation .ppt 15Dec2004_DuPont-EPA Meeting Agenda .pdf DuPont Presentation, 10 November 2004 : `~'r USEPA DuPont Ongoing Research Summary 9Nov2004_NOCBI .ppt IC US10 Nov2004_EPA_DuPont Ongoing Research Summary_CBI-RVVR .ppt / ~ 60? p . 47 DuPont Presentation, 17 June 2004 : DuPont Consumer Article Exposure & Risk Characterization (FINAL VERSION) , nbe DuPont Risk Assessment - Article Presentation USEPA 17June2004 .pdf ~ DuPont Risk Assessment - Article Presentation for EPA_17June2004 .ppt DuPont Presentation, 30 April 2004 (FINAL CBI VERSION) '4, 0 1 niiPont PMA RPdur.tinn R, DuPnnt PFC)A RPCiurtinn RPnnrt I NON-CBI VERSION 11 0 DuPont PFnA RPriurtinn RPnnrt 043f1(1 nuPnnt RPnnrt mAnriI9M4 n U .S . EPA-OPPT S DuPont Presentation, 16 March 2004 (FINAL : CBI VERSION) Rrand PrPSPntatinn I JS FPA 16 Ma U .S . EPA-OPPT S DuPont Presentation, 25 November 2002 0 (FINAL : CBI VERSION) 95Nnv9009 l1SFPA niiPnnt PS lJnclat P a (FINAL : NON-CBI VERSION) 25Nnv2 009 t1SFPA DuPnnt PS tlnriatP F Copy of the Non-CBI Document in the Public Record (AR-226) 95Nnvqn(lq DuPont PrPSantatinn to l JSF F U .S . EPA-OPPTS DuPont Presentation, 17 December 2001 / Z~ p . 48 17 DEC 2001 DUPONT 17nar.?nf11 r)iiPnnt FPA C ;RI (CBI VERSION : 17 Dec 2001 RCB ) ~~ . ~ 1 17 DEC 2001 DUPONT 17f)Pr2(101 f]uPnnt FPA Nnn ( ;Rl(NON-CBI VERSION : 17 Dec 2001 RCB) 17Dec2001 EPA Presentation with Post Meeting Corrections/Updates 91 nPrg(l(11 DuPont FPA PrPSantatinn Cony w P U.S. EPA-OPPTS DuPont Presentation, 17 December 2001, RESUBMITTED 12 FEB, 2002 ;, ,. 0 17r)ar2n(11 DuPont FPA Nnn(RI 19Fahg(11 17nPr.?M1 DuPont FPA CRI 1?FPh2n(1 M USFPA UnrlatPtf PrPSantatinn C ;nvPr I Ptt a U .S . EPA-OPPTS TRP Presentation ( 20 JUN 2000) 20 June 2000 Mtg TRP mtg_w EPA Chart: U .S . EPA-OPPTS TRP Presentation (23 OCT 2000) 0 EPA Review Presentation OCT23_2000_fina U .S . EPA-OPPTS Presentations DuPont & TRP ( 21 Feb 2001) ( DO NOT MAKE ANY CHANGES ) FPA RaviPw D l1PC)NT PrPSPntatinn FFR 9 1 FPA FII F TRP PraGantatinn FFR : (CHARTS GIVEN TO THE EPA-OPPTS on 21 FEB 2001 non-CBI Versions of the Above) 0 0 PPA RPViPw DUPONT PrPSantatinn FFR91 gfl(11 FINA FPA FII F TRP PrP .qantatinn FFR?1 ?001 no r ~zg p . 49 ,~`. . U .S . EPA-ORD, Duluth, MN, 7 June 2001 ( R . C . Buck) 7Jun2001 EPA_ORD Discussion . Miscellaneous Presentations to and by U .S . EPA . s August 3 . 2000 EPA Staff Presentation on PFOS 226-0619 .pd f SPI-FMG Presentation to U .S . EPA 23 April 2002 Final No CBI Ap ri l 23 EPA Presentatior . .. .. .. .. .. . .. .. .. .. .. .. .. Edit Histo ry: Rev. Editor Edit Date 70 . Robe rt C Buck 03/08/2007 03 :48 :37 PM 69 . Robert C Buck 03/07/2007 04 :32 :26 PM 68 . Robe rt C Buck 03/07/2007 07 :53:07 AM 67 . Robe rt C Buck 03/07/2007 07 :48 :16 AM 66 . Robe rt C Buck 03/07/2007 07 :47 :5 9 AM Only past five edits are shown i~ ~ p . 50 COPY RESTRICTED DOCUMEN T DOCUMENT REMOVED Document Number in Index : 1 5 Document Description : Biodegradation studies of fluorotelomer-based polymers in soil s Comment: DuPont has provided a Read Only copy of this document to EPA for inclusion in the AR- 226 printed paper file . The document may be viewed (but not copied) from the EPA public printed paper copy file located at the EPA Public Reading Room . Information from the EPA web-site on the Public Reading Room is provided below . Note that this information reflects information on EPA's website as of the date of this letter . Readers should verify with EPA that the information has not changed . The EPA/DC Public Reading Room is located in the EPA Headquarters West Building, Room 3334, located at 1301 Constitution Ave ., NW, Washington, DC . The EPA/DC Public Reading Room hours of operation are 8 :30 a .m. to 4 :30 p .m. Eastern Standard Time, Monday through Friday, excluding federal holidays . Please visit the EPA/DC website (http ://www .epa .gov/epahome/dockets .htm) for the latest status concerning the Public Reading Room and public access to docket materials . Pollution Prevention and Toxics Docket Email : oppt .ncic(&epa .go v Phone Number : 202-566-0280 Fax Number : 202-566-9744 Mail Code : 2822T DuPont Contact information to request author permission to copy the document Contact Information : Dr . Robert Buck 302-892-8935 robert.c .buck@usa .dupont .com ~~~ ~ ~~ p . 51 COPY RESTRICTED DOCUMEN T DOCUMENT REMOVED Document Number in Index : 1 6 Document Description : Biodegradation studies of fluorotelomer-based polymers in activated sludge, soil, and sediment s Comment: DuPont has provided a Read Only copy of this document to EPA for inclusion in the AR- 226 printed paper file . A printed copy of the document may be viewed (but not copied) from the EPA public printed paper copy file located at the EPA Public Reading Room . Information from the EPA web-site on the Public Reading Room is provided below . Note that this information reflects information on EPA's website as of the date of this letter. Readers should verify with EPA that the information has not changed . The EPA/DC Public Reading Room is located in the EPA Headquarters West Building, Room 3334, located at 1301 Constitution Ave ., NW, Washington, DC . The EPA/DC Public Reading Room hours of operation are 8 :30 a .m . to 4 :30 p .m . Eastern Standard Time, Monday through Friday, excluding federal holidays . Please visit the EPA/DC website (http ://www .epa .gov/epahome/dockets .htm) for the latest status concerning the Public Reading Room and public access to docket materials . Pollution Prevention and Toxics Docket Email : oppt .ncic ~~epa .go v Phone Number : 202-566-0280 Fax Number : 202-566-9744 Mail Code : 2822T DuPont Contact information to request author permission to copy the document : Contac t Information : Dr . Robert Buck 302-892-8935 robert .c .buck@usa .dupont .com 1.31