Document xxM46DOOQ9M49NKNYbRBq0Lm
AR226-2808
DuPont-11561
TRADE SECRET
Study Title H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Laboratory Project ID: DuPont-11561
Author: Carol Finlay, B.A.
Study Completed on: April 7, 2003
# Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
Work Request Number:
Service Code Number:
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Reviewed by:
Paul M. Hinderliter, PhJD. Research Toxicologist
O l - ^ r -3 Date
Issued by Study Director:
Card! Hnlay, B.A/J Staff Toxicologist
M ' p v ' l
Date
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
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TABLE OF CONTENTS
Page
CERTIFICATION.............................................................................................................................. 2
LIST OF TABLES.............................................................................................................................. 4
LIST OF FIGURES............................................................................................................................ 4
LIST OF APPENDICES....................................................................................................................5
STUDY INFORMATION..................................................................................................................6
STUDY PERSONNEL.....................................
8
S U M M A R Y .......................................................................................................................
9
INTRODUCTION..............................................................................................................................11
MATERIALS AND METHODS.........................................................................
11
A. Test Substance and Positive Controls....................................................................................11
B. Test Species.............................................................................................................................H
C. Animal Husbandry..................................................................................................................11
D. Quarantine and Pretest............................................................................................................12
E. Study Design........................... ...............................................................................................
F. Assignment to Groups and Study Start..................................................................................13
G. Dosing Material Preparation and Administration.................................................................13
H. Body Weights.......................................................................................................................... I 4
I. Mortality and Clinical Observations.........................................................
I4
J. Collection and Analysis o f Blood, Livers, and F at...............................................................14
K. Treatment of Fluorine Data.................................................................................................... 13
L. Statistical Analyses................................................................................................................. 16
RESULTS AND DISCUSSION...................................................................................................... 17 A. In-Life Toxicology..................................................................................................................I 7 B. Liver Weights.......................................................................................................................... I 7 C. Fluorine D ata.......................................................................................................................... I 8
C O N C L U S IO N S .............................................................................................
19
RECORDS AND SAMPLE STORAGE.......................................
29
TABLES................................................................................................................................. :........... 21
FIGURES............................................................................................................................................ 28
APPENDICES................................................................................................................................... 39
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LIST OF TABLES
Page
...............................22
z . a tt am D n n v u n c r r i u r r : a tmq
....................................................................................................... ...............................23
D. xic*AT o n n v ax m r r\/rn ? w n m w T ^
........................................................... ...............................24
4. aire a m d i rrr r r T T rvD T M T ? T P V R i
......................................................... ...............................25
5. M E A N B L O O D F L U O R IN E C O N C E N T R A T IO N N O R M A L IZ E D T O D O S E ........................ ...............................25
o. Tt T2 AXT T T\rCD ET TT/"VDTTvlT? T P\/ThT
...............................2 6........................................................................
7. M E A N L IV E R F L U O R IN E C O N C E N T R A T IO N N O R M A L IZ E D T O D O S E .......................... ...............................26
o. A/TC A M C A T m n r D T M R T C A /T T
........................................................................................... ...............................27
9. M E A N FA T F L U O R IN E C O N C E N T R A T IO N N O R M A L IZ E D T O D O S E ................................. ...............................27
LIST OF FIGURES
Page
1. MEAN BODY WEIGHTS........................................................................................................................................... 29
2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE C O N T R O L ....................................................................................................................................................................... 30
3. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS.....................................................................................................................................................................31
4. MICROMOLAR EQUIVALENTS IN RAT BLOOD................................................................................................32
5. NORMALIZED H-24750 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A 10-DAY ORAL GAVAGE
34
6. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO D O SE.....................................................35
7. COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24750 AND NEGATIVE CONTROL....................................................................... 36
8. MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE.......................................................... 37
9. COMPARISON OF MEAN BLOOD, MEAN LIVER, AND MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE...................................................................................................... 38
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LIST OF APPENDICES
Page
A. APPENDIX A INDIVIDUAL BODY WEIGHTS...................................................................................................... 40
B. APPENDIX B INDIVIDUAL CLINICAL OBSERVATIONS...............................................
50
C. APPENDIX C TERMS AND CALCULATIONS FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS................................................................................................................................................... 64
D. APPENDIX D INDIVIDUAL FLUORINE LEVELS IN BLOOD........................................................................... 68
E. APPENDIX E INDIVIDUAL FLUORINE LEVELS IN LIVER............................................................................. 75
F. APPENDIX F INDIVIDUAL FLUORINE LEVELS IN F A T ..................................................................................79
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION Test Substance
9th Collective Nomenclature: Svnonvms/Codes:
Submitter's Notebook Numberfsr Haskell Number: 24750
CAS Registry Number:! Purity:
Known Impurities: Positive Control
Substance Tested: Potassium perfluoroalkyl sulfonate Synonyms/Codes: H-24019
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Haskell Number: 24019 Composition1
Known Impurities:
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STUDY INFORMATION (Continued) Positive Control
Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt Svnonvms/Codes: H-24020
Perfluorooctanoate, ammonium salt APFO Ammonium perfluorooctanoate
Haskell Number: 24020 -
Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A.
Study Tnitiated/Completed: October 10, 2002 / (see report cover page)
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STUDY PERSONNEL
Study Director: Carol Finlay, B.A. Management: Scott E. Loveless, Ph.D.
Primary Technician: James C. Mackay II
Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T.
Toxicology Report Preparation: Sean M. Callaghan, B.A. Management: Nancy S. Selzer, M.S.
Laboratory Veterinarian: Thomas W. Mayer, D.V.M., A.C.L.A.M.
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
SUMMARY
The objective of this study was to evaluate the potential for H-24750 to be absorbed and to accumulate in a mammalian system. Two groups o f 5 male rats each were dosed by gavage with 5 mg/kg/day o f H-24750 for 10 consecutive days. One group was sacrificed on Day 10, and the other group was maintained without dosing for an additional 84 days. Blood was collected on test days 1, 5, 10,13, 24, 52, and 94. Livers and fat were collected at sacrifice. Body weights and clinical signs were recorded throughout the dosing and recovery periods. Additionally, a negative control of deionized water, and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-24750. Blood, liver, and fat samples were analyzed for total fluorine content to determine test substance absorption and retention in these tissues.
No deaths occurred in rats treated with the test substance, H-24750. Several clinical signs and body weight effects were evident as a result o f dosing with H-24750. One rat exhibited red nasal discharge during the recovery period, two exhibited hair loss during the recovery period, and one exhibited hair loss during the dosing and recovery period. The mean body weight gains of rats dosed with H-24750 were lower than the negative control rats during the dosing and recovery periods. Mean body weight gain o f rats dosed with the positive control materials was similar during the dosing period to the weight gain o f rats dosed with the test substance. During the recovery period, the mean body weight gain o f rats dosed with the positive control materials was higher than the weight gain of rats dosed with the test substance.
For the purpose of comparing absorption and retention of the test substance to the positive controls, all doses were normalized on a molar basis. The total fluorine in each tissue was considered as a percentage of the fluorine delivered in the administered dose, and all units were expressed as pM equivalents. As a measure o f total dose in the blood, an area under the curve (estimated to infinity) was calculated for each test material. The AUCINF/D for the fluorine component of H-24750 was 4.83x 105, compared to AUCINF/D values of 5.22xl05 and 8.15xl04 for H-24019 and H-24020, respectively. This parameter indicated a total dose in blood for H-24750 that was similar to or greater than the positive controls.
The mean relative liver weight (liver/body weight) o f rats dosed with H-24750 was 62% higher than the negative control rats on day 10. By day 94, the mean relative liver weight o f rats dosed with H-24750 was 23% higher than the negative control rats. The mean relative liver weight of rats dosed with H-24019 was 41% higher and for H-24020 was 93% higher on day 10 than the negative control group. By day 94, the mean relative liver weights o f rats dosed with H-24019 and H-24020 were similar to the negative control group. Therefore, liver weights of rats dosed with the test substance were higher than negative controls at both the end o f the dosing period and at the end of the recovery period.
The concentration of fluorine in the livers from rats dosed with H-24750 was 5518.80 pM equivalents on day 10 and 1476.01 pM equivalents on day 94. The mean pM equivalent concentration o f fluorine in the livers from rats dosed with the test substance was slightly higher
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on days 10 and 94 than the value for the positive control H-24019. The mean jiM equivalent concentration of fluorine in the livers from rats dosed with the test substance was approximately 6x higher (end o f dosing) or approximately 86x higher (end o f recovery) than the concentration in the positive control H-24020. Therefore, rats treated with H-24750 absorbed and retained considerably more fluorine in the liver than rats treated with H-24019 or H-24020.
The fluorine concentration in the fat from rats dosed with the test substance was 99.15 pM equivalents on day 10. There was no detectable fluorine by day 94. The fluorine concentration of the positive control H-24019 was approximately 2x higher than H-24750 on day 10. The fluorine concentration o f the test substance was approximately 2x higher on day 10 than the positive control H-24020. By day 94, there was no detectable fluorine in the fat from rats dosed with the positive control, H-24020. Therefore, some absorption of fluorine occurred in fat from rats dosed with the test substance.
Under the conditions o f this study, fluorine was evident in the blood during the period of dosing with H-24750. Not ail o f the fluorine was eliminated by the end of the recovery period from the blood or liver. The amounts of fluorine in the livers from rats dosed with the test substance were higher than levels in the blood and fat. Fluorine levels in the blood and liver were comparable or higher than levels in rats dosed with H-24019. Fluorine levels in the liver and fat were considerably higher than levels in rats dosed with the positive control H-24020.
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INTRODUCTION
The objective of this study was to define the potential of H-24750 to be absorbed and to bioaccumulate in a mammalian system, as indicated by analytical determination of total fluorine in blood, liver, and fat. The test substance was compared to 2 positive controls that were materials previously shown to bioaccumulate in mammals.
The daily dosage of 5 mg/kg for the test substance was selected based on existing toxicity information and the results of a rangefinding study. Groups o f five rats were dosed at 0 (control) or 20 mg/kg for 5 consecutive days. The rats dosed with the test substance had an overall body weight loss of 37 grams compared to a weight gain 28 grams for the control rats. The dosage of 5 mg/kg was chosen for the study and was expected to produce less than a 10% difference in mean body weight over 10 days when compared to the negative control.
MATERIALS AND METHODS
A. Test Substance and Positive Controls
The test substance, H-24750, was supplied by the sponsor as a white solid. The positive controls, H-24019 and H-24020, were supplied by the sponsor as white solids. The test substance and positive controls appeared to be stable under the conditions o f the study. No evidence of instability, such as a change in color or physical state, was observed.
B. Test Species
Male Crl:CD(SD)IGS BR rats were received from Charles River Laboratories, Inc., Raleigh, North Carolina. The Crl:CD(SD)IGS BR rat was selected on the basis o f extensive experience with this strain and its suitability with respect to longevity, hardiness, and low incidence of spontaneous diseases.
C. -Animal Husbandry
-
1. Housing Environment
Rats were housed singly in stainless steel, wire-mesh cages suspended above cage boards. Animal rooms were maintained on an approximate 12-hour light/dark cycle (fluorescent light) and at a temperature o f 23 1C and a relative humidity of 50 10%.
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2. Feed and Water
Tap water was provided ad libitum. All rats were fed PMI Nutrition International, LLC Certified Rodent LabDiet 5002 chow. The feed is guaranteed by the manufacturer to meet specified nutritional requirements and to be free of specified contaminants.
3. Identification
Each rat was assigned a unique identification number which was recorded on a card affixed to the cage. The last 3 digits of the number were tattooed on the tail of each rat.
4. Health Monitoring Program
As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to ensure that contaminant levels are below those that would be expected to impact the scientific integrity of the study:
Water samples are analyzed for total bacterial counts, and the presence o f coliforms, lead, and other contaminants.
Feed samples are analyzed for total bacterial, spore, and fungal counts.
Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers.
Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations o f key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity o f the study.
The animal health and environmental monitoring program is administered by the attending laboratory anim al veterinarian. Evaluation o f these data did not indicate any conditions that affected the validity o f the study.
D. Quarantine and Pretest
_
Upon arrival at Haskell Laboratory, the rats were removed from shipping cartons and quarantined for 6 days. The rats were weighed 3 times during the pretest period and examined daily for any clinically apparent signs of disease or injury. The rats were observed daily for mortality and signs of illness, injury, or abnormal behavior.
On the bases of acceptable body weight gains and freedom from clinically apparent signs of
disease or injury, the rats were released from quarantine by the laboratory animal veterinarian or
designee.
.
.
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
E. Study Design
Dosage Number o f
Substance
Vehicle
(mg/kg) Animals
Negative Control
Deionized water Not applicable
0
10
Positive Controls
H-24019
Acetone/Com Oil 10
10
H-24020
Acetone/Com Oil 20
10
Test Substance
H-24750
Deionized Water 5
10
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F. Assignment to Groups and Study Start
After the quarantine period, the rats were selected on the bases o f adequate body weight gain, freedom from any clinical signs of disease or injury. The selected rats were arbitrarily assigned to each group.
After assignment to groups, each rat was housed individually. The rats were 7 weeks of age at the time o f dosing. Dosing began on test day 1.
Rats that were not assigned to the study were released for other laboratory purposes, or were sacrificed by carbon dioxide asphyxiation and discarded without pathology evaluation.
G. Dosing Material Preparation and Administration
1. Test Substance
H-24750 was suspended in deionized water. The amount o f test substance each rat received was based on the body weight collected on each day of dosing and the suspension concentration. The rats were dosed at a volume of 1 mL/100 g of body weight. The mixture was stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity.
2. Positive Controls
It was necessary to dissolve H-24019 and H-24020 in acetone before suspending them in com oil. The ratio of acetone to com oil was 20:80. The amount each rat received was based on the body weight collected on each day of dosing and the suspension concentration. The rats were dosed at a volume 1 mL/100 g o f body weight. The dosing preparations were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity.
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H-24750: Biopersistence Screening _ 10-Dose Oral Gavage Study in Rats
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3. Negative Control
Deionized water was chosen as the negative control. The rats were dosed at a volume of 1 mL/100 g of body weight. These rats were dosed in a separate room from the rats dosed with the test substance or positive controls.
H. Body Weights All rats were weighed on each day of dosing and weekly during the recovery period.
I. Mortality and Clinical Observations
Cage-site examinations to detect moribund or dead rats and abnormal behavior and appearance among rats were conducted at least once daily throughout the study. At every weighing, each rat was individually handled and examined for abnormal behavior and appearance.
J. Collection and Analysis of Blood, Livers, and Fat
At time points selected for blood sampling other than sacrifice days, approximately 1 mL of blood was collected into EDTA tubes from the orbital sinus of each rat. Rats designated for sacrifice were euthanized by carbon dioxide anesthesia and exsanguination and blood, livers, and fat were collected according to the following schedules:
Group Dosing Days Tissue Collected
Sampling Time
I 1-10 Blood
Test day 1 (2 hours post dosing)
I 1-10 Blood
Test day 5 (2 horns post dosing)
I 1-10 Blood, Liver, and Fat Test day 10 at sacrifice (2 hours post dosing)
m 1-10 Blood
Test day 13
m 1-10 Blood
Test day 24
in 1-10 Blood
Test day 52
m 1-10 Blood, Liver, and Fat Test day 94
As much blood as possible was collected into EDTA tubes at sacrifice. The livers and fat were weighed. The fat weights were used for the calculation of fluorine levels. The liver weights were used for the calculation o f fluorine levels and for the calculation of liver weight relative to total body weight. The blood from all rats was refrigerated and the livers were frozen. The livers and fat were appropriately packaged and shipped refrigerated to Jackson Laboratory, Deepwater, New Jersey where they were analyzed for total fluorine. The liver and fat samples remained frozen during shipping.
The total fluorine content o f the blood and tissue samples was determined using a Wickbold torch combustion method, followed by analysis with a fluoride ion selective electrode. The samples were decomposed or volatilized in the presence of wet oxygen and swept through an oxy-hydrogen flame in a closed quartz apparatus. The combustion products were collected in an
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aqueous absorbing solution and analyzed with a fluoride ion selective electrode. Kinetic analysis of the data received (ppm F in each sample) was performed by Haskell Laboratory personnel for evaluation of fluorine biopersistence.
K. Treatment of Fluorine Data
Since the test substance and positive controls had different toxicity profiles, it was not possible to administer a uniform mg/kg dose for all test materials. Properly conducted kinetic comparisons in situations with varied doses required the use of a normalized dose. The dose-normalization was conducted in pmolar units to accommodate test material molecular weight differences and was adjusted to an arbitrary normalized dose o f 0.1 mmoles/kg.
H-24019 and H-24020 were used as positive controls and a dosing diluent was used as the negative control. The percent of fluorine and molecular weight o f the test substance and positive controls were used as provided by the sponsor. The measured total fluorine values were used as received (ppm F) from Jackson Laboratory. For the Wickbold torch method, the background fluorine level is 0.2 ppm. This is the limit o f detection (LOD) o f this method and was subtracted from each sample. The limit of quantification (LOQ) for this method is 0.5 ppm, and any values listed as less than 0.5 ppm were excluded from further treatment.
Since the dosages and molecular weight for positive controls and test material differed, the data were converted from a mass to a molar basis. In addition, the data were standardized to a single reference dose to allow comparison between compounds o f differing fluorine content. The doses were first converted from a mg of test material basis to millimoles of fluorine. Raw fluoride ion data (ppm F) were then normalized to a 0.1 millimole dose of active component. Finally the molar dosage and normalized concentration were combined to yield the pmolar (pM) equivalents o f active component in the tissue. Detailed calculations can be found in Appendix C. The pM equivalents can be compared across compounds provided the considerations listed in Appendix C are observed.
Noncompartmental analysis was conducted on fluorine data derived from rats dosed with H-24750 and the positive controls using WinNonlin Version 4.0 software (Pharsight Corp, Mountain View, CA). WinNonlin software provided a means of computing derived pharmacokinetic parameters from experimental data. All analysis was calculated using the pM equivalent in blood data.
The maximum observed concentration in blood was Cmax (pM equivalent). Biopersistence was assessed by quantifying terminal elimination blood half-life (T>/2, days). The points included in determination of the T>/2were selected manually and included only points after apparent log-linear elimination was achieved. Internal exposure was determined by calculating the blood areaunder-the-curve (AUC). AUC, which is simply the integral of blood concentration over time, is the most common means for expressing internal dose. With the calculated half-life, the AUC can be extrapolated to infinity (AUCINF) to reflect the elimination of the compound. AUCINF normalized to dose (AUCINF/D) can be used to compare the relative exposures of different compounds and dosages.
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
L. Statstica! Analyses
Descriptive statistics (e.g. mean, standard deviation) were used.
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RESULTS AND DISCUSSION
A. In-Life Toxicology (Tables 1-2, Figure 1, Appendices A-B)
No deaths occurred in rats dosed with the test substance, H-24750. One rat exhibited red nasal discharge during the recovery period, two exhibited hair loss during the recovery period, and one exhibited hair loss during the dosing and recovery period. The mean body weight gains o f rats dosed with H-24750 were lower than the negative control rats during the dosing and recovery periods. Mean body weight gain o f rats dosed with the test substance was comparable during the dosing period but lower during the recovery period than the weight gain o f rats dosed with the positive control materials.
B. Liver Weights (Table 3, Figures 2, 3, and 7)
1. Test Substance
The mean relative liver weight (liver/body weight) o f rats dosed with H-24750 was 62% higher than the negative control rats on day 10. By day 94, the mean relative liver weight o f rats dosed with H-24750 was 23% higher than the negative control rats.
2. Positive Controls
The mean relative liver weight of rats dosed with one of the positive controls, H-24019, was 15% lower at day 10 than the liver weight o f rats dosed with H-24750. By day 94, the mean relative liver weight of rats dosed with H-24019 was 15% lower than the liver weight o f rats dosed with H-24750.
The mean relative liver weight o f rats dosed with the other positive control, H-24020, was 16% higher at day 10 than the liver weight o f rats dosed with H-24750. By day 94, the mean relative liver weight of rats dosed with H-24020 was 21% lower than the liver weight of rats dosed with H-24750.
The mean relative liver weight of rats dosed with H-24019 was 38% higher at day 10 than the negative control group. By day 94, the mean relative liver weight of rats dosed with H-24019 was similar to the negative control group. The mean relative liver weight of rats dosed with H24020 was 88% higher on day 10 than the negative control group. By day 94, the weights were similar.
Therefore, the relative liver weights o f rats dosed with the test substance, H-24750, were higher than negative controls both at the end of the dosing period and at the end of the recovery period.
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C. Fluorine Data (Tables 4-9, Figures 4-9, Appendices C-F)
1. Factors Influencing Interpretation of Analysis
The data used in the kinetic analysis were derived from a limited screen, and therefore several caveats and considerations are important. A couple of considerations of particular importance are (1) a single dose was used and kinetics may or may not be linear, (2) the kinetics apply only to blood, (3) steady-state may not have been achieved, and (4) the sample size is low and may impact calculation o f the terminal half-life. A more complete list o f considerations is shown in Appendix C.
2. Positive Controls
The positive controls were H-24019 and H-24020. The H-24019 and H-24020 normalized pM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state (Figure 4A and4B). The Cmax for H-24019 was 1043.08 54.57 pM equivalents (mean S.D.) with a terminal half-life o f 42.2 days. The Cmax for H-24020 was 541.45 50.37 pM equivalents (mean S.D.) with a terminal half-life of 15.1 days. For each o f the positive controls, blood was sampled at 7 time points throughout the study, with only 4 of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between positive controls and the test substance. The AUCINF/D for the fluorine component was 5.22 x 105 for H-24019 and 8.15 x 104 for H-24020.
The concentrations o f fluorine in the livers on day 10 from rats dosed with the positive control materials were 4802.15 and 866.67 pM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1292.92 and 17.10 pM equivalents.
The concentrations o f fluorine in the fat on day 10 from rats dosed with the positive control materials were 194.46 and 57.25 pM equivalents for H-24019 and H-24020, respectively. By day 94, the concentration of fluorine in the fat from rats dosed with H-24019 was 15.38 pM equivalents. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020.
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ~
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3. Test Substance
The H-24750 normalized uM equivalents in rat blood rose rapidly and did not reach steady-state (Figure 4C). The Cmax for H-24750 was 974.25 201.73 uM equivalents (Mean SD) with a terminal half-life of 31.1 days. Blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between H-24750 and positive controls. The AUCINF/D for the fluorine component o f H-24750 was 4.83xl05 as compared to AUCINF/D values of 5.22x10s and 8.15xl04 for H-24019 and H-24020, respectively (Figure 4C).
Levels o f total fluorine in livers from rats dosed with the test substance, H-24750, were higher than the levels in livers from rats dosed with the positive control materials. The total fluorine concentration in the liver from rats dosed with H-24750 was 5518.80 pM equivalents at day 10 and 1476.01 pM equivalents at day 94. The liver concentrations in rats dosed with H-24750 were somewhat higher on days 10 and 94 than the concentrations in rats dosed with the positive control material H-24019. The liver concentrations in rats dosed with H-24750 were approximately 6x higher (day 10) and 86x higher (day 94) than the concentrations in rats dosed with the positive control material H-24020.
Levels of total fluorine in fat from rats dosed with the test substance were lower than the levels in fat from rats dosed with the positive control material H-24019. However, the level on day 10 was higher in rats dosed with the test substance than in rats dosed with the other positive control, H-24020. The fluorine concentration in the fat from rats dosed with the test substance was 99.15 pM equivalents on day 10. There was no detectable fluorine on day 94. The fluorine concentration of one of the positive control materials, H-24019, was approximately 2x higher than H-24750 on day 10. The fluorine concentration o f the test substance was approximately 2x higher on day 10 than the other positive control material, H-24020. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020.
The uM equivalents in the liver of rats treated with H-24750 were higher than levels in the blood and fat.
-
A
CONCLUSIONS
Rats dosed for 10 consecutive days with 5 mg/kg H-24750 exhibited no mortality. Clinical signs
and body weight effects were observed. Liver weights were affected in rats dosed with the test
substance. A steady-state for fluorine in the blood was not achieved during the 10-day dosing
period with the test substance. Dose-adjusted area under the curve (AUCINF/D) for the positive
control H-24019 was similar to the AUCINF/D for the test substance. Dose-adjusted area under
the curve (AUCINF/D) for the test substance was 6x the AUCENF/D for the positive control H-
24020.
.
ompany Sanitized. Does not contain TSCA CBI -19-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
Under the conditions of this study, there was considerable absorption and retention o f fluorine in the blood following dosing with H-24750. Administration o f the test substance, H-24750, to male rats for 10 consecutive days resulted in absorption and retention of fluorine in the liver. There was some absorption and little retention of fluorine in the fat. Levels in the fat in rats dosed with the test substance were lower than levels in the positive control material H-24019 but higher than levels in the positive control material H-24020.
RECORDS AND SAMPLE STORAGE
Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware.
Company Sanitized. Does not contain TSCA CBf -20-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
TABLES
Company Sanitized. Does not contain TSCA CBf -21 -
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
TABLE 1
MEAN BODY WEIGHTS
Negative Control DAYS ON TEST Deionized Water
MEAN BODY WEIGHTS (g)
Positive Controls
H-24019
H-24020
1
189.6
184.4
184.1
2
195.5
189.2
187.8
3
204.6
199.5
197.8
4
211.7
206.4
204.8
5
222.1
216.2
212.5
6
225.0
222.0
216.0
7
235.2
229.4
223.6
8
242.3
233.7
226.9
9
248.3
240.4
234.2
10
257.0
246.5
243.0
17
293.3
290.1
297.4
24
325.5
313.3
338.6
31
359.3
348.4
381.9
38
380.9
370.8
404.9
45
407.4
403.2
434.4
52
420.5
422.8
460.6
59
436.4
439.6
483.1
66 - - -
67
452.8
455.6
502.7
73
472.6
480.1
525.6
80
489.6
496.5
542.1
87
497.7
512.2
546.2
94
511.7
524.6
570.5
Test Substance H-24750
189.2 195.3 - 203.0 213.3 224.5 231.8 239.2 244.5 249.1 248.4 237.4 247.9 265.2 279.5 305.2 330.0 342.3 369.6
-
390.5 413.1 423.5 434.6
- Indicates the animals were not weighed
Company Sanitized. Does not contain TSCA CB! - 22-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
TABLE 2
MEAN BODY WEIGHT GAINS DOSING
DAYS ON TEST
MEAN BODY WEIGHT GAINS (g)
Negative Control
Positive Controls
Test Substance
Deionized Water H-24019
H-24020
H-24750
1-5 32.5 31.8 28.4 35.3 5-10 34.9 30.3 30.5 23.9
1-10 67.4 62.1 58.9 59.2
RECOVERY
DAYS ON TEST
MEAN BODY WEIGHT GAINS (g)
Negative Control
Positive Controls
Test Substance
Deionized Water H-24019
H-24020
H-24750
10-17 17-24 24-52 52-94
36.3 43.6 54.4 -11.1
32.2 23.2 41.2 10.5
95.0
109.5
122.0
82.1
91.2
101.8
109.9
104.6
10-94
254.7
278.1
327.5
186.2
Company Sanitized. Does not contain TSCA CBi - 23-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
TABLE 3
MEAN BODY AND LIVER WEIGHTS
DAYS ON TEST
NEGATIVE CONTROL - DEIONIZED WATER
WEIGHT fe)
Absolute
Mean Relative Liver Weight
Body Weight
Liver Weight
(Liver/Body Weight)
10
258.8
10.814
94
511.7
17.868
0.042 0.035
DAYS ON TEST
10 94
POSITIVE CONTROL - H-24019
WEIGHT (g)
Absolute
Mean Relative Liver Weight
Body Weight
Liver Weight
(Liver/Body Weight)
243.9 524.6
14.205 19.296
0.058 0.037
DAYS ON TEST
10 94
POSITIVE CONTROL - H-24020_________________________
__________________ WEIGHT (g)_________________________
Absolute______________ Mean Relative Liver Weight
Body Weight
Liver Weight
(Liver/Body Weight)
243.1 570.5
19.174 19.590
0.079 0.034
DAYS ON TEST
TEST SUBSTANCE - H-24750
WEIGHT (g)
. Absolute
Mean Relative Liver Weight
Body Weight
Liver Weight
(Liver/Body Weight)
10
260.9
17.785
94
434.6
18.841
0.068 0.043
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-24-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
TABLE 4
MEAN BLOOD FLUORINE LEVELS
DAYS ON TEST
MEAN FLUORINE LEVELS (ppm)
Negative Control
Positive Controls
Test Substance
Deionized Water H-24019
H-24020
H-24750
1
0.60a
2.60 (0.1)b
9.40 (2.4)
3.14 (0.32)
5
C
31.32 (1.1)
74.92 (7.0)
17.13 (2.22)
10
1.10a
68.00 (3.5)
61.76 (5.5)
30.44 (1.22)
13
c
53.98 (1.2)
29.52 (4.9)
34.40 (7.08)
24
0.5a
39.62 (3.4)
11.18 (2.9)
27.60 (4.21)
52
c
23.56 (2.1)
2.26 (1.1)
9.30 (2.78)
94
c
12.60 (1.2)
0.85d (0.1)
6.24 (0.46)
a One o f 5 values. Four o f the values were below the limit o f quantification (LOQ) or non-detectable. b Standard deviation (S.D.) is in parentheses, c All values were below the LOQ or non-detectable. d Mean o f 2 o f the 5 values. Three o f the values were below the LOQ.
TABLE 5
MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE
MEAN FLUORINE CONCENTRATION (pM F Equivalents)
_____________ Positive Controls_________________ Test Substance
DAYS ON TEST
H-24019_____________ H-24020_____________ H-24750
1
36.92 ( l.l ) a
66.67 (17.5)
83.70 (9.07)
5
478.77 (17.6)
541.45 (50.4)
482.39 (63.24)
10
1043.08 (54.6)
446.09 (39.7)
861.42 (34.76)
13
- 827.38 (19.0)
212.46 (35.6)
974.25 (201.73)
24
606.46 (53.1)
79.57 (20.8)
780.68 (120.05)
52
359.38 (32.7)
14.93 (7.7)
259.15 (79.34)
94
190.77 (19.2)
4.71b (0.5)
172.14 (13.23)
a Standard deviation (S.D.) is in parentheses. b Mean o f 2 o f the 5 values. Three of the values were below the limit of quantification (LOQ).
Company Sanitized. Doss not contain TSCA CBI - 25-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
TABLE 6
MEAN LIVER FLUORINE LEVELS
DAYS ON TEST
MEAN FLUORINE LEVELS (ppm)
Negative Control
Positive Controls
Test Substance
Deionized Water H-24019
H-24020
H-24750
10
0.90 (0.2)a 312.34 (19.7) 119.80 (3.5)
193.91 (7.17)
94
0.78 (0.0)
84.24 (7.4)
2.56 (1.3)
52.01 (5.26)
a Standard deviation (S.D.) is in parentheses.
TABLE 7
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
DAYS ON TEST
MEAN FLUORINE CONCENTRATION (pM F Equivalents)
Positive Controls
Test Substance
H-24019
H-24020
H-24750
10
4802.15 (303.8)a
866.67 (25.5)
5518.80 (204.34)
94
1292.92 (114.1)
17.10 (9.8)
1476.01 (149.92)
a Standard deviation (S.D.) is in parentheses.
Company Sanitized. Does net contain TSCA CBl
-26-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
MEAN FAT FLUORINE LEVELS
DAYS ON TEST
MEAN FLUORINE LEVELS (ppm)
Negative Control
Positive Controls
Test Substance
Deionized Water H-24019
H-24020
H-24750
. 10 94
a
12.84 (1.7)b
8.10 (1.1)
3.68 (0.6)
a 1.20 (0.3)
a
a
a All values were below the limit of quantification (LOQ) or non-detectable. b Standard deviation (S.D.) is in parentheses.
TABLE 9
MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE
DAYS ON TEST
10 94
MEAN FLUORINE CONCENTRATION (gM F Equivalents)
Positive Controls
Test Substance
H-24019
H-24020
H-24750
194.46 (26.8)a 15.38 (4.1)
57.25 (7.8) b
99.15 (17.1) _b
a Standard deviation (S.D.) is in parentheses. b All values were below the limit o f quantification (LOQ) or non-detectable.
Company Sanitized. Does no! contain TSCA CBI - 27-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
FIGURES
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-28-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 1 MEAN BODY WEIGHTS
w D u P o n t-11561
Company Sanitized. Does not contain TSCA CBf -29-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROL
D uP ont-11561
Mean Relative Liver Wt. (%of Body Wt.)
T est Days -30-
Company Sanitized. Does not contain TSCA CBI
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 3 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
D uPont-11561
Mean Relative Liver Wt. (%of Body Weight)
10 94 T est Days
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
FIGURE 4 MICROMOLAR EQUIVALENTS IN RAT BLOOD
A . N o r m s lise d R a t B l o o d H -2 4 0 1 9 pM E q u i v a l e n t s R e s u l t i n g f r o m a 1 0 - D a y O r a I G a v a g e
Micromolar (pM) equivalents of H-24019 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation.
B. No rm a l l i e d Rat B l o o d H -24020 y M E q u i v a l e n t s R e s u lt in g from a 10-D a y O ra l G a v a g e
Micromolar (iM) equivalents o f H-24020 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation.
Company Sanitized. Does not contain TSCA CBI - 32-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
C. Normalized Rat Blood H-24750 uM Equivalents Resulting from a 10-Day Oral Gavage
20 30 40 50 60 70 80 Time (days)
Micromolar (pM) equivalents of H-24750 (test substance) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation.
Company Sanitized. Does not contain TSCA CBI
-33-
m9 H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ____________________________________________________________________________________________ DuPont-11561 FIGURE 5 NORMALIZED H-24750 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A 10-DAY ORAL GAVAGE
-34-
Rat Blood AUCINF/C
ri
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 6 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
D uPont-11561
Day 10
Test Days -35-
Day 94
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats____________________________________________________________________________________________ DuPont-11561
FIGURE 7 COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION
FOR H-24750 AND NEGATIVE CONTROL
V>
'i>aad5>
*OD
CD
c(O)
caro>
Day 10
Day94
Test Days
Deionized Water (Negative Control) H H-24750 (Test Substance)
-36-
Day 10
Day 94
Test Days
Company Sanitized. Does not contain TSCA CBi
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 8 MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE
DuPont-11561
H-24019 (Positive Control) HJ H-24020 (Positive Control) H-24750 (Test Substance)
Day 10
T est Days
Day 94 ' rtnpany Sanitized. Does not contain S C A CB
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 9 COMPARISON OF MEAN BLOOD, MEAN LIVER, AND MEAN FAT FLUORINE
CONCENTRATION NORMALIZED TO DOSE
DuPont-11561
Company Sanitized. Does not contain TS CA CBI
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D u P o n t-11561
# APPENDICES
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX A Individual Body Weights
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H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
INDIVIDUAL BODY WEIGHTS
EXPLANATORY NOTES ABBREVIATIONS:
SD - sacrificed by design
DuPont-11561
Company Sanitized. Does not contain TSCA CB! -41 -
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
.
ANIMAL NUMBER
646932 646933 646934 646935 646936
Day 1
197.1 191.9 182.6 203.0 196.8
Day 2
202.1 195.4 188.9 208.9 201.4
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
Day 3
TEST DAY
Day 4
Day 5
Day 6
212.9 205.0 195.4 218.0 208.6
221.5 213.2 207.0 228.1 204.0
233.6 226.5 213.9 239.7 216.7
234.1 219.3 215.9 243.0 218.6
Day 7
242.4 232.9 224.2 257.2 229.9
ANIMAL NUMBER
Day 10
646932 646933 646934 646935 646936
264.7 254.4 248.6 278.0 248.1
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
D uPont-11561
Day 8
251.1 239.5 230.9 265.7 234.9
Day 9
256.1 246.4 238.1 270.5 240.3
Company Sanitized. Does not contain T S C A CB1
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
646937 646938 646939 646940 646941
Day 1
184.5 163.7 193.3 191.5 191.6
Day 2
186.8 173.8 204.2 197.1 195.9
ANIMAL NUMBER
Day 10
646937 646938 646939 646940 646941
249.8 237.2 277.9 244.5 266.3
Day 17
282.6 270.4 329.7 271.5 312.4
ANIMAL NUMBER
Day 73
646937 646938 646939 646940 646941
504.3 . 417.4 540.1 407.3 493.7
Day 80
520.5 431.0 568.9 424.0 503.5
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
Day 3
GROUP III
TEST DAY
Day 4
Day 5
Day 6
199.0 180.9 214.6 203.8 207.9
205.9 189.3 221.1 209.8 216.8
213.6 200.6 234.4 218.5 223.1
221.0 204.1 240.7 222.1 231.2
Day 7
229.6 215.9 248.9 229.3 241.3
Day 24
322.3 296.5 372.8 291.3 344.8
TEST DAY
Day 31
Day 38
366.1 325.8 415.2 317.9 371.6
395.0 342.9 443.2 332.8 390.7
Day 45
425.0 367.5 475.2 356.3 412.8
Day 52
443.9 373.7 484.1 367.0 433.6
Day 87
533.7 432.1 578.1 430.8 513.8
TEST' DAY Day 94
545.7 450.6 586.9 449.4 525.8
SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94
D uPont-11561
Day 8
238.7 221.8 259.2 232.0 249.1
Day 9
241.4 228.3 267.3 237.5 257.0
Day 59
461.7 378.7 505.7 380.3. 455.8
Day 67
478.7 401.3 516.4 392.5 475.3
Company Sanitized. Does not contain TS C A CB1
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
646910 646911 646912 646913 646914
Day 1
184.3 184.4 180.5 186.4 185.5
Day 2
185.3 187.1 185.9 189.7 189.6
m
H-24019 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
Day 3
TEST DAY
Day 4
Day 5
Day 6
197.3 195.7 195.9 198.1 198.8
204.8 202.0 202.5 205.7 203.8
212.7 211.7 211.9 221.8 215.4
216.3 217.2 217.7 225.0 222.7
ANIMAL NUMBER
Day 10
646910 646911 646912 646913 646914
241.1 236.6 241.9 257.0 243.0
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
Day 7
226.4 225.2 227.2 228.6 226.7
D uPont-11561
Day 8
232.3 225.6 232.4 238.4 231.5
Day 9
237.5 231.9 238.5 245.8 238.8
Company Sanitized. Does not contain TSCA CB1 -44-
F
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
646915 646916 646917 646918 646919
Day 1
182.7 185.0 187.8 175.5 192.0
Day 2
188.3 192.7 194.3 181.9 196.9
w
H-24019 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP III
Day 3
TEST DAY
Day 4
Day 5
Day 6
200.1 205.5 205.8 189.8 207.9
206.3 211.6 210.4 197.6 218.9
211.1 220.0 221.8 207.0 228.3
220.6 226.9 227.5 212.4 234.0
Day 7
225.8 239.5 233.9 220.2 240.8
DuPont-11561
Day 8
231.1 240.1 235.8 224.1 245.8
Day 9
236.8 249.3 244.3 228.2 252.9
ANIMAL NUMBER
Day 10
646915 646916 646917 646918 646919
240.4 257.0 250.2 237.1 260.2
Day 17
283.4 310.0 280.3 270.6 306.1
Day 24
302.9 334.4 308.2 292.7 328.5
TEST DAY
Day 31
Day 38
326.7 383.2 339.7 323.2 369.2
337.9 411.9 362.8 350.8 390.4
Day 45
371.1 450.3 387.9 379.4 427.3
Day 52
399.7 469.8 409.0 396.7 439.0
Day 59
412.5 494.8 420.0 418.8 451.7
Day 67
433.1 511.8 441.5 433.8 457.6
ANIMAL NUMBER
Day 73
646915 646916 646917 646918 646919
461.2 544.2 457.5 448.1 489.5
Day 80
475.0 565.5 464.7 472.5 504.8
Day 87
498.1 578.5 479.5 480.8 524.2
TEST' DAY Day 94
513.3 590.9 495.7 500.3 522.9
SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94
Company Sanitized. Does not contain TSCA CB1 -45-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in R a ts____________________________________________________________________________________________ DuPont-11561
H-24020 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
ANIMAL NUMBER
Day 1
Day 2
Day 3
TEST DAY
Day 4
Day 5
Day 6
Day 7
Day 8
Day 9
646921 646922 646923 646924 646925
204.5 178.3 185.9 188.7 172.2
204* 9
180.9 185.1 194.6 176.0
215.7 188.9 196.0 209.5 183.9
222.2 197.4
202.3 212.6 193.4
233.3 197.7 203.5 230.4 202.7
238.8 202.4 204.9 225.9 202.7
244.3 202.4 219.1 236.2 215.1
244.9 204.0 224.9 236.5 221.5
252.2 207.7 231.7 250.9 228.7
ANIMAL NUMBER
Day 10
646921 646922 646923 646924 646925
262.6 218.6 240.6 259.1 234.8
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
ontpany Sanitized. Does not contain TSCA CB1
j %
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in R a ts_________________________________________________
H-24020 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
ANIMAL NUMBER
Day 1
Day 2
Day 3
GROUP III
TEST DAY
Day 4
Day 5
Day 6
646926 646927 646928 646929 646930
182.0 172.2 190.4 178.6 187.9
187.8 178.8 195.3 182.5 191.6
197.0 191.1 204.5 190.7 201.0
203.6 195.1 212.3 198.3 210.4
206.0 206.6 222.1 203.5 219.6
213.2 213.2 230.1 204.2 224.4
ANIMAL NUMBER
Day 10
646926 646927 646928 646929 646930
250.8 243.3 254.1 225.5 240.3
Day 17
297.2 306.2 310.6 280.9 292.3
Day 24
349.3 355.1 349.4 323.2 316.2
TEST DAY
Day 31
Day 38
383.8 404.0 392.8 376.0 352.7
412.1 424.8 422.4 396.8 368.4
Day 45
432.1 463.2 443.5 426.4 406.8
ANIMAL NUMBER
Day 73
646926 646927 646928 646929 646930
535.3 551.2 540.3 502.6 498.5
Day 80
551.9 574.5 557.0 509.6 517.6
Day 87
564.4 541.3 568.3 526.7 530.3
TEST DAY Day 94
580.6 602.5 581.9 544.0 543.4
SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94
Day 7
222.9 218.2 236.1 214.6 226.7
Day 52
467.8 484.0 464.8 450.2 436.0
w DuPont-11561
Day 8
229.9 228.7 235.8 210.3 232.6
Day 9
239.7 233.1 243.4 219.2 235.3
Day 59
493.2 511.7 492.2 463.8 454.7
Day 67
509.5 527.2 518.6 484.2 474.2
Company Sanitized. Does not contain TSCACW
-47-
11
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
Day 1
665547 665548 665549 665550 665553
194.5 191.6 190.1 193.0 193.4
Day 2
200.9 195.8 197.4 196.6 199.0
li
H -2 4 7 5 0 (TEST SUBSTANCE)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
Day 3
TEST DAY
Day 4
. Day 5
Day 6
210.4 202.5 201.0 204.9 209.0
225.1 211.7 218.3 218.8 216.4
238.6 225.5 230.0 230.0 228.4
243.3 231.3 233.2 241.4 235.1
Day 7
252.1 238.6 245.4 248.4 247.3
DuPont-11561
Day 8
259.9 244.3 252.2 259.5 255.1
Day 9
267.0 246.4 263.1 266.2 262.7
ANIMAL NUMBER Day 10
665547 665548 665549 665550 665553
274.1 253.9 253.2 264.0 259.5
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
Company Sanitized. Does not contain TSCA CBf
-48-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
Day 1
665554 665555 665558 665559 665560
175.0 184.9 187.8 189.2 192.9
ANIMAL NUMBER
Day 10
665554 665555 665558 665559 665560
209.4 254.7 229.5 239.5 246.4
ANIMAL NUMBER Day 73
665554 665555 665558 665559 665560
330.1 387.9 345.4 418.3 470.9
Day 2
180.0 194.4 192.0 196.6 200.2
Day 17
212.0 247.7 229.7 243.9 253.5
Day 80
355.1 412.1 366.1 441.6 490.5
H-24750 (TEST SUBSTANCE)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP III.
Day 3
TEST DAY
Day 4
Day 5
Day 6
187.8 201.6 201.9 204 .2 206.5
195.2 208.7 211.1 213.4 214.1
204.6 217.6 221.9 222.9 225.8
210.8 230.9 226.5 230.8 234.7
Day 24
TEST DAY
Day 31
Day 38
Day 45
220.3 255.3 224.6 257.8 281.7
232.1 275.0 229.8 271.8 317.3
234.0 281.7 227.1 305.8 348.7
249.4 316.0 235.4 336.1 389.1
Day 87
TEST DAY Day 94
368.3 425.9 373 .1 451.0 499.3
380.2 431.8 381.2 467.1 512.5
SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94
Day 7
212.5 240.0 229.0 240.7 238.0
Day 52
270.5 340.3 259.9 370.2 408.9
#
DuPont-11561
Day 8
209.7 247.4 234.4 243.6 239.3
Day 59
290.5 353.6 280.3 380.4 406.8
Day 9
211.3 247.0 235.6 246.8 244.6
Day 67
312.8 369.3 316.6 400.6 448.7
Company Sanitized. Does not contain TSCA CBI -49-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX B Individual Clinical Observations
Company Sanitized. Does not contain TSCA CBI -50-
H-24750; Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________________________________________________________________________ DuPont-11561
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I
Animal Observation
Days
646932 General observation, No Abnormality Detected
1
Eye Observations, Exophthalmus, Left
2-9
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Discharge, Eye left, Black
10
Sacrificed by design
10
646933 General observation, No Abnormality Detected
1-5
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Eye Observations, Dark, Left
6-10
Discharge, Eye left, Black
6-7
Swollen Observations, Mouth
10
Sacrificed by design
10
646934 General observation, No Abnormality Detected
1-5,10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Discharge, Eye Jeft, Black
6-9
Sacrificed by design
10
-51 -
Company Sanitized. Does not contain TSCA CBI
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ,
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I (Continued)
Animal Observation
Days
646935 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Sacrificed by design
10
646936 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Sacrificed by design
10
D uPont-11561
Company Sanitized. Does not contain TSCA CBI -52-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP III
Animal Observation
Days
646937 General observation, No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Left
13
Eye Observations, Bled via Orbital for Clin Path, Right
24,52
Sacrificed by design
94
646938 General observation, No Abnormality Detected
1-10,17-45
Eye Observations, Enophthalmus, Right
80-94
Eye Observations, Exophthalmus, Right
52-73
Eye Observations, Bled via Orbital for Clin Path, Bilateral
13,52
Eye Observations, Bled via Orbital for Clin Path, Right
24
Eye Observations, Corneal Opacity, Right
80-94
Discharge, Eye right, Black
59
Sacrificed by design
94
646939 General observation, No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Left
13
Eye Observations, Bled via Orbital for Clin Path, Right
24,52
Sacrificed by design
94
#
DuPont-11561
-53-
C o m p a n y Sanitized. Does not contain TSCA CB1
J
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP III (Continued)
Animal Observation
Days
646940 General observation, No Abnormality Detected
1-10,17-80,94
Eye Observations, Bled via Orbital for Clin Path, Left
13
Eye Observations, Bled via Orbital for Clin Path, Right
24,52
Discharge, Nose, Black
87
Sacrificed by design
94
646941 General observation, No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Bilateral
52
Eye Observations, Bled via Orbital for Clin Path, Left
13
Eye Observations, Bled via Orbital for Clin Path, Right
24
V Sacrificed by design
94
#
D u P o n t-11561
Company Sanitized. Does net contain TSCA CB1
-54-
m
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24019 (POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I
Animal Observation
Days
646910 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Sacrificed by design
10
646911 General observation, No Abnormality Detected
1,10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Eye Observations, Dark, Left
2-9
Sacrificed by design
10
646912 General observation, No Abnormality Detected ?
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1
Eye Observations, Bled via Orbital for Clin Path, Right
5
Sacrificed by design
10
646913 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Sacrificed by design
10
646914 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Sacrificed by design
10
m DuPont-11561
Company Sanitized. Does net contain TSCA CB1 -55-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal 646915
646916
646917
Observation
GROUP III
General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left Discharge, Eye right, Red
Hair Loss, Forelimb, Right
Hair Loss, Forepaw, Right
Sacrificed by design
General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design
General observation, No Abnormality Detected
Eye Observations, Bledvia Orbital for Clin Path, Left Eye Observations, Bledvia Orbital for Clin Path, Right Sacrificed by design
Days 1,17-38,73 13,24,52 2-10 80 45-67,87-94 94 1-10,17-94 24 13 52 94 1-10,17-94 13,24 52 94
D uPont-11561
Company Sanitized. Does not contain TS CA CBI
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24019 (POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP III (Continued)
Animal 646918
Observation General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left Eye Observations, Bledvia Orbital for Clin Path, Right
Days 1-10,17-94 13,24 52
Sacrificed by design
94
646919
General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left
1-10,17-94 13,24
Eye Observations, Bledvia Orbital for Clin Path, Right
52
Sacrificed by design
94
D uP ont-11561
ompany Sanitized. Does not contain TSCA CBI -57-
i m
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________
H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal 646921
GROUP I Observation General observation, No Abnormality Detected
Days 1,5-10
646922 646923
Eye Observations, Bled via Orbital for Clin Path, Left Eye Observation^, Dark, Left Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design General observation, No Abnormality Detected Eye Observations, Exophthalmus, Left Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Eye Observations, Dark, Left Sacrificed by design
1,5 2-4 10 1-10 1 5 10 1,7-10 4-6 1 5 2-6 10
DuPont-11561
Ccmpany Sanitized. Does not contain TSCACBl -58-
%
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24020 (POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I (Continued)
Animal Observation
Days
646924 General observation, No Abnormality Detected
1-10
Eye Observations, Bledvia Orbital for Clin Path, Left
1
Eye Observations, Bledvia Orbital for Clin Path, Right
5
Sacrificed by design
10
646925 General observation, No Abnormality Detected
1-10
Eye Observations, Bledvia Orbital for Clin Path, Left
1
Eye Observations, Bledvia Orbital for Clin Path, Right
5
Sacrificed by design
10
D u P o n t-11561
Company Sanitized. Does not contain TSCA CBI
m
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24020(POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal Observation
GROUP III.
Days
646926 General observation, No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Bilateral
52
Eye Observations, Bled via Orbital for Clin Path, Left
13,24
646927
Sacrificed by design General observation, No Abnormality Detected
94 1-10,17-94
646928
Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design General observation, No Abnormality Detected
52 13,24 94 1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Left
13,24,52
Sacrificed by design
94
#
DuPont-11561
Company Sanitized. Doss not contain TS C A CBI
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24020(POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP III (Continued)
Animal Observation
Days
646929 General observation, No Abnormality Detected
1-10,17-31
Eye Observations, Bled via Orbital for Clin Path, Left
13,24
Eye Observations, Bled via Orbital for Clin Path, Right
52
Hair Loss, Forelimb, Bilateral
52-73
Hair Loss, Neck, Left
73
Hair Loss, Neck, Ventral
38-67,80-94
Wound, Superficial, Face
59
Sacrificed by design
94
646930 General observation, No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Left
13,24
Eye Observations, Bled via Orbital for Clin Path, Right
52
Sacrificed by design
94
D u P o n t-11561
-61 -
Company Sanitized. Does not contain TSCA CBl
mtasa --
m
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24750 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I
Animal Observation
Days
665547 General observation, No Abnormality Detected
1-109
Eye Observations, Bled via Orbital for Clin Path, Right
1,5
Sacrificed by design
10
665548 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Right
1,5
Sacrificed by design
10
665549 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Right
1,5
Sacrificed by design
10
665550 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Right
1,5
Sacrificed by design
10
665553 General observation, No Abnormality Detected
1-10
Eye Observations, Bled via Orbital for Clin Path, Right
1,5
Sacrificed by design
10
w DuPont-11561
-62-
Company Sanitized. Does not contain TSCA CB1
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
H-24750 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP III
Animal 665554
Observation General observation, No Abnormality Detected
Days 1-94
665555
Eye Observations, Bled via Orbital for Clin Path, Right General observation, No Abnormality Detected
24.52 1-10
Eye Observations, Closed
38-94
Eye Observations, Exophthalmus, Right
17-31
Eye Observations, Bled via Orbital for Clin Path, Left Discharge, Nose, Red
24.52 45
665558
Hair Loss, Face, Right Comments General observation, No Abnormality Detected
31-45 17-94 1-17,87-94
Eye Observations, Bled via Orbital for Clin Path, Right
24.52
Hair Loss, Forelimb, Bilateral
24-80
665559
Hair Loss, Forepaw, Bilateral General observation, No Abnormality Detected
24-66 1-94
665560
Eye Observations, Bled via Orbital for Clin Path, Right General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right
24.52 1-4,17-94 24.52
Hair Loss, Shoulder, Bilateral
9-10
Hair Loss, Shoulder, Left
5-Sompany Sanitized. Does not contain T S C A CB1
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX C Terms and Calculations Factors Influencing Interpretation of Kinetic Analysis
Company Sanitized. Does not contain TSCA CB1 -64-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
Terms: Active % Active
Mol Wt Active Formulation Dose % F in Active
Mol Wt F
TERMS AND CALCULATIONS
Fluorine containing compound The % of formulation that is made up o f fluorine containing components The molecular weight of the fluorine containing components (g/mole) The mg of formulation given per kg o f animal body weight The % fluorine in the fluorine containing components of the formulation (weight basis) The molecular weight o f fluorine g/mol
Compound Calculations:
Dose Active (mg/kg)
The mg o f fluorine containing compound administered per kg of animal body weight. = (% active/100) x Formulation Dose
Dose Active (mmole/kg) The mmole of fluorine containing compound administered per kg of animal body weight = dose [mg/kg] / Mol Wt Active [mg/mmol]
Dose F (mg/kg)
The mg Fluorine administered per kg o f animal body weight = (% F in active/100) x Dose Active [mg/kg]
Dose F (mmol/kg)
The mmole of fluorine administered per kg of animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol]
Molar Ratio (Active/F) The moles of fluorine containing compound per mole o f fluorine = Dose Active [mmol/kg] / Dose F [mmol/kg]
Formulation Dose Normalization Factor
The formulation dose that would be required to administer the amount of active needed to achieve the normalized dose = (Normalized dose o f Active [mmol/kg] / Dose Active [mmol/kg])
x Formulation Dose
Company Sanitized. Doss not contain TSCA CBI
-65-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppm F
The ppm fluoride measured
Individual Animal Calculations:
ppm F minus Bkg 0.2 ppm
The ppm fluoride measured minus the background fluoride measured in control animal. In this case the value was established at 0.2 ppm.
ppm f normalized to 0.1 mmol/kg Dose
The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead of the actual active dose. This assumes linearity between administered dose and blood fluorine levels, but is needed because different doses o f active were used in the
study. = (0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in blood
minus background)
pmolar equivalents o f active
The pmolar [pmol/L] concentration o f fluorine containing compound based on the ppm fluorine normalized to 0.1 mmol/kg active dose. This assumes that all fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm --1 mg/L = (Normalized ppm [mg/L] fluorine / Mol Wt F [mg/mmol]) x
molar ratio active/F [mmol active/mmol F] x 1000 pmol/mmol
!
Company Sanitized. Does not contain TSCA CBI - 66-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ....
DuPont-11561
FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS
Considerations:
- The data used for kinetic analysis was from a limited screen and extrapolation should be done cautiously.
- Sample size is low - Analytical data used without validation - Steady-state not reached - Terminal phase may not be reached - Some compounds are mixtures of fluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - Each compound may have very different potency for producing toxicity
Assumptions: (May or may not bejustified in all cases)
- Fluorine concentrations are linear with respect to dose - Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of
individual components - Background Fluorine is 0.2 ppm - % F data is the % Fluorine of the active (Fluorine containing component(s) in the formulation) - Molecular weight is the molecular weight o f the active component in the formulation
Company Sanitized. Does not contain TSCA CB1
-67-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX D Individual Fluorine Levels in Blood
Company Sanitized. Does nc.contain TSCA CB1 -68-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):_________________ _
10 0.020 0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):
6.5
_______________
Rat Number
Test ppm F Day in Sample Blood
Group I 646910 646911 646912 646913 646914
1 1 1 1 1
2.6 2.7 2.5 2.6 2.6
ppm F in Blood
Minus Bkg 0.2 ppm
2.4 2.5 2.3 2.4 2.4
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
11.93 12.43 11.43 11.93 11.93
pmolar Equivalents of Active in
Blood
36.92 38.46 35.38 36.92 36.92
Group I 646910 646911 646912 646913 646914
5 5 5 5 5
30.1 30.3 31.3 32.2 32.7
29.9 30.1 31.1 32.0 32.5
148.60 149.60 154.57 159.04 161.53
460.00 463.08 478.46 492.31 500.00
Group I
646910 10 71.5
646911
10 70.5
646912
10 66.9
646913
10 62.5
646914 10 68.6
71.3 70.3 66.7 62.3 68.4
354.36 349.39 331.50 309.63 339.95
1096.92 1081.54 1026.15 958.46 1052.31
Company Sanitized. Does ne t contain TSCA CBI -69-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test ppm F Day in Sample Blood
Group i n 646915 646916 646917 646918 646919
13 13 13 13 13
55.0 55.3 52.2 53.8 53.6
ppm F in Blood
Minus Bkg 0.2 ppm
54.8 55.1 52.0 53.6 53.4 .
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
272.36 273.85 258.44 266.39 265.40
DuPont-11561
pmolar Equivalents of Active in
Blood
843.08 847.69 800.00 824.62 821.54
Group III 646915 646916 646917 646918 646919
Group III
646915 646916 646917 646918 646919
24 24 24 24 24
52 52 52 52 52
38.9 43.8 37.6 35.4 42.4
23.2 20.3 25.9 24.9 23.5
38.7 43.6 37.4 35.2 42.2
23.0 20.1 25.7 24.7 23.3
192.34 216.69 185.88 174.94 209.73
114.31 99.90 127.73 122.76 115.80
595.38 670.77 575.38 541.54 649.23
353.85 309.23 395.38 380.00 358.46
Group III 646915 646916 646917 646918 646919
94 94 94 94 94
13.3 11.7 12.9 11.0 14.1
13.1 11.5 12.7 10.8 13.9
65.11 57.16 63.12 53.68 69.08
201.54 176.92 195.38 166.15 213.85
Company Sanitized. Doss nc" contain TSCA CB1 - 70-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24020
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
% F in Active: Mol Wt. F (g/mol):
69 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg) :
13.8
Rat Number
Test ppm F Day in Sample Blood
Group I
646921
1 10.6
646922 1 10.7
646923 1 9.2
646924 1 5.3
646925 1 11.2
ppm F in Blood
Minus Bkg 0.2 ppm
10.4 10.5 9.0 5.1 11.0
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
22.15 22.37 19.17 10.86 23.43
fimolar Equivalents of Active in
Blood
75.36 76.09 65.22 36.96 79.71
Group I 646921 646922 646923 646924 646925
5 5 5 5 5
78.7 83.2 77.0 66.1 69.6
78.5 83.0 76.8 65.9 69.4
167.21 176.79 163.58 140.37 147.82
568.84 601.45 556.52 477.54 502.90
Group I
646921
10 63.0
646922 10 69.1
646923 10 60.3
646924 10 62.5
646925 10 53.9
62.8 68.9 60.1 62.3 53.7
133.76 146.76 128.01 132.70 114.38
455.07 499.28 435.51 451.45 389.13
Comoany Sanitized. Docs not contain TSCA CBl -71 -
H-24750: Biopersistenc Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test ppm F Day in Sample Blood
Group III 646926 646927 646928 646929 646930
13 13 13 13 13
34.8 30.0 23.7 33.7 25.4
ppm F in Blood
Minus Bkg 0.2 ppm
34.6 29.8 23.5 33.5 25.2
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
73.70 63.47 50.06 71.36 53.68
DuPont-11561
pmolar Equivalents of Active in
Blood
250.72 215.94 170.29 242.75 182.61
Group III 646926 646927 646928 646929 646930
24 24 24 24 24
11.9 10.4 8.1 15.7 9.8
11.7 10.2 7.9 15.5 9.6
24.92 21.73 16.83 33.02 20.45
84.78 73.91 57.25 112.32 69.57
Group m
646926
52
2.5
2.3
4.90 16.67
646927
52
1.7
1.5 '
3.20
10.87
646928
52
1.3
1.1
2.34 7.97
i
646929
52
4.0
3.8
8.09 27.54
646930
52
1.8
1.6
3.41 11.59
Group in 646926 646927 646928 646929 646930
94 94 94 94 94
0.8 <0.5 <0.5 0.9 <0.5
0.6 *
*
0.7 *
* Below LOQ (Limit of Quantification)
1.28 4.35 ** **
1.49 5.07 **
Company Sanitized. Does not contain TSCA CB!
-72-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24750
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
514 5 100
% F in Active: Mol Wt. F (g/mol):
70.2 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
5 0.010 0.185
Molar Ratio (Active/F): 0.053
Dose F (mg/kg):
3.5
Rat Number
Test Day Sample
ppm F in
Blood
Group I
665547 1 3.01
665548
1 3.25
665549 1 2.68
665550 1 3.21
665553
1 3.54
ppm F in Blood
Minus Bkg 0.2 ppm
2.8 3.1 2.5 3.0 3.3
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
28.89 31.36 25.50 30.95 34.34
fimolar Equivalents of Active in
Blood
80.06 86.89 70.66 85.75 95.16
Group I
665547 5 19.27
665548
5 15.37
665549
5 19.83
665550
5 15.47
665553
5 15.72
19.1 15.2 19.6 15.3 15.5
196.07 155.97 201.83 157.00 159.57
543.30 432.19 559.26 435.04 442.17
Group I 665547 665548 665549 665550 665553
.
10 10 10 10 10
30.45 28.9 30.09 30.44 32.3
30.3 28.7 29.9 30.2 32.1
311.02 295.08 307.32 310.92 330.04
861.82 817.66 851.57 861.54 914.53
Company Sanitized Does not contain TSCA CBI
-73
*
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test ppm F Day in Sample Blood
Group III 665554 665555 665558 665559 665560
13 13 13 13 13
46.92 31.35 32.95 30.2 30.56
ppm F in Blood
Minus Bkg 0.2 ppm
46.7 31.2 32.8 30.0 30.4
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
480.36 320.27 336.72 308.45 312.15
DuPont-11561
limolar Equivalents of Active in
Blood
1331.05 887.46 933.05 854.70 864.96
Group III 665554 665555 665558 665559 665560
24 24 24 24 24
31.31 25.29 32.36 26.8 22.25
31.1 25.1 32.2 26.6 22.1
319.86 257.97 330.66 273.49 226.71
886.32 714.81 916.24 757.83 628.21
Group III 665554 665555 665558 665559 665560
52 52 52 52 52
11.09 7.64 12.96 8.88 5.91
10.9 7.4 12.8 8.7 5.7
111.97 76.50 131.19 89.24 58.71
310.26 211.97 363.53 247.29 162.68
Group III 665554 665555 665558 665559 665560
94 94 94 94 94
6.3 6.28 6.13 6.9 5.6
6.1 6.1 5.9 6.7 5.4
62.72 62.51 60.97 68.89 55.52
173.79 . 173.22
168.95 190.88 153.85
Company Sanitized. Does not contain TSCA CB1 -74-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX E Individual Fluorine Levels in Liver
I
C otnoan,r Sanfi^pd. Dna* r ' ' r>ta!r*TROA CPI -75-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
10 0.020 0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):
6.5
Rat Number
Test ppm F Day in Sample Liver
Group I
646910
10 320.9
646911
10 334.0
646912
10 318.6
646913
10 281.6
646914
10 306.6
ppm F in Liver
Minus Bkg 0.2 ppm
320.7 333.8 318.4 281.4 306.4
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
1593.88 1658.99 1582.45 1398.56 1522.81
pmolar Equivalents of Active in
Liver
4933.85 5135.38 4898.46 4329.23 4713.85
Group III 646915 646916 646917 646918 646919
94 94 94 94 94
85.6 75.9 87.5 78.0 94.2
85.4 75.7 87.3 77.8 94.0
424.44 376.23 433.88 386.67 467.18
1313.85 1164.62 1343.08 1196.92 1446,15
Company Sanitized. Does not contain TSCA CB? -76-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24020
DuPont-11561
Given: . Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
% F in Active: Mol Wt. F (g/mol):
69 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat Number
Test ppm F Day in Sample Liver
Group I
646921
10 118.3
646922
10 122.7
646923
10 114.3
646924
10 121.6
646925
10 122.1
ppm F in Liver
Minus Bkg 0.2 ppm
118.1 122.5 114.1 121.4 121.9
-
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
251.55 260.93 243.03 258.58 259.65
pmolar Equivalents of Active in
Liver
855.80 887.68 826.81 879.71 883.33
Group III 646926 646927 646928 646929 646930
94 94 94 94 94
1.8 1.2 2.2 4.7 2.9
1.6 1.0 2.0 4.5 2.7
3.41 11.59 2.13 7.25 4.26 14.49 9.59 32.61 5.75 19.57
-77-
anU 'T O ^ D n a c nn? a n n ta in
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24750
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
514 5 100
% F in Active: Mol Wt. F (g/mol):
70.2 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
5 0.010 0.185
Molar Ratio (Active/F): 0.053
Dose F (mg/kg):
3.5
Rat Number
Test ppm F Day in Sample Liver
Group I
665547
10 186.4
665548
10 189.7
665549
10 200.5
665550
10 190.3
665553
10 202.6
Group III 665554 665555 665558 665559 665560
94 94 94 94 94
57.33 49.65 57.97 48.41 46.68
ppm F in Liver
Minus Bkg 0.2 ppm
186.2 189.5 200.3 190.1 202.4
57.1 49.5 57.8 48.2 46.5
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
1914.54 1948.47 2059.10 1954.84 2081.30
587.39 508.43 593.97 495.68 477.89
pmolar Equivalents of Active in
Liver
5305.13 5399.15 5705.70 5416.81 5767.24
1627.64 1408.83 1645.87 1373.50 1324.22
Company Sanitized. Does nof confato TSCA
-78-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX F Individual Fluorine Levels in Fat
- 79-
Jompany Sanitized. Does not contain TSCA CB1
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg): ______________
10 0.020 0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):
6.5
_________________________
Rat Number
Test ppm F Day in Sample Fat
Group I 646910 646911 646912 646913 646914
10 10 10 10 10
14.4 14.9 10.9 11.6 12.4
ppm F in Fat
Minus Bkg 0.2 ppm
14.2 14.7 10.7 11.4 12.2
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
pmolar Equivalents of Active in
Fat
70.57 73.06 53.18 56.66 60.63
218.46 226.15 164.62 175.38 187.69
Group III 646915 646916 646917 646918 646919
94 94 94 94 94
1.3 1.1 1.3 0.8 1.5
1.1 0.9 1.1 0.6 1.3
5.47 16.92 4.47 13.85 5.47 16.92 2.98 9.23 6.46 20.00
^irtroany Sanitized. Does no} contain Tfir a
-80-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ___
Data for H-24020
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
% F in Active: Mol Wt. F (g/mol):
69 19
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat Number
Test ppm F Day in Sample Fat
Group I 646921 646922 646923 646924 646925
10 10 10 10 10
6.2 8.5 8.7 8.8 8.3
ppm F in Fat
Minus Bkg 0.2 ppm
6.0 8.3 8.5 8.6 8.1
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
12.78 17.68 18.11 18.32 17.25
pmolar Equivalents of Active in
Fat
43.48 60.14 61.59 62.32 58.70
Group III
646926 94 ND
ND
ND
ND
646927 94 ND
ND
ND
ND
646928 94 ND
ND
ND
ND
646929 94 ND
ND
ND
ND
646930 94 ND
ND
ND
ND
ND Non-detectable.
Company Sanitized. Does no! contain TSCA CB1
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24750
DuPont-11561
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
514 5 100
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
5 0.010 0.185
% F in Active: Mol Wt. F (g/mol):
___ /V.2 19
Molar Ratio (Active/F): Dose F (mg/kg):
.. 0.053 3.5
Rat Number
Test Day Sample
ppm F in Fat
Group I 665547 10 4.21 665548 10 3.71 665549 10 2.79 665550 10 4.24 665553 10 3.45
Group III 665554 665555 665558 665559 665560
94 94 94 94 94
<.5 <.5 <.5 <.5 <5
ppm F in Fat
Minus Bkg 0.2 ppm
4.0 3.5 2.6 4.0 3.3
* * * * *
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
41.23 36.09 26.63 41.54 33.42
* * * *
limolar Equivalents of Active in
Fat
114.25 100.00 73.79 115.10 92.59
* * * * *
* Below LOQ (Limit o f Quantification)
ompany Sanitized. Does nof contain TSCACBI
- 82-
