Document yp8MDNqm1vpNvQrOpJ0Qd2Lj6
AR226-2808
TRADE SECRET
Study Title H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ED: DuPont-11561
AUTHOR: CarolFinlay,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
^B^ WORKREQUEST NUMBER:
. ^^^--*
SERVICE CODE NUMBER:^B||||\
Company Sanitized. Does not contain TSCA CBI
Page 1 of 82
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Reviewed by: Issued by Study Director;
'
/
Paul M- Hinderliter, PbD. Research Toxicologist
C<dL46"L \\'\SLlL^
CWlFinlay.B.A}
Staff Toxicologist
01-^r -3
Date
^.^r^3 Date
Company Sanitized. Does not contain TSCA CBf
7 -
-
TABLE OF CONTENTS
Page
CERTIFICATION........................................................................................................................^
LIST OF TABLES .........................................................................................................................4 LIST OF FIGURES .......................................................................................................................4 LIST OF APPENDICES...............................................................................................................5 STUDY INFORMATION .............................................................................................................6 STUDY PERSONNEL...................................................................................................................8
SUMMARY...................................................................................................................................^
INTRODUCTION........................................................................................................................11
MATERIALS AND METHODS................................................................................................11
A. Test Substance and Positive Controls................................................................................! 1 B. Test Species .......................................................................................................................11
C. Animal Husbandry.............................................................................................................11 D. Quarantine and Pretest....................................................................................................... 12 E. Study Design......................................................................................................................13 F. Assignment to Groups and Study Start..............................................................................13 G. Dosing Material Preparation and Administration.............................................................. 13 H. BodyWeights.....................................................................................................................l4 I. Mortality and Clinical Observations..................................................................................14 J. Collection and Analysis of Blood, Livers, and Fat............................................................ 14 K. Treatment of Fluorine Data................................................................................................15 L. Statistical Analyses............................................................................................................16 RESULTS AND DISCUSSION ..................................................................................................17 A. h-Life Toxicology............................................................................................................. 17 B. Liver Weights.....................................................................................................................17 C. Fluorine Data.....................................................................................................................18 CONCLUSIONS ....:.........................................................................................:...........................19 RECORDS AND SAMPLE STORAGE ....................................................................................20
TABLES........................................................................................................................................21
FIGURES......................................................................................................................................28
APPENDICES ..............................................................................................................................39
Company Sanitized. Does not contain TSCA CBt
3-
LIST OF TABLES
Page
1. MEAN BODY WEIGHTS...................................................................................................................................22 2. MEAN BODY WEIGHT GAINS........................................................................................................................23 3. MEAN BODY AND LIVER WEIGHTS............................................................................................................. 24 4. MEAN BLOOD FLUORINE LEVELS...............................................................................................................25 5. MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE............................................... 25 6. MEAN LIVER FLUORINE LEVELS................................................................................................................. 26 7. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE ................................................. 26 8. MEAN FAT FLUORINE LEVELS..................................................................................................................... 27 9. MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE...................................................... 27
LIST OF FIGURES
Page
1. MEAN BODY WEIGHTS...................................................................................................................................29 2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE
CONTROL...........................................................................................................................................................30
3. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE
31 CONTROLS.........................................................................................................................................................
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 DOSE................................................. 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
Company Sanitized. Doss not contain TSCA CBI
.4-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
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 FAT............................................................................ 79
Company Sanitized. Does no? contain TSCA CBT 5-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION
Test Substance 9th Collective Nomenclature: Synonyms/Codes:
Submitter's Notebook NumberfsY Haskell Number: 24750
DuPont-11561
Known Impurities Positive Control
Substance Tested: Potassium perfluoroalkyi sulfonate Synonyms/Codes: H-24019
Haskell Number: 24019 Compositioni
Known Impurities:
Company Sanitized. Does not contain TSCA CBT
-6-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
STUDY INFORMATION (Continued)
Positive Control Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt Synonyms/Codes: H-24020
C-8 Perfluorooctanoate, ammonium salt APFO Ammonium perfluorooctanoate Haskell Number: 24020 Compositiom
Known Impurities:
Sponsor:
E.I. du Pont de Nemours and Company Wilmington, Delaware 19898
U.S.A.
Study Initiated/Completed: October 10, 2002 / (see report cover page)
Company Sanitized. Does not contam TSCA CBI
7-
H-24750: Biopersistence Screening jO-Dose Oral Gavage Study in Rats
__ ___ ________DuPont-11561
STUDY PERSONNEL
Study Director: Carol Finlay, B.A. Management: Scott E. Loveless, Ph.D.
Primary Technician: James C. Mackay n
Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanfiy, 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.
Company Sanitized. Does not contam TSCA CBI
-8-
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 of 5 male rats each were dosed by gavage with 5 mg/kg/day of 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 ofdeionized 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 of 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 of rats dosed with the positive control materials was similar
during the dosing period to the weight gain of rats dosed with the test substance. During the
recovery period, the mean body weight gain of 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 uM equivalents. As a measure of 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 of5.22xl05 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) of rats dosed with H-24750 was 62% higher than the negative control rats on day 10. By day 94, the mean relative liver weight of 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 of 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 of 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 aM equivalents on day 10 and 1476.01 aM equivalents on day 94. The mean uM equivalent concentration of fluorine in the livers from rats dosed with the test substance was slightly higher
Company Sanitized. Does not contain TSCA CBl
-9-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
on days 10 and 94 than the value for the positive control H-24019. The mean uM equivalent concentration of fluorine in the livers from rats dosed with the test substance was approximately 6x higher (end of dosing) or approximately 86x higher (end of 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 uM 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 of 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 of this study, fluorine was evident in the blood during the period of dosing with H-24750. Not all of 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.
Company Sanitized. Does no? contain TSCA CBT 10-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
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 of 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 of 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)IGSBR rat was selected on the basis of 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 of 23 1C and a relative humidity of 50 10%.
-ll -
Company Sanitized. Does. not contain TSCA CBT /'
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
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 ofcoliforms, 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 of 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 of the study.
The animal health and environmental monitoring program is administered by the attending
laboratory animal veterinarian. Evaluation of these data did not indicate any conditions that affected the validity of 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.
_^_ Comoaw Sanitized. Does nol conlain TSCA CBl
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
E.
Study Design
Substance
Negative Control Deionized water
Positive Controls
H-24019 H-24020 Test Substance H-24750
Vehicle
Not applicable Acetone/Corn Oil Acetone/Corn Oil
Deionized Water
Dosage (mg/kg)
0
10 20
5
Number of
Animals
10
10 10
10
DuPont-11561
F.
Assignment to Groups and Study Start
After the quarantine period, the rats were selected on the bases of 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 of 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 of 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 corn oil. The ratio of acetone to corn 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 of body weight. The dosing preparations were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity.
Company Sanitized. Does "ol contain TSCA CBI 13-
H-24750: Biopersistence Screening
_
10-Dose Oral Gavage Study in Rats
DuPont-11561
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 I I I m m m
m
Dosing Days 1-10 1-10 1-10 1-10 1-10 1-10 1-10
Tissue Collected Blood Blood Blood, Liver, and Fat Blood Blood Blood Blood, Liver, and Fat
Sampling Time Test day 1 (2 hours post dosing) Test day 5 (2 hours post dosing) Test day 10 at sacrifice (2 hours post dosing) Test day 13 Test day 24 Test day 52 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 of 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 of 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
Comoqnv Sanitized. Does not contain TSCA CB\
_^_
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
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 umolar units to accommodate test material molecular weight differences and was adjusted to an arbitrary normalized dose of 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 of 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 of detection (LOD) of 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, m addition, the data were standardized to a single
reference dose to allow comparison between compounds of 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 umolar (u.M) equivalents
of active component in the tissue. Detailed calculations can be found in Appendix C. The uM
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 phannacokinetic parameters from experimental data. All analysis was calculated using the uM equivalent in blood data.
The maximum observed concentration in blood was Cmax (uM equivalent). Biopersistence was
assessed by quantifying terminal elimination blood half-life (T%, days). The points included in
determination of the T-/, were 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.
Comoany Sanitized. Does not contain TSCA CBT
-^-
H-24750: Biopersistence Screening 10-DoseOralGavage Study in Rats
L.
Statistical Analyses
Descriptive statistics (e.g. mean, standard deviation) were used.
DuPont-11561
Comnnv Sanitized. Does not confqm TSCA CBV
-16-
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 of rats dosed with H-24750 were lower than the negative control rats during the dosing and recovery periods. Mean body weight gain of rats dosed with the test substance was comparable during the dosing period but lower during the recovery period than the weight gain of 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) of rats dosed with H-24750 was 62% higher than the negative control rats on day 10. By day 94, the mean relative liver weight of 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 of 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 of rats dosed with
H-24750.
The mean relative liver weight of rats dosed with the other positive control, H-24020, was 16% higher at day 10 than the liver weight of 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 of 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.
Comoanv Sanitized. Docs not cona!n TBRA R%
nv
H-24750: Biopersistence Screening 10-DoseOral Gavage Study in Rats
DuPont-11561
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 of the terminal half-life. A more complete list of 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 uM
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 uM equivalents
(mean S.D.) with a terminal half-life of 42.2 days. The Cmax for H-24020 was 541.45 50.37 uM equivalents (mean S.D.) with a terminal half-life of 15.1 days. For each of 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 of fluorine in the livers on day 10 from rats dosed with the positive control materials were 4802.15 and 866.67 uM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1292.92 and 17.10 uM equivalents.
The concentrations of fluorine in the fat on day 10 from rats dosed with the positive control materials were 194.46 and 57.25 uM 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 uM equivalents. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020.
-18- Company Sanitized. Does not contain TSCA CBf
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
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 of H-24750 was 4.83xl05 as compared to AUCINF/D values of5.22xl05 and 8.15xl04 for H-24019 and H-24020, respectively (Figure 4C).
Levels of 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 uM equivalents at day 10 and 1476.01 uM 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 uM 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 of 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.
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 AUCINF/D for the positive control H24020.
company Sanitized. Doss not contain TSCA CBI
-19-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
Under the conditions of this study, there was considerable absorption and retention of fluorine in the blood following dosing with H-24750. Administration of 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.
TABLES
Company Sanffeed. Does not contain TSCA CBI -21 -
H-24750: Biopersistence Screening 10-DoseOralGavage Study in Rats
DuPont-11561
TABLE 1
MEAN BODY WEIGHTS
DAYS ON TEST
Negative Control 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 CBI 22-
H-24750: Biopersistence Screening 10-DoseOral Gavage Study in Rats
DuPont-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 5-10
32.5 34.9
31.8 30.3
28.4 30.5
35.3 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 32.2 95.0 91.2
43.6 23.2 109.5 101.8
54.4 41.2 122.0 109.9
-11.1 10.5 82.1 104.6
10-94
254.7
278.1
327.5
186.2
Company Sanitized. Does not contain TSCA CB3
-23-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
TABLE 3
DAYS ON TEST
10 94
MEAN BODY AND LIVER WEIGHTS
NEGATIVE CONTROL - DEIONIZED WATER
WEIGHT (g)
Absolute
Mean Relative Liver Weight
Body Weight
Liver Weight
(Liver/Body Weight)
258.8 511.7
10.814 17.868
0.042 0.035
DAYS ON TEST
POSITIVE CONTROL - H-24019
WEIGHT (g)
Absolute
Body Weight
Liver Weight
Mean Relative Liver Weight (Liver/Body Weight)
10
243.9
14.205
0.058
94
524.6
19.296
0.037
-
DAYS ON TEST
10 94
POSITIVE CONTROL - H-24020
WEIGHT (g)
Absolute
Body Weight
Liver Weight
Mean Relative Liver Weight (Liver/Body Weight)
243.1 570.5
19.174 19.590
0.079 0.034
DAYS ON TEST
10 94
TEST SUBSTANCE - H-24750
WEIGHT (g)
Absolute
Body Weight
Liver Weight
Mean Relative Liver Weight (Liver/Body Weight)
260.9 434.6
17.785 18.841
0.068 0.043
Company Sanitized. Doas not contain TSCA CBI
-24-
H-24750: Biopersistence Screening 10-DoseOral Gavage Study in Rats
DuPont-11561
TABLE 4
MEAN BLOOD FLUORINE LEVELS
DAYS ON TEST
MEAN I ^LUORDsFELEVE]LS (ppm)1
Negative Contn3l Deionized Wate;r
Positive Controls
H-2^?19
H-2-1020
Test Suibstance H-2-1750
1
0.60s1
2.60 (0.1)"
9.40 (2.4)
3.14 (0.32)
5
c
31.32 (1.1)
74.92 (7.0)
17.13 (2.22)
10
1.10'
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
0.5s
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)
0.85'1 (0.1)
6.24 (0.46)
a One of 5 values. Four of the values were below the limit of 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 of 2 of the 5 values. Three of the values were below the LOQ.
TABLE 5
MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE
DAYS ON TEST
MEAN FLUORINE CONCENTRATION (uM F Equivalents)
Positive Controls
Test Substance
H-24019
H-24020
H-24750
1
36.92 (I.I)3
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)
4.7113 (0.5)
172.14 (13.23)
a Standard deviation (S.D.) is in parentheses. b Mean of 2 of the 5 values. Three of the values were below the limit of quantification (LOQ).
Company Sanitized. Doss not contain TSCA CBI
-25-
TABLE 6
MEAN LIVER FLUORINE LEVELS
DAYS ON TEST
MEAN FLUORINE LEVELS (ppm)
Negative Control
Positive Controls
Deionized Water
H-24019
H-24020
Test Substance H-24750
10
0.90 (0.2)' 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 (uM F Equivalents)
Positive Controls
Test Substance
H-24019
H-24020
H-24750
10
4802.15 (303.8)'
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 SanitizccL Does net contain TSCA CBt
-26-
H-24750: Biopersistence Screening 10-DoseOral Gavage Study in Rats
DuPont-11561
TABLE 8
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
-a
12.84 (I.?)"
8.10 (1.1)
3.68 (0.6)
94 . -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
MEAN FLUORINE CONCENTRATION (uM F Equivalents)
Positive Controls
Test Substance
H-24019
H-24020
H-24750
10
194.46 (26.8)"
57.25 (7.8)
99.15 (17.1)
94
15.38 (4.1)
-b
-"
a Standard deviation (S.D.) is in parentheses. b All values were below the limit of quantification (LOQ) or non-detectable.
Company Sanitized. Does no? contain TSCA CB1 -27-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
FIGURES
Comnany Sanitized. Does not contain TSCA CB!
-28-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
300 T
FIGURE 1 MEAN BODY WEIGHTS
-29-
Company Sanitized. Does n
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROL
-< >> 0) "0
> o
;% m
&
0.03
Test Days
-30-
Cffltlpsmy Sanitized. Does not co
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 3 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
Test Days 31 -
^-anpany SanFffzed. Does not c
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
FIGURE 4 MICROMOLAR EQUIVALENTS IN RAT BLOOD
A. Normalized Ral Blood H-24019 |iU Equivalents Resulting from a 10-Day Oral Gavage
Micromolar (uM) 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. Normalized Rat Blood H-2 4020 uM Equivalents Re culling from a 10-Day Oral Gavage
Micromolar (pM) equivalents ofH-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 CB1 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
MOO 1200 1000
600
-100 200
Micromolar (uM) 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.
;ompany Sanitized. Doas not contain TSCA CB1 33
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
600000
FIGURE 5
NORMALIZED H-24750 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A 10-DAY ORAL GAVAGE
500000
400000
300000
CD
200000
100000
03 AUCINF/D
H-24020 8.15&-04
34-
. ompany Sanitized. Does n
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 6 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Day 10
Test Days
-35-
Day 94
Company Sanitized. Dees no
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 7 COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRA
FOR H-24750 AND NEGATIVE CONTROL
Day 10 Test Days Day 94
D Deionized Water (Negative Control) H-24750 (Test Substance)
36-
Day 10
Day
Test Days
company Sanitized. Does
FT""
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 8 MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE
Day 10
Test Days
-37-
Day 94
:mpany Sanitized. Does n
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
-38-
Company Sanitized. Does n
APPENDICES
39-
APPENDIX A Individual Body Weights
Company Sanitized. Does not contain TSCA CB1 .40-
INDIVIDUAL BODY WEIGHTS
ABBREVIATIONS: SD - sacrificed by design
EXPLANATORY NOTES
Company Sanitized. Does not contain TSCA CBI
-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
TE ST 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
ANIMAL NUMBER
646932 646933 646934 646935 646936
Day 10
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
Day 7 242 .4 232 .9 224 .2 257 .2 229 .9
-42-
Company Sanitized. Does
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
ANIMAL NUMBER
646937 646938 646939 646940 646941
Day 10 249 .8 237 .2 277 .9 244 .5 266 .3
ANIMAL NUMBER
646937 646938 646939 646940 646941
Day 73 504 .3 . 417 .4 540 .1 407 .3 493 .7
Day 2
186.8 173.8 204.2 197.1 195.9
Day 17
282 .6 270 .4 329 .7 271 .5 312 .4
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
TE ST 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 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 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 3D test day 94 SD test day 94 SD test day 94 SD test day 94
Day 7
229. 6 215. 9 248. 9 229. 3 241. 3
Day 52 443 .9 373 .7 484 .1 367 .0 433 .6
-43-
;ompany Sanitized. Doe
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
H-24019 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (9) OF MALE RATS
GROUP I
Day 3
TE ST 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
646910 646911 646912 646913 646914
Day 10
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
-44-
Company Sanitized. Does n
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
646915 646916 646917 646918 646919
Day 1 182 .7 IBS .0 187 .8 175 .5 192 .0
Day 2 188 .3 192 .7 194 .3 181 .9
196 .9
AMIMAL NUMBER
646915 646916 646917 646918 646919
Day 10 240 .4 257 .0 250 .2 237 .1 260 .2
Day 17 283 .4 310 .0 280 .3 270 .6 306 .1
ANIMAL NUMBER
646915 646916 646917 646918 646919
Day 73 461 .2 544 .2 457 .5 448 .1 489 .5
Day 80 475 .0 565 .5 464 .7 472 .5 504 .8
H-24019 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
Day 3
GROUP III
TE ST 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
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 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
.45-
"ompany Sanitized. Doe
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
646921 646922 646923 646924 646925
Day 1 204 .5 178 .3 185 .9 188 .7 172 .2
Day 2
204 9 180. 9 185. 1 194. 6 176. 0
H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
Day 3
TES T DAY
Day 4
Day 5
Day 6
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
ANIMAL NUMBER
646921 646922 646923 646924 646925
Day 10
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
Day 7 244 .3 202 .4 219 .1 236 .2 215 .1
-46-
"ompany Sanitized. D
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL NUMBER
646926 646927 646928 646929 646930
Day 1 182 .0 172 .2 190 .4 178 .6 187 .9
ANIMAL NUMBER
646926 646927 646928 646929 646930
Day 10 250 .8 243 .3 254 .1 225 .5 240 .3
ANIMAL NUMBER
646926 646927 646928 646929 646930
Day 73
535 .3 551 .2 540 .3 502 .6 498 .5
Day 2 187 .8 178 .8 195 .3 182 .5 191 .6
Day 17 297 .2 306 .2 310 .6 280 .9 292 .3
Day 80 551 .9 574 .5 557 .0 509 .6 517 .6
H-24020 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (9) OF MALE RATS
Day 3
GROUP III
TE ST DAY
Day 4
Day 5
Day 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
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
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
-47-
Company Sanitized. Do
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24750 (TEST SUBSTANCE)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
ANIMAL
T EST DAY
NUMBER
Day 1
Day 2
Day 3
Day 4
. Day 5
Day 6
Day 7
D
665547
194 .5
200.9
210 .4
225 .1
238 .6
243 .3
252 .1
2
665548
191 .6
195.8
202 .5
211 .7
225 .5
231 .3
238 .6
2
665549
190 .1
197.4
201 .0
218 .3
230 .0
233 .2
245 .4
2
665550
193 .0
196.6
204 .9
218 .8
230 .0
241 .4
248 .4
2
665553
193 .4
199.0
209 .0
216 .4
228 .4
235 .1
247 .3
2
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
.48-
Company Sanitized. Does
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24750 (TEST SUBSTANCE)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP III,
ANIMAL
TEI3SST DAY
NUMBER
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
D
665554
175 .0
180 .0
187 .8
195.2
204 .6
210 .8
212 .5
2
665555
184 .9
194 .4
201 .6
208.7
217 .6
230 .9
240 .0
2
665558
187 .8
192 .0
201 .9
211.1
221 .9
226 .5
229 .0
2
665559
189 .2
196 .6
204 .2
213.4
222 .9
230 .8
240 .7
2
665560
192 .9
200 .2
206 .5
214.1
225 .8
234 .7
238 .0
2
ANIMAL NUMBER
Day 10
Day 17
Day 24
TES3T DAY
Day 31
Day 38
Day 45
Day 52
665554 665555 665558 665559 665560
209 .4 254 .7 229 .5 239 .5 246 .4
ANIMAL NUMBER
Day 73
212 .0 247 .7 229 .7 243 .9 253 .5 Day 80
220 .3 255 .3 224 .6 257 .8 281 .7
Day 87
232 .1 275 .0 229 .8 271 .8 317 .3
234 .0 281 .7 227 .1 305 .8 348 .7
Day 94 TEST DAY
249 .4 316 .0 235 .4 336 .1 389 .1
270 .5 340 .3
259 .9 370 .2 408 .9
665554 665555 665558 665559 665560
330 .1 387 .9 345 .4 . 418 .3 470 .9
355 .1 412 .1 366 .1 441 .6 490 .5
368 .3 425 .9 373 .1 451 .0 499 .3
380 .2 431 .8 381 .2 467 .1 512 .5
3D test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94
-49-
Company Sanitized. Does no
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX B Individual Clinical Observations
.50-
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
Animal Observation 646932 General observation, No Abnormality Detected
Eye Observations, Exophthalmus, Left Eye Observations, Bled via Orbital for Clin Path, Left
Discharge, Eye left. Black
Sacrificed by design 646933 General observation. No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Dark, Left
Discharge, Eye left. Black
Swollen Observations, Mouth Sacrificed by design 646934 General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left .
Discharge, Eye 3-eft, Black Sacrificed by design
Days
1
2-9 1,5
10 10
1-5 1,5 6-10 6-7
10 10
1-5,10 1,5 6-9
10
.51 -
"ompany Sanitized. Does no
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 646935
646936
Observation General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left
Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design
Days
1-10 1,5
10
1-10 1,5
10
52-
^ompany Sanitized. Do
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal 646937 646938
646939
Observation
GROUP III
General observation. No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left
Eye Observations, Bled via Orbital for Clin Path, Ri9ht
Sacrificed by design
General observation. No Abnormality Detected
Eye Observations, Enophthalmus, Right
Eye Observations, Exophthalmus, Right
Eye Observations, Bled via Orbital for Clin Path, Bilateral
Eye Observations, Bled via Orbital for Clin Path, Right
Eye Observations, Corneal Opacity, Right
Discharge, Eye right. Black
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
Days
1-10,17-94
13
24,52
94
1-10,17-45 80-94 52-73 13,52
24 80-94 59 94
1-10,17-94
13
24,52
94
.53-
Company SanWzed. DOGS
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal 646940
646941
Observation
GROUP III (Continued)
General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left
Eye Observations, Bled via Orbital for Clin Path, Right
Discharge, Nose, Black
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
Days
1-10,17-80,94
13
24,52
87 94
1-10,17-94
52 13 24 94
.54-
Company Sanitized. Does
-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 646910 646911
646912
646913 646914
Observation General observation. No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, 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, Bled via Orbital for Clin Path, Left Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design
Days
1-10 1,5
10
1,10 1,5 2-9
10
1-10
1
5
10
1-10 1,5
10
1-10 1,5
10
.55-
Company Sanitized. Do
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
Dischar9e, 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, Bled via Orbital for Clin Path, Left
Eye Observations, Bled via 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
.56-
lompany Sanitized. Does
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 646918
646919
H-24019 (POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP III (Continued)
Days
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
General observation. No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right
13,24
52
Sacrificed by design
94
-57-
'ompany SanHlzed. Does n
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I
Animal 646921
646922
646923
Observation
General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left
Eye Observations, 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
Days
1,5-10 1,5 2-4
10 1-10
1 5
10
1,7-10 4-6
1 5
2-6
10
.58-
;cmpany Sanitized. Does
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 646924
646925
H-24020 (POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I (Continued)
Observation
Days
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
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
.59-
"ompany Sanitized. Does
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 646926
646927
646928
H-24020(POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP III.
Days
General observation. No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left
52
13,24
Sacrificed by design
94
General observation. No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left
52
13.24
Sacrificed by design
,
94
General observation. No Abnormality Detected
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Left
13,24,52
Sacrificed by design
94
.60-
Company Sanitized. Does not
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 646929
646930
H-24020(POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP III (Continued)
Days
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
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
-61-
Cntttpany Sanitized. Does n
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24750 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I
Animal 665547 665548 665549 665550 665553
Observation General observation. No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design General observation. No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design General observation. No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by.design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design
Days
1-109 1,5
10
1-10 1,5
10
1-10 1,5
10
1-10 1,5
10
1-10 1,5
10
.62-
Company Sanitized. Does n
H-24750: Biopersistence Screening 10-DoseOralGavage Study in Rats
Animal 665554 665555
665558
665559 665560
H-24750 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP III
Days
General observation. No Abnormality Detected
1-94
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
Hair Loss, Face, Right
31-45
Comments
17-94
General observation. No Abnormality Detected
1-17,87-94
Eye Observations, Bled via Orbital for Clin Path, Right Hair Loss, Forelimb, Bilateral Hair Loss, Forepaw, Bilateral General observation, No Abnormality Detected
24,52 24-80 24-66 1-94
Eye Observations, Bled via Orbital for Clin Path, Right
General observation. No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right
Hair Loss, Shoulder, Bilateral
Hair Loss, Shoulder, Left
24,52 1-4,17-94 24,52 9-10 5 -company Sanitized.
-63-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-11561
APPENDIX C
Terms and Calculations Factors Influencing Interpretation of Kinetic Analysis
-64-
fompany Sanitized. Does not contain TSCA CB1
f
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 of fluorine containing
components
The molecular weight of the fluorine containing components (g/mole) The mg of formulation given per kg of animal body weight The % fluorine in the fluorine containing components of the formulation (weight basis) The molecular weight of fluorine g/mol
Compound Calculations:
Dose Active (mg/kg)
The mg of 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 of animal body weight = (% F in active/I 00) 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 of 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 of Active [mmol/kg] / Dose Active [mmol/kg])
x Formulation Dose
Company Sanitized. Does no{ contain TSCA CBI
-65-
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppmF
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.
ppmF
normalized to 0.1 mmol/kgDose
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 of active were used in the
study.
= (0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in blood minus background)
amolar equivalents of
active
The pmolar [umol/L] concentration of 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 CB1 -66-
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 offluorinated 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 be justified 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 of the active component in the formulation
Company Sanitized. Does not contain TSCA CB1 -67-
APPENDIX D Individual Fluorine Levels in Blood
Company Sanitized. Does nc';.con?a?n TSCA CBI
-68-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-11561
Given:
Mol Wt. Active (g/mole):
497
Formulation Dose (mg/kg):
10
% Active (F Containing) in Formulation: 100
% F in Active:
65
Mol Wt. F (g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
10
0.020 0.342
Molar Ratio (Active7F): Dose F (mg/kg):
0.059 6.5
Rat Number
Test Day Sample
ppm I r in
Blood
Group I
646910
1
2.6
646911
1
2.7
646912
1
2.5
646913
1
2.6
646914
1
2.6
ppm F in Blood
Minus Bkg 0.2 ppm
2.4 2.5 2.3 2.4 2.4
Group I
646910
5
646911
5
646912
5
646913
5
646914
5
30.1 30.3 31.3 32.2 32.7
29.9 30.1 31.1 32.0 32.5
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
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
11.93 12.43 11.43 11.93 11.93
148.60 149.60 154.57 159.04 161.53
354.36 349.39 331.50 309.63 339.95
umolar Equivalents of Active in
Blood
36.92 38.46 35.38 36.92 36.92
460.00 463.08 478.46 492.31 500.00
1096.92 1081.54 1026.15 958.46 1052.31
Company Sanitized. Does nc- contam TSCA CBI
-69-
,?'
H-24750: Biopersistence Screening 10-DoseOral Gavage Study in Rats
Rat Number
Test Day Sample
ppmF
in
Blood
Group m
646915
13
646916
13
646917
13
646918
13
646919
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
Group m
646915
24
646916
24
646917
24
646918
24
646919
24
Group III
646915
52
646916
52
646917
52
646918
52
646919
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
Group III
646915
94
13.3
13.1
646916
94
11.7
11.5
646917
94
12.9
12.7
646918
94
11.0
10.8
646919
94
14.1
13.9
ppm F in B lood Normalize)[lt0
0.1 mmoles,/kg Dose
272.36 273.85 258.44 266.39 265.40
192,34 216.69 185.88 174.94 209.73
114.31 99.90 127.73 122.76 115.80
65.11 57.16 63.12 53.68 69.08
DuPont-11561
(imolar Equivalents of Active in
Blood
843.08 847.69 800.00 824.62 821.54
595.38 670.77 575.38 541.54 649.23
353.85 309.23 395.38 380.00 358.46
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-2-:t020
DuPont-11561
Given:
MolW t. Active (g/mole):
426
Formulation Dose (mg/kg):
20
% ActrVG (F Conta.ining) in I 'ormulation: 100
% F in Active:
69
Mol Wt. F (g/mol):
19
Calculated1Values: Dose A ctive (mg/kg): Dose A ctive (mmcle/kg): DoseF (mmol/kg);
20 0.04 7 0.72 6
Molar Ratio (Active/F): Dose F (mg/teg):
0.065 13.8
Rat Number
Group I
646921 646922 646923 646924 646925
Test Day Sample
ppmF
in Blood
1
10.6
1
10.7
1
9.2
1
5.3
1
11.2
ppm F in Blood
Minus Bkg 0.2 ppm
10.4 10.5 9.0 5.1 11.0
Group I
646921
5
78.7
646922
5
83.2
646923
5
77.0
646924
5
66.1
646925
5
69.6
78.5 83.0 76.8 65.9 69.4
Group I
646921
10
63.0
62.8
646922
10
69.1
68.9
646923
10
60.3
60.1
646924
10
62.5
62.3
646925
10
53.9
53.7
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
umolar Equivalents of Active in
Blood
22.15 22.37 19.17 10.86 23.43
75.36 76.09 65.22 36.96 79.71
167.21 176.79 163.58 140.37 147.82
133.76 146.76 128.01 132.70 114.38
568.84 601.45 556.52 477.54 502.90
-
455.07 499.28 435.51 451.45 389.13
Comoany Sanitized. Docs not contain TSCA CBi
-71-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test Day Sample
ppmF
in
Blood
Group in
646926
13
646927
13
646928
13
646929
13
646930
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
Group III
646926
24
11.9
11.7
646927
24
10.4
10.2
646928
24
8.1
7.9
646929
24
15.7
15.5
646930
24
9.8
9.6
Group HI
646926
52
2.5
646927
52
1.7
646928
52
1.3
646929
52
4.0
646930
52
1.8
2.3
1.5
1.1
3.8 1.6
Group in
646926
94
0.8
0.6
646927
94
0.5
*
646928
94
0.5
*
646929
94
0.9
0.7
646930
94
0.5
*
Below LOQ (Limit of Quantification)
ppm F in I ilood Normalizedto 0.1 mmoleis/kg
Dose
DuPont-11561
umolar Equivalents of Active in
Blood
73.70 63.47 50.06 71.36 53.68
250.72 215.94 170.29 242.75 182.61
24.92 21.73 16.83 33.02 20.45
84.78 73.91 57.25 112.32 69.57
4.90 3.20 2.34 8.09 3.41
16.67 10.87 7.97 27.54 11.59
1.28
* *
1.49
*
-
4.35
*
*
5.07
*
Company Sanitizad. Does ."of contain TSCA CB! 72-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-247;50
DuPont-11561
Given:
MoPtVt. Active (g/imole):
514
Formulation Dose (nig/kg):
5
%Active (F Containing) in Fo'rmulation: 100
% F in Active: Mol Wt. F (g/mc)!):
70.2 19
Calculateed Values: Dose Active (mg/lq0: Dose Active (mmole/kg): Dose F (mmol/kg):
5
0.010 0.185
Molar Ratio (Acttive/F): Dose F (mg/kg):
0.053 3.5
Rat Number
Test Day Sample
ppmF
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
pmolar 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
10
30.45
665548
10
28.9
665549
10
30.09
665550
10
30.44
665553
10
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-
f
H-24750: Biopersistence Screening 10-DoseOral Gavage Study in Rats
Rat Number
Group m 665554 665555 665558 665559 665560
Test Day Sample
ppmF
in Blood
13
46.92
13
31.35
13
32.95
13
30.2
13
30.56
ppm F in Blood
Minus Bkg 0.2 ppm
46.7 31.2 32.8 30.0 30.4
ppm F in B lood Normalize'dto 0.1 mmole;s/kg
Dose
480.36 320.27 336.72 308.45 312.15
DuPont-11561
pmolar Equivalents of Active in
Blood
1331.05 887.46 933.05 854.70 864.96
Group III
665554
24
665555
24
665558
24
665559
24
665560
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
52
11.09
10.9
665555
52
7.64
7.4
665558
52
12.96
12.8
665559
52
8.88
8.7
665560
52
5.91
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
94
6.3
6.1
665555
94
6.28
6.1
665558
94
6.13
5.9
665559
94
6.9
6.7
665560
94
5.6
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 nol contain TSCA CB1
-74-
APPENDIX E Individual Fluorine Levels in Liver
C^mi'"1" SanWmd. H-w^ not r;Anqtr> TRf*A riRI -75-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-240 19
DuPont-11561
Given:
Mol Wt. Active (g/mole):
497
Formulation Dose (mg/kg):
10
% Active (F Containing) in Fcrmulation: 100
% F in Actr/e:
65
Mol Wt. F (,g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
10 0.020 0.342
Molar Ratio (Active/F): Dose F (mg/kg):
0.059 6.5
Rat Number
Test Day Sample
ppm F in
Liver
Group I 646910 646911 646912 646913 646914
10
320.9
10
334.0
10
318.6
10
281.6
10
306.6
ppm F in Liver
Minus Bkg 0.2 ppm
320.7 333.8 318.4 281.4 306.4
Group III
646915
94
646916
94
646917
94
646918
94
646919
94
85.6 75.9 87.5 78.0 94.2
85.4 75.7 87.3 77.8 94.0
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
1593.88 1658.99 1582.45 1398.56 1522.81
424.44 376.23 433.88 386.67 467.18
Umolar Equivalents of Active in
Liver
4933.85 5135.38 4898.46 4329.23 4713.85
1313.85 1164.62 1343.08 1196.92 1446.15
Gomoany SanKFzed. Does not conla.-n T8CA CB?
-76-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-240:20
DuPont-11561
Given:
MoPNt. Active (g/imole):
426
Formulation Dose (mg/kg):
20
%Ac tive (F Containing) in Fcrmulation: 100
% F in Active:
69
Mol Wt. F (g/mc)!):
19
Calculat<id Values:
Dose Active (mg/k{?): Dose Active (mmol e/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Addve/F):
Dose F (mg/kg):
0.065 13.8
Rat Number
Test Day Sample
ppmF
in 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
Group II I
646926
94
1.8
1.6
646927
94
1.2
1.0
646928
94
2.2
2.0
646929
94
4.7
4.5
646930
94
2.9
2.7
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
251.55 260.93 243.03 258.58 259.65
3.41 2.13 4.26 9.59 5.75
umolar Equivalents of Active in
Liver
855.80 887.68 826.81 879.71 883.33
11.59 7.25 14.49 32.61 19.57
-77-
^nin-mv t;anW,o,( rnoe ,,,,{ Mntam TttC'A fW
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24750
DuPont-11561
Given:
Mol Wt. Active (g/mole):
514
Formulation Dose (mg/kg):
5
% Active (F Containing) in Formulation: 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): Dose F (mg/kg):
0.053 3.5
Rat Number
Test Day Sample
ppmF
in 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
94
665555
94
665558
94
665559
94
665560
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
-^panySan,,,zed.Doe3nofcon,arnTSCACBI
-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-240 19
DuPont-11561
Given:
Mol Wt. Active (g/:mole):
497
Formulation Dose (mg/kg):
10
% Active (F Contairung) in Fc)rmulation: 100
% F in Actih/e:
65
Mol Wt. P (,g/mol ):
19
Calculated Values: Dose Active (mg/k{?): Dose Active (mmol e/kg): Dose F (mmol/kg):
10 0.020 0.342
Molar Ratio (Active/F): Dose F (mg/kg):
0.059 6.5
Rat Number
Group I 646910 646911 646912 646913 646914
Test Day Sample
ppmF
in Fat
10
14.4
10
14.9
10
10.9
10
11.6
10
12.4
ppm F in Fat
Minus Bkg 0.2 ppm
14.2 14.7 10.7 11.4 12.2
Group III
646915
94
1.3
1.1
646916
94
1.1
0.9
646917
94
1.3
1.1
646918
94
0.8
0.6
646919
94
1.5
1.3
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
70.57 73.06 53.18 56.66 60.63
5.47 4.47 5.47 2.98 6.46
umolai Equivale nts of Active in
Fat
218.46 226.15 164.62 175.38 187.69
16.92 13.85 16.92 9.23 20.00
-80- ^^,wn^,^,_,^
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-240:10
DuPont-11561
Given:
Mol Wt. Active (g/mole):
426
Formulation Dose (mg/kg):
20
% Active (F Containing) in Fcrmulation: 100
% F ir.lActr/e:
69
MolV/t. F (,g/mc>1):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Active/F): Dose F (mg/kg):
0.065 13.8
Rat Number
Group I 646921 646922 646923 646924 646925
Test Day Sample
ppm F in
Fat
10
6.2
10
8.5
10
8.7
10
8.8
10
8.3
ppm F in Fat
Minus Bkg 0.2 ppm
6.0 8.3 8.5 8.6
8.1
Group III
646926
94
ND
ND
646927
94
ND
ND
646928
94
ND
ND
646929
94
ND
ND
646930
94
ND
ND
ND Non-detectable.
ppm F in Fat Normalizfedto 0.1 mmol(is/kg
Dose
12.78 17.68 18.11 18.32 17.25
(iroolar Equivalents of Active in
Fat
43.48 60.14 61.59 62.32 58.70
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
company Sanitized. Does no? contain TSCA CBI 81-
H-24750: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24750
DuPont-11561
Given:
Mol Wt. Active (g/mole):
514
Formulation Dose (mg/kg):
5
% Active (F Containing) in Formulation: 100
%Fin Active: MolW't. P (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): 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
94
<5
665555
94
<5
665558
94
<5
665559
94
<5
665560
94
<5
ppm F in Fat
Minus Bkg 0.2 ppm
4.0 3.5 2.6 4.0 3.3
* * * * *
Below LOQ (Limit of Quantification)
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
41.23 36.09 26.63 41.54 33.42
* * * * *
umolar Equivalents of Active in
Fat
114.25 100.00 73.79 115.10 92.59
* * * * *
-82-
Company Sanitized-"Dooee<s?nno^{ cont. ain TSCACB1