Document RJRmma90Xb3BKM1qxgV3xN9wn
AR226-3193
TRADE SECRET
Study Title H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID: DuPont-6542
AUTHOR: Carol Finlay, B.A. STUDY COMPLETED ON: November 26, 2001
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:BBBB^
SERVICE CODE NUMBER:BHB
Page 1 of 83 iPftnmanu Cfanitf-arl n<^oi nft />rk---?ain TC?'A t^tti
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats '
DuPont-6542
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Reviewed by::
^ 0 - ^ M ^ i ^^'77T^Tau-l H;iTnd~erl.itTer,,P,h.TD->.
- Postdoctoral Fellow
2/-/^- Z^l Date
Reviewed by:
D.A.B.T.Date C^idcd C //Judith C. Stadler,MPh^.DL., dU^______
v
Director
^-^J-ZOOf
Issued by Study Director:
C.aJ[^C^arol ^JinOlayb,B[.aA.^^,________
Staff Scientist
1^ -iOnO.^O /
Date
-2-
Company Sanitized. Doss not cor.iai.-i TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542 i
TABLE OF CONTENTS
Page
CERTIFICATION.......................................................................................................................^
LIST OF TABLES........................................................................................................................4 LIST OF FIGURES........ LIST OF APPENDICES.
STUDY INFORMATION............................................................................................................^ STUDY PERSONNEL..................................................................................................................8
SUMMARY...................................................................................................................................^ INTRODUCTION....................................................................................................................... 11
MATERIALS AND METHODS............................................................................................... 11
A. Test Substance and Positive Controls............................................................................... 11 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. Body Weights.................................................................................................................... 14 I. Mortality and Clinical Observations................................................................................. 14 J. Collection and Analysis of Blood, Livers, and Fat........................................................... 14 K. Treatment of Fluorine Data............................................................................................... 15 L. Statistical Methods............................................................................................................ 15
RESULTS AND DISCUSSION................................................................................................. 16
A. In-Life Toxicology............................................................................................................ 16 B. Liver Weights.................................................................................................................... 16 C. Fluorine Data..................................................................................................................... 17 CONCLUSIONS......................................................................................................................... 18
RECORDS AND SAMPLE STORAGE................................................................................... 19
TABLES....................................................................................................................................... 20 FIGURES..................................................................................................................................... 27
APPENDICES.............................................................................................................................38
-3-
Ccmpany Sanitized. Does not con?2in TSCA CBS
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
LIST OF TABLES
Page
1. MEAN BODY WEIGHTS...................................................................................................................................21 2. MEAN BODY WEIGHT GAINS .......................................................................................................................22 3. MEAN BODY AND LIVER WEIGHTS .............................................................................................................23 4. MEAN BLOOD FLUORINE LEVELS................................................................................................................24 5. MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE................................................24 6. MEAN LIVERFLUORINE LEVELS..................................................................................................................25 7. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................25 8. MEAN FAT FLUORINE LEVELS......................................................................................................................26 9. MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE......................................................26
LIST OF FIGURES
Page
1. MEAN BODY WEIGHTS...................................................................................................................................28 2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE
CONTROL...........................................................................................................................................................29
3. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE
CONTROLS.........................................................................................................................................................30
4. MICROMOLAR EQUIVALENTS IN RAT BLOOD..........................................................................................31 5. NORMALIZED H-24944 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A
10-DAY ORAL GAVAGE...................................................................................................................................33 6. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................34 7. COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE
CONCENTRATION FOR H-24944 AND NEGATIVE CONTROL ..................................................................35 8. MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE......................................................36 9. COMPARISON OF MEAN BLOOD, MEAN LIVER, AND MEAN FAT FLUORINE
CONCENTRATION NORMALIZED TO DOSE................................................................................................37
-4-
Ccmpany Sanitised. DGSP roS contain TSCA CBF
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in
Rats_________________________[__________DuPont-6542
LIST OF APPENDICES
Page
A.
INDIVIDUAL
BODY
WEIGHTS
39 .......................................................................................................................
B. INDIVIDUAL CLINICAL OBSERVATIONS...................................................................................................^
C. TERMS AND CALCULATIONS FACTORS INFLUENCING INTERPRETATION OF KINETIC
ANALYSIS...........................................................................................................................................................65
D. INDIVIDUAL FLUORINE LEVELS IN BLOOD ..............................................................................................69 E. INDIVIDUAL FLUORINE LEVELS IN LIVER ................................................................................................76 F. INDIVIDUAL FLUORINE LEVELS IN FAT.....................................................................................................80
-5-
Company Sanitized. Doss not contain TSCA CBI
TEST SUBSTANCE:
STUDY INFORMATION
Substance Tested:
Synonyms/Codes
Haskell Number: 24944 Composition:
Known Impurities:
POSITIVE CONTROL:
Substance Tested: Potassium perfluoroalkyi sulfonate Synonyms/Codes:
Haskell Number: 24019 Composition
Known Impurities:
-6-
jCompany
SasiSiszea.
,,
uo-is
>.-.
----_*---:-
rc"r-'A"
- -fv.1.
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
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 Composition
Known Impurities
Sponsor:' E. I. du Font de Nemours and Company Wilmington, Delaware 19898 U.S.A.
Study Initiated/Completed: May 22, 2001 / (see report cover page) In-Life Initiated/Completed: May 22, 2001 / August 23, 2001
-7-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY PERSONNEL
Study Director: Management:
Primary Technician:
Carol Finlay, B.A.
Judith C. Stadler, Ph.D., D.A.B.T. James C. Mackay n
Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T.
Toxicology Report Preparation: Wanda F. Dinbokowitz
Laboratory Veterinarian: William Singleton, D.V.M., A.C.L.A.M
DuPont-6542
-8Company Sanitized. Doe'- nci conSa'n T^A r.R?
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
SUMMARY
The objective of this study was to evaluate the potential for H-24944, when administered by gavage, to be absorbed and to accumulate in a mammalian system. Two groups of 5 male Crl:CD(SD)IGS BR rats each were exposed to 1000 mg/kg/day of H-24944 for 10 consecutive days. Blood was collected from the orbital sinus of 5 rats (group I) approximately 2 hours after dosing on test days 1 and 5. Approximately 2 hours after the last dose, these rats were euthanized and blood, livers, and fat were collected. Blood was collected from the orbital sinus of the remaining 5 rats (group US) on test days 13, 24, and 52. These rats were euthanized on test day 94 and blood, livers, and fat were collected. Body weights and clinical signs were recorded on each day of dosing and then weekly during the recovery period. Additionally, a negative control, 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-24944.
No deaths occurred. No test substance-related clinical signs were observed.
Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive controls or negative control. This difference resulted in differences in mean body weights on test day 1. Accounting for the age difference at study start and the expected rate of body weight gain, the mean body weights and mean body weight gains of the rats dosed with H-24944 were comparable to the negative control and equal to or greater than the positive controls.
The mean relative liver weights (liver/body weight) of rats dosed with the test substance, H-24944, were similar to the weights of the negative control rats on day 10 and day 94. The mean relative liver weight of rats dosed with the positive control H-24019 was 38% higher at day 10 than the negative control group. The mean relative liver weight of rats dosed with the positive control H-24020 was 88% 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.
/-'-'
A steady-state for fluorine levels in whole blood was not achieved during 10 consecutive days of dosing with 1000 mg/kg H-24944. An area under the curve (estimated to infinity) was calculated and normalized for fluorine content for the test substance and each positive control. The
AUCINF/D for the fluorine component of the test substance, H-24944, was 6.64xl02 compared to AUCINF/D values of 5.22xl05 and 8.15xl04 for H-24019 and H-24020, respectively.
The concentration of fluorine in the livers from rats dosed with the test substance, H-24944, was
23.22 /AM equivalents on day 10 and 3.42 ^iM equivalents on day 94. On day 10, mean pM
equivalent concentrations of fluorine in the livers from rats dosed with the positive control materials were approximately 210-fold (H-24019) and 40-fold (H-24020) greater than the fluorine concentration in livers from rats treated with the test substance. By day 94, the concentrations were approximately 380x and 5x the fluorine concentration in rats treated with H-24944.
-9-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
The fluorine concentration in the fat from rats dosed with the test substance was 7.79 fi.M
equivalents on day 10. On day 94, the concentration was 1.50 ^M equivalents. The fluorine
concentrations of the positive controls were approximately 25x or 7x higher than H-24944 on day 10. By day 94, the /zM equivalent concentration of fluorine in the fat from rats dosed with the positive control H-24019 was approximately lOx higher than H-24944. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020.
The pM equivalents of fluorine in the liver and fat of animals dosed with the test substance were
higher than levels in the blood.
Under the conditions of this study, there was limited absorption and no retention of fluorine in the blood in rats dosed with H-24944. Administration of the test substance to male rats for 10 consecutive days resulted in some absorption and retention of fluorine in the liver; however, there was no evidence of increased relative liver weight, either at the end of dosing or following the recovery period. Fluorine levels in blood and liver were lower than levels in rats dosed with the positive control materials, H-24019 and H-24020. Some retention of fluorine in the fat was evident. Fluorine levels in the fat from rats dosed with the test substance were lower than the levels from rats dosed with the positive control H-24019 and were lower on day 10 than the levels in fat from rats dosed with the positive control H-24020, but higher on day 94.
- 10-
Company Sanitized. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
INTRODUCTION
The objective of this study was to define the potential of H-24944 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 1000 mg/kg for the test substance was selected based on available toxicity data. The dosage of 1000 mg/kg was selected for the limit dosage for this project. The limit dosage of 1000 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-24944, was supplied by the sponsor as a light brown liquid. 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)IGS BR rat was selected on the basis of extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, 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%.
2.
Feed and Water
Tap water was provided ad libitum. All rats were fed PMI Nutrition International, Inc. Certified Rodent LabDiet 5002 chow. The feed is guaranteed by the manufacturer to meet specified nutritional requirements and to be free of specified contaminants.
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in
Rats___________________________________DuPont-6542
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.
Animal 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 of 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 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 designee of the laboratory animal
veterinarian.
t)
- 12-
Company Sanitized. Does not contain TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
E.
Study Design
Substance
Negative Control Deionized water
Positive Controls H-24019
H-24020 Test Substance
H-24944
Vehicle
Not applicable Acetone/Corn Oil Acetone/Corn Oil
Not applicable
Dosage (mg/kg)
0
10 20
1000
Number of Animals
10
10 10
10
F.
Assignment to Groups and Study Start
After the quarantine period, the rats were selected on the bases of adequate body weight gain and 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 or 8 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-24944 was dosed as received. The amount of test substance each rat received was based on the body weight collected on each day of dosing and the test substance density of 1.21 g/mL. The test substance 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 of 1 mL/100 g of body weight. The dosing preparations were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity.
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.
IP
Company Sanitized. Does not contain TSCA CB?
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
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 only 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 while shipped.
The total fluorine content of the samples was determined by 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 aqueous absorbing solution and analyzed with a fluoride ion selective electrode. Kinetic analysis of the data reviewed (ppm F in each sample) was performed by Haskell Laboratory personnel for evaluation of fluorine biopersistence.
Company Sanitized. Does c; contain TSCf. -"H]
H-24944: Biopersistence Screening
10-Dose Oral Gavage Study in
DuPont-6542
Rats____________________________________
K. Treatment of Fluorine Data
Since the test substance and positive control materials 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 dosenormalization was conducted in ^imolar 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. In 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 ^molar (fiM) equivalents of active component in the tissue. Detailed calculations can be found in Appendix C. The /iM 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-24944 and the positive controls using WinNonlin Version 3.1 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 /iM
equivalent in the data.
The maximum observed concentration in blood was Cmax (/AM 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. AUCENF normalized to dose (AUCINF/D) can be used to compare the relative exposures of different
compounds and dosages.
L.
Statistical Methods
Descriptive statistics (e.g. mean, standard deviation) were used.
-15-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
RESULTS AND DISCUSSION
A.
In-Life Toxicology
(Tables 1-2, Figure 1, Appendices A-B)
No deaths occurred. No test substance-related clinical signs were observed in rats dosed with the test substance, H-24944. A swollen mouth was observed during the dosing period in a negative control rat. One negative control rat exhibited black nasal discharge during the recovery period. Hair loss was observed in rats dosed with H-24019 and H-24020, and a wound was observed in a rat dosed with H-24020. Ocular discharge, dark eyes, comeal opacity, enophthalmus, and exophthalmus observed in several rats are considered to be a result of orbital sinus bleeding.
Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive controls or negative control. This difference resulted in differences in mean body weights on test day 1. The rats dosed with the test substance, H-24944, were older and heavier in weight than the positive and negative control rats. Accounting for the age difference at study start and the expected rate of body weight gain, the mean body weights and mean body weight gains of the rats dosed with H-24944 were comparable to the negative control and equal to or greater than the
positive controls.
B.
Liver Weights
(Table 3, Figures 2, 3, and 7)
1.
Test Substance
The mean relative liver weights (liver/body weight) of rats dosed with the test substance, H-24944, were similar to the weights of the negative control group on day 10 and day 94.
2.
Positive Controls
The mean relative liver weight of rats dosed with one of the positive controls, H-24019, was 35% higher at day 10 than the liver weight of rats dosed with the test substance, H-24944. By day 94, the weights were similar.
The mean relative liver weight of rats dosed with the other positive control, H-24020, was 84% higher at day 10 than the liver weight of rats dosed with H-24944. By day 94, the weights were
similar.
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
-16-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in
Rats___________________________________DuPont-6542 H-24020 was 88% higher on day 10 than the negative control group. By day 94, the weights
were similar.
Therefore, liver weights of rats dosed with the test substance, H-24944, were not affected.
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 dosage 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 [JLM
equivalents in rat blood continued to rise throughout the dosing period and may not have reached
steady-state (Figures 4A and 4B). The Cmax for H-24019 was 541.45 50.37 ^M equivalents
(Mean SD) with a terminal half-life of 42.2 days. The Cmax for H-24020 was
R
1043.08 54.57 /xM equivalents (Mean SD) with a terminal half-life of 15.1 days. For each of
the positive controls, 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 positive controls and the test substance. The AUCINF/D for the
fluorine component was 5.22xl05 for H-24019 and 8.15xl04 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 pM equivalents for H-24019 and H-24020, respectively. By
day 94, the concentrations were 1292.92 and 17.10 /xM 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 fJiM 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 fiM equivalents. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020.
3.
Test Substance
The H-24944 normalized ^M equivalents in rat blood rose rapidly and did not reach steady-state (Figure 4C). The Cmax for H-24944 was 4.65 0.76 juM equivalents (Mean SD) with a terminal
half-life of 5.1 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
- 17-
Company Sanitized. Does nol c'-mlam TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
taken into account when using the derived terminal half-life for comparative purposes. Since the day 52 and 94 blood values were less than the limit of quantification, only three points were available for the terminal half-life calculation. The total internal exposure resulting from a
normalized dose was described by AUCINF/D and was the basis for comparison between H-24944 and the positive controls. The AUCINP/D for the fluorine component of H-24944 was 6.64 xlO2 as compared to AUCINP/D values of 5.22xl05 and 8.15xl04 for H-24019 and
H-24020, respectively.
Levels of total fluorine in livers from rats dosed with the test substance, H-24944, were lower 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-24944 was 23.22 {iM equivalents at day 10 and
3.42 fiM equivalents at day 94. For the positive control H-24019, the liver concentrations were
approximately 210x higher (day 10) and approximately 380x higher (end of study) than H-24944. For H-24020, the liver concentrations were approximately 40x higher (end of dosing) and 5x higher (end of study) than H-24944.
Levels of total fluorine in fat from rats dosed with the test substance were generally lower than the levels in fat from rats dosed with the positive control materials. The fluorine concentration in the fat from rats dosed with the test substance was 7.79 on day 10 and 1.50 /xM equivalents on day 94. The fluorine concentration in the fat from rats dosed with the positive controls were
approximately 25x or 7x higher than H-24944 on day 10. By day 94, the ^M equivalent concentration of fluorine in the fat from rats dosed with the positive control H-24019 was
approximately lOx higher than H-24944. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020.
The pM equivalents of fluorine in the liver and fat of animals dosed with the test substance were
higher than levels in the blood.
CONCLUSIONS
Rats dosed for 10 consecutive days with 1000 mg/kg H-24944 exhibited no mortality, body weight effects, liver weight effects, or test substance-related clinical signs. A steady-state for fluorine in the blood was not achieved during the 10-day dosing period with the test substance. Dose-adjusted areas under the curve (AUCDMF/D) for positive controls, H-24019 and H-24020, were approximately 785x and 125x the AUCINF/D for the test substance.
Under the conditions of this study, there was limited absorption and no retention of fluorine in the blood following dosing with H-24944. Administration of the test substance, H-24944, to male rats for 10 consecutive days resulted in some absorption and retention of fluorine in the liver; however, there was no evidence of increased relative liver weight, either at the end of dosing or following the recovery period. Fluorine levels in blood and liver in rats dosed with H-24944 were lower than the levels in rats dosed with the positive control materials, H-24019 and H-24020. Some retention of fluorine in the fat was evident. Fluorine levels in the fat from rats dosed with the test substance were lower than the levels in fat from rats dosed with the
- 18-
Company Sanitized. Does not contain TS<-A ci3(
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
positive control H-24019 and were lower on day 10 than the levels in fat from rats dosed with the positive control H-24020, but higher on day 94.
RECORDS AND SAMPLE STORAGE
All original records will be retained at Haskell Laboratory, E. I. du Pont de Nemours and Company, Newark, Delaware or at Iron Mountain Records Management, 200 Todds Lane, Wilmington, Delaware.
-19-
Company Sanitized. Does not contain TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
TABLES
-20-
Company Sanitized. Does not contain TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TABLE 1 MEAN BODY WEIGHTS (g)
Test Days 1 2 3 4 5 6 7 8 9 10 17
24 31 38 45 52 59 66 67 73 80 87 94
Negative Control Deionized Water
189.6 195.5 204.6 211.7 222.1 225.0 235.2 242.3 248.3 257.0 293.3 325.5 359.3 380.9 407.4 420.5 436.4
-
452.8 472.6 489.6 497.7 511.7
Positive Controls
H-24019
H-24020
184.4
184.1
189.2
187.8
199.5
197.8
206.4
204.8
216.2
212.5
222.0
216.0
229.4
223.6
233.7
226.9
240.4
234.2
246.5
243.0
290.1
297.4
313.3
338.6
348.4
381.9
370.8
404.9
403.2
434.4
422.8
460.6
439.6
483.1
-
-
455.6 480.1 496.5 512.2 524.6
502.7 525.6 542.1 546.2 570.5
Indicates the animals were not weighed.
DuPont-6542
Test Substance H-24944 248.2 256.0 264.6 271.3 280.3 289.7 296.0 304.3 310.9 316.9 353.9 384.9 409.6 429.6 450.2 476.2 495.1 512.1
-
521.2 540.2 556.6 572.9
<P
21Company Sanfced. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
TABLE 2 MEAN BODY WEIGHT GAINS (g)
Test Days
1-5 5-10
1-10
Negative Control Deionized Water
32.5 34.9
67.4
Dosing
Positive Controls
Test Substance
H-24019______H-24020______H-24944
31.8
28.4
32.1
30.3
30.5
36.6
62.1
58.9
68.7
Test Days 10-17 17-24 24-52 52-94
10-94
Negative Control Deionized Water
36.3 32.2 95.0 91.2
254.7
Recovery
Positive Controls .
Test Substance
H-24019______H-24020______H-24944
43.6
54.4
37.0
23.2
41.2
31.0
109.5
122.0
91.3
101.8
109.9
96.7
278.1
327.5
256.0
-22-
Company Sanitized. Does not contain TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
Test Days 10 94
TABLE 3
MEAN BODY AND LIVER WEIGHTS (g)
DEIONIZED WATER (NEGATIVE CONTROL)
Absolute
Body Weight
Liver Weight
258.8
10.814
511.7
17.868
Mean Relative Liver Weight (Liver/Body Weight)
0.042 0.035
Test Days 10 94
H-24019 (POSITIVE CONTROL)
Absolute
Body Weight
Liver Weight
243.9
14.205
524.6
19.296
Mean Relative Liver Weight (Liver/Body Weight)
0.058 0.037
Test Days
.10 94
H-24020 (POSITIVE CO NTROL)
Absolute
Body Weight
Liver Weight
243.1
19.174
570.5
19.590
Mean Relative Liver Weight (Liver/Body Weight)
0.079 0.034
Test Days 10 94
H-24944 (TEST SUBSTANCE)
Absolute________
Body Weight
Liver Weight
326.4
14.098
572.9
19.564
Mean Relative Liver Weight (Liver/Body Weight)
0.043 0.034
-23-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
TABLE 4 MEAN BLOOD FLUORINE LEVELS
Test Days
1 5 10 13
24 52 94
Negative Control Deionized Water
(ppm) 0.603
c
I.IO3
c
0.5"
c
c
Positive (controls
H-24019
H-24020
(ppm)
(ppm)
2.60 (0.1)"
9.40 (2.4)
31.32 (1.1)
74.92 (7.0)
68.00 (3.5)
61.76 (5.5)
53.98 (1.2)
29.52 (4.9)
39.62 (3.4)
11.18 (2.9)
23.56 (2.1) 12.60 (1.2)
2.26 (1.1) 0.85'1 (O.I)
Test Substance
H-24944 (ppm)
0.76 (0.2) 3.48 (1.1) 4.74 (0.7) 3.32 (2.5) 0.88 (0.1)
c
c
a One of 5 values. Four of the values were below the limit of quantification (LOQ) or non-detectable. b Standard deviation 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
Test Days
1 5 10 13
24 52 94
Positive Controls
H-24019
H-24020
IJLMF Equivalents 36.92 (I.I)-1
/iM F Equivalents
66.67 (17.5)
478.77 (17.6)
541.45 (50.4)
1043.08 (54.6)
446.09 (39.7)
827.38 (19.0)
212.46 (35.6)
606.46 (53.1)
79.57 (20.8)
359.38 (32.7) 190.77 (19.2)
14.93 (7.7) 4.711' (0.5)
Test Substance
H-24944
^M F Equivalents
0.57 (0.2) 3.36 (1.2) 4.65 (0.8) 3.20 (2.5) 0.70 (0.2)
c
c
Standard deviation is in parentheses.
Mean of 2 of the 5 values. Three of the values were below the limit of quantification (LOQ). All values were below the LOQ.
-24-
Company Sanitized. Does nol contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TABLE 6 MEAN LIVER FLUORINE LEVELS
Test Days
10
94
Negative Control Deionized Water (ppm) 0.90 (0.2)'1 0.78 (0.0)
Positive Controls
H-24019 (ppm)
312.34 (19.7) 84.24 (7.4)
H-24020 (ppm)
119.80 (3.5) 2.56 (1.3)
a Standard deviation is in parentheses.
DuPont-6542
Test Substance
H-24944 (ppm)
22.86 (2.3) 3.54 (1.1)
TABLE 7
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Positive Controls
Test
H-24019
H-24020
Days_____^M Equivalents
10
4802.15 (303.8)
IJiM Equivalents 866.67 (25.5)
94
1292.92 (114.1)
17.10 (9.8)
a Standard deviation is in parentheses.
Test Substance H-24944
(JiM Equivalents 23.22 (2.3) 3.42 (1.1)
-25-
Company Sanitized. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TABLES MEAN FAT FLUORINE LEVELS
Test Days
10 94
Negative Control Deionized Water (ppm)
a
a
Positive Controls
H-24019 (ppm)
12.84 (I.?)" 1.20 (0.3)
H-24020 (ppm)
8.10 (1.1)
a
All values were below the limit of quantification (LOQ) or non-detectable.
Standard deviation is in parentheses.
DuPont-6542
Test Substance
H-24944 (ppm)
7.80 (1.7) 1.66 (0.2)
TABLE 9
MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE
Positive Controls
Test
H-24019
H-24020
Days_____pM. Equivalents_____/xM Equivalents
10
194.46 (26.8)-
57.25 (7.8)
94
15.38 (4.1)
b
Standard deviation is in parentheses. All values were non-detectable.
Test Substance H-24944
/zM Equivalents 7.79 (1.7) 1.50 (0.2)
-26-
Company Sanitized. Does not contain TSCA ("
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
FIGURES
|
-27-
Company Sanitized. Does not contain TSCA CBf
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 1 MEAN BODY WEIGHTS (g)
Test Days
-28-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROL
Test Days -29-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 3 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
Test Days
.30-
FIGURE 4 MICROMOLAR EQUIVALENTS IN RAT BLOOD
A. Normalized Rat Blood H-24019^M Equivalents Resulting from a 10-Day Oral Gavage
S 600 }
Micromolar (/iM) equivalents ofH-24019 (positive control) in rat blood resulting from a 10-day oral gavage
exposure. Values are means and error bars are standard deviation. ^.Normalized Rat Blood H-24020f/M Equivalents Resulting from a 10-Day Oral Gavage
TIB l Id.?.)
Micromolar (/iM) 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.
|
-31Company Sanitized. Does not contain TSCA CK
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
C. Normalized Rat Blood H-24944 pW Equivalents Resulting from a 10-Day Oral Gavage
Limit of Quantification
/
10
20
30
40
50
60
70
80
90
100
Time (days)
Micromolar (liM) equivalents of H-24944 (test substance) in rat blood resulting from a 10-day oral gavage
exposure. Values are means and error bars are standard deviation. Test days 52 and 94 values were below the limit of quantification.
-32-
Company Sanitized. Doss not contain TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
600000
FIGURE 5
NORMALIZED H-24944 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A 10-DAY ORAL GAVAGE
500000
400000
e
300000
200000
100000
EIAUCINF/D
H-24020
H
8.15E+04
6.
33-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 6 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
-34-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE? COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTR
FOR H-24944 AND NEGATIVE CONTROL
Day 10
Day 94
DuylO
D Deionized Water (Negative Control) H-24944 (Test Substance)
-35-
.y
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURES MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE
Day 10
Day 94
^
H-24944: 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
Blood / Liver / Fat
Blood / Liver / Fat
-37-
Blood / Liver
APPENDICES
-38-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX A Individual Body Weights
-39-
Company Sanitized. Does not cinta!rs TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
INDIVIDUAL BODY WEIGHTS
ABBREVIATIONS: SD - sacrificed by design
EXPLANATORY NOTES
DuPont-6542
t)
-40-
Company Sanitized. Doss not contain TSCA CBI
H-24944: 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
Day 3
GROUP I_
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
646932 646933 646934 646935 646936
Day 10
264.7
254.4 248.6
278.0
248,. 1
SD test day 10 3D test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
-41-
H-24944: 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
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 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 SD 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
-42-
1-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
AMNIMAL 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 (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
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
-43.
H-24944: 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
9
H-24019 (POSITIVE CONTROL)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
Day 3
GROUP III
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
ANIMAL 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
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
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
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
-44-
H-24944: 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
TEST 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
-45-
H-24944: 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 (g) OF MALE RATS
Day 3
GROUP III
TEST 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
-46-
H-24944; Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL
NUMBER
648905 648906 648907 648908 648909
Day 1 242 .8 266 .4 258 .6 251 .8 258 .0
Day 2
245.0 281.9 263.6 259.0 266.3
H-24944 (TEST SUBSTANCE)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
GROUP I
Day 3
TEST DAY
Day 4
Day 5
Day 6
254.1 294.5 274.2
266.4 275.9
259.1
302.7 283.8 275.3 287.8
270.9 304.6 293.8 289.3 294.6
274.1 314.7 299.4 298.6 300.7
ANIMAL
NUMBER
648905 648906 648907 648908 648909
Day 10
305 .9 343 .3 324 .6 327 .1 331 .2
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
Day 7 284 .1 324 .6 308 .7 307 .7 310 .5
-47-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
ANIMAL
NUMBER
648910 648911 648912 648913 648914
Day 1 238 .8 255 .5 234 .4 231 .8 243 .7
ANIMAL NUMBER
648910 648911 648912 648913 648914
Day 10 307 .2 326 .3 319 .5 277 .7 306 .4
ANIMAL NUMBER
648910 648911 648912 648913 648914
Day 73 513 .0 545 .1 572 .1 449 .0 526 .6
Day 2
248.4 263.5 243.5 241.3 247.9
Day 17
363.7 373.2 377.1 312.0 343.3
Day 80
532.5 561.3 594.8 470.5 542.1
H-24944 (TEST SUBSTANCE)
INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS
Day 3
GROUP III
TEST DAY
Day 4
Day 5
Day 6
257 .6 266 .1 254 .1 246 .1 257 .0
250.1 274.1
265.0 250.2
265.1
260.6 281.5 276.2 259.7 271.7
281 .3 291 .4 287 .6 267 .8 281 .2
Day 24 396 .3 404 .2 413 .3 334 .5 376 .2
TEST DAY
Day 31
Day 38
411.4 435.5 438.7
357.6 404.6
421.4 457.6 470.6 374.1
424.5
Day 45
422 .3 478 .7 496 .3 392 .3 461 .6
Day 87 549 .9 579 .0 617 .1 490 .0 547 .1
TEST DAY Day 94
564.6 SD test day 94 589.9 SD test day 94 640.6 SD test day 94 505.2 SD test day 94 564.2 SD test day 94
Day 7
280. 4 303 . 3 288. 2 268. 3 283. 9
Day 52
454. 6 492. 9 530. 7 417. 1 485. 7
-48-
APPENDIX B Individual Clinical Observations
-49-
Company Sanitized. Does not contain TSCA CBt
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I
Animal 646932
646933
646934
Observation
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
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
General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left
Discharge, Eye left. 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
-50-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 646935
646936
DEIONIZED WATER (NEGATIVE 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 Sacrificed by design
General observation, No Abnormality Detected
1,5
10
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
1,5
Sacrificed by design
10
-51
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 646937 646938
646939
DEIONIZED WATER (NEGATIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP III
Days
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, 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
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
-52-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DEIONIZED WATER (NEGATIVE CONTROL)
Animal 646940
646941
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP III (Continued)
Days
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 Discharge, Nose, Black
24,52
87
Sacrificed by design
94
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
1-10,17-94
52 13 24 94
-53-
^
H-24944: 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
-54-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24019 (POSITIVE CONTROL)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal Observation
GROUP III
Days
646915 General observation, No Abnormality Detected
1,17-38,73
Eye Observations, Bled via Orbital for Clin Path, Left
13,24,52
Discharge, Eye right. Red Hair Loss, Forelimb, Right
2-10
80
Hair Loss, Forepaw, Right
45-67,87-94
646916
Sacrificed by design General observation, No Abnormality Detected
94
1-10,17-94
Eye Observations, Bled via Orbital for Clin Path, Bilateral
24
Eye Observations, Bled via Orbital for Clin Path, Left
13
0
Eye Observations, Bled via Orbital for Clin Path, Right
52
0
o
Sacrificed by design
0)
< w
646917 General observation. No Abnormality Det:ected
0)
^
Eye Observations, Bled via Orbital for Clin Path, Left
N'
(D
P-
Eye Observations, Bled via Orbital for Clin Path, Right
0
0 CD
Sacrificed by design
U)
3
0
0 0
3"
^
0 >
0
S
-55-
94
1-10,17-94 13,24
52 94
^l/H-24944: 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 Sacrificed by design
13,24
52 94
-56-
H-24944: 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
-57-
H-24944: 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 Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations. Bled via Orbital for Clin Path, Right Sacrificed by design
1-10
1 5
10
-58-
H-24944: 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
52
Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design
13,24
94
General observation. No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design
1-10,17-94
52
13,24
94
General observation. No Abnormality Detected
1-10,17-94
Eye Observations. Bled via Orbital for Clin Path, Left Sacrificed by design
13,24,52
94
-59-
H-24944: 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 Eye Observations, Bled via Orbital for Clin Path, Right Hair Loss, Forelimb, Bilateral
13,24
52
52-73
Hair Loss, Neck, Left
73
Hair Loss, Neck, Ventral Wound, Superficial, Face Sacrificed by design
38-67,80-94
59 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 Sacrificed by design
13,24
52 94
-60-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Animal 648905
648906 648907 648908
H-24944 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Observation
GROUP I.
Days
General observation. No Abnormality Detected
1-4
Eye Observations, Bled via Orbital for Clin Path, Left
5
Eye Observations, Bled via Orbital for Clih Path, Right
1
Discharge, Eye right. Black Sacrificed by design
5-10
10
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
1-10
5 1
10
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
1-10
5 1
10
1-10
Eye Observations, Bled via Orbital for Clin Path, Left
5
Eye Observations. Bled via Orbital for Clin Path, Right
1
Sacrificed by design
10
-61-
H-24944: Biopersistence Screening IQ-Dose Oral Gavage Study in Rats
Animal 648909
H-24944 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
GROUP I (Continued)
Observation
Days
General observation. No Abnormality Detected
1-6
Eye Observations, Bled via Orbital for Clin Path, Left
5
Eye Observations, Bled via Orbital for Clin Path, Right
1
Discharge, Eye right, Black Sacrificed by design
7-10
10
-62
-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H-24944 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal Observation
GROUP III
Days
648910 General observation. No Abnormality Detected
1-10,17-94
648911
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
13,24,52 24,52
94
1-10,17-94 13,24,52 24,52
94
648912 General observation. No Abnormality Detected
0
0
Eye Observations, Bled via Orbital for Clin Path, Left
T3
01
Eye Observations, Bled via Orbital for Clin Path, Right
<
w fi)
Sacrificed by design
=.
R"
648913 General observation. No Abnormality Detected
(0
a.
0
Eye Observations, Bled via Orbital for Clin Path, Left
0
<0
If)
Eye Observations, Bled via Orbital for Clin Path, Right
"s
2. fl
Sacrificed by design
.3
3
--
W
3"
-Vi)
0 >
0 00
-63-
1-10,17-94 13,24,52 24,52
94
1-10,17-94 13,24,52 24,52
94
^ pH-24944: Biopersistence Screening
10-Dose Oral Gavage Study in Rats
_____
^
__
w H-24944 (TEST SUBSTANCE)
INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS
Animal Observation
GROUP III (Continued)
Days
648914
General observation, No Abnormality Detected
Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right
1-10,17-94 13,24,52 24,52
Sacrificed by design
94
-64.
APPENDIX C
Terms and Calculations Factors Influencing Interpretation of Kinetic Analysis
-65-
Company Sanitized. QQS-. n"( conta'n TSCA CP>
B^
'--y
^
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 rnmole 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/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/P)
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
-66-
Comoany Sanitized. 0"--^ ""< co^a'n TSCA CW
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppmF
The ppm fluoride measured
Individual Animal Calculations:
ppm F minus Bkg 0.2 ppm
ppmF
normalized to 0.1 mmol/kgDose
The ppm fluoride measured minus the background fluoride measured in control animal. In this case the value was established at 0.2 ppm.
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)
/imolar equivalents of
active
The /xmolar [^mol/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 /imol/mmol
-67-
Company Sanitized. Uoes not contain TSCA CCT
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
_____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 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
-68- Company Sanitized. Does not contain TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX D Individual Fluorine Levels in Blood
-69-
^-.-..^v Sanitized. Does not contain TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24D19
DuPont-6542
Given:
MoHiVt. Active (g/mole):
497
Form ulation Dose (mg/kg):
10
%Ac tive (F Contai ning) in Fc)rmulation: 100
% F in Active:
65
MolWt.F(g/mc)!):
19
Calculat)id Values: Dose Active (mg/k{I): Dose Active (mmol e/kg): Dose F (mmol/kg):
10 0.020 0.342
Molar Ratio (Acitive/F): Dose F (mg/kg):
0.059 6.5
Rat Number
Test Day Sample
ppmF
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
30.1
646911
5
30.3
646912
5
31.3
646913
5
32.2
646914
5
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
^molar 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 not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test Day Sample
ppmF
in Blood
Group III
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 III
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
ppniiFinB lood
NO]rmalizcidto 0.1 mmoles,/kg
Dose
DuPont-6542
/imolar Equivalents of Active in
Blood
272.36 273.85 258.44 266.39 265.40
843.08 847.69 800.00 824.62 821.54
192.34 216.69 185.88 174.94 209.73
595.38 670.77 575.38 541.54 649.23
114.31 99.90 127.73 122.76 115.80
353.85 309.23 395.38 380.00 358.46
65.11 57.16 63.12 53.68 69.08
201.54 176.92 195.38 166.15 213.85
-71 -
Company Sanitized. Does not contain TSCA CBt
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-240 20
DuPont-6542
Given:
MoPWt. Active (g/imole):
426
Form ulation Dose (mg/kg):
20
%Ac tive (F Contai ning) in Fc>rmulation: 100
% F in Active:
69
Mol Wt. F (g/mc)!):
19
Calculatied Values: Dose Active (mg/lq^ Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Acltive/F): Dose F (mg/kg):
0.065 13.8
Rat Number
Test Day Sample
ppmF
in 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
Group I
646921
5
646922
5
646923
5
646924
5
646925
5
78.7 83.2 77.0 66.1 69.6
78.5 83.0 76.8 65.9 69.4
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
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
^molar 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
568.84 601.45 556.52 477.54 502.90
133.76 146.76 128.01 132.70 114.38
455.07 499.28 435.51 451.45 389.13
-72-
Company Sanitized. Does not conta'n TSCA CB,
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test Day Sample
ppmF
in
Blood
Group III
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 III
646926
52
2.5
2.3
646927
52
1.7
1.5
646928
52
1.3
1.1
646929
52
4.0
3.8
646930
52
1.8
1.6
Group III
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 Bflood
Normalize dto
0.1 nunoles/kg Dose
73.70 63.47 50.06 71.36 53.68
24.92 21.73 16.83 33.02 20.45
4.90 3.20 2.34 8.09 3.41.
1.28
* *
1.49
*
DuPont-6542
^molar Equivaleiits of Active in
Blood
250.72 215.94 170.29 242.75 182.61
84.78 73.91 57.25 112.32 69.57
16.67 10.87 7.97 27.54 11.59
4.35
* *
5.07
*
D
-73-
Company Sanitized. Does not contain TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-249-44
DuPont-6542
Given:
MoPtVt. Active (g/mole): Formulation Dose (mg/kg): %Ac;tive (F Containing) in F'ormulation:
3150 1000 40
% F in Active:
Mol Wt. F (g/moI):
24.4
19
Calculatied Values: Dose Active (mg/k{?): Dose Active (mmol e/kg): Dose F (mmol/kg):
400 0.127 5.137
Molar Ratio (Act:ive/F): Dose F (mg/kg):
0.025 97.6
Rat Number
Test Day Sample
ppmF
in Blood
Group I
648905
1
0.5
648906
1
1.0
648907
1
0.7
648908
1
0.8
618909
1
0.8
ppm F in Blood
Minus Bkg 0.2 ppm
0.3 0.8 0.5 0.6 0.6
Group I
648905
5
1.9
1.7
648906
5
4.3
4.1
648907
5
3.2
3.0
648908
5
3.2
3.0
618909
5
4.8
4.6
Group I
648905
10
4.2
4.0
648906
10
5.6
5.4
648907
10
4.6
4.4
648908
10
5.4
5.2
618909
10
3.9
3.7
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
fimolw Equivalents of Active in
Blood
0.24 0.63 0.39 0.47 0:47
0.31 0.82 0.51 0.61 0.61
1.34 3.23 2.36 2.36 3.62
1.74 4.20 3.07 3.07 4.71
3.15 4.25 3.47 4.10 2.91
4.10 5.53 4.51 5.33 3.79
.74-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test Day Sample
ppmF
in
Blood
Group III
648910
13
7.6
648911
13
3.0
648912
13
1.4
648913
13
2.0
648914
13
2.6
ppm F in Blood
Minus Bkg 0.2 ppm
7.4 2.8 1.2 1.8 2.4
Group III
648910
24
1.1
0.9
648911
24
0.8
0.6
648912
24
0.7
0.5
648913
24
0.9
0.7
648914
24
0.9
0.7
Group III
648910
52
<0.5
*
648911
52
<0.5
*
648912
52
<0.5
*
648913
52
<0.5
*
648914
52
<0.5
*
Group III
648910
94
<0.5
*
648911
94
<0.5
*
648912
94
<0.5
*
648913
94
<0.5
*
648914
94
<0.5
*
Below LOQ (Limit of Quantification)
ppm F in B;](>od Normalize d to 0.1 mmoleS/kg
Dose
DuPont-6542
^molar Equivalents of Active in
Blood
5.83 2.21 0.95 1.42 1.89
7.58 2.87 1.23 1.84 2.46
0.71 0.47 0.39 0.55 0.55
* * * * *
0.92 0.61 0.51 0.72 0.72
* * * * *
*
*
*
*
*
*
*
*
*
*
-75-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX E Individual Fluorine Levels in Liver
-76-
Company Sanitized. Does not contain TSCA CBt
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Datafc)rH-24019
Given:
MolWt:. Active (g/mole):
49';1
Formuliation Dose (mg/kg):
10
% Actn/e (F Contalining) in Fc>rmulation: 10C)
% F in Iactive:
Mol Wt F(g/nnol):
DuPont-6542
65 19
Calculated Values: Dose Aictive (mg/h-g): Dose Ai=tive (mmc>le/kg): DoseF (mmol/kg)
10 0.0:20 0.3-42
Molar R atio (A.ctive/F): Dose F (;mg/kg):
0.059 6.5
Rat Number
Group I 646910 646911 646912 646913 646914
Test Day Sample
ppmF
in
Liver
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
NormalizedIto 0.1 mmolesyikg
Dose
1593.88 1658.99 1582.45 1398.56 1522.81
424.44 376.23 433.88 386.67 467.18
/imolar 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
\)
-77Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-2<4020
DuPont-6542
Given:
Mol Wt. Active (g/mole):
426
Formulation Dose (mg/kg):
20
% Active (F Contalining) in Fc)rmulation: 100
%Fin Active:
69
MolWt.F(g/inol):
19
Calculated Values: Dose Active (mg/l<^g): Dose Active (mmcle/kg): Dose F (mmol/kg);
20
0.04 7 0.72 6
Molar I latio (/LCtive/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 III
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 L iver Normalize!I to 0.1 mmoles/kg
Dose
/imolar Equivalents of Active in
Liver
251.55 260.93 243.03 258.58 259.65
855.80 887.68 826.81 879.71 883.33
3.41 2.13 4.26 9.59 5.75
11.59 7.25 14.49 32.61 19.57
78-
Company Sanitized. Does not confab TSCA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24944
DuPont-6542
Given; Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
3150 1000 40
% F in Active:
Mol Wt. F (g/mol):
24.4 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
400 0.127 5.137
Molar Ratio (Active/F): Dose F (mg/kg):
0.025 97.6
Rat Number
Test Day Sample
ppmF
in
Liver
Group I
648905
10
26.1
648906
10
23.5
648907
10
23.4
648908
10
20.7
648909
10
20.6
Group III
648910
94
5.3
648911
94
3.9
648912
94
3.0
648913
94
3.1
648914
94
2.4
ppm F in Liver
Minus Bkg 0.2 ppm
25.9 23.3 23.2 20.5 20.4
5.1 3.7 2.8 2.9 2.2
ppm Fin Liver
Normalized to 0.1 mmoles/kg
Dose
/imolar Equivalents of Active in
Liver
20.40 18.35 18.27 16.14 16.07
26.54 23.87 23.77 21.00 20.90
4.02 2.91 2.21 2.28 1.73
5.23 3.79 2.87 2.97 2.25
79Company Sanitized. Does not contain TSCA CBt
H-24944: Biopersistence Screening _10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX F Individual Fluorine Levels in Fat
-80-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-6542
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 (Active/F): Dose F (mg/kg):
0.059 6.5
Rat Number
Group I 646910 646911 646912 646913 646914
Test Day Sample
ppm F in Fat
10
14.4
10
14.9
10
10.9
10
11.6
10
12.4
Group III
646915
94
1.3
646916
94
1.1
646917
94
1.3
646918
94
0.8
646919
94
1.5
ppm F in Fat
Minus Bkg 0.2 ppm
14.2 14.7 10.7 11.4 12.2
1.1 0.9 1.1 0.6 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
fimolar Equivalents of Active in
Fat
218.46 226.15 164.62 175.38 187.69
16.92 13.85 16.92 9.23 20.00
81Company Sanitized. Does not contain TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-240 20
DuPont-6542
Given:
MolWt. Active (g/mole):
426
Formul ation Dose (mg/kg):
20
% Acti've (F Containing) in Fcirmulation: 100
% F in Active:
69
Mol Wt. F (g/mc)!):
19
Calculated1Values: Dose A ctive (mg/kj?): Dose A ctive (mmol e/kg): DoseF (mmol/kg):
20 0.047 0.726
Molar Ratio (Acltive/F): Dose F (mg/kg):
0.065 13.8
Rat Number
Group I 646921 646922 646923 646924 646925
Test Day Sample
ppmF
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 Normalized to 0.1 mmoles/kg
Dose
12.78 17.68 18.11 18.32 17.25
/imolar 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
82Company Sanitized. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24944
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
3150 1000 40
% F in Active:
MolWt.F(g/mol):
24.4 19
Calculated Values: Dose Active (mg/kg):
Dose Active (mmole/kg): Dose F (mmol/kg):
400 0.127 5.137
Molar Ratio (Active/F): Dose P (mg/kg):
0.025 97.6
Rat Number
Test Day Sample
ppmF
in
Fat
Group I
648905
10
648906
10
648907
10
648908
10
648909
10
5.5 10.2 7.3 7.7 8.3
Group III
648910
94
1.6
648911
94
1.6
648912-
94
1.5
648913
94
1.6
648914
94
2.0
ppm F in Fat
Minus Bkg 0.2 ppm
5.3 10.0 7.1 7.5 8.1
1.4 1.4 1.3 1.4 1.8
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
4.17 7.88 5.59 5.91 6.38
1.10 1.10 1.02 1.10 1.42
/imolar Equivalents of Active in
Fat
5.43 10.25 7.27 7.68 8.30
1.43 1.43 1.33 1.43 1.84
-83-
Company Sanifoed. Does not contain TSCA CBI