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TRADE SECRET
Study Title H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Laboratory Project ID: DuPont-6542
AR226-3193
` DuPont-6542
Author: Carol Finlay, B.A.
Study Completed on: November 26,2001
Perform ing L aboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
Work Request Number:
Service Code Number:
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H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats t
DuPont-6542
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Reviewed by:
d /x i. O
Judith C. Stadler, Ph.D., D.A.B.T. D irec to r
Issued by Study Director:
C aro lrin lay , B.A. Staff Scientist
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H-24944: Biopersistence Screening
TABLE OF CONTENTS
Page
C E R T IF IC A T IO N .................................................................................................................................2
LIST O F T A B L E S ................................................................................................................................. 4
LIST O F F IG U R E S ................................................................................................................ ............. 4
LIST O F A PPEN D IC ES...................................................................
5
STUDY IN FO R M A TIO N ..............................................................
6
STUDY PE R SO N N E L ...........................................
8
SUM M ARY...........................................................................
9
IN T R O D U C TIO N .................................................................................................................................11
M ATERIA LS AND M E T H O D S .......................................................................................................11
A. Test Substance and Positive Controls...................................................................................... 11
B. Test Species.................................................................................................................................11
C. Animal H usbandry.....................................................................................................................11
D. Quarantine and P retest.............................................................................................................12
E. Study D esign........................ .............................................................................................. . 13
F. Assignment to Groups and Study S tart...................
13
G. Dosing Material Preparation and Administration...................................................................13
H. Body W eights............................................................................................................................. 14
I. Mortality and Clinical Observations........................................................................................ 14
J. Collection and Analysis of Blood, Livers, and F a t................................................................ 14
K. Treatment of Fluorine D ata.......................................................................................................15
L. Statistical M ethods.....................................................................................................................15
RESULTS AND D ISC U SSIO N ...........................
16
A. In-Life Toxicology.....................................................................................................................16
B. Liver W eights............................................................................................................................. 16
C. Fluorine D ata.............................................................................................................................. 17
C O N C L U S IO N S...................................................................................................................................18
RECO RD S AND SAM PLE S T O R A G E .......................................................................................... 19
T A B LE S..................................................................................................................................................20
F IG U R E S ................................................................................................................................................27
A P P E N D IC E S.......................................................................................................................................38
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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 LIVER FLUORINE 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................................................................................................................................i......... 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
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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..........................................................................................................49 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
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H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TEST SUBSTANCE:
STUDY INFORMATION
Substance Tested:
Svnonvms/Codes1
Haskell Number: 24944 Composition:
Known Impurities:
POSITIVE CONTROL:
DuPont-6542
3
Haskell Number: 24019
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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
Svnonvms/Codes: H-24020
C-8 Perfluorooctanoate, ammonium salt APFO Ammonium perfluorooctanoate Haskell Number: 24020 Composition
Known Impurities
Sponsor: ' E. I. du Pont 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
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STUDY PERSONNEL Study Director: Carol Finlay, B.A.
Management: Judith C. Stadler, Ph.D., D.A.B.T. Primary Technician: James C. Mackay II
Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T.
Toxicology Report Preparation: Wanda F. Dinbokowitz
Laboratory Veterinarian: William Singleton, D.V.M., A.C.L.A.M
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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 HI) 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 AUCESTF/D for the fluorine component of the test substance, H-24944, was 6.64x10 compared to AUCINF/D values of 5.22x10s 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 fiM equivalents on day 10 and 3.42 fiM equivalents on day 94. On day 10, mean xM 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.
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The fluorine concentration in the fat from rats dosed with the test substance was 7.79 /xM equivalents on day 10. On day 94, the concentration was 1.50 fiM equivalents. The fluorine concentrations of the positive controls were approximately 25x or 7x higher than H-24944 on day 10. By day 94, the nM 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 juM 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.
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H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
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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.
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H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
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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. Q uarantine 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.
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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/Com Oil Acetone/Com 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 com oil. The ratio of acetone to com oil was 20:80. The amount each rat received was based on the body weight collected on each day of dosing and the suspension concentration. The rats were dosed at a volume 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.
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H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
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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.
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H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
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 /molar 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 (/M) equivalents of active component in the tissue. Detailed calculations can be found in Appendix C. The /M 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 /M
equivalent in the data.
The maximum observed concentration in blood was Cmax (/M 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.
L. Statistical Methods Descriptive statistics (e.g. mean, standard deviation) were used.
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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
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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 juM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state (Figures 4A and 4B). The for H-24019 was 541.45 50.37 fxM equivalents (Mean SD) with a terminal half-life of 42.2 days. The Cmaxfor H-24020 was 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 8.15x10 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 fxM 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 /xM 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 fxM 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 xM equivalents in rat blood rose rapidly and did not reach steady-state (Figure 4C). The Cmax for H-24944 was 4.65 0.76 /xM 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
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H-24944: Biopersistence Screening i0-Dose Oral Gavage Study in Rats
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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 AUCINF/D for the fluorine component of H-24944 was 6.64 xlO2 as compared to AUCINF/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 fiM 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 fiM 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 fiM 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 fiM 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 (AUCINF/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 t s c a OBf
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 CBI
H-24944: Biopersistence Screening 10-Pose Oral Gavage Study in Rats
DuPont-6542
TABLES
- 20-
Company Sanitized. Does not contain TSC 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
502.7
480.1
525.6
496.5
542.1
512.2
546.2
524.6
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
\
\y
-21
Company Sanifced. Dono not contain TSCA CB!
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
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
H-24019
H-24020
31.8 28.4
30.3 30.5
62.1 58.9
DuPont-6542
Test Substance H-24944 32.1 36.6 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 .
H-24019
H-24020
43.6 54.4
23.2 41.2
109.5
122.0
101.8
109.9
278.1
327.5
Test Substance H-24944 37.0 31.0 91.3 96.7
256.0
- 22-
Company Sanitized. Does not contain TSCA CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
TABLE 3
MEAN BODY AND LIVER WEIGHTS (g)
DEIONIZED WATER (NEGATIVE CONTROL)
Test Days
10 94
_____________ Absolute_____________
Mean Relative
Body Weight
Liver Weight Liver Weight (Liver/Body Weight)
258.8 511.7
10.814 17.868
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 CONTROL)
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
TABLE 4
MEAN BLOOD FLUORINE LEVELS
DuPont-6542
Negative Control
Positive Controls
Test Substance
Test Deionized Water
H-24019
H-24020
H-24944
Days________ (ppm)_____________ (ppm) ________ (ppm)____________ (PPm)
1 0.60a 5C 10 1.10a 13 c 24 0.5a 52 c 94 c
2.60 (0.1)b 31.32 (1.1) 68.00 (3.5) 53.98 (1.2) 39.62 (3.4) 23.56 (2.1) 12.60 (1.2)
9.40 (2.4) 74.92 (7.0) 61.76 (5.5) 29.52 (4.9) 11.18 (2.9)
2.26 (1.1) 0.85d (0.1)
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 o f 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 o f 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
/xM F Equivalents
ixM F Equivalents
36.92 (l.l)a
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)
14.93 (7.7)
190.77 (19.2)
4.71b (0.5)
Test Substance H-24944
fiM F Equivalents 0.57 (0.2) 3.36 (1.2) 4.65 (0.8) 3.20 (2.5) 0.70 (0.2)
C
a Standard deviation is in parentheses. b Mean of 2 o f the 5 values. Three o f the values were below the limit o f quantification (LOQ). c All values were below the LOQ.
- 24-
Company Sanitized. Does not contain TSCA CB!
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TABLE MEAN LIVER FLUORINE LEVELS
Test Days
10 94
Negative Control Deionized Water (ppm) 0.90 (0.2)a 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 Substance
Test H-24019
H-24020
H-24944
Pays_______ /M Equivalents________ tM Equivalents_________ fiM Equivalents
10
4802.15 (303.8)a
866.67 (25.5)
23.22 (2.3)
94________ 1292.92 (114.1)_________ 17.10 (9.8)____________3.42 (1.1)
a Standard deviation is in parentheses.
-25 -
Company Sanitized. Does not contain TSC CB1
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TABLE 8 MEAN FAT FLUORINE LEVELS
Test Days
10 94
Negative Control
Positive Controls
Deionized Water
H-24019
H-24020
(ppm)_____________(ppm) _________ (Ppm)
~J
12.84 (1.7)b
8.10 (1.1)
-a 1.20 (0.3)__________ -a
a All values were below the limit of quantification (LOQ) or non-detectable. b 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
Test Days
10 94
Positive Controls
Test Substance
H-24019
H-24020
H-24944
/M Equivalents_______ nM Equivalents_______ fiM Equivalents
194.46 (26.8)a
57.25 (7.8)
7.79 (1.7)
15.38 (4.1)_______________^ ________________1-50 (0-2)
a Standard deviation is in parentheses, b All values were non-detectable.
- 26-
Company Sanitized. Does not contain TSCA C
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
FIGURES
- 27-
Company Sanitized. Does not contain TSCA CB
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 1 MEAN BODY WEIGHTS (g)
DuPont-6542
ao
3
TQ3)
3
th
03 3
N
a.
ao
3O oo3 Ef 3* H<O/>
O
-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
DuPont-6542
Mean Relative Liver Wt.
Test Days
o
3
S5'. H(O>/>I o
CD - 2 9 -
t
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats________________________________
DuPont-6542
FIGURE 3
COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS
H -24019 (Positive C ontrol) H -24020 (Positive C ontrol)
H -24944 (Test Substance)
Cornpany Sanitized. Does not contain TSCA CB'
Test Days -30-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
FIGURE 4 MICROMOLAR EQUIVALENTS IN RAT BLOOD
A. N o r m a li z e d R a l B l o o d H-24019 /;M E q u i v a l e n t s R es u lt in g from a 10-Day O r a l G a v a g e
M icromolar (/xM) 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.
M icromolar (/xM) equivalents of H-24020 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation.
- 31 -
Company Sanitized. Does not contain TSCA CSV
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
C. Normalized Rat Blood H-24944 //M Equivalents Resulting from a 10-Day Oral Gavage
Micromolar Q i M ) equivalents o f 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 o f quantification.
- 32-
Company Sanitized. Doss not contain TSC CBI
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
(P
DuPont-6542
500000
400000
z
ao 30Do)
o3< OOT3 3 0 0 0 0 0
5
</>
CD
3. 2 0 0 0 0 0 N<QD.
ao IwD 1 0 0 0 0 0 3o--w oo3 --)i
5'
-o>f Q A U C I N F / D o
03
H-24019 5.22E+05
**." **'"^**1
H -24020 8.15E + 04
-33-
H-24944 6.64E+02
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 6 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
DuPont-6542
Company Sanitized. Does not contain TSC CBl
- 34-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_______________________
DuPont-6542
FIGURE 7
COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24944 AND NEGATIVE CONTROL
Company Sanitized. Doss not contain TSCA CE
Deionized Water (Negative Control) H-24944 (Test Substance)
- 35-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_________________________________ ___________________________________________________________________ DuPont-6542
FIGURE 8 MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE
- 36-
Company Sanitized. Does not contain TSCA CBI
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
DuPont-6542
Company Sanitized. Does not contain TSCA CBt
- 37-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDICES
D
__________________________________________ - 38 -
Company Sanitized. Does not contain TSC CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX A Individual Body Weights
- 39-
Company Sanitized. Does not contain TSC CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
INDIVIDUAL BODY WEIGHTS
EXPLANATORY NOTES ABBREVIATIONS:
SD - sacrificed by design
DuPont-6542
- 40-
Company Sanitized. Does not contain TSCA CB!
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
D E IO N IZ E D WATER (NEGATIVE CONTROL)
IN D IV ID U A L 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
Day 10
646932 646933 646934 646935 646936
264.7 254.4 248.6 278.0 248.1
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
DuPont-6542
Day 8
251.1 239.5 230.9 265.7 234.9
Day 9
256.1 246.4 238.1 270.5 240.3
Company Sanitized. Does not contain TSCA CEJl
-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
Day 10
646937 646938 646939 646940 646941
249.8 237.2 277.9 244.5 266.3
ANIMAL NUMBER
Day 73
646937 646938 646939 646940 646941
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
D E IO N IZ E D WATER (NEGA TIVE CONTROL)
IN D IV ID U A L BODY W EIGHTS ( g ) OF MALE RATS
Day 3
GROUP I I I
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
DuPont-6542
Day 8
238.7 221.8 259.2 232.0 249.1
Day 9
241.4 228.3 267.3 237.5 257.0
Day 59
461.7 378.7 505.7 380.3 455.8
Day 67
478.7 401.3 516.4 392.5 475.3
Company Sanitized. Does not contain TSCA CBI
-42-
-24944: 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 - 2 4 0 1 9 (P O S IT IV E CONTROL)
IN D IV ID U A L BODY WEIGHTS ( g ) OF MALE RATS
Day 3
GROUP I
TEST DAY
Day 4
Day 5
Day 6
197.3 195.7 195.9 198.1 198.8
204.8 202.0 202.5 205.7 203.8
212.7 211.7 211.9 221.8 215.4
216.3 217.2 217.7 225.0 222.7
ANIMAL NUMBER
Day 10
646910 646911 646912 646913 646914
241.1 236.6 241.9 257.0 243.0
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
Day 7
226.4 225.2 227.2 228.6 226.7
DuPont-6542
Day 8
232.3 225.6 232.4 238.4 231.5
Day 9
237.5 231.9 238.5 245.8 238.8
Company Sanitized. Does not contain TSCA
-43O00
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats____________________________
H-24019 (POSITIVE CONTROL) I N D I V I D U A L BO D Y W E I G H T S (g) OF M A L E RATS
ANIMAL NUMBER
Day 1
Day 2
Day 3
GROUP III
TEST DAY
Day 4
Day 5
Day 6
646915 646916 646917 646918 646919
182.7 185.0 187.8 175.5 192.0
188.3 192.7 194.3 181.9 196.9
200.1 205.5 205.8 189.8 207.9
206.3 211.6 210.4 197.6 218.9
211.1 220.0 221.8 207.0 228.3
220.6 226.9 227.5 212.4 234.0
Day 7
225.8 239.5 233.9 220.2 240.8
DuPont-6542
Day 8
231.1 240.1 235.8 224.1 245.8
Day 9
236.8 249.3 244.3 228.2 252.9
ANIMAL NUMBER
Day 10
646915 646916 646917 646918 646919
240.4 257.0 250.2 237.1 260.2
Day 17
283.4 310.0 280.3 270.6 306.1
Day 24
302.9 334.4 308.2 292.7 328.5
TEST DAY
Day 31
Day 38
326.7 383.2 339.7 323.2 369.2
337.9 411.9 362.8 350.8 390.4
Day 45
371.1 450.3 387.9 379.4 427.3
Day 52
399.7 469.8 409.0 396.7 439.0
Day 59
412.5 494.8 420.0 418.8 451.7
Day 67
433.1 511.8 441.5 433.8 457.6
ANIMAL NUMBER
Day 73
646915 646916 646917 646918 646919
461.2 544.2 457.5 448.1 489.5
Day 80
475.0 565.5 464.7 472.5 504.8
Day 87
498.1 578.5 479.5 480.8 524.2
TEST DAY Day 94
513.3 590.9 495.7 500.3 522.9
SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94
fV'rnoany Sanitized. Does not contain TSCA OP3f
-44-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________________________________________________________________________________DuPont-6542
H - 2 4 0 2 0 (P O S IT IV E CONTROL)
IN D IV ID U A L BODY WEIGHTS ( g ) OF MALE RATS
ANIMAL NUMBER
Day 1
Day 2
Day 3
GROUP I
TEST DAY
Day 4
Day 5
Day 6
Day 7
Day 8
Day 9
646921 646922 646923 646924 646925
204.5 178.3 185.9 188.7 172.2
204.9 180.9 185.1 194.6 176.0
215.7 188.9 196.0 209.5 183.9
222.2 197.4 202.3 212.6 193.4
233.3 197.7 203.5 230.4 202.7
238.8 202.4 204.9 225.9 202.7
244.3 202.4 219.1 236.2 215.1
244.9 204.0 224.9 236.5 221.5
252.2 207.7 231.7 250.9 228.7
ANIMAL NUMBER
Day 10
646921 646922 646923 646924 646925
262.6 218.6 240.6 259.1 234.8
SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10
TEST DAY
Sanitized D09S not contain TSCA CBS Company Sanitize^. uo
-45-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________________________________
H-24020 (POSITIVE CONTROL)
IND I V I D U A L BODY W E I G H T S (g) OF M A L E RATS
ANIMAL NUMBER
Day 1
Day 2
Day 3
GROUP III
TEST DAY
Day 4
Day 5
Day 6
646926 646927 646928 646929 646930
182.0 172.2 190.4 178.6 187.9
187.8 178.8 195.3 182.5 191.6
197.0 191.1 204.5 190.7 201.0
203.6 195.1 212.3 198.3 210.4
206.0 206.6 222.1 203.5 219.6
213.2 213.2 230.1 204.2 224.4
Day 7
222.9 218.2 236.1 214.6 226.7
DuPont-6542
Day 8
229.9 228.7 235.8 210.3 232.6
Day 9
239.7 233.1 243.4 219.2 235.3
ANIMAL NUMBER
Day 10
Day 17
Day 24
TEST DAY
Day 31
Day 38
Day 45
Day 52
Day 59
Day 67
646926
250.8
646927
243.3
rt
646928
254.1
o
646929
225.5
3
X3
646930
240.3
D3<)
297.2 306.2 310.6 280.9 292.3
349.3 355.1 349.4 323.2 316.2
383.8 404.0 ' 392.8 376.0 352.7
412.1 424.8 422.4 396.8 368.4
ANIMAL NUMBER
Day 73
Day 80
Day 87
TEST DAY Day 94
432.1 463.2 443.5 426.4 406.8
467.8 484.0 464.8 450.2 436.0
493.2 511.7 492.2 463.8 454.7
509.5 527.2 518.6 484.2 474.2
646926 646927 646928 646929 646930
535.3 551.2 540.3 502.6 498.5
551.9 574.5 557.0 509.6 517.6
564.4 541.3 568.3 526.7 530.3
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
Sanitized. Doss not contain TSCA CB1
-46-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_________________________________
H -2 4 9 4 4 (TEST SUBSTANCE)
IN D IV ID U A L BODY W EIGHTS ( g ) OF MALE RATS
ANIMAL NUMBER
Day 1
Day 2
Day 3
GROUP I
TEST DAY
Day 4
Day 5
Day 6
648905 648906 648907 648908 648909
242.8 266.4 258.6 251.8 258.0
245.0 281.9 263.6 259.0 266.3
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
Day 7
284.1 324.6 308.7 307.7 310.5
ANIMAL NUMBER
Day 10
648905 648906 648907 648908 648909
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
DuPont-6542
Day 8
290.0 334.6 312.4 314.5 322.7
Day 9
296.8 341.5 317.1 323.0 326.7
O m nany Sanitized. Does not contain TSCA CBI
-47-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats________________________________ _________
ANIMAL NUMBER
Day 1
Day 2
H -2 4 9 4 4 (TEST SUBSTANCE)
IN D IV ID U A L BODY W EIGHTS ( g ) OF MALE RATS
Day 3
GROUP I I I
TEST DAY
Day 4
Day 5
Day 6
648910 648911 648912 648913 648914
238.8 255.5 234.4 231.8 243.7
248.4 263.5 243.5 241.3 247.9
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
ANIMAL NUMBER
Day 10
648910 648911 648912 648913 648914
307.2 326.3 319.5 277.7 306.4
Day 17
363.7 373.2 377.1 312.0 343.3
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
ANIMAL NUMBER
Day 73
648910 648911 648912 648913 648914
513.0 545.1 572.1 449.0 526.6
Day 80
532.5 561.3 594.8 470.5 542.1
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.e 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
DuPont-6542
Day 8
291.9 307.1 302.3 275.0 292.7
Day 9
299.5 315.8 312.4 279.8 296.6
Day 59
476.8 518.6 555.3 422.3 502.5
Day 66
501.2 534.0 572.3 436.4 516.5
Company Sanitized. Does not contain TSC
o SI
-48-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX B Individual Clinical Observations
- 49Company Sanitized. Doe: no! contain TSCA CB
Company Sanitized. Does not contain TSCA CBt
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats__________________________
D E IO N IZ E D WATER (NEGA TIVE CONTROL)
A nim al 646932
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
GROUP I
O b serv atio n
Days
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1
Eye O b s e rv a tio n s , E xophthalm us, L e ft
2-9
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
1 ,5
D isc h a rg e , Eye l e f t , B lack
10
S a c rific e d by d esig n
10
646933
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, L e ft Eye O b se rv a tio n s, D ark, L e ft D isc h a rg e , Eye l e f t , B lack S w o lle n O b s e rv a tio n s , M outh
1-5 1 ,5 6-10 6-7 10
S a c rific e d by d esig n
10
646934
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e te c te d
1 -5 ,1 0
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft D isch arg e, Eye l e f t , B lack S a c rific e d by d esig n
1 ,5 6-9 10
-50-
DuPont-6542
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D E IO N IZ E D WATER (NEGATIVE CONTROL) IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
A nim al 646935
GROUP I ( C o n t i n u e d ) O b serv atio n G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
Days 1-10
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft S a c rific e d by d esig n
1 ,5 10
646936
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1-10
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
1 ,5
S a c rific e d by d esig n
10
DuPont-6542
Company Sanitized. Does not contain TSCA CB1
-51 -
Company Sanffizecf. Does not contain TSCA
H-24944: Biopersistence Screening 10-Dose ()ral Gavage Study in Rats
D E IO N IZ E D WATER (NEGATIVE CONTROL)
A nim al 646937
646938
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
GROUP I I I
O b serv atio n
Days
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
L eft
13
R ight
2 4 ,5 2
S a c rific e d by d esig n
94
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -4 5
Eye O b se rv a tio n s , E nophthalm us, R ig h t
80-94
Eye O b s e rv a tio n s , E xophthalm us, R ig h t
52-73
B ila te ra l
1 3 ,5 2
R ight
24
Eye O b serv a tio n s, C orneal O p acity , R ight
80-94
646939
D isc h a rg e , Eye r ig h t, B lack S a c rific e d by d esig n G e n e ra l o b s e r v a tio n . No A b n o rm a lity D e te c te d
L eft
59 94 1 -1 0 ,1 7 -9 4 13
S a c rific e d by d esign
R ight
2 4 ,5 2 94
-52-
DuPont-6542
H-24944: Biopersistence Screening 10-Dose ( )ral Gavage Study in Rats
A nim al 646940
646941
D E IO N IZ E D WATER (NEGA TIVE CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
GROUP I I I ( C o n t i n u e d )
O b serv atio n
D ays
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -8 0 ,9 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
13
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
2 4 ,5 2
D isch arg e, N ose, B lack
87
S a c rific e d by d esig n
94
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , B ila te r a l
52
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
13
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
24
S a c rific e d by d esig n
94
0
DuPont-6542
Company Sanitized. Does not contain TSCA CB1
- 53-
Company Sanitized. Does not contain TSC CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats________________________________________________________________________
H - 2 4 0 1 9 (P O S IT IV E CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
GROUP I
A nim al O b s e rv a tio n
Days
646910
G e n e ra l o b s e r v a tio n , No A b n o rm a lity D e te c te d
1-10
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft S a c rific e d by d esig n
1 ,5 10
646911
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e t e c te d
1 ,1 0
Eye O b se rv a tio n s , B led v ia O rb ita l fo r C lin P ath , L e ft
1 ,5
Eye O b s e rv a tio n s , D ark, L e ft
2-9
S a c rific e d by d esig n
10
646912
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e t e c te d
1-10
Eye O b se rv a tio n s , B led v ia O rb ita l fo r C lin P ath , L e ft
1
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
5
S a c rific e d by d esig n
10
646913
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e te c te d
1-10
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
1 ,5
S a c rific e d by d esig n
10
646914
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e te c te d
1-10
Eye O b se rv a tio n s, B led v ia O rb ita lfo r C lin P ath , L e ft
1 ,5
S a c rific e d by d esig n
10
-54-
DuPont-6542
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
H - 2 4 0 1 9 (P O S IT IV E CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
A nim al O b s e rv a tio n
GROUP I I I
Days
646915
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 ,1 7 -3 8 ,7 3
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
1 3 ,2 4 ,5 2
D isc h a rg e , Eye r i g h t. Red
2-10
H air L oss, F o relim b , R ight
80
H a ir L oss, Forepaw , R ig h t
4 5 -6 7 ,8 7 -9 4
S a c rific e d by d esig n
94
646916
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , B ila te r a l
24
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , L e ft
13
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , R ig h t
52
S a c rific e d by d esig n
94
646917
G e n e r a l o b s e r v a t i o n . No A b n o r m a lity D e t e c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , L e ft
1 3 ,2 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
52
S a c rific e d by d esig n
94
DuPont-6542
Company Sanitized. Does not contain TSCA CIS
- 55-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_______________________________
H - 2 4 0 1 9 (P O S IT IV E CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
GROUP I I I ( C o n t i n u e d )
A nim al O b s e rv a tio n
Days
646918
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b serv a tio n s,
B ledv ia O rb ita l fo r C lin P ath , L e ft
1 3 ,2 4
Eye O b se rv a tio n s,
B ledv ia O rb ita l fo r C lin P ath , R ig h t
52
S a c rific e d by d esig n
94
646919
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , L e ft
1 3 ,2 4
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , R ig h t
52
S a c rific e d by d esig n
94
DuPont-6542
Company Sanitized. Does not
o
o
3
U is"
a>OWH
o
-56-
Company Sanitized. Does not contain TSC CBf
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in R a ts __________________________________________
H - 2 4 0 2 0 (P O S IT IV E CONTROL) IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
A nim al O b s e rv a tio n
GROUP I
Days
646921
G e n e ra l o b s e r v a tio n , No A b n o rm a lity D e te c te d
1 ,5 -1 0
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft Eye O b se rv a tio n s , D ark, L e ft S a c rific e d by d esig n
1 ,5 2-4 10
646922
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft Eye O b se rv a tio n s , B led v ia O rb ita l fo r C lin P ath , R ig h t S a c rific e d by d esig n
1-10 1 5 10
646923
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 ,7 -1 0
Eye O b s e rv a tio n s , E xophthalm us, L e ft Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
4-6 1
Eye O b se rv a tio n s, B ledv ia O rb ita l fo r C lin P ath , R ig h t Eye O b s e rv a tio n s , D ark, L e ft S a c rific e d by d esig n
5 2-6 10
-57-
DuPont-6542
%
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
#
H - 2 4 0 2 0 (P O S IT IV E CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
GROUP I ( C o n t i n u e d )
A nim al O b s e rv a tio n
D ays
646924
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1-10
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
1
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
5
S a c rific e d by d esig n
10
646925
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1-10
Eye Eye
O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
1 5
S a c rific e d by d esig n
10
ii)
DuPont-6542
Company Sanitized. Does not contain TSCA CBI
- 58-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats___________________________
A nim al 646926
646927
H - 2 4 0 2 0 ( P O S I T I V E CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
GROUP I I I
O b serv atio n
Days
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, B ila te r a l
52
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, L e ft
1 3 ,2 4
S a c rific e d by d esig n
94
G e n e ra l o b s e r v a tio n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, B ila te r a l Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, L eft
52 1 3 ,2 4
646928
S a c rific e d by d esign G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, L e ft S a c rific e d by d esig n
94 1 -1 0 ,1 7 -9 4 1 3 ,2 4 ,5 2 94
DuPont-6542
Company Sanitized. Does not contain TSCA COT
-59-
Oo 3>03o<) (W/)
3,
(aD.
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
A nim al 646929
646930
H - 2 4 0 2 0 ( P O S I T I V E CONTROL)
IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
O b serv atio n
GROUP I I I ( C o n t i n u e d )
D ays
G e n e ra l o b s e r v a tio n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -3 1
L eft
1 3 ,2 4
R ight
52
H a ir L oss, F o relim b , B ila te r a l
52-73
H air L oss, Neck, L eft
73
H air L oss, Neck, V e n tra l Wound, S u p e r f ic ia l, Face
3 8 -6 7 ,8 0 -9 4 59
S a c rific e d by d esig n
94
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e te c te d
1 -1 0 ,1 7 -9 4
L eft
1 3 ,2 4
R ight
52
S a c rific e d by d esign
94
DuPont-6542
Does not containTSCA
-60-
O
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________________________________________________________________________________DuPont-6542
A nim al O b s e rv a tio n
H -2 4 9 4 4 (TEST SUBSTANCE) IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
GROUP I . D ays
648905
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1-4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
5 1
D isch arg e, Eye r ig h t, B lack
5-10
S a c rific e d by d esig n
10
648906
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
1-10 5
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ight
1
648907
S a c rific e d by d esig n G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ight S a c rific e d by d esig n
10 1-10 5 1 10
648908
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e t e c te d
1-10
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft Eye O b se rv a tio n s , B led v ia O rb ita l fo r C lin P ath , R ig h t S a c rific e d by d esig n
5 1 10
Company Sanitized. Does not contain TSCA CBt
-61 -
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
A nim al 648909
H -2 4 9 4 4 (TEST SUBSTANCE)
IN D IV ID U A L C L IN IC A L OBSERVATIONS I N MALE RATS
O b serv atio n
GROUP I ( C o n t i n u e d )
Days
G e n e r a l o b s e r v a t i o n , NoA b n o r m a lity D e t e c te d
1-6
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , L e ft
5
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
1
D isc h a rg e , Eye r ig h t,B lack
7-10
S a c rific e d by d esig n
10
DuPont-6542
Company Sanitized. Does
3o
fa *
oo3 ST 5' -cos=nt ogj
-62-
Company Sanitized. Does not contain TSCA
i.L l-'
H-24944: Biopersistence Screening
10-Dose Oral Gavage Study in Rats
A nim al 648910
648911
648912
648913
H -2 4 9 4 4 (TEST SUBSTANCE)
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
GROUP I I I
O b serv atio n G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
D ays 1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s , B led v ia O rb ita l fo r C lin P ath , L e ft Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, R ight S a c rific e d by d esign
1 3 ,2 4 ,5 2 2 4 ,5 2 94
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b serv a tio n s, Eye O b serv a tio n s,
B led v ia O rb ita l fo r C lin P ath, L e ft B led v ia O rb ita l fo r C lin P ath, R ight
1 3 ,2 4 ,5 2 2 4 ,5 2
S a c rific e d by d esign
94
G e n e ra l o b s e r v a t i o n , No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, L e ft
1 3 ,2 4 ,5 2
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, R ight
2 4 ,5 2
S a c rific e d by d esign
94
G e n e ra l o b s e r v a t i o n . No A b n o rm a lity D e te c te d
1 -1 0 ,1 7 -9 4
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, L e ft
1 3 ,2 4 ,5 2
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath, R ight
2 4 ,5 2
S a c rific e d by d esign
94
Oon
- 63-
#
DuPont-6542
H-24944: Biopersistence Screening 10-Dose ( iral Gavage Study in Rats
A nim al 648914
H -2 4 9 4 4 (TEST SUBSTANCE)
IN D IV ID U A L C L IN IC A L OBSERVATIONS IN MALE RATS
GROUP I I I ( C o n t i n u e d )
O b serv atio n
Days
G e n e r a l o b s e r v a t i o n , No A b n o r m a lity D e t e c te d
1 -1 0 ,1 7 -9 4
Eye O b s e rv a tio n s , B led v ia O rb ita l fo r C lin P ath , L e ft
1 3 ,2 4 ,5 2
Eye O b se rv a tio n s, B led v ia O rb ita l fo r C lin P ath , R ig h t
2 4 ,5 2
S a c rific e d by d esig n
94
Company Sanitized. Does not contain TSCA CD
-64-
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX C
Terms and Calculations Factors Influencing Interpretation of Kinetic Analysis
- 65-
Company Sanitized, D o s' not contain TSC CPI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats______________________________________________ '__________DuPont-6542
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/100) x Dose Active [mg/kg]
Dose F (mmol/kg)
The mmole of fluorine administered per kg of animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol]
Molar Ratio (Active/F) The moles of fluorine containing compound per mole 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. Do--.? not conia:n TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppm F
The ppm fluoride measured
Individual Animal Calculations:
ppm F minus Bkg 0.2 ppm
The ppm fluoride measured minus the background fluoride measured in control animal. In this case the value was established at 0.2 ppm.
ppm F normalized to 0.1 mmol/kg Dose
The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead of the actual active dose. This assumes linearity between administered dose and blood fluorine levels, but is needed because different doses of active were used in the study.
= (0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in blood minus background)
/molar equivalents of active
The /molar [/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 /mol/mmol
-67-
Company Sanitized. Does not contain TSCACB1
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 bejustified in all cases)
- Fluorine concentrations are linear with respect to dose - Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of
individual components - Background Fluorine is 0.2 ppm - % F data is the % Fluorine of the active (Fluorine containing component(s) in the formulation) - Molecular weight is the molecular weight of the active component in the formulation
- 68-
Company Sanitized. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-6542
APPENDIX D Individual Fluorine Levels in Blood
- 69-
r ------- Sanitized. Does not contain TS CA CBl
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
10 0.020 0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):
6.5
Rat Number
Test ppm F Day in Sample Blood
Group I 646910 646911 646912 646913 646914
1 1 1 1 1
2.6 2.7 2.5 2.6 2.6
ppm F in Blood
Minus Bkg 0.2 ppm
2.4 2.5 2.3 2.4 2.4
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
/imolar Equivalents of Active in
Blood
11.93 12.43 11.43 11.93 11.93
.
36.92 38.46 35.38 36.92 36.92
Group I 646910 646911 646912 646913 646914
5 5 5 5 5
30.1 30.3 31.3 32.2 32.7
29.9 30.1 31.1 32.0 32.5
148.60 149.60 154.57 159.04 161.53
460.00 463.08 478.46 492.31 500.00
Group I
646910 10 71.5
646911
10 70.5
646912 10 66.9
646913 10 62.5
646914 10 68.6
71.3 70.3 66.7 62.3 68.4
354.36 349.39 331.50 309.63 339.95
1096.92 1081.54 1026.15 958.46 1052.31
- 70-
Company Sanitized. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test ppm F Day in Sample Blood
Group III 646915 646916 646917 646918 646919
13 13 13 13 13
55.0 55.3 52.2 53.8 53.6
ppm F in Blood
Minus Bkg 0.2 ppm
54.8 55.1 52.0 53.6 53.4
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
DuPont-6542
/molar 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
Group III 646915 646916 646917 646918 646919
Group III
646915 646916 646917 646918 646919
24 24 24 24 24
52 52 52 52 52
38.9 43.8 37.6 35.4 42.4
23.2 20.3 25.9 24.9 23.5
38.7 43.6 37.4 35.2 42.2
23.0 20.1 25.7 24.7 23.3
192.34 216.69 185.88 174.94 209.73
114.31 99.90 127.73 122.76 115.80
595.38 670.77 575.38 541.54 649.23
353.85 309.23 395.38 380.00 358.46
Group III 646915 646916 646917 646918 646919
94 94 94 94 94
13.3 11.7 12.9 11.0 14.1
13.1 11.5 12.7 10.8 13.9
65.11 57.16 63.12 53.68 69.08
201.54 176.92 195.38 166.15 213.85
-71 -
Company Sanitized. Does not contain TSCA CBt
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24020
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
% F in Active: Mol Wt. F (g/mol):
69 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat Number
Test ppm F Day in Sample Blood
Group I 646921 1 10.6 646922 1 10.7 646923 1 9.2 646924 1 5.3 646925 1 11.2
ppm F in Blood
Minus Bkg 0.2 ppm
10.4 10.5 9.0 5.1 11.0
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
22.15 22.37 19.17 10.86 23.43
pmolar Equivalents of Active in
Blood
75.36 76.09 65.22 36.96 79.71
Group I 646921 646922 646923 646924 646925
5 5 5 5 5
78.7 83.2 77.0 66.1 69.6
78.5 83.0 76.8 65.9 69.4
167.21 176.79 163.58 140.37 147.82
568.84 601.45 556.52 477.54 502.90
Group I 646921 10 63.0 646922 10 69.1 646923 10 60.3 646924 10 62.5 646925 10 53.9
62.8 68.9 60.1 62.3 53.7
133.76 146.76 128.01 132.70 114.38
455.07 499.28 435.51 451.45 389.13
- 72-
Company Sanitized. Does not contain TSCA CB.
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test ppm F Day in Sample Blood
Group III 646926 646927 646928 646929 646930
13 13 13 13 13
34.8 30.0 23.7 33.7 25.4
ppm F in Blood
Minus Bkg 0.2 ppm
34.6 29.8 23.5 33.5 25.2
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
73.70 63.47 50.06 71.36 53.68
DuPont-6542
/molar Equivalents of Active in
Blood
250.72 215.94 170.29 242.75 182.61
Group III 646926 646927 646928 646929 646930
24 24 24 24 24
11.9 10.4 8.1 15.7 9.8
11.7 10.2 7.9 15.5 9.6
24.92 21.73 16.83 33.02 20.45
84.78 73.91 57.25 112.32 69.57
Group III 646926 646927 646928 646929 646930
52 52 52 52 52
2.5 1.7 1.3 4.0 1.8
2.3 1.5 1.1 3.8 1.6
4.90 3.20 2,34 8.09 3.41.
16.67 10.87 7.97 27.54 11.59
Group III 646926 646927 646928 646929 646930
94 94 94 94 94
0.8 <0.5 <0.5 0.9 <0.5
0.6 * *
0.7 *
Below LOQ (Limit of Quantification)
1.28 4.35 ** **
1.49 5.07 **
- 73-
Company Sanitized. Does not contain 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): 0.025
Dose F (mg/kg):
97.6
Rat Number
Test ppm F Day in Sample Blood
Group I 648905 648906 648907 648908 618909
1 1 1 1 1
0.5 1.0 0.7 0.8 0.8
ppm F in Blood
Minus Bkg 0.2 ppm
0.3 0.8 0.5 0.6 0:6
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
gmolar Equivalents of Active in
Blood
0.24 0.31 0.63 0.82 0.39 0.51 0.47 0.61 0!47 0.61
Group I 648905 648906 648907 648908 618909
5 5 5 5 5
1.9 4.3 3.2 3.2 4.8
1.7 4.1 3.0 3.0 4.6
1.34 1.74 3.23 4.20 2.36 3.07 2.36 3.07 3.62 4.71
Group I 648905 648906 648907 648908 618909
10 10 10 10 10
4.2 5.6 4.6 5.4 3.9
4.0 5.4 4.4 5.2 3.7
3.15 4.10 4.25 5.53 3.47 4.51 4.10 5.33 2.91 3.79
- 74-
Company Sanitized. Does not contain TSCA CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Rat Number
Test ppm F Day in Sample Blood
Group III 648910 648911 648912 648913 648914
13 13 13 13 13
7.6 3.0 1.4 2.0 2.6
ppm F in Blood
Minus Bkg 0.2 ppm
7.4 2.8 1.2 1.8 2.4
ppm F in Blood Normalized to 0.1 mmoles/kg
Dose
5.83 2.21 0.95 1.42 1.89
DuPont-6542
/molar Equivalents of Active in
Blood
7.58 2.87 1.23 1.84 2.46
Group III 648910 648911 648912 648913 648914
24 24 24 24 24
1.1 0.8 0.7 0.9 0.9
0.9 0.6 0.5 0.7 0.7
0.71 0.92 0.47 0.61 0.39 0.51 0.55 0.72 0.55 0.72
Group III 648910 648911 648912 648913 648914
52 52 52 52 52
<0.5 <0.5 <0.5 <0.5 <0.5
***
* .*
*
***
*
*.
*
* * -*
Group III 648910 648911 648912 648913 648914
94 94 94 94 94
<0.5 <0.5 <0.5 <0.5 <0.5
* * * * *
* * * * *
* * * * *
Below LOQ (Limit of Quantification)
- 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
Data fo r H-24019
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values:
Dose Active (mg/kg):
10 Molar Ratio (Active/F): 0.059
Dose Active (mmole/kg):
0.020
Dose F (mg/kg):
6.5
Dose F (mmol/kg):____________________ 0.342_________________________________
Rat Number
Test ppm F Day in Sample Liver
Group I
646910
10 320.9
646911
10 334.0
646912
10 318.6
646913
10 281.6
646914
10 306.6
ppm F in Liver
Minus Bkg 0.2 ppm
320.7 333.8 318.4 281.4 306.4
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
1593.88 1658.99 1582.45 1398.56 1522.81
/molar Equivalents of Active in
Liver
4933.85 5135.38 4898.46 4329.23 4713.85
Group III 646915 646916 646917 646918 646919
94 94 94 94 94
85.6 75.9 87.5 78.0 94.2
85.4 75.7 87.3 77.8 94.0
424.44 376.23 433.88 386.67 467.18
1313.85 1164.62 1343.08 1196.92 1446.15
- 77-
Company Sanitized. Does not contain TSCA CBf
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24020
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
% F in Active: Mol Wt. F (g/mol):
69 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat Number
Test Day Sample
ppm F 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
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
limolar 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
Group III 646926 646927 646928 646929 646930
94 94 94 94 94
1.8 1.2 2.2 4.7 2.9
1.6 1.0 2.0 4.5 2.7
3.41 11.59 2.13 7.25 4.26 14.49 9.59 32.61 5.75 19.57
- 78-
Company Sanitized. Does not contain TSCA CBf
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
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
400 0.127 5.137
% F in Active: Mol Wt. F (g/mol):
24.4 19
Molar Ratio (Active/F): 0.025
Dose F (mg/kg):
97.6
Rat Number
Test ppm F Day in Sample 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 648911 648912 648913 648914
94 94 94 94 94
5.3 3.9 3.0 3.1 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 F in Liver Normalized to 0.1 mmoles/kg
Dose
20.40 18.35 18.27 16.14 16.07
4.02 2.91 2.21 2.28 1.73
/tmolar Equivalents of Active in
Liver
26.54 23.87 23.77 21.00 20.90
5.23 3.79 2.87 2.97 2.25
79-
Company Sanitized. Does not contain TSCA CB l
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 TSC CBI
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data fo r H-24019
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values:
Dose Active (mg/kg):
10 Molar Ratio (Active/F): 0.059
Dose Active (mmole/kg):
0.020
Dose F (mg/kg):
6.5
Dose F (mmol/kg):____________________ 0.342_____________________ ___________
Rat Number
Test ppm F Day in Sample Fat
Group I 646910 646911 646912 646913 646914
10 10 10 10 10
14.4 14.9 10.9 11.6 12.4
ppm F in Fat
Minus Bkg 0.2 ppm
14.2 14.7 10.7 11.4 12.2
Group III 646915 646916 646917 646918 646919
94 94 94 94 94
1.3 1.1 1.3 0.8 1.5
1.1 0.9 1.1 0.6 1.3
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
limolar 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
- 81 -
Company Sanitized. Does not contain TSCA CBi
H-24944: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24020
DuPont-6542
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
% F in Active: Mol Wt. F (g/mol):
69 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):
20 0.047 0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat Number
Test ppm F Day in Sample Fat
Group I 646921 646922 646923 646924 646925
10 10 10 10 10
6.2 8.5 8.7 8.8 8.3
ppm F in Fat
Minus Bkg 0.2 ppm
6.0 8.3 8.5 8.6 8.1
Group III 646926 646927 646928 646929 646930
94 94 94 94 94
ND ND ND ND ND
ND ND ND ND ND
ND Non-detectable.
ppm F in Fat Normalized to 0.1 mmoles/kg
Dose
/tmolar Equivalents of Active in
Fat
12.78 17.68 18.11 18.32 17.25
43.48 60.14 61.59 62.32 58.70
ND ND ND ND ND ND ND ND ND ND
-82-
Company Sanitized. Does not contain TSCA CB
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): 0.025
Dose F (mg/kg):
97.6
Rat Number
Test Day Sample
ppm F in Fat
Group I 648905 648906 648907 648908 648909
10 10 10 10 10
5.5 10.2 7.3 7.7 8.3
Group III 648910 648911 648912648913 648914
94 94 94 94 94
1.6 1.6 1.5 1.6 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
/tmolar 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 Sanitized. Does not contain TSCA CBI
