Document 8R7yqzO2yJQoQ6QVqOvdJJeOK
TRADE SECRET
Study Title H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Supplement No. 1
Laboratory Project ID: DuPont-2926
AR226-3216
AUTHOR: CarolPinlay,B.A.
STUDY COMPLETED ON: September 5, 2000
SUPPLEMENT NO. 1 COMPLETED ON: January 11, 2002
PERFORMING LABORATORY:
E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
WORK REQUEST NUMBER:
SERVICE CODE NUMBER: K
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CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained
from this study.
Reviewed by:
Paul M. Hhideriiter, PhJ5.
Postdoctoral Fellow
o^J^-w^
Date
Reviewed by:
/V^J^g6<^/,i.J^^ciULuO Judith C. Stadler.PhJ).. D.A.B .T. Director
/O-TAfJ-760-^
Date
Issned by Study Director:
tfLAbL tiC^L/liA_________
Caroimnlay.BAj Staff Scientist
J_L_A^L-^ ^ Wate
^te-.^
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TABLE OF CONTENTS
Page
CERTIFICATION......................................................................................................................,.^
LIST OF TABLES .........................................................................................................................4 LIST OF FIGURES .......................................................................................................................4 LIST OF APPENDICES...............................................................................................................4 STUDY INFORMATION .............................................................................................................5 STUDY PERSONNEL...................................................................................................................7 REASON FOR SUPPLEMENT NO. 1........................................................................................8
SUMMARY...................................................................................................................................^
MATERIALS AND METHODS................................................................................................10 RESULTS AND DISCUSSION ..................................................................................................10
A. Liver Weights.....................................................................................................................10
1. Test substance.........................................................................................................................................10 2. Positive Controls .................................................................................................................................... 10
B. Liver Fluorine Data............................................................................................................II
1. Positive Controls ....................................................................................................................................11 2. Test Substance........................................................................................................................................11
CONCLUSIONS..........................................................................................................................11
TABLES........................................................................................................................................12
FIGURES......................................................................................................................................16
APPENDICES ..............................................................................................................................22
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LIST OF TABLES
Page 1. MEAN BODY AND LIVER WEIGHTS.............................................................................................................13 2. MEAN LIVER FLUORINE LEVELS..................................................................................................................15 3. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................15
LIST OF FIGURES
Page 1. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE
CONTROLS.........................................................................................................................................................17
2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE
CONTROLS.........................................................................................................................................................18
3. COMPARISON OF RELATIVE LIVER WEIGHT ANDMEAN LIVER FLUORINE CONCENTRATION FOR H-24002 AND NEGATIVE CONTROLS................................................................ 19
4. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................20 5. COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION
NORMALIZED TO DOSE..................................................................................................................................21
LIST OF APPENDICES
Page A. INDIVIDUAL FINAL BODY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY
WEIGHT..............................................-............................................................................................................^
B. INDIVIDUAL FLUORINE LEVELS..................................................................................................................34
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STUDY INFORMATION TEST SUBSTANCE:
Substance Tested:
Synonyms/Codes: H-24002
Haskell Number: 24002 Composition
Known Impurities: Unknown
POSITIVE CONTROL:
Known Impurities: Unknown
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STUDY INFORMATION (Continued) POSITIVE CONTROL:
Synonyms/Codes: H-24020
Haskell Number: 24020 Composition:
Known Impurities: Unknown
Sponsor:
E. I. du Pont de Nemours and Company Wilmington, Delaware 19898
U.S.A.
Study Initiated/Completed: April 30, 1999 / (see report cover page)
In-Life Initiated/Completed: July 11, 1999/October 12, 1999
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
STUDY PERSONNEL
Study Director: Management:
Primary Technician:
Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. Richard P. Mathena
Liver 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: Wanda L. West, D.V.M., A.C.L.A.M.
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
REASON FOR SUPPLEMENT NO. 1
The purpose of this supplement is to present the results from analysis of selected liver samples from rats treated with the test substance (H-24002), 2 positive control (H-24019 and H-24020),
and 2 negative controls (corn oil and corn oiVacetone).
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
SUMMARY
The objective of this study was to evaluate the potential for H-24002, when administered by gavage, to be absorbed and to accumulate in a mammalian system. Six groups of 5 male Crl:CD(SD)IGS BR rats each were given 75 mg/kg/day of H-24002. The test substance was administered to one group of 5 rats for 5 consecutive days and to 5 groups for 10 days. Approximately 2 hours after the first dose, blood was collected from the orbital sinus of each rat
from the 5-dose group. On selected days (5,10,13,24,52,94) 5 rats per group were euthanized
and the blood and livers were collected. Livers were weighed and those collected on study days 10 and 94 were analyzed for total fluorine. This supplement includes liver weight data and the results of total fluorine analyses in liver. Additionally, 2 negative controls, corn oil and corn oiVacetone (80:20), and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-24002.
The mean relative liver weights (liver/body weight) of rats dosed with the test substance, H-24002, were similar to the negative controls. On day 10, the mean relative liver weight of rats dosed with the positive control H-24019 was up to 27% higher than the negative controls. By day 94, the mean relative liver weight was up to 28% higher than the negative controls. In rats given H-24020, the mean relative liver weight was up to 75% higher than the negative controls. By day 94, the weights were similar.
The concentration of fluorine in the livers from rats dosed with the test substance, H-24002, was
696.43 fiM equivalents on day 10 and 15.58 /xM equivalents on day 94. On day 10, mean ^M
equivalent concentrations of fluorine in the livers from rats dosed with the positive control materials were approximately 7-fold (H-24019) and 2-fold (H-24020) greater than the fluorine concentration in livers from rats treated with the test substance. By day 94, the concentration in the livers from rats treated with the positive control H-24019 was approximately 70x fluorine concentration in rats treated with H-24002. The concentrations of fluorine on day 94 were similar in rats treated with the positive control H-24020 and the test substance, H-24002.
The {JIM equivalents of fluorine in the livers from rats treated with the test substance, H-24002, were higher than levels in the blood.
Under the conditions of this study, there was some absorption and retention of fluorine in the livers from rats dosed with H-24002. Fluorine levels in the livers from rats dosed with the test substance were lower than levels in rats dosed with the positive control H-24019. Fluorine levels in the livers from rats dosed with the test substance were lower on day 10 than levels in rats dosed with the positive control H-24020. On day 94, the fluorine levels were similar. Liver weights were not affected in rats dosed with the test substance.
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
MATERIALS AND METHODS
Blood and livers were collected on days 5, 10,13, 24, 52, and 94. The livers were weighed. Only the test day 10 and 94 livers were analyzed for total fluorine. These time points provided sufficient data to determine the accumulation and persistence of fluorine in the liver. See the original report for details of the analysis.
RESULTS AND DISCUSSION
A. Liver Weights (Tables 1, Figures 1-3, Appendix A)
As with body weights, comparison of absolute liver weights was complicated by the fact that there was a difference in age on test day 1 between rats dosed with the test substance and those dosed with the H-24019 positive control or the corn oil:acetone negative control. Therefore, liver weights relative to body weight are presented for comparison.
1.
Test Substance
The mean relative liver weights of rats dosed with the test substance, H-24002, were similar to the negative controls.
2.
Positive Controls
The mean relative liver weight of rats dosed with one of the positive controls, H-24019, was 12% higher on day 10 than the liver weight of rats dosed with the test substance, H-24002. On days 13, 24, and 52 (recovery), the mean relative liver weights of rats dosed with H-24019 were 22 or 33% higher than the liver weights of rats dosed with H-24002. By test day 94 (end of recovery), the mean relative liver weight of rats dosed with H-24019 was 21% higher than the liver weight of rats dosed with H-24002. The mean relative liver weight of rats dosed with H-24020, the other positive control, was 54% higher on day 10 than the liver weight of rats dosed with H-24002. On days 13 and 24 (recovery), the mean relative liver weights of rats dosed with H-24020 were 33 or 24% higher than the liver weights of rats dosed with H-24002. On days 52 and 94, the weights were similar.
The mean relative liver weight of rats dosed with H-24019 was up to 27% higher at day 10 than the negative control groups. By day 94, the mean relative liver weight of rats dosed with H-24019 was up to 28% higher than the negative controls. The mean relative liver weight of rats dosed with H-24020 was up to 75% higher at day 10 than the negative controls. The mean relative liver weights of rats dosed with H-24020 began to decrease during the recovery period and were similar by test day 94 to the negative controls.
Therefore, liver weights of rats dosed with the test substance, H-24002, were not affected.
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
B. Liver Fluorine Data
(Tables 2-3, Figures 3-5, Appendix B)
1.
Positive Controls
The positive controls were H-24019 and H-24020. The concentrations of fluorine in the livers on day 10 from rats dosed with the positive control materials were 5045.85 and 1127.25 ^iM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15
and 10.72 pM. equivalents.
2.
Test Substance
Levels of total fluorine in livers from rats dosed with the test substance, H-24002, were generally 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-24002 was 696.43 /AM equivalents at day 10 and 15.58 /xM equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 7x higher (day 10) and approximately 70x higher (end of study) than H-24002. For H-24020, the liver concentration was approximately 2x higher than H-24002 by the end of dosing. The concentrations were similar by the end of recovery.
The pM. equivalents of fluorine in the livers from rats dosed with the test substance, H-24002, were higher than levels in the blood.
CONCLUSIONS
Under the conditions of this study, there was some absorption and retention of fluorine in the livers from rats dosed with H-24002. Fluorine levels in the livers from rats dosed with the test substance were lower than levels in rats dosed with the positive control H-24019. Fluorine levels in the livers from rats dosed with the test substance were lower on day 10 than levels in rats dosed with the positive control H-24020. On day 94, the fluorine levels were similar. Liver weights were not affected in rats dosed with the test substance.
%
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TABLES
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DuPont-2926 Supplement No. 1
Test Days
5 10 13
24 52 94
TABLE 1
MEAN BODY AND LIVER WEIGHTS (g)
CORN OIL (NEGATIVE CONTROL)
Absclute
Body Weight
Liver Weight
259.3
12.620
311.1
15.861
322.9
14.906
391.2
16.635
464.7
17.240
557.8
20.118
Mean Relative Liver Weight(% of Body Weight)
0.049 0.051 0.046 0.043 0.037 0.036
Test Days 5 10
13
24 52 94
CORN OELACETONE (NEGATIVE CONTROL)
Absolute
Body Weight
Liver Weight
313.4
13.632
356.5
15.597
377.6
17.715
423.5
17.172
527.6
21.275
567.8
18.447
Mean Relative Liver Weight(% of Body Weight)
0.043 0.044 0.047 0.041 0.040 0.032
Test Days
5 c 10 13
24 52 94
H-24019 (POSITIVE CONTROL)
___Bo_d_y_W__ei_gAhtb_s__o_luLitvee_r_W_e_ig_h_t __
i3r>0<6:.6c.
1ie5.4A5C6.
310.1
17.336
344.8
22.049
387,9
21.313
438.1
19.826
597.9
24.455
Mean Relative Liver Weight(% of Body Weight)
0f\.0r\5cr0\ 0.056 0.064 0.055 0.045 0.041
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TABLE 2 (Continued)
MEAN BODY AND LIVER WEIGHTS (g)
Test Days
5 c 10 13
24 52 94
H-24020 (POSITIVE CONTROL)
__B_o_d_y_W_e_igAhbt_s_o_l_uLtivee_r_W_e_ig_h_t __
^2A4 4^ . 8o
1i i7.f0\5e ^2
284.4
21.944
289.1
21.780
370.3
20.619
456.7
17.713
538.7
18.205
Mean Relative Liver Weight(% of Body Weight)
0f\.0f\7if0\ 0.077 0.075 0.056 0.039 0.034
Test Days 5 10
13 24 52
94
H-24002 (TEST SUBSTANCE)
__B_o_d_y_W_e_i_gAhtb_s__o_lLuitvee_r _W_e_ig_h_t __
266.5
13.716
308.1
15.379
322.1
15.490
371.9
16.724
532.7
19.962
593.9
20.154
Mean Relative Liver Weight(% of Body Weight)
0.051 0.050 0.048 0.045 0.037 0.034
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TABLE 2 MEAN LIVER FLUORINE LEVELS
Negative Controls
Positive Controls
Test Substance
Corn
Test Corn Oil
Oil:Acetone
H-24019
H-24020
H-24002
""LO<02<0.5328.18 Days (ppm)_____(ppm)_______(ppm)____(ppm)_______(ppm) (19.3)13155.76 (29.2)125.4 (16.3)
94
0.6"______1"______77.56 (10.1)
1.68 (0.3)
3.0 (1.19)
a One value. Four of the values were below the level of detection (LOD). b Standard deviation is in parentheses.
TABLE 3
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Days
10 94
Positive Controls
Test Substance
H-24019
H-24020
H-24002
JU.MF Equivalents 5045.85 (296.91)'
iiM F Equivalents____juM F Equivalents
1127.25 (211.37)
696.43 (90.76)
1190.15 (156.05)
10.72 (2.14)______15.58 (6.61)
a Standard deviation is in parentheses.
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
FIGURES
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 1 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROLS
0.06
0.05
0.04
i:-
&
U
0
BS
.0
0,03
0.02
0.01
0.00
Test Day
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
Test Day
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURES COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN UVER FLUORINE CONCENTRA
FOR H-24002 AND NEGATIVE CONTROLS
Day 10
Day 13
Day 94
Day 10
II Corn Oil (Negative Control)
Corn Oil:Acetone (Negative Control)
D H-24002 (Test Substance)
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 4 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
6000 5000 4000 s 3000 2000 1000
Day 10
Day 94
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 5 COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZE
Blood / Liver
Blood / Liver
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Blood / Liver
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
APPENDICES
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
APPENDIX A Individual Final Body Weights, Liver Weights, and Relative Liver to Body Weight
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN OIL (NEGATIVE (20NTROL)
INDI\^IDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g ) , AND RELATIVE LI\n3R TO BODY WEIG;HT OF MALE RATS
Animal
Number 627789
627801
627805
627810
627818
Final
Body Wt. 247.7
240.1
283.7
265.0
260.2
GROUP I
Liver Wt.
11.428 12.216 14.081 13.395 11.981
Relative Liver to Body Wt. 0.046
0.051
0.050
0.051
0.046
Test Day
5 5 5 5 5
Animal Number 627793
627797
627799
627800 627808
Final
Body Wt. 283.5
324.2
294.7
331.3
321.8
GROUP II
Liver Wt. 15.292 15.803 14.285 16.344 17.580
Relative Liver to Body Wt. 0.054
0.049
0.048
0.049
0.055
Test Day
10
10 10
10 10
Animal Number 627796 627806
627807
627809
627816
Final
Body Wt.
322.7
320.3
305.2
336.2
330.2
GROUP III
Liver Wt. 13.404 15.413
14.935 14.967 15.811
Relative Liver
to Body Wt. 0.042 0.048 0.049 0.045 0.048
Test Day
13 13 13 13
13
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
0 ORN OIL (NEGATIVE; <CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g) ' AND
RELATIVE LIVER TO BODY WEIG;HT OF MALE RATS
Animal
Number 627791 627794 627804
627812
627817
Final
Body Wt.
365.6
416.7
378.1
388.4
407.3
GROUP IV
Liver Wt. 15.371 19.105 15.326 16.775 16.600
Relative Liver to Body Wt.
0.042 0.046 0.041 0.043 0.041
Test Day
24 24
24 24 24
Animal Number 627792
627798 627813
627814
627815
Final
Body Wt. 496.8
517.3
416.8
386.9
505.9
GROUP V
Liver Wt. 16.359
19.293 15.234 14.358 20.958
Relative Liver
to Body Wt. 0.033
0.037
0.037
0.037
0.041
Test Day
52
52 52
52 52
Animal Number 627795 627811
627819
627820
627821
Final
Body Wt.
485.0
511.6
653.1
618.0
521.5
GROUP VI
Liver Wt. 16.649.
17.405 21.828 26.766 17.943
Relative Liver to Body Wt.
0.034
0.034
0.033
0.043
0.034
Test Day
94 94 94 94
94
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN <3IL:ACETONE (NEGAT IVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g) l AND
RELATIVE LIVER TO BODY WEIGHT OP MALE RATS
Animal
Number 625361
625363
625364
625380
625383
Final
Body Wt. 349.7
286.7
312.0
305.5
313.1
GROUP I
Liver Wt.
16.800 11.236 13.845 13.253 13.026
Relative Liver to Body Wt. 0.048
0.039
0.044
0.043
0.042
Test Day
5 5 5 5 5
Animal
Number 625355
625362
625367
625368
625378
Final
Body Wt. 333.6
332.9
363.8
386.7
365.4
GROUP II
Liver Wt. 14.409
14.363 16.139 17.745 15.328
Relative Liver
to Body Wt. 0.043
0.043
0.044
0.046
0.042
Test Day
10 10 10 10 10
Animal
Number 625357
625358
625370
625384
625385
Final
Body Wt. 405.6
340.7
407.0
357.1
377.6
GROUP III
Liver Wt. 20.091 12.954 21.654 16.520 17.355
Relative Live:r to Body Wt. 0.050
0.038
0.053
0.046
0.046
Test Day
13 13
13 13 13
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H-24002; Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN lOIL:ACETONE (NEGAT:IVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g) / AND
RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 625354
625365
625371
625376
625381
Final
Body Wt. 396.1
459.6
447.3
432.9
381.8
GROUP IV
Liver Wt.
16.383 18.313 17.859 19.168 14.135
Relative Liver to Body Wt.
0.041
0.040
0.040
0.044
0.037
Test Day
24 24 24 24 24
Animal Number 625356
625359
625360
625372
625374
Final
Body Wt. 533.7
530.3
555.4
506.2
512.3
GROUP V
Liver Wt. 20.498 22.901 23.312 21.287 18.378
Relative Live:r to Body Wt. 0.038
0.043
0.042
0.042
0.036
Test Day
52
52 52
52 52
Animal
Number 625353
625369
625373
625375
625377
Final
Body Wt. 565.3
514.8
673.7
588.8
496.6
GROUP VI
Liver Wt.
18.655 17.379 20.073 18.869 17.261
Relative Live:r to Body Wt.
0.033 0.034 0.030 0.032 0.035
Test Day
94 94 94 94
94
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H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24 019 (POSITIVE (:ONTROL) INDIVI DUAL FINAL BOI3Y WEIGHTS (g), LIVER WEIGHTS (g ) , AND
RELATIVE LIV ER TO BODY WEK3HT OF MALE RATS
Animal
Number 625322 625323
625336
625343
625349
Final
Body Wt. 329.0
298.0
318.1
294.0
293.7
GROUP I
Liver Wt. 16.409 14.830 16.545 13.809 15.685
Relative Liver to Body Wt. 0.050
0.050
0.052
0.047
0.053
Test Day
5 5
5 5
5
Animal Number 625320
625330
625332
625341
625350
Final
Body Wt. 312.1
310.1
318.3
325.6
284.3
GROUP II
Liver Wt. 18.845 16.343
17.528 18.254 15.709
Relative Liver to Body Wt.
0.060
0.053
0.055
0.056
0.055
Test Day
10 10 10 10
10
Animal Number 625321
625327
625342 625345
625348
Final
Body Wt.
371.2
355.3
324.4
336.9
336.4
GROUP III
Liver Wt. 22.322
22.259 20.718 24.403 20.541
Relative Liver to Body Wt. 0.060
0.063 0.064 0.072 0.061
Test Day
13 13 13 13 13
-28- Company Sanitized. Does not contain TSCA CBf
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-241019 (POSITIVE CONTROL)
INDIV?IDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g ) , AND
RELATIVE LI\rER TO BODY WEI'GHT OF MALE RATS
Animal Number 625325
625328
625338
625340
625352
Final
Body Wt. 381.3
423.9
339.9
419.0
375.3
GROUP IV
Liver Wt. 21.479 22.100 18.018 23.131 21.835
Relative Liver to Body Wt. 0.056
0.052
0.053
0.055
0.058
Test Day
24 24 24 24 24
Animal Number 625324
625329
625333
625334
625344
Final
Body Wt. 447.4
498.1
427.4
426.5
391.0
GROUP V
Liver Wfc. 20.900
20.911 20.508 19.144 17.666
Relative Liver to Body Wt.
0.047
0.042
0.048
0.045
0.045
Test Day
52 52 52
52 52
Animal Number 625326
625335
625346
626347
625351
Final
Body Wt. 577.5
579.5
556.9
638.9
636.8
GROUP VI
Liver Wt. 23.749 21.548 23.945 27.416 25.615
Relative Liver to Body Wt.
0.041
0.037
0.043
0.043
0.040
Test Day
94 94 94 94
94
-29-
Company Sanitized. Does not contain TSCA CBf
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-241020 (POSITIVE (:ONTROL)
INDI\riDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g ) , AND RELATIVE LI\FER TO BODY WEK3HT OF MALE RATS
Animal Number 627756
627767
627771
627773
627777
Final
Body Wt. 237.5
248.6
239.6
262.3
236.0
GROUP I
Liver Wt. 15.360 19.033 16.968 17.629 16.268
Relative Liver to Body Wt.
0.065
0.077
0.071
0.067
0.069
Test Day
5 5
5 5
5
Animal Number 627768
627774
627776
627782
627785
Final
Body Wt. 293.5
338.2
254.4
268.2
267.9
GROUP II
Liver Wt. 22.536 27.332
18.821 20.172 20.857
Relative Liver to Body Wt. 0.077
0.081
0.074
0.075
0.078
Test Day
10 10 10
10 10
Animal
Number 627758
627763
627764
627787
627788
Final
Body Wt. 308.7
285.8
306.7
234.9
309.4
GROUP III
Liver Wt. 26.021 22.123 21.526 16.498 22.730
Relative Liver to Body Wt. 0.084
0.077
0.070
0.070
0.073
Test Day
13 13
13 13
13
-30-
Company SanRlzed. Does not contain T8CA CBi>
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-2'1020 (POSITIVE CONTROL)
INDI\riDUKL FINAL BODY WEIGHTS (g),, LIVER WEIGHTS (gr), AND RELATIVE Ll\/ER TO BODY WEI GHT OF MALE RATS
Animal Number 627759
627761
627770
627779
627786
Final
Body Wt.
380.1
359.4
379.8
406.2
326.1
GROUP IV
Liver Wt. 22.707 19.579 19.540 24.858 16.410
Relative Liver to Body Wt. 0.060
0.054
0.051
0.061
0.050
Test Day
24 24 24 24 24
Animal Number 627757
627760
627765
627772
627781
Final
Body Wt.
436.7
456.5
455.6
513.5
421.0
GROUP V
Liver Wt. 16.899 18.240 16.505 20.225 16.695
Relative Liver
to Body Wt. 0.039
0.040
0.036
0.039
0.040
Test Day
52
52 52 52 52
Animal Number 627762
627769
627775
627778
627783
Final
Body Wt. 528.6
609.6
552.8
445.3
557.2
GROUP VI
Liver Wt. 17.732 19.324 17.296 15.628 21.044
Relative Liver to Body Wt. 0.034
0.032
0.031
0.035
0.038
Test Day
94 94
94 94 94
-31-
Company Sanitized. Does not contain TSCA CB1
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-:24002 (TEST SUB;STANCE)
IND][VIDUAL FINAL BO DY WEIGHTS (g), LIVER WEIGHTS (g ) , AND RELATIVE L^/ER TO BODY WEK3HT OF MALE RATS
Animal
Number 628192
628195
628207
628211
628212
Final
Body Wt. 251.3
279.5
269.9
250.9
281.1
GROUP I
Liver Wfc. 13.034 14.705 14.429 12.800 13.611
Relative Liver to Body Wt.
0.052
0.053
0.053
0.051
0.048
Test Day
5 5
5 5
5
Animal Number 628187
628188
628199
628209
628217
Final
Body Wt. 314.4
328.1
315.3
299.3
283.4
GROUP II
Liver Wt. 14.111 16.885 16.606 15.575
13.716
Relative Liver to Body Wt.
0.045
0.051
0.053
0.052
0.048
Test Day
10 10 10
10 10
Animal Number 628190
628202
628204
628206
628214
Final
Body Wt. 314.2
318.6
361.0
320.0
296.9
GROUP III
Liver Wt. 14.735 16.595 18.015 15.690 12.414
Relative Liver to Body Wt.
0.047 0.052 0.050 0.049 0.042
Test Day
13 13 13
13
13
-32- jCpmpanySanHIzed. Does not contain TSCA CB
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-;'4002 (TEST SUBSTANCE) INDIVI DUAL FINAL BODY WEIGHTS (g),, LIVER WEIGHTS (g ) , AND
RELATIVE LI\TER TO BODY WE I GHT OF MALE RATS
Animal Number 628197
628198
628201
628203
628216
Final
Body Wt. 345.6
378.8
362.8
423.4
348.9
GROUP IV
Liver Wt.
13.523 18.369 15.533 20.489 15.704
Relative Liver to Body Wt.
0.039
0.048
0.043
0.048
0.045
Test Day
24
24 24 24
24
Animal
Number 628191
628196
628208
628210
628215
Final
Body Wt.
498.3
565.9
543.3
542.0
514.0
GROUP V
Liver Wt. 17.776 21.754 20.901 20.735 18.644
Relative Liver to Body Wt.
0.036
0.038
0.038
0.038
0.036
Test Day
52 52 52 52 52
Animal Number 628194
628200
628213
628218
628219
Final
Body Wt. 578.2
556.4
609.7
633.8
591.2
GROUP VI
Liver Wt. 21.190 16.602 21.672 22.255 19.050
Relative Liver to Body Wt.
0.037 0.030 0.036 0.035 0.032
Test Day
94 94
94 94 94
^
-33-
:Companf Sanitized. Does net contain TSCA CBI
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
APPENDIX B Individual Fluorine Levels
-34.
Company Sanitized. Does not contain TSCA CBI
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
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 [mmbl/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
-35-
Company Sanitized. Does not contain T8CA CBI
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppm F in liver
The ppm fluoride measured in liver
Individual Animal Calculations:
ppm F in Liver minus Bkg 0:2 ppm
ppm F in Liver normalized to 0.1 mmol/kgDose
The ppm fluoride measured in liver minus the background fluoride measured in control animal liver. 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 liver 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)
ju,molar equivalents of active in liver
The ^.molar [^mol/L] concentration of fluorine containing compound
in liver based on the ppm fluorine in liver normalized to 0.1 mmol/kg active dose. This assumes that all liver fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm -
Img/L = (Normalized ppm [mg/L] fluorine in liver / Mol Wt F
[mg/mmol]) x molar ratio active/F [mmol active/mmol F] x 1000 ^mol/mmol
-36-
Company Sanitized. Does not contain TSCA CR
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)
- Liver 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
-37-
Company Sanitized. Does not contain TSCA CP
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24019
DuPont-2926 Supplement No. 1
Given:
Mol Wt. Active (g/mole):
497
Formulation Dose (mg/kg):
10
% Active (F Containing) in Formulation: 100
%F in Aictive:
65
Mol Wt. F (g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
10 0.020 49.7
Dose F (mmol/kg):
0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):_____6.5
Rat Number
Test Day Sample
ppmF
in
Liver
Group II
625320
10
319.4
625330
10
355.4
625332
10
338.4
625341
10
305.0
625350
10
322.7
Group VI
625326
94
625335
94
625346
94
625347
94
625351
94
79.8 89.6 73.8 62.5 82.1
ppm F in Liver
Minus Bkg 0.2 ppm
319.2 355.2 338.2 304.8 322.5
79.6 89.4 73.6 62.3 81.9
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
1586.42 1765.34 1680.85 1514.86 1602.83
395.61 444.32 365.79 309.63 407.04
/imolar Equivalents of Active in
Liver
4910.77 5464.62 5203.08 4689.23 4961.54
1224.62 1375.38 1132.31 958.46 1260.00
-38- jCompany Sanitized. Does not contain TSCA C91
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24020
DuPont-2926 Supplement No. 1
Given:
Mol Wt. Active (g/mole):
426
Formulation Dose (mg/kg):
20
% Active (F Containing) in Formulation: 100
%F in Act ive:
69
Mol Wt.F (g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
20 0.047 42.6
Dose F (mmol/kg):
0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):_____13.8
Rat Number
Test Day Sample
ppmF
in
Liver
Group II
627768
10
627774
10
627776
10
627782
10
627785
10
182.5 109.0 174.0 147.5 165.8
Group VI
627778
94
1.9
627783
94
1.2
627762
94
1.9
627775
94
1.6
627769
94
1.8
ppm F in Liver
Minus Bkg 0.2 ppm
182.3 108.8 173.8 147.3 165.6
1.7 1.0 1.7 1.4 1.6
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
388.30 231.74 370.19 313.75 352.73
3.62 2.13 3.62 2.98 3.41
/imolar Equivalents of Active in
Liver
1321.01 788.41
1259.42 1067.39 1200.00
12.32 7.25 12.32 10.14 11.59
-39
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24002
DuPont-2926 Supplement No. 1
Given:
Mol Wt. Active (g/mole):
570
Formulation Dose (mg/kg):
75
% Active (F Containing) in Formulation: 47
% Fin Active:
51
Mol Wt. F (g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg):
Formulation Dose Normalization Factor:
35.25 0.062 121.3
Dose F (mmol/kg):
0.946
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):_____17.9775
Rat Number
Test Day Sample
ppmF
in
Liver
Group II
628209
10
628187
10
628199
10
628188
10
628217
10
99.6 128.5 123.4 131.4 144.1
Group VI
628218
94
5.1
628213
94
2.3
628194
94
2.3
628200
94
2.6
628219
94
2.7
ppm F in Liver
Minus Bkg 0.2 ppm
99.4 128.3 123.2 131.2 143.9
4.9
2.1 2.1
2.4 2.5
ppm F in L iver
Normalize*ito
0.1 mmoles/kg Dose
160.73 207.46 199.22 212.15 232.69
7.92 3.40 3.40 3.88 4.04
/imolar Equivalents of Active in
Liver
552.91 713.67 685.30 729.80 800.45
27.26 11.68 11.68 13.35 13.91
40-
.CompanySanitized. Does not contain TSCA fw