Document 0LaGD59MMzJzXME6m5vaqq3NV
TRADE SECRET Study Title
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Supplement No. 1
Laboratory Project ID: DuPont-2926
DuPont-2926 Supplement No. 1
AR226-3216
AUTHOR: Carol Finlay, 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
W ork Request Number:
Service Code Number:
Page 1 of 40
Company Sanitized. D oes not contain TSCA CBI
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Reviewed by: f r r * ~ ~ * * * * % l ' ' Paul M. Hmderliter, Ph.D. Postdoctoral Fellow
Reviewed by:
^ Z )u jL c d C Jc tm o U b u O / 7 Judith C. Stadier. PhX., D.A.B.T. ' / Director
Issued by Study Director:
t( L A b t iu fo iA u , Carolfenlay.B.A^
Staff Scientist
Date
/D -JT fA -2 O O . Dale
11' (i)atc
s-VF**--
-2-
Company Sanitized. D oes not contain TSCA CB)
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_______
DuPont-2926 Supplement No. 1
TABLE OF CONTENTS
Page
CERTIFICATION............................................................................................................................. -2
LIST OF TABLES................................................................................................................................. 4
LIST OF FIGURES............................................................................................................................... 4
LIST OF APPENDICES...................................................................................................................... 4
STUDY INFORM ATION.................................................................................................................... 5
STUDY PERSONNEL.................................................................................
?
REASON FOR SUPPLEMENT NO. 1 ..............................................................................................8
SUMMARY............................................................................................................................................. 9
MATERIALS AND M ETHODS...................................................................................................... 10
RESULTS AND DISCUSSION.........................................................................................................10
A. Liver Weights.............................................................................................................
10
1. Test substance................................................................................................................................................ 10
2. Positive Controls.............................................
10
B. Liver Fluorine D ata................................................................................................................... 11
1. Positive Controls...........................................................................................................................................11
2. Test Substance............................................................................................................................................... 11
C O N C L U S IO N S ...................................................................................................................................11
TABLES.......................................................................................................................
12
FIGURES................................................................................................................................................16
APPENDICES.......................................................................................................................................22
-3 Company Sanitized. D oes not contain TSCA CBl
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
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................................................................................................................................................................. I7
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 D OSE.........................................................................................................................................21
LIST OF APPENDICES
Page
A. INDIVIDUAL FINAL BODY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY W EIGHT...................................................................................................................................................................... 23
B. INDIVIDUAL FLUORINE LEVELS........................................................................................................................34
-4Company Sanitized. D oes no! contain TSCA CBf
H-24002: Biprsistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION TEST SUBSTANCE:
Substance Tested:,
Haskell Number: 24002
Known Impurities: Unknown POSITIVE CONTROL:
Substance Tested:
DuPont-2926 Supplement No. 1
Known Impurities: Unknown
-5 Company Sanitized. Does not contain TSCA
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORM ATION (Continued) POSITIVE CONTROL:
Substance Testei
DuPont-2926 Supplement No. 1
Haskell Number: 24020
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-LifeInitiated/Completed: July 11, 1999/O ctober 12, 1999
-6-
Company Sanitized. Does not eontain TSCA CBi
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
STUDY PERSONNEL Study Director: Carol Finlay, B.A.
Management: Judith C. Stadler, Ph.D., D.A.B.T. Primary Technician: 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.
-7 Company Sanitized. Doe* ho! certain TSCA C tl
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 (com oil and com oil/acetone).
.
%
-8-
Company Sanitized. D oes not contain TSCACBl
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 ,1 0 ,1 3 ,2 4 ,5 2 ,9 4 ) 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, com oil and com oil/acetone (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 /xM equivalents on day 10 and 15.58 /xM equivalents on day 94. On day 10, mean jxM 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 fiM 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.
-9 Company Sanitized. D oes not contain TSCA cr|
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 com 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.
1 0- -
Coropany Sanitized. D oes not contain TSCA CBI
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 fiM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15 and 10.72 fiM 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 fiM equivalents at day 10 and 15.58 fiM 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 ptM 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.
-11 -
.Company SanRIzed. D oes not contain TSCA CB1
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TABLES
- 12Oompany Sanitized. D oes no! contain TSCA CBI
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
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)
Absolute
Body Weight
Liver Weight
259.3 311.1
12.620 15.861
322.9 391.2
14.906 16.635
464.7 557.8
17.240 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 OIL:ACETONE (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 10 13 24 52 94
H-24019 (POSITIVE CONTROL)
Absolute
Body Weight
Liver Weight
306.6
15.456
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)
0.050 0.056 0.064 0.055 0.045 0.041
-13-
Company Sanitized. D oes not contain TSCA CB
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
Test Days 5 10 13 24 52 94
TABLE 2 (Continued)
MEAN BODY AND LIVER WEIGHTS (g)
H-24020 (POSITIVE CONTROL)
Absolute
Body Weight
Liver Weight
244.8
17.052
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)
0.070 0.077 0.075 0.056 0.039 0.034
Test Days 5 10 13 24 52 94
H-24002 (TEST SUBSTANCE)
Absolute
Body Weight
Liver Weight
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
1 4- -
Company Sanitized. D oes not contain TSCA CBI
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
TABLE 2 MEAN LIVER FLUORINE LEVELS
Test Days
10 94
Negative Controls
Com
Com Oil Oil: Acetone
(ppm)
(ppm)
<0.2 <0.5
0.6a l a
Positive Controls
H-24019 (ppm)
328.18 (19.3)b 77.56 (10.1)
H-24020 (ppm)
155.76 (29.2) 1.68 (0.3)
a One value. Four of the values were below the level of detection (LOD). b Standard deviation is in parentheses.
Test Substance
H-24002 (ppm)
125.4 (16.3) 3.0 (1.19)
TABLE 3
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Pays
10 94
Positive Controls
Test Substance
H-24019
H-24020
H-24002
/iM F Equivalents iM F Equivalents______ fiM F Equivalents
5045.85 (296.9l) a 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.
-15-
Company Sanitized. D oes not contain TSCA CB!
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
FIGURES
- 16-
jCompany Sanitized. D oes not contain TSCA CBS
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_________________________________________________
DuPont-2926 Supplement No. 1
0.060.05-
FIGURE 1
COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE CONTROLS
tUCorn Oil (Negative Control) HCorn OihAcetone (Negative Control) H-24002 (Test Substance)
j 0.04-
V '-'U"2'
3* i3Va $2a 0.03-
0 .0 2 -
0.01
0.00-
T est D ay
Company SanRIzed. Does notconfain TSCA
-17-
H-24002: Biopersistence Screening
DuPont-2926
10-Dose Oral Gavage Study in Rats______________________________________________________________________________________________ Supplement No. 1
FIGURE 2
COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS
CompanySanitized. Does no!contain TSCACRi
-18-
.1.
H-24002: Biopersistence Screening
DuPont-2926
10-Dose Oral Gavage Study in Rats______________________________________________________________________________________________Supplement No. 1
FIGURE 3
COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN OVER FLUORINE CONCENTRATION FOR H-24002 AND NEGATIVE CONTROLS
CompanySanitized. Does notcontain TSCACB1
El Com Oil (Negative Control) Com Oil:Acetone (Negative Control) H-24002 (Test Substance)
- 19-
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats__________________________________________________________________
FIGURE 4 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
DuPont-2926 Supplement No. 1
6000 5000
GjUH-24019 ( Positi ve Control) H-24020 (Positive Control) H-24002 (Test Substance)
ipanySanitized. Does notcontain TSCACBl
D ay 10
-20-
D ay 94
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
FIGURE 5 COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
pM Equivalent of Fluorine
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
APPENDICES
2 2- -
Company Sanitized. D oes not contain TSCA CBf
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
2 3- -
Company Sanitized. D oes not contain TSCA CB1
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627789
627801
627805
627810 627818
Final Body W t .
247.7
240.1
283.7
265.0
260.2
GROUP I
Liver W t . 11.428 12.216 14.081 13.395 11.981
Relative Liver to Body W t . 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 W t .
283.5
324.2
294.7
331.3
321.8
GROUP II
Liver W t . 15.292 15.803 14.285 16.344 17.580
Relative Liver to Body W t . 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 W t .
322.7
320.3
305.2
336.2 330.2
GROUP III
Liver W t . 13.404 15.413 14.935 14.967 15.811
Relative Liver to Body W t . 0.042
0.048
0.049
0.045
0.048
Test Day 13 13 13 13 13
- 24-
Company Sanitized. Does not contain TSCA CEt
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g) , AND RELATIVE LIVER TO BODY WEIGHT 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 W t . 15.371 19.105 15.326 16.775 16.600
Relative Liver to Body W t . 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 W t . 16.359 19.293 15.234 14.358 20.958
Relative Liver to Body W t . 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 W t .
485.0
511.6
653.1
618.0
521.5
GROUP VI
Liver W t . 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 : 94 94 94 94 94
- 25Company Sanitized. D oes not contain TSCA CB
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN OIL:ACETONE (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS ( g ) , LIVER WEIGHTS ( g ) , AND RELATIVE LIVER TO BODY WEIGHT OF 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 W t . 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 W t .
333.6
332.9 363.8
386.7
365.4
GROUP II
Liver W t . 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 W t .
405.6 340.7
407.0
357.1
377.6
GROUP III
Liver W t . 20.091 12.954 21.654 16.520 17.355
Relative Liver to Body W t . 0.050
0.038
0.053 0.046
0.046
Test Day 13 13 13 13 13
- 26-
Company Sanitized. D oes not contain TSCA CB'
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
CORN OIL:ACETONE (NEGATIVE 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 W t .
396.1 459.6
447.3
432.9
381.8
GROUP IV
Liver W t . 16.383 18.313 17.859 19.168 14.135
Relative Liver to Body W t . 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 W t .
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 Liver to Body W t . 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 W t .
565.3
514.8
673.7
588.8
496.6
GROUP VI
Liver W t . 18.655 17.379 20.073 18.869 17.261
Relative Liver to Body W t . 0.033
0.034
0.030
0.032
0.035
Test Day 94 94 94 94 94
2 7- -
Company Sanitized. D oes not contain TSCA CBF
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24019 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT 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 W t . 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 W t . 18.845 16.343 17.528 18.254 15.709
Relative Liver to Body W t . 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 W t .
371.2
355.3
324.4
336.9 336.4
GROUP III
Liver W t . 22.322 22.259 20.718 24.403 20.541
Relative Liver to Body W t . 0.060
0.063
0.064
0.072
0.061
Test Day 13 13 13 13 13
2 8- -
Company Sanitized. D oes not contain TSCA CBf
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24019 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT 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 W t . 21.479 22.100 18.018 23.131 21.835
Relative Liver to Body W t . 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 W t .
447.4
498.1 427.4
426.5
391.0
GROUP V
Liver W t . 20.900 20.911 20.508 19.144 17.666
Relative Liver to Body W t . 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 W t .
577.5 579.5
556.9
638.9
636.8
GROUP VI
Liver W t . 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
2 9- -
jCompany Sanitized. D oes not contain TSCA CBf
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24020 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627756
627767
627771
627773
627777
Final Body W t .
237.5
248.6
239.6
262.3
236.0
GROUP I
Liver W t . 15.360 19.033 16.968 17.629 16.268
Relative Liver to Body W t . 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 W t .
293.5
338.2 254.4
268.2
267.9
GROUP II
Liver W t . 22.536 27.332 18.821 20.172 20.857
Relative Liver to Body W t . 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 W t .
308.7
285.8
306.7
234.9
309.4
GROUP III
Liver W t . 26.021 22.123 21.526 16.498 22.730
Relative Liver to Body W t . 0.084 0.077
0.070 0.070
0.073
Test Day 13 13 13 13 13
- 30-
Company Sanitized. D oes not contain TSCA CB
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24020 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627759
627761
627770
627779
627786
Final Body W t .
380.1 359.4
379.8
406.2
326.1
GROUP IV
Liver W t . 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 W t .
436.7
456.5 455.6
513.5
421.0
GROUP V
Liver W t . 16.899 18.240 16.505 20.225 16.695
Relative Liver to Body W t . 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 W t .
528.6 609.6
552.8 445.3
557.2
GROUP VI
Liver W t . 17.732 19.324 17.296 15.628 21.044
Relative Liver to Body W t . 0.034
0.032
0.031
0.035
0.038
Test Day 94 94 94 94 94
-31 Company Sanitized. D oes not contain TSCA CBl
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24002 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 628192
628195
628207
628211
628212
Final Body W t .
251.3
279.5
269.9
250.9
281.1
GROUP I
Liver W t . 13.034 14.705 14.429 12.800 13.611
Relative Liver to Body W t . 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 W t .
314.4
328.1 315.3
299.3
283.4
GROUP II
Liver W t . 14.111 16.885 16.606 15.575 13.716
Relative Liver to Body W t . 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 W t .
314.2
318.6
361.0
320.0
296.9
GROUP III
Liver W t . 14.735 16.595 18.015 15.690 12.414
Relative Liver to Body W t . 0.047
0.052
0.050
0.049
0.042
Test : 13 13 13 13 13
3 2- -
jDompany Sanitized. D oes not contain TSCA CBi
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
H-24002 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT 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 W t . 13.523 18.369 15.533 20.489 15.704
Relative Liver to Body W t . 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 W t .
498.3
565.9
543.3
542.0
514.0
GROUP V
Liver W t . 17.776 21.754 20.901 20.735 18.644
Relative Liver to Body W t . 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 W t .
578.2
556.4
609.7
633.8
591.2
GROUP VI
Liver W t . 21.190 16.602 21.672 22.255 19.050
Relative Liver to Body W t . 0.037
0.030
0.036
0.035
0.032
Test Day 94 94 94 94 94
- 33-
[Company 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
DuPont-2926
10-Dose Oral Gavage Study in Rats____________________________________________________ Supplement No. 1
Terms: Active % Active
Mol W t 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
-35-
Company Sanitized. Does not contain TSCA CBs
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
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.
ppm F in Liver 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 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)
/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 -
1 mg/L = (Normalized ppm [mg/L] fluorine in liver / Mol W t F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x 1000 nmol/mmol
- 36-
Company Sanitized. Does not contain TSCA CR
H-24002: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2926 Supplement No. 1
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)
- 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): 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): Formulation Dose Normalization Factor:
10
Dose F (mmol/kg):
0.342
0.020 Molar Ratio (Active/F): 0.059
49.7_____ Dose F (mg/kg):________ 6.5
Rat Number
Test Day Sample
ppm F 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 625335 625346 625347 625351
94 94 94 94 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
molar Equivalents o f 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 CB1
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): 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): 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
ppm F in
Liver
Group II 627768 627774 627776 627782 627785
10 10 10 10 10
182.5 109.0 174.0 147.5 165.8
Group VI 627778 627783 627762 627775 627769
94 94 94 94 94
1.9 1.2 1.9 1.6 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
% F in Active: Mol Wt. F (g/mol):
51 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 99.6
628187
10 128.5
628199
10 123.4
628188
10 131.4
628217
10 144.1
Group VI 628218 628213 628194 628200 628219
94 94 94 94 94
5.1 2.3 2.3 2.6 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 Liver Normalized to 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
Hmolar 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-
.Company Sanitized. Does not contain TSCA cm
