Document jYjbeg0Xar0rzganYyK1Y9xZ
AR226-3210
DuPont-2921 Supplement No. 1
TRADE SECRET Study Title
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Supplement No. 1
Laboratory Project ID: DuPont-2921
AUTHOR: Carol Finlay, B.A.
Study Completed on: August 29,2000
Supplement No . 1 Completed on: December 13,2001
Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
W ork Request Number:
Service Code Number:
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Reviewed by:
'aul M. Hinderliter, Ph.D. Postdoctoral Fellow
Reviewed by:
Judith C. Stadler, Ph.D., D A B .T . Director
Issued by Study Director:
t l A C A j ^
Carolfinlay, B.A. Q
Staff Scientist
13-j3rc-2-",f
Date
Date
Date
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
TABLE OF CONTENTS
Page
CERTIFICATION.............................................................................................................................. ...2
LIST OF TABLES................................................................................................................................. 4
LIST OF FIGURES............................................................................................................................... 4
LIST OF APPENDICES............................................................................................................... 4
STUDY INFORMATION.................................................................................................................... 5
STUDY PERSONNEL...........................................................................................................................7
REASON FOR SUPPLEMENT NO. 1 ......................................................................................... . ..8
S U M M A R Y ....................... ......................................................................................................................9
MATERIALS AND M ETHODS...................................................... ................................................10
RESULTS AND DISCUSSION.........................................................................................................10 A. Liver D ata...................................................................................................................................10
1. Test Substance................................................................................................................................................................ 10 2. Positive C ontrols............................................................................................................................................................10
B. Liver Fluorine D ata....................................................................................................................10
1. Positive C ontrols............................................................................................................................................................11 2. Test Substance..................................................................... ........................................................ .................................11
CONCLUSIONS...................... ............................................................................................................11
TABLES............................................................................................................................................ .....12
FIGURES...............................................................................................................
16
APPENDICES..............................................................
22
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
LIST OF TABLES
Page
1. M EA N BO DY A N D LIVER W EIGHTS.......... .................................................................................................................. ,.13 2. M EAN LIVER FLUORINE LEVELS..................................................................................................................................... 15 3. M EAN LIVER FLUORINE CONCENTRATION NORMALIZED TO D O SE.......................................................... 15
LIST OF FIGURES
Page
1. COMPARISON OF M EAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE C ONTROLS.............. '...............'.................................................................................................................................................... 17
2. COMPARISON OF M EAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE A ND POSITIVE C ONTROLS.....................................................................................................................................................................................18
3. COMPARISON OF RELATIVE LIVER WEIGHT A N D MEAN LIVER FLUORINE CONCENTRATION FOR H-23925 A N D NEGATIVE CONTROLS............................................
19
4. M EAN LIVER FLUORINE CONCENTRATION NORMALIZED TO D O SE ...........................................................20
5. COMPARISON OF M EA N BLOOD AND M EAN LIVER FLUORINE CONCENTRATION NORMALIZED TO D O S E .........................................................................................................................................................21
LIST OF APPENDICES
Page
A. INDIVIDUAL FINAL BO DY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY W EIG H T.......................................................................................................................................................................................... 23
B. INDIVIDUAL FLUORINE L E V E L S...................................................................................................................................... 34
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STUDY INFORMATION TEST SUBSTANCE:
DuPont-2921 Supplement No. 1
Known Impurities: None POSITIVE CONTROL:
Known Impurities: Unknown
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORM ATION (Continued) POSITIVE CONTROL:
DuPont-2921 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-Life Initiated/Completed: May 16, 1999 / August 17,1999
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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.
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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-23925), 2 positive controls (H-24019 and H-24020), and 2 negative controls (com oil and com oil/acetone).
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
SUMMARY
The objective of this study was to evaluate the potential for H-23925, 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 200 mg/kg/day of H-23925. The test substance was administered to one group of 5 rats for 5 consecutive days and to 5 groups for 10 days. 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 livers. 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 and analyzed as described for H-23925.
The mean relative liver weight (liver/body weight) of rats dosed with the test substance, H-23925, was up to 25% higher at day 10 than the mean relative liver weights of the negative control groups. By day 94, the mean relative liver weight of rats dosed with the test substance was 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 liver fluorine concentration of rats dosed with the test substance, H-23925, was 888.28 /xM equivalents on day 10 and 69.47 f i M 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 6-fold (H-24019) and 1.3-fold (H-24020) greater than the /xM equivalent fluorine concentration in livers from rats treated with the test substance. On day 94, the /xM equivalent concentration of H-23925 in the liver was 17-fold lower than the /xM equivalent concentration of H-24019 but 6x higher than the /xM equivalent concentration of H-24020.
The /xM equivalents o f fluorine in the livers from rats treated with the test substance were higher . than levels in the blood.
Under the conditions of this study, there was some absorption and retention of fluorine in the liver from rats dosed with H-23925. Fluorine levels in the liver in rats dosed with H-23925 were lower than levels in rats dosed with the positive control material H-24019. Fluorine levels in the liver in rats dosed with H-23925 were lower on day 10 than the levels in rats dosed with the positive control H-24020 but were higher on day 94. Liver weights were elevated in rats dosed with the test substance, H-23925, at the end of the dosing period, but not at the end of the recovery period.
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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 (Table 1; Figures 1-3; Appendix A)
As with the 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 positive or negative controls. The rats dosed with the test substance, H-23925, were older and heavier in weight than the com oil negative control and H-24020 positive control rats, but younger and lighter in weight than the com oil/acetone negative control and H-24019 positive control rats. Therefore, liver weights relative to total body weight are presented for comparison.
1. Test Substance
The mean relative liver weight of rats dosed with the test substance, H-23925, was up to 25% higher at day 10 than the mean relative liver weights of the negative control groups. By day 94, the mean relative liver weight of rats dosed with the test substance was 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 similar on day 10 to the mean relative liver weight of rats dosed with the test substance. At day 94, the mean relative liver weight of rats dosed with H-24019 was 21% higher than the mean relative liver weight of rats dosed with the test substance. The mean relative liver weight of rats dosed with the other positive control, H-24020, was 40% higher on day 10 than the mean relative liver weight of rats dosed with the test substance. The mean relative liver weights of rats dosed with H-24020 began to decrease during the recovery period and were similar on test days 52 and 94 to the mean relative liver weights of rats dosed with the test substance.
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
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
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.
WdghtS
d0Sed With the test substance- H-23925, were elevated at the end
w e ih , h T f pen0d bUt " 0t at the end of ,he recove,> period. The mean relative liver weight by the end of recovery in rats dosed with the test substance was lower than in rats dosed
with the positive control H-24019 but similar to rats dosed with the positive control H-24020.
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 y 10 from rats dosed with the positive control materials were 5045.85 and 1127 25 M
r
`"
*- --
rions were 1,90.15
2. Test Substance
The total fluorine concentration in the liver from rats dosed with H-23925 was 888.28 uM equivalents at day 10 and 69.47 f i M equivalents at day 94. For the positive control H-24019 the
nfT the e n / f i T
apProximately 6x hiSher (day 10) and approximately 17x higher (end F a H"24020' the liver concentration was approximately 1.3x higher at
H-23925
wTM "
TM
" ** dosed with
ta S "
" in ,he livers from rats dosed wi,h ,he test substance--
CONCLUSIONS
Under the conditions of this study, there was some absorption and retention of fluorine in the
?tSlnwe fthm i OSed Wlt5 H' 23925- Flurine levels in the liver in rats dosed with H-23925 were er than levels;in rats dosed with the positive control material H-24019. Fluorine levels in the hver m rats dosed wiAH-23925 were lower on day 10 than the levels in rats dosed with the positive control H-24020 but were higher on day 94. Liver weights were elevated in rats dosed
recove^ period5813" 06' H ~2 3 9 2 5 ' * the end f the dosing period' but not at the end of the
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DuPont-2921 Supplement No. 1
TABLES
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DuPont-2921 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
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 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
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
Test Days 5 10 13 24 52 94
TABLE 1 (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-23925 (TEST SUBSTANCE)
Absolute
Body Weight
Liver Weight
293.4
14.905
339.4
18.613
321.8
22.310
408.0
22.171
480.3
20.056
538.9
18.451
Mean Relative Liver Weight(% of Body Weight)
0.051 0.055 0.069 0.054 0.042 0.034
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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 o f the values were below the level o f detection (LOD). b Standard deviation is in parentheses. - Indicates no value for this day.
Test Substance
H-23925 (PPm )______
283.56 (78.2) 22.36 (8.7)
TABLE 3
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Pays
10 94
Positive Controls
Test Substance
H-24019
H-24020
H-23925
f i M F Equivalents x M F Equivalents______f i M F Equivalents
5045.85 (296.91)3 1127.25 (211.37)
888.28 (245.18)
1190.15 (156.05)
10.72 (2.14)_______ 69.47 (27.36)
a Standard deviation is in parentheses.
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
FIGURES
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H-23925: Biopersistence Screening
DuPont-2921
10-Dose Oral Gavage Study in Rats________________________________________________________________________________________ Supplement No. 1
0.07
0.06
0.05 0.04
5* I5 M avt 0.03
ca 0.02
0.01
0 .0 0 -
FIGURE 1 COM PARISON OF M EA N RELATIVE LIVER W EIGHTS:
TEST SUBSTANCE A ND NEGATIVE CONTROLS HslCorn Oil ( N eg ativ e C o n t r o l) B C orn Oil:Acetone (Negative Control) H-23925 (Test Substance)
Test Day
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H-23925: Biopersistence Screening
DuPont-2921
10-Dose Oral Gavage Study in Rats________________________________________________________________________________________ Supplement No. 1
FIGURE 2
CO M PARISO N OF M EA N RELATIVE LIVER W EIGHTS: TEST SU B ST A N C E A N D POSITIVE CONTROLS
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5 10 13 24 52 94 Test Day
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DuPont-2921 Supplement No. 1
FIGURE 3
COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-23925 AND NEGATIVE CONTROLS
MeanRelativeUver tW. (Organ:BodyWl)
EH Com Oil (Negative Control) Com Oil:Acetone (Negative Control) H-23925 (Test Substance)
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 4 M EAN LIVER FLUORINE CONCENTRATION NORM ALIZED TO DO SE
#
DuPont-2921 Supplement No. 1
6000
5000
4000
ae tOo' to
sa.
2000
1000
QH-24019 (Positive Control) H-24020 (Positive Control) H-23925 (Test Substance)
D ay 10
D ay 94
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________________________
#
DuPont-2921 Supplement No. 1
FIGURE 5 COM PARISON OF M EA N BLOOD A N D M EAN LIVER FLUORINE CONCENTRATION NORM ALIZED TO D O SE
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DuPont-2921 Supplement No. 1
APPENDICES
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DuPont-2921 Supplement No. 1
APPENDIX A Individual Final Body Weights, Liver Weights, and Relative Liver to Body Weight
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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
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DuPont-2921 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 W t .
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 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 W t .
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 W t . 0.034
0.034
0.033
0.043
0.034
Test Day 94 94
94 94 94
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DuPont-2921 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 W t . 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 W t . 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
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DuPont-2921 Supplement No. 1
CORN O I L :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 W t . 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
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DuPont-2921 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 W t . 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 W t .
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
- 28-
Sompany Sanitized. Does no! contain TSCA CBI
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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 W t .
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 W t . 0.041
0.037
0.043
0.043
0.040
Test Day 94 94 94
94 94
- 29Pompany Sanitised. Does no! contain TSCA r*"
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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 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 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
- 30Company Sanitized. Does not contain TSCA CBi
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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 Wt. 22.707
19.579 19.540 24.858 16.410
Relative Liver to Body W t . 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. Does not contain TSCA CBf
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-23925 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 626399
626407
626426
626427
626428
Final Body W t .
307.9
256.1
298.6
321.7
282.9
GROUP I
Liver W t . 15.381 13.622 14.857 16.687 13.978
Relative Liver to Body Wt. 0.050
0.053
0.050
0.052
0.049
Test Day 5 5
5 5 5
Animal Number 626396
626397
626403
626406
626421
Final Body W t .
342.7
335.7
337.0
333.8
347.7
GROUP II
Liver W t . 19.454 17.422 18.141 17.180 20.866
Relative Liver to Body W t . 0.057
0.052
0.054
0.051
0.060
Test Day 10
10 10
10 10
Animal Number 626402
626404
626409
626417
626418
Final Body W t .
345.6
277.7
320.3
300.4
364.9
GROUP III
Liver Wt. 21.310 21.304 21.848
23.841 23.248
Relative Liver to Body W t . 0.062
0.077
0.068
0.079
0.064
Test Day 13
13 13
13 13
- 32Company SanJIIzd. Does Wotcontain TSCA CBI
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-23925 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 626405
626414
626419
626423
626423
Final Body W t .
417.7
402.5
395.6
427.7
396.6
GROUP IV
Liver W t . 22.726
23.389 22.530 21.703 20.507
Relative Liver to Body W t . 0.054
0.058
0.057
0.051
0.052
Test Day 24 24 24 24 24
Animal Number 626411
626415
626416
626422
626425
Final Body W t .
430.1
440.0
508.3
509.7
513.6
GROUP V
Liver W t . 18.450 19.086 20.697 21.089 20.960
Relative Liver to Body W t . 0.043
0.043
0.041
0.041
0.041
Test Day 52 52
52 52 52
Animal Number 626398
626401
626408
626410
626413
Final Body W t .
505.5
455.6
493.4
542.4
697.5
GROUP VI
Liver W t . 17.470 16.843 15.781 18.412 23.750
Relative Liver to Body Wt. 0.035
0.037
0.032
0.034
0.034
Test Day 94
94 94 94 94
-33-
Cotnpany Sanitized. Does no*contain TSCA CB
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
APPENDIX B Individual Fluorine Levels
- 34-
Company Sanitized. Does notcontain TSCA CBI
H-23925: Biopersistence Screening
DuPont-2921
10-Dose Oral Gavage Study in Rats_______________________________________________________ Supplement No. 1
TERMS AND CALCULATIONS
Terms: Active % Active
Mol W t Active Formulation Dose % F in Active
Mol W t F
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
m
-35-
Oompany Sanitized. Does notcontain TSCA CBl
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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)
ixmolar 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 Wt F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x 1000 /mol/mmol
-36Company Sanitized. Does not contain TSCA CB)
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 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 CW
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24019
DuPont-2921 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 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
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 III 625321 625327 625342 625345 625348
13 13 13 13 13
255.9 275.3 277.9 260.4 275.6
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
255.7 275.1 277.7 260.2 275.4
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
1270.83 1367.25 1380.17 1293.19 1368.74
395.61 444.32 365.79 309.63 407.04
gmolar Equivalents of Active in
Liver
4910.77 5464.62 5203.08 4689.23 4961.54
3933.85 4232.31 4272.31 4003.08 4236.92
1224.62 1375.38 1132.31 958.46 1260.00
-38-
Company Sanitized. Does not co n ta in t s c a C 5 i
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24020
DuPont-2921 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
10 182.5
627774
10 109.0
627776
10 174.0
627782
10 147.5
627785
10 165.8
Group III 627758 627764 627787 627763 627788
13 13 13 13 13
61.5 40.5 33.3 51.3 45.2
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
61.3 40.3 33.1 51.1 45.0
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
130.57 85.84 70.50 108.84 95.85
3.62 2.13 3.62 2.98 3.41
/molar Equivalents o f Active in
Liver
1321.01 788.41 1259.42 1067.39 1200.00
444.20 292.03 239.86 370.29 326.09
12.32 7.25 12.32 10.14 11.59
-39-
Company Sanitized. Does not contain TSCA CBI
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-23925
DuPont-2921 Supplement No. 1
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
669.5 200 29
% F in Active: Mol Wt. F (g/mol):
55 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
58 0.087 230.9
Dose F (mmol/kg):
1.679
Molar Ratio (Active/F): 0.052
Dose F (mg/kg):
31.9
Rat Number
Test ppm F Day in Sample Liver
Group II 626396 626397 626403 626406 626421
10 10 10 10 10
415.6 250.3 220.7 291.4 239.8
Group VI 626398 626401 626408 626410 626413
94 94 94 94 94
20.1 17.6 24.1 36.4 13.6
ppm F in Liver
Minus Bkg 0.2 ppm
415.4 250.1 220.5 291.2 239.6
19.9 17.4 23.9 36.2 13.4
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
479.50 288.69 254.53 336.14 276.57
22.97 20.09 27.59 41.79 15.47
/molar Equivalents of Active in
Liver
1302.19 784.01 691.22 912.85 751.10
62.38 54.55 74.92 113.48 42.01
- 40-
Company Sanitized. Does not contain TSCA
