Document RpVDLjoEvyz6Bbb55mZ0yaKJV
TRADE SECRET Study Title
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Supplement No. 1
Laboratory Project ID: DuPont-2924
AR226-3203
D uPont-2924 Supplem ent N o. 1
Author: Carol Finlay, B.A.
S t u d y C o m p l e t e d o n : August 3,2000
Supplement No. 1 Completed on: December 12,2001
P e r f o r m in g L a b o r a t o r y : 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 R eq u est Num ber
S e r v ic e C o d e N u m b e r I ( ( B B F 1
Page 1 o f 40
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
C E R T IF IC A T IO N
We, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study.
Reviewed by:
aul M. Hinderliter, Ph.D. Postdoctoral F ellow
(J Judith C. Stadler, Ph.D., D.A.B.T.
D irector
Issued by Study D irector:
Carol Bj&ilay, B-A. d Staff Scientist
D ate
Elate
V L A L-'IMS
Date
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
TABLE OF CONTENTS
Page
C E R T IF IC A T IO N LIST OF TABLES1< LIST OF FIGURES a**************
....2 .... 4 ****************************4
L IST OF A PPEN D ICES__a
STUDY IN FO R M A TIO N .... >ei
STUDY PERSONNEL
7
****************************** '
R F SfyN FOR. SUPPLEM ENT NXL 1 .............................................. STIMMARY
.................... . 8 9> *
MATERIALS AND M ETHODS a*******************************************10
RESULTS AND D ISCU SSIO N ...
# * * * * * * * * * * * * * * * '
10
A. Liver W eights...............................................
^
1. T est S ubstance........................................................................................................................................................
2. Positive C o n tro ls.................. ...... ................................................................................................................. ...... .
B. Liver Fluorine D ata............................................................................................................. 11
1. Positive C o n tro ls..................................................................................................................................................... *j
2. T est S ubstance.........................................................................................................................................................."
CONCLUSIONS T A B L E S ..............
a *
.......11
12 a 1
F IG U R E S .
.16
A PPENDICES.................
a***<
.22
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Stndy in Rats
DuPont-2924 Supplement No. 1
LIST OF TABLES
Page
1. M EAN BODY AND LIV ER W EIG H TS........................... .............................................................................................. 13 2. M EAN LIV ER FLU O RIN E LEVELS............................................................................................................................... 15 3. M EAN LIV ER FLU O RIN E CONCENTRATION NORM ALIZED TO D O SE....................................................... 15
LIST OF FIGURES
Page
1. COM PARISON O F M EAN RELATIVE LIVER W EIGHTS: TEST SUBSTANCE AND NEGATIVE CO N TRO LS.........................
17
2. COM PARISON O F M EAN RELATIVE LIVER W EIGHTS: TEST SUBSTANCE AND POSITIV E CO N TRO LS............................................................................................................................................................................ 18
3. COM PARISON OF RELATIVE LIVER W EIGHT AND M EAN LIV ER FLUORINE CONCENTRATION FO R H -24001 AND NEGATIVE CO N TRO LS........................................................................19
4. M EAN LIV ER FLU O RIN E CONCENTRATION NORM ALIZED TO D O SE........................................................20
5. COM PARISON O F M EAN BLOOD AND M EAN LIV ER FLU O RIN E CONCENTRATION NORM ALIZED TO D O S E ..................................................................................................................................................21
LIST OF APPENDICES
Page
A . INDIVIDUAL FIN A L BODY W EIGHTS, LIVER W EIGHTS, AND RELATIVE LIV ER TO BODY W E IG H T ................................................................................................................................................................................ 23
B . IND IV ID U AL FLU O RIN E L E V E L S.............................................................................................................................. 34
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION TEST SUBSTANCE:
Substance Tested:
mm.Synonvms/Codes: H-24001 , -- I
Haskell Number: 24001
DuPont-2924 Supplement No. 1
Known Impurities: Unknown
Known Impurities: Unknown
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION (Continued) POSITIVE CONTROL:
Substance Tested:
DuPont-2924 Supplement No. 1
Known Impurities: unknown Sponsor: E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A.
Study Tnitiated/Completed: April 30,1999 / (see report cover page) Tn-Life Tnitiated/Completed: May 30,1999 / August 30,1999
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
REASON FOR SUPPLEMENT NO. 1
The purpose o f this supplement is to present the results from analysis o f selected liver samples from rats treated with the test substance (H-24001), 2 positive controls (H-24019 and H-24020), and 2 negative controls (com oil and com oil/acetone).
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Stady in Rats
DuPont-2924 Supplement No. 1
SUMMARY
The objective o f this study was to evaluate the potential for H-24001, when administered by gavage, to-be absorbed and to accumulate in a mammalian system. Six groups o f 5 male Crl:CD(SD)IGS BR rats each were given 1000 mg/kg/day o f H-24001. The test substance was administered to one group o f 5 rats for 5 consecutive days and to 5 groups for 10 days. 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 o f 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-24001.
The mean relative liver weight (liver/body weight) o f rats dosed with the test substance, H-24001, was up to 25% higher than the negative controls on day 10. By day 94, the weights were similar. On day 10, the mean relative liver weight o f 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 at day 10 than the negative controls. By day 94, the
weights were similar.
The concentration o f fluorine in the livers from 101.28 jaM equivalents on day 10 and 5.14 /xM
rats dosed with the equivalents on day
test 94.
substance, H-24001, On day 10, the mean
wfiMas
equivalent concentrations o f fluorine in the livers from rats dosed with the positive control
materials were approximately 50-fold (H-24019) and 11-fold (H-24020) greater than the fluorine
concentration in livers from rats treated with the test substance. By day 94, the concentrations
were approximately 230x and 2x the fluorine concentration in rats treated with H-24001.
Levels o f total fluorine in livers from rats dosed with the test substance, H-24001, were higher than levels in the blood.
Under the conditions o f this study, there was some absorption and retention o f fluorine in the liver from rats dosed with H-24001. Fluorine levels in the liver were significantly lower than levels in rats dosed w ith the positive control materials, H-24019 and H-24020. Liver weights were elevated in rats dosed with the test substance, H-24001, at the end o f the dosing period, but
not at the end o f the recovery period.
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
MATERIALS AND METHODS
Blood and livers were collected on days 5,10,1 3 ,2 4 , 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 o f fluorine in the liver. See the original report for details o f the analysis.
RESULTS AND DISCUSSION
A. Liver W eights (Tables 1, Figures 1-3, Appendix A)
As w ith 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-24001, 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 body weight are presented for comparison.
1. Test Substance
The mean relative liver weight o f rats dosed with the test substance, H-24001, was up to 25 ^ higher than the negative controls on day 10. On day 13, the mean relative liver weight o f rats dosed with the test substance was up to 26% higher than the negative controls. The weights were s im ila r throughout the remainder o f the recovery period.
2. Positive Controls
The mean relative liver weight o f rats dosed with one o f the positive controls, H-24019, was similar on day 10 to the liver weight o f rats dosed with the test substance, H-24001. On days 13, 24, and 52 (recovery), the mean relative liver weights o f rats dosed with H-24019 were 10-20% higher than the liver weights o f rats dosed with H-24001. By test day 94 (end o f recovery), the mean relative liver weight o f rats dosed with H-24019 was 24% higher than the liver weight o f rats dosed with H-24001. The mean relative liver weight o f rats dosed with H-24020, the other positive control, was 40% higher on day 10 than the liver weight o f rats dosed with H-24001. On days 13 and 24 (recovery), the mean relative liver weights o f rats dosed w ith H-24020 were 29 or 22% higher than the liver weights o f rats dosed with H-24001. On days 52 and 94, the weights were similar.
The mean relative liver weight o f 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 o f rats dosed with
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
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 o f rats dosed with H-24020 began to decrease during the recovery period an d were similar by test day 94 to the negative controls.
Therefore^ liver weights o f rats dosed with the test substance, H-24001, were elevated at the end o f the 10-day dosing period. The mean relative liver weight by the end o f recovery in rats dosed with the test substance was significantly 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 D ata (Tables 2 - 3 , Figures 3 - 5 , Appendix B)
1. Positive Controls
The day
positive 10 from
controls were H-24019 and rats dosed with the positive
H-24020. The concentrations o f fluorine in control materials were 5045.85 and 1127.25
thiMe .livers
on
equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15
and 10.72 /M equivalents.
2. Test Substance
Levels o f total fluorine in livers from rats dosed with the test substance, H-24001, were lower than the levels in livers from rats dosed with the positive control materials H-24019 and
H10-214.20820x.MTehqeutiovtaallenfltusoartindeayco1n0caenndtra5t.i1o4n fiinMtheeqluivivearlefrnotms artadtsaydo9s4e.dFwoirththHe -p2o4s0it0i1vewcaosntrol
H-24019, the liver concentrations were approximately 50x higher (day 10) and approximately 23Ox higher (end o f study) than H-24001. For H-24020, the liver concentrations were approximately 1lx higher (end o f dosing) and approximately 2x higher (end o f study) than
H-24020.
The fiM equivalents o f fluorine in the livers from rats dosed with the test substance, H-24001,
were higher than levels in the blood.
CONCLUSIONS
Under the conditions o f this study, there was some absorption and retention o f fluorine in the liver from rats dosed with H-24001. Fluorine levels in the liver were significantly lower than levels in rats dosed with the positive control materials, H-24019 and H-24020. Liver weights were elevated in rats dosed with the test substance, H-24001, at the end o f the dosing period, but not at the end of the recovery period.
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
TABLES
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TABLE 1
MEAN BODY AND LIVER WEIGHTS (g)
' CORN OIL (NEGATIVE CONTROL)
Test Days
5 10 13 24 52 94
_____________ Absolute_____________
Mean Relative
Body Weight
Liver Weight Liver Weight(% o f Body Weight)
259.3 311.1 322.9 391.2 464.7 557.8
12.620 15.861 14.906 16.635 17.240 20.118
0.049 0.051 0.046 0.043 0.037 0.036
Test Days 5 10 13 24 52 94
CORN OEL: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(% o f 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
Bodv Weight
Liver Weight
306.6
15.456
310.1
17.336
344.8
22.049
387.9 438.1 597.9
21.313 19.826 24.455
Mean Relative
Liver Weight(% o f Body Weight) 0.050 0.056 0.064 0.055 0.045 0.041
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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
244.8 284.4
17.052 21.944
289.1
21.780
370.3
20.619
456.7
17.713
538.7
18.205
Mean Relative Liver Weight(% o f Body Weight)
0.070 0.077 0.075 0.056 0.039 0.034
Test Days 5 10 13 24 52 94
H-24001 (TEST SUBSTANCE)
Absolute
Body Weight
Liver Weight
278.0
15.171
338.9 321.3 388.5
18.703 18.573 17.789
471.6 570.5
17.979 19.109
Mean Relative
Liver Weight(% o f Body Weight)
0.055
0.055
0.058
0.046
0.038
'
0.033
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
TABLE 2
MEAN LIVER FLUORINE LEVELS
Test
Days 10 94
Negative Controls Com
Com Oil Oil:Acetone
(ppm)_______ ( m l__ <0.2 <0.5
0.6a l a
Positive Controls
Test Substance
H-24019
(ppm ) 328.18 (19.3)D 77.56 (10.1)
H-24020
H-24001
(ppm)__________ (BBS)______ 155.76 (29.2) 212.88 (10.6)
1.68 (0.3)
1.70 (1.70)
a O ne value. Four o f the values w ere below the level o f detection (LO D ). b Standard deviation is in parentheses.
TABLE 3
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Days
10 94
Positive Controls
Test Substance.
nM
H-24019 F Equivalents
uM
H-24020 F Equivalents
M
H-24001 F Equivalents____
5045.85 (296.9l)a 1127.25 (211.37)
101.28 (5.1)
1190.15 (156.05)
10.72 (2.14)__________ 5.14 (0.8)_______
a Standard deviation is in parentheses.
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
FIGURES
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 1 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROLS
0 n*
DuPont-2924 Supplement No. 1
Mean Relative Liver Wt. (% of Body Wt.)
5 10 13 24 52 Test Day
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
DuPont-2924 Supplement No. 1
Mean Relative Liver Wt. (% of Body Wt.)
Test Day
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
FIGURE 3
COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24001 AND NEGATIVE CONTROLS
Mean Relative Liver W t (ergam body weight)
HI C om O il (N egative C ontrol) Com O il:A cetone (N egative C ontrol) H -24001 (T est Substance)
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 4 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
DuPont-2924 Supplement No. 1
fiM F Equivalents
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
FIGURE 5 COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
pM Equivalent of Fluoriene
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
APPENDICES
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
APPENDIX A Individual Final Body W eights, Liver W eights, and Relative Liver to Body W eight
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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 W t . 0.034
0.034
0.033
0.043
0.034
Test Day 94 94 94 94 94
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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 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 Wt.
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 Wt. 0.050
0.038
0.053
0.046
0.046
Test Day 13 13 13 13 13
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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 Liver 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 W t .
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 Liver 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-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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
- 2 8 - Company Sanitized- Does not contain TSCA CBI
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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 Wt. 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- not contain TSCACB1 Cotnpany Sanitized. Des
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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 Sanitized. D oes not contain TSCA CBI
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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 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 ISC A CBI
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rate
DuPont-2924 Supplement No. 1
H-24001 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g) , AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 626904
626905
626915
626923
626930
Final Body Wt.
300.9
274.2
275.4
280.5
258.8
GROUP I
Liver Wt. 16.166 15.170 13.453 15.305 15.760
Relative Liver to Body Wt. 0.054
0.055
0.049
0.055
0.061
Test Day 5 5 5 5 5
Animal Number 626900
626909 626912
626919
626921
Final Body Wt.
331.0 398.2
322.2
307.6
335.6
GROUP II
Liver Wt. 19.442 16.358 20.381 17.815 19.518
Relative Liver to Body Wt. 0.059
0.041
0.063
0.058
0.058
Test Day 10 10 10 10 10
Animal Number 626901 626907
626910
626913
626914
Final Body Wt.
301.9
345.5 334.4
282.7
342.1
GROUP III
Liver Wt. 17.784 20.760 18.960 15.419 19.944
Relative Liver to Body Wt. 0.059 0.060
0.057
0.055
0.058
Test Day 13 13 13 13 13
-32- M P ^ S a n iU re d .D o e s not contain TSCA.CB1
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
H-24001 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 626897
626903
626916
626925
626926
Final Body Wt.
379.3
404.0
382.7
390.7
385.7
GROUP IV
Liver Wt. 16.158 18.340 17.460 18.534 18.451
Relative Liver to Body Wt. 0.043
0.045
0.046
0.047
0.048
Test Day 24 24 24 24 24
Animal Number 626898
626902
626908
626918
626922
Final Body Wt.
467.6
500.1
523.4
433.2
433.6
GROUP V
Liver Wt. 17.675 18.708 21.200 16.491 15.822
Relative Liver to Body Wt. 0.038
0.037
0.041
0.038
0.036
Test Day 52 52 52 52 52
Animal Number 626906
626911
626920
626924
626927
Final Body Wt.
674.6
506.9
527.1
549.5
594.5
GROUP VI
Liver Wt. 23.015 16.909 18.046 19.028 18.546
Relative Liver to Body Wt. 0.034
0.033
0.034
0.035
0.031
Test Day 94 94 94 94 94
- 3 3 - Company Sanlteod. Does noi contain IS C A CBS
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
APPENDIX B Individual Fluorine Levels
-34-
tscacbi
Sani r e d ..a ^ otcaatsla
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
T erm s: Active % Active
TERMS AND CALCULATIONS
Mol Wt Active Formulation Dose % F in Active
Mol Wt F
The mg o f formulation given per kg o f animal body weight The molecular weight of fluorine g/mol
Com pound 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 o f
animal body weight = dose [mg/kg] / Mol Wt Active [mg/mmol]
Dose F (mg/kg)
The mg Fluorine administered per kg o f animal body weight = (% F in active/100) x Dose Active [mg/kg]
Dose F (mmol/kg) -
The mmole of fluorine administered per kg o f animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol]
Molar Ratio (Active/F) The moles o f fluorine containing compound per mole o f 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 o f Active [mmol/kg] / Dose Active
[mmol/kg]) x Formulation Dose
Company Sanitized. Does not contain TSCA CBI
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
__________________________ TERMS AND CALCULATIONS
Individual Animal M easurem ent:
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/kgDose
The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead o f the actual active dose. This assumes linearity between administered dose and liver fluorine levels, but is needed because different doses o f active were used in the study.
= ( 0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in liver
minus background)
/molar equivalents o f active in liver
The /molar [/mol/L] concentration o f 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 /moi/mmol
Company Sanitized. Does not contain TSCA CB1
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS
Considerations:
- The used for kinetic analysis was from a limited screen and extrapolation should be done Qautiously.
- Sample size is low - Analytical data used without validation - Steady-state not reached - Terminal phase may not be reached - Some compounds are mixtures o f 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 o f the active (Fluorine containing component(s) in the formulation)
- Molecular weight is the molecular weight o f the active component in the formulation
- 37Company Sanitized. Does not contain TSCACBi
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H -24019
DuPont-2924 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
C alculated 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
R at N um ber
Test Day S am ple
ppm F in
ppm F
L iv er
in M inus Bkg
Liver______ 0.2 ppm
Group II
625320
10 319.4
625330
10 355.4
625332
10 338.4
625341
10 305.0
625350
10 322.7
319.2 355.2 338.2 304.8 322.5
Group VI 625326 625335 625346 625347 625351
94 94 94 94 94
79.8 89.6 73.8 62.5 82.1
79.6 89.4 73.6 62.3 81.9
ppm F in Liver Norm alized to 0.1 mmoles/kg
Dose
molar E q u iv ale n ts of Active in
L iv er
1586.42 1765.34 1680.85 1514.86 1602.83
4910.77 5464.62 5203.08 4689.23 4961.54
395.61 444.32 365.79 309.63 407.04
1224.62 1375.38 1132.31 958.46 1260.00
- 38-
C o m p ass ganitLssd. Does notcontain TSCA CB1
H-24001 : Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24020
DuPont-2924 Supplement No. 1
G iven: 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
C alculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
20 0.047 42.6
Dose F (mmol/kg):
0.726
M olar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat N um ber
Test ppm F Day in Sample Liver
Group II
627768
10 182.5
627774
10 109.0
627776
10 174.0
627782
10 147.5
627785
10 165.8
ppm F in Liver
M inus Bkg 0.2 ppm
182.3 108.8 173.8 147.3 165.6
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
388.30 231.74 370.19 313.75 352.73
gm olar E q u iv ale n ts of Active in
Liver
1321.01 788.41 1259.42 1067.39 1200.00
Group VI 627778 627783 627762 627775 627769
94 94 94 94 94
1.9 1.2 1.9 1.6 1.8
1.7 1.0 1.7 1.4 1.6
3.62 12.32 2.13 7.25 3.62 12.32 2.98 10.14 3.41 11.59
- 3 9 - Sanitized. D oes not contain TSCACB1 Company
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
D ata for H-24001
DuPont-2924 Supplement No. 1
G iven: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
60 1000 35
% F in Active: Mol Wt. F (g/mol):
60 19
C alculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
350 0.583 171.4
Dose F (mmol/kg):
11.053
Molar Ratio (Active/F): 0.053
Dose F (mg/kg):________ 210
Rat N um ber
Test Day S am ple
ppm F in
ppmF
Liver
in M inus Bkg
Liver______ 0.2 ppm
Group II
626904
10 202.8
626905
10 223.6
626915
10 203.2
626923
10 224.5
626930
10 210.3
202.6 223.4 203.0 224.3 210.1
Group VI 626906 626911 626920 626924 626927
94 94 94 94 94
10.7 9.1 13.7 11.2 10.3
10.5 8.9 13.5 11.0 10.1
ppm F in Liver Norm alized to 0.1 mmoles/kg
Dose
pm oiar E q u iv ale n ts of Active in
Liver
34.73 38.30 34.80 38.45 36.02
.
96.48 106.38 96.67 106.81 100.05
1.80 5.00 1.53 4.24 2.31 6.43 1.89 5.24 1.73 4.81
-40Company Sanitized. D oes not contain TSCA CBi
