Document ENom05ZZYjk5KkG8KkJdn4Z0
. 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
DuPont-2924 Supplement No. 1
ia^
AUTHOR: Carol Finlay, B.A.
STUDY COMPLETED ON: August 3,2000
SUPPLEMENT No. 1 COMPLETED ON: December 12,2001
PERFORMING LABORATORY:
E.I. du Font de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
WORK REQUEST NUMBER:||B||B||
SERVICE CODE NUMBERJuy i
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained
from this study.
R^ewed by:
T ^ ^ ^ / / ^ : P&ul UM. Fffrimnd)c-rMlitMer-, PI h.D.
'^IThillPostdoctoral Fellow
Reviewed by:
Qf^d^ ^fet^^ G
(I
Judith C.Stadler.Hi.D-.DAB.T.
Director
Issued by Study Director: ____(_ ^JLACa^roCl Fip^laCy,UBL.A-.ILi^(l
StaffScientist
ii-D^-t^ Date
oy-OfeC-2<3a/
Date
i-ifaZ- U^Ofli 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
CERTIFICATION..............:.........................................................................................................!
LIST OFTABLES........................................................................................................................4 LIST OFFIGURES......................................................................................................................4
LISTOFAPPENDICES...............................................................................................................4
STUDYINFORMATION.............................................................................................................5
STUDYPERSONNEL..................................................................................................................7
REASON FOR SUPPLEMENT NO. 1....................................................................................... 8
SUMMARY..................................................................................................................................9
MATERIALS AND METHODS ............................................................................................... 10
RESULTS AND DISCUSSION................................................................................................. 10
A.
Liver
10 Weights...................................................................................................................
1. Test Substance........................................................................................................................................10
2. Positive Controls....................................................................................................................................10
B. Liver Fluorine Data........................................................................................................... 11
1. Positive Controls ....................................................................................................................................11
2. Test Substance........................................................................................................................................ 11
11 CONCLUSIONS.........................................................................................................................
TABLES....................................................................................................................................... 12
16 FIGURES.....................................................................................................................................
APPENDICES.............................................................................................................................22
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
LIST OF TABLES
Page
1. MEAN BODY AND LIVER WEIGHTS.............................................................................................................13
2. MEANLIVERFLUORINELEVELS..................................................................................................................15
3. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE.................................................. 15
LIST OF FIGURES
Page
1. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE CONTROLS .........................................................................................................................................................17
2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS .........................................................................................................................................................18
3. COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24001 AND NEGATIVE CONTROLS ................................................................19
4. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................20 5. COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION
NORMALIZED TO DOSE ..................................................................................................................................21
LIST OF APPENDICES
Page
A. INDIVIDUAL FINAL BODY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY
WEIGHT..............................................................................................................................................................23
B. INDIVIDUAL FLUORINE LEVELS..................................................................................................................34
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION
TEST SUBSTANCE:
Substance Tested: fif---lBB--iB--r-'
Svnonvms/Codes: Haskell Number:
i r^ H-24001
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: ]j|
Synonyms/Codes: .
DuPont-2924 Supplement No. 1
Known Impurities: iJnknown
Sponsor:
E. I. du Font de Nemours and Company Wihmngton, Delaware 19898
U.S.A.
Study Initiated/Completed: April 30,1999 / (see report cover page)
m-Life Initiated/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: Management:
Primary Technician:
Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. Richard P. Mathena
Liver Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T.
Toxicology Report Preparation: Wanda F. Dinbokowitz
Laboratory Veterinarian: Wanda L. West, D.V.M., A.C.L.A.M.
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 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-24001), 2 positive controls (H-24019 and H-24020),
and 2 negative controls (corn oil and corn oil/acetone).
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Stady m Rats
DuPont-2924 Supplement No. 1
SUMMARY
The objective of 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 of 5 male Crl:CD(SD)IGSBR rats each were given 1000 mg/kg/day of H-24001. 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 liver. Additionally, 2 negative controls, corn oil and corn 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) of 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 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 at day 10 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-24001, was 101.28 pM equivalents on day 10 and 5.14 piM equivalents on day 94. On day 10, the mean pM equivalent concentrations of 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 of total fluorine in livers from rats dosed with the test substance, H-24001, 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-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 me end of 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
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 positive or negative controls. The rats dosed with the test substance, H-24001, were older and heavier in weight than the corn oil negative control and H-24020 positive control rats, but younger and lighter in weight than the corn 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 of rats dosed with the test substance, H-24001, was up to 25% higher than the negative controls on day 10. On day 13, me mean relative liver weight of rats
dosed with the test substance was up to 26% higher than the negative controls. The weights were similar throughout the remainder of the recovery period.
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 liver weight of rats dosed with the test substance, H-24001. On days 13, 24, and 52 (recovery), the mean relative liver weights of rats dosed with H-24019 were 10-20% higher than the liver weights of rats dosed with H-24001. By test day 94 (end of recovery), the mean relative liver weight of rats dosed with H-24019 was 24% higher than the liver weight of rats dosed with H-24001. The mean relative liver weight of rats dosed with H-24020, the other positive control, was 40% higher on day 10 than "the liver weight of rats dosed with H-24001. On days 13 and 24 (recovery), the mean relative liver weights of rats dosed with H-24020 were 29 or 22% higher than the liver weights of rats dosed with H-24001. 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
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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 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 me test substance, H-24001, were elevated at the end of the 10-day dosing period. The mean relative liver weight by the end of 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 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 ^M
equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15
and 10.72 /iM equivalents.
2.
Test Substance
Levels of 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
H-24020. The total fluorine concentration in the liver from rats dosed with H-24001 was 101.28 jllM equivalents at day 10 and 5.14 ^M equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 50x higher (day 10) and approximately
230x higher (end of study) than H-24001. For H-24020, the liver concentrations were approximately 1 Ix higher (end of dosing) and approximately 2x higher (end of study) than
H-24020.
The fiM equivalents of fluorine in the livers from rats dosed with the test substance, H-24001,
were higher than levels in the blood.
CONCLUSIONS
Under the conditions of this study, there was some absorption and retention of 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 of the dosing period, but not at the end of the recovery period.
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DuPont-2924 Supplement No. 1
TABLES
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DuPont-2924 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-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
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-24001 (TEST SUBSTANCE)
Absolute
Body Weight
Liver Weight
278.0
15.171
338.9
18.703
321.3
18.573
388.5
17.789
471.6
17.979
570.5
19.109
Mean Relative Liver Weight(% of 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
Negative Controls
Positive Controls
Test Substance
Test Corn Oil
Corn OiLAcetone
H-24019
H-24020
H-24001
K)<0.20.5328.18 Days 94
(ppm)_____(ppm)______(ppm) ____(ppm)______(ppm) (19.3)"155.76 (29.2)212.88 (10.6)
0.6"______^______77.56 (10.1)
1.68 (0.3)
1.70 (1.70)
a One value. Four of the values were below tfae level of detection (LOD).
b Standard deviation is in parentheses.
TABLES
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Days
10 94
Positive Controls
Test Substance
H-24019
H-24020
H-24001
pM F Equivalents
5045.85 (296.91)"
;nM F Equivalents___jttM F Equivalents
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
0.06
0.05-+
0.04-4 ^
ItP
i 0.03^
^
s 0.024
0.014
0.00
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study m Rats
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
Test Day 18-
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 3 COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTR
FOR H-24001 AND NEGATIVE CONTROLS
Day 10
Day 94
Day 10
H Corn Oil (Negative Control) Corn OihAcetone (Negative Control) D H-24001 (Test 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
6000
5000
4000
er 3000
fc.
s a.
2000
1000
BH-24019 (P BH-24020 (P OH-24001 (T
Day 10
Day 94
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURES COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZE
6000
5000
S 4000
'8
3000
a o" y
S 2000
1000
Blood / Liver
Blood / Liver
Blood / Liver
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H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
APPENDICES
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
APPENDIX A Individual Final Body Weights, Liver Weights, and Relative Liver to Body Weight
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H-24001: Biopersistaace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
COWF OIL (NEGATIVE CONTROL)
innm/TDUAL FINAL BC>DY WEIGHTS (g), LIVER WEIGHTS (g ) , AMD RELATIVE LFTER TO BODY WEK3HT 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: Biopersisteace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
CORKF OIL (NEGATIVE CONTROL)
INDFyiDUAL FINAL BCIDY WEIGHTS (g) , LIVER WEIGHTS (g; , AMD RELATIVE LT\fEB. TO BODY WEI:GHT OF MALE RATS
.
Animal Number 627791 627794 627804 627812 627817
Final
Body Wt. 365.6
416.7
378.1
388.4
407.3
GROUP IV
Liver Wt. 15.371 19.105
15.326 16.775 16.600
Relative Liver to Body Wt. 0.042
0.046
0.041
0.043
0.041
Test Day
24 24 24 24 24
Animal Number 627792
627798
627813
627814 627815
Final
Body Wt.
496.8
517.3
416.8
386.9
505.9
GROUP V
Liver Wt.
16.359 19.293 15.234 14.358 20.958
Relative Liver to Body Wt.
0.033
0.037 0.037 0.037 0.041
Test Day
52
52 52 52
52
Animal
Number 627795
627811
627819
627820
627821
Final
Body Wt.
485.0
511.6
653.1
618.0
521.5
GROUP VI
Liver Wt.
16.649 17.405 21.828 26.766 17.943
Relative Liver to Body Wt. 0.034
0.034
0.033
0.043
0.034
Test Day
94 94 94 94 94
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DuPont-2924 Supplement No. 1
CORN 0 IL:ACETONE (NEGA'.PIVE CONTROL)
1W>I\/IDUAL FINAL :BODY WEIGHTS (g),, LIVER WEIGHTS (9r), AND
RELATIVE I jIVER 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 Wt. 14.409 14.363 16.139 17.745 15.328
Relative Liver to Body Wt. 0.043
0.043
0.044
0.046
0.042
Test Day
10 10 10 10 10
Animal Number 625357
625358 625370
625384
625385
Final
Body Wt. 405.6
340.7
407.0
357.1
377.6
GROUP III
Liver Wt. 20.091 12.954 21.654 16.520 17.355
Relative 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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CoB(ipa"y
H-24001: Biopersistaace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
CORN OIL:.ACETONE (NEGA1'IVE CONTROL)
INDI\UDUKL FINAL BOD-r WEIGHTS (g), LIVER WEIGHTS (gr), AMD
RELATIVE LIVE!R TO BODY WEII3HT 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 Wt. 565.3
514.8
673.7
588.8
496.6
GROUP VI
Liver Wt.
18.655 17.379 20.073 18.869 17.261
Relative 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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DuPont-2924 Supplement No. 1
H-2'1019 (POSITIVE iCONTROL) INDI\riDUMj FINAL BOiDY WEIGHTS (g), LIVER WEIGHTS (gl1 , AND
RELATIVE LB/ER TO BODY WEIiGHT OF MALE RATS
Animal
ISTuinber
625322 625323
625336 625343
625349
Final
Body Wt. 329.0
298.0
318.1
294.0
293.7
GROUP I
Liver Wt.
16.409 14.830 16.545 13.809 15.685
Relative Liver to Body Wt.
0.050 0.050 0.052 0.047 0.053
Test Day
5 5
5 5
5
Animal
Number 625320
625330
625332
625341
625350
Final
Body Wt. 312.1
310.1
318.3
325.6
284.3
GROUP II
Liver Wt. 18.845 16.343
17.528 18.254 15.709
Relative Liver to Body Wt. 0.060 0.053 0.055
0.056 0.055
Test Day
10 10 10 10 10
Animal
Number 625321
625327
625342
625345
625348
Final
Body Wt.
371.2
355.3
324.4
336.9
336.4
GROUP III
Liver Wt. 22.322 22.259 20.718
24.403 20.541
Relative Liver
to Body Wt. 0.060 0.063 0.064 0.072 0.061
Test Day
13 13
13 13 13
-28-
Company Sanitized. Does not contain TSCA CBI
H-24001: Biopersistelace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
H-2'1019 (POSITIVE CONTROL)
IMDI\H:DUAL FINAL BOIDY WEIGHTS (g) , LIVER WEIGHTS (g), AND RELATIVE LB/ER TO BODY WE3:GHT OF MALE RATS
Animal Number 625325
625328
625338
625340
625352
Final
Body Wt. 381.3
423.9
339.9
419.0
375.3
GROUP IV
Liver Wt. 21.479
22.100 18.018 23.131 21.835
Relative Liver to Body Wt. 0.056
0.052
0.053
0.055
0.058
Test Day
24 24 24 24 24
Animal
Number 625324
625329
625333
625334
625344
Final
Body Wt. 447.4
498.1
427.4
426.5
391.0
GROUP V
Liver 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 Wfc. 0.041
0.037
0.043
0.043
0.040
Test Day
94 94 94 94 94
-29-
S,n^.Ooesn.----..nT^CB.
coinpwy
H-24001: Biopersisteace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
H-2.4020 (POSITIVE 'CONTROL)
INDFVIDUAL FINAL BCIDY WEIGHTS (g) , LIVER WEIGHTS (ar), AND RELATIVE LF/ER TO BODY WEHGHT 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 Wfc. 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. Does not contain TSCA CBI
H-24001: Biopersiste:nee Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
H- 24020 (POSITIVE CONTROL) INDB/IDUAL FINAL I iODY WEIGHTS (g)., LIVER WEIGHTS (g ) , AND
RELATIVE L IVER TO BODY WEIGHT OP MALE RATS
.
Animal Nuinber 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
Nuinber 627762
627769
627775 627778
627783
Final
Body Wt. 528.6
609.6
552.8
445.3
557.2
GROUP VI
Liver Wt.
17.732 19.324 17.296 15.628 21.044
Relative Liver to Body Wt. 0.034
0.032
0.031
0.035
0.038
Test Day
94 94 94 94 94
-31- Company Sanitized. Does not contain TSCA, CBI
H-24001: Biopersiste:ace Screening 10-Dose Oral Gavage Study in Rate
DuPont-2924 Supplement No. 1
H-:24001 (TEST SUBSTANCE)
INDI-^7IDUAL FINAL BC1DY WEIGHTS (g) , LIVER WEIGHTS (g)' , AND RELATIVE LFi/ER TO BODY WEIiGHT 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- c^------00--------1"^61
H-24001: Biopersistaace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
H-:24001 (TEST SUBSTANCE)
iND:n/TDUAL FINAL BOIDY WEIGHTS (g) ,, LIVER WEIGHTS (g ) , AMD RELATIVE LF7ER TO BODY WEI GHT 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
-33-
Company Sanitized. Does not contain TSCA CBI
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
APPENDIX B Individual Fluorine Levels
-34- .^s^."-^"3^8"06'
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
Terms: Active % Active
TERMS AND CALCULATIONS
Mol Wt Active
Formulation Dose
% Fin Active
MolWtF
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/I 00) 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/P)
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-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2924 Supplement No. 1
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppm F in liver
The ppm fluoride measured in liver
Individual Animal Calculations:
ppm F in Liver minus Bkg 0.2 ppm
ppm P in Liver normalized to 0.1 mmol/kgDose
The ppm fluoride measured in liver minus the background fluoride measured in control animal liver. In this case the value was established at 0.2 ppm.
The ppm fluoride minus background that would be expected if the
active dose was 0.1 mmol/kg instead of the actual active dose. This assumes linearity between administered dose and liver fluorine levels, but is needed because different doses of active were used in the study.
= ( 0.1 [mmpl/kg] / Active dose [mmol/kg]) x (ppm F in liver
minus background)
punolar equivalents of
active in liver
The /nnolar [j^mol/L] concentration of fluorine containing compound
in liver based on the ppm fluorine in liver normalized to 0.1 mmol/kg active dose. This assumes that all liver fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm -
Img/L = (Normalized ppm [mg/L] fluorine in liver / Mol Wt F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x
1000 ^mol/mmol
-36- Company Sanitized. Does not contain TSCA 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 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 offluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - Each compound may have very different potency for producing toxicity
Assumptions: (May or may not be justified in all cases)
- Liver fluorine concentrations are linear with respect to dose - Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of
individual components - Background Fluorine is 0.2 ppm
- % F data is the % Fluorine of the active (Fluorine containing components) in the formulation) - Molecular weight is the molecular weight of the active component in the formulation
Company Sanitized. Does not contain TSCACB!
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):
497
Formulation Dose (mg/kg):
10
% Active (F Containing) in Formulation: 100
%F in Active:
65
Mol Wt. F (g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg):
Formulation Dose Normalization Factor:
10 0.020 49.7
Dose F (mmol/kg):
0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):_____6.5
Rat Number
Test Day Sample
ppmF
in
Liver
Group II
625320
10
625330
10
625332
10
625341
10
625350
10
319.4 355.4 338.4 305.0 322.7
ppmF in Liver
Minus Bkg 0.2 ppm
319.2 355.2 338.2 304.8 322.5
Group VI
625326
94
625335
94
625346
94
625347
94
625351
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 Normalized to 0.1 mmoles/kg
Dose
/imolar Equivalents of Active in
Liver
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
.38Cosapsas Sanitized. Does nQicordaia TSCA CBi
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24020
DuPont-2924 Supplement No. 1
Given:
Mol Wt. Active (g/mole):
426
Formulation Dose (mg/kg):
20
% Active (F Containing) in Formulation: 100
% P in Active:
69
MolWt.F(g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
20 0.047 42.6
Dose F (mmol/kg):
0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):_____13.8
Rat Number
Test Day Sample
ppmF
in Liver
Group II
627768
10
627774
10
627776
10
627782
10
627785
10
182.5 109.0 174.0 147.5 165.8
ppm F in Liver
Minus Bkg 0.2 ppm
182.3 108.8 173.8 147.3 165.6
Group VI
627778
94
1.9
1.7
627783
94
1.2
1.0
627762
94
1.9
1.7
627775
94
1.6
1.4
627769
94
1.8
1.6
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
fimolar Equivalents of Active in
Liver
388.30 231.74 370.19 313.75 352.73
1321.01 788.41
1259.42 1067.39 1200.00
3.62 2.13 3.62 2.98 3.41
12.32 7.25
12.32 10.14 11.59
.39-
Sanilteed.DoesnotoonlamTSCACBl
Csiapany
H-24001: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24001
DuPont-2924 Supplement No. 1
Given:
Mol Wt. Active (g/mole):
60
% Fin Active:
60
Formulation Dose (mg/kg):
1000
MolWt.F(g/mol):
19
% Active (F Containing) in Formulation: 35
Calculated 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 Number
Test Day Sample
ppmF
in Liver
Group II
626904
10
202.8
626905
10
223.6
626915
10
203.2
626923
10
224.5
626930
10
210.3
ppmF in
Liver Minus Bkg
0.2 ppm
202.6 223.4 203.0 224.3 210.1
Group VI
626906
94
10.7
10.5
626911
94
9.1
8.9
626920
94
13.7
13.5
626924
94
11.2
11.0
626927
94
10.3
10.1
ppm F in Liver Normalized to
0.1 mmoles/kg Dose
fimolar Equivalents 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 1.53 2.31 1.89 1.73
5.00 4.24 6.43 5.24 4.81
-40- Company Sanitized. Does not contain TSCA CBI