Document edJK23n3mK9Dv1RR0gXeQJv4
AR226-3210
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 Font de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
WORK SERV
REQUEST ICE CODE
N N
U U
M M
B B
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l.ER:
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Page 1 of 40
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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:
^ ^ ^ J ^ /PaalM. Hinderliter. Ph.D.
Postdoctoral Fellow
/J-Ac-.Z-l
Date
Reviewed by:
C,L.cLc^ C J^3^
Director
g^)^-20^/ Date
Issued by Study Director:
(_/JL/J^C'JLIA.i Carol ^mlay,B.Af.LQU,______
Staff Scientist
\^-^ U ^/U^\
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........................................................................................................................^
LIST OF TABLES .........................................................................................................................4 LIST OF FIGURES .......................................................................................................................4 LIST OF APPENDICES...............................................................................................................4 STUDY INFORMATION .............................................................................................................5 STUDY PERSONNEL...................................................................................................................? REASON FOR SUPPLEMENT NO. 1 ........................................................................................8 SUMMARY ....................................................................................................................................9 MATERIALS AND METHODS ................................................................................................10 RESULTS AND DISCUSSION ..................................................................................................10
A. Liver Data..........................................................................................................................10
1. Test Substance........................................................................................................................................ 10 2. Positive Controls.................................................................................................................................... 10
B. Liver Fluorine Data............................................................................................................10
1. Positive Controls ....................................................................................................................................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. MEAN BODY AND LIVER WEIGHTS............................................................................................................. 13 2. MEAN LIVER FLUORINE LEVELS..............................................................................................-........-.......15 3. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE.................................................. 15
LIST OF FIGURES
Page
1. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE
CONTROLS.........................................................................................................................................................17
2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE
CONTROLS.........................................................................................................................................................18
3. COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-23925 AND NEGATIVE CONTROLS................................................................ 19
4. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................20 5. COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION
NORMALIZED TO DOSE..................................................................................................................................21
LIST OF APPENDICES
Page
A. INDIVIDUAL FINAL BODY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY
WEIGHT.............................................................................................................................................................^
B. INDIVIDUAL FLUORINE LEVELS ..................................................................................................................34
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION TEST SUBSTANCE:
sm 11 mi i Ti ii 11[flB^BBJ Synonyms/Codes: _ H-23925
Haskell Number: 23925
Composition:MM||
Known Impurities: None
POSITIVE CONTROL:
DuPont-2921 Supplement No. 1
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Known Impurities: Unknown empany Sanitized. Does not contain TSCA RR
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION (Continued) POSITIVE CONTROL;
Synonyms/Codes: H-24020
DuPont-2921 Supplement No. 1
Haskell Number: 24020 Composition:
Known Impurities: Unknown
Sponsor:
E. I. du Font 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: 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-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 (corn oil and corn oiVacetone).
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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, 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 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 ^.M equivalents on day 94. On day 10, mean pM 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 ffM 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 JU.Mequivalent concentration of
H-24019 but 6x higher than the /xM equivalent concentration of H-24020.
The /zM equivalents of 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 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 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
-10CompanySanitized. Does not contain TSCA CB(
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.
Therefore, liver weights of rats dosed with the test substance, H-23925, were elevated at the end of the 10-day dosing period but not at the end of the recovery 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 day 10 from rats dosed with the positive control materials were 5045.85 and 1127.25 p.M equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15
and 10.72 fiM equivalents.
2.
Test Substance
The total fluorine concentration in the liver from rats dosed with H-23925 was 888.28 jiiM equivalents at day 10 and 69.47 fjiM equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 6x higher (day 10) and approximately 17x higher (end of study) than H-23925. For H-24020, the liver concentration was approximately 1.3x higher at the end of dosing. On day 94, the concentration was approximately 6x higher in rats dosed with H-23925 than in rats dosed with H-24020.
The pM equivalents of fluorine in the livers from rats dosed with the test substance 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-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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TABLES
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
TABLE 1 MEAN BODY AND LIVER WEIGHTS (g)
DuPont-2921 Supplement No. 1
Test Days
5 10 13
24 52 94
Body Weight 259.3 311.1 322.9 391.2 464.7 557.8
Liver Weight 12.620 15.861 14.906 16.635 17.240 20.118
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 OBLACETONE (NEGATIVE CONTROL)
___Bo_d_y_W_e_i_gAhtb_s__o_lLuitvee_r _W_e_ig_h_t __
i n313.4A
356.5
i1^3.(6W32
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)
n0.r0u4-13 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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Test Days 5 10
13
24 52 94
TABLE 1 (Continued)
MEAN BODY AND LIVER WEIGHTS (g)
H-24020 (POSITIVE CONTROL)
Abs olute
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)
___Bo_d_y_W_e_i_gAhtb_s__o_lLuitvee_r _W_e_ig_h_t __
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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TABLE 2 MEAN LIVER FLUORINE LEVELS
Negative Controls
Positive Controls
Test Substance
Corn
Test Corn Oil
OiLAcetone
H-24019
H-24020
H-23925
Days (ppm)_____(ppm)_______(ppm)
(ppm)_______(ppm)
K)<02<0.5328.181(19.3)''155.76 (29.2)283.56 (78.2)
94
O.S8______I3______77.56 (10.1)
1.68 (0.3)
22.36 (8.7)
a One value. Pour of the values were below the level of detection (LOD). b Standard deviation is in parentheses.
Indicates no value for this day.
%
TABLE 3
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Days
10
94
Positive Controls
Test Substance
H-24019
H-24020
H-23925
/zM F Equivalents 5045.85 (296.91)3
[JI.MF Equivalents___pM F Equivalents
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 10-Dose Oral Gavage Study in Rats
-^-
0.07
0.064
0.05
1s &
S >
o
i
^
as
-o
0.04 0.03
0.02
0.01
0.00
FIGURE 1 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROLS
Test Day
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
^
FIGURE 2
COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS
Test Day
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURES COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRA
FOR H-23925 AND NEGATIVE CONTROLS
Day 10
Day 13
Day 94
Day 10
D
Eg Corn Oil (Negative Control) Corn Oil:Acetone (Negative Control) D H-23925 (Test Substance)
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 4 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Day 10
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Day 94
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURES COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZE
Blood / Liver
Blood / Liver
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Blood / Liver
APPENDICES -22- iRpmpanySanHIzed. Does not contain TSCA CRt
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
^
^y
APPENDIX A Individual Final Body Weights, Liver Weights, and Relative Liver to Body Weight
23Company Sanitized. Does not contain TSCA CBI
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDIV'IDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g ) , AND
RELATIVE LI\rER TO BODY WE]:GHT 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
-24Company Sanmzed. Does no! contain TSCA CW
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDIV^IDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g0 ,
RELATIVE LI\^ER TO BODY WE][GHT OF MALE RATS
AND
Animal
Number 627791 627794
627804
627812
627817
Final
Body Wt. 365.6
416.7
378.1
388.4
407.3
GROUP IV
Liver Wt. 15.371 19.105 15.326 16.775 16.600
Relative Liver to Body Wt. 0.042
0.046
0.041
0.043
0.041
Test Day
24 24
24 24 24
Animal Number 627792
627798
627813
627814
627815
Final
Body Wt. 496.8
517.3
416.8
386.9
505.9
GROUP V
Liver Wt. 16.359 19.293 15.234 14.358 20.958
Relative Liver to Body Wt. 0.033
0.037
0.037
0.037
0.041
Test Day
52
52 52
52 52
Animal Number 627795 627811
627819 627820
627821
Final
Body Wt.
485.0
511.6
653.1
618.0
521.5
GROUP VI
Liver Wt. 16.649
17.405 21.828 26.766 17.943
Relative Liver to Body Wt. 0.034
0.034
0.033
0.043
0.034
Test Day
94 94 94 94
94
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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
CORN OIL :ACETONE (NEGA1'IVE CONTROL)
INDIV^DUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (gr), AND
RELATIVE LI\?ER TO BODY WEK3HT 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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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
CORN OIL :ACETONE (NEGA^"I'VE CONTROL)
INDIVI DUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (g ) , AND RELATIVE LI\KR TO BODY WEIiGHT 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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H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-241019 (POSITIVE (:ONTROL)
INDIV'IDVAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g ) , AND RELATIVE LI\^ER TO BODY WEK3HT OF MALE RATS
Animal Number 625322
625323
625336
625343
625349
Final
Body Wt. 329.0
298.0
318.1
294.0
293.7
GROUP I
Liver Wt. 16.409 14.830 16.545 13.809 15.685
Relative Liver to Body Wt. 0.050
0.050
0.052
0.047
0.053
Test Day
5
5 5 5 5
Animal Number 625320
625330
625332
625341
625350
Final
Body Wt.
312.1
310.1
318.3
325.6
284.3
GROUP II
Liver Wt. 18.845
16.343 17.528 18.254 15.709
Relative Liver to Body Wt. 0.060
0.053
0.055
0.056
0.055
Test Day
10 10 10
10 10
Animal Number 625321
625327
625342
625345
625348
Final
Body Wt. 371.2
355.3
324.4
336.9
336.4
GROUP III
Liver Wt. 22.322 22.259 20.718 24.403 20.541
Relative Liver to Body Wt. 0.060
0.063
0.064
0.072
0.061
Test Day
13 13
13 13 13
-28- CompanySanHized. Does not contain TSCA CBI
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-241019 (POSITIVE C'ONTROL) INDIVriDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g ) , AND
RELATIVE LI\^ER TO BODY WEIG;HT 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- pompanySanitized. Does not contain TSCA r*"
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-2'1020 (POSITIVE (CONTROL) INDIVFIDUAL FINAL BOOY WEIGHTS (g), LIVER WEIGHTS (g ) , AND
RELATIVE LP/ER TO BODY WEK3HT OF MALE RATS
Animal Number 627756 627767
627771 627773
627777
Final
Body Wt.
237.5
248.6
239.6
262.3
236.0
GROUP I
Liver Wt.
15.360 19.033 16.968 17.629 16.268
Relative Liver to Body Wt. 0.065
0.077
0.071
0.067
0.069
Test Day
5 5 5 5 5
Animal Number 627768
627774
627776
627782
627785
Final
Body Wt. 293.5
338.2
254.4
268.2
267.9
GROUP II
Liver Wt. 22.536 27.332
18.821 20.172 20.857
Relative Liver to Body Wt.
0.077
0.081
0.074
0.075
0.078
Test Day
10 10 10
10
10
Animal Number 627758
627763
627764
627787
627788
Final
Body Wt. 308.7
285.8
306.7
234.9
309.4
GROUP III
Liver Wt. 26.021
22.123 21.526 16.498 22.730
Relative Liver to Body Wt. 0.084
0.077
0.070
0.070
0.073
Test Day
13 13
13 13 13
-30- Company Sanitized. Does not contain TSCA CBi
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-2'1020 (POSITIVE CONTROL)
;
INDIVODUAL FINAL BOCY WEIGHTS (g),, LIVER WEIGHTS (g ) ,
RELATIVE LI\/ER TO BODY WE I GHT OF MALE RATS
AND
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-
Compwiy Sanitized. Does not contain TSCA CBl
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 Wt. 307.9
256.1
298.6
321.7
282.9
GROUP I
Liver Wt 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
Final
Body Wt.
342.7
GROUP II
Liver Wfc 19.454
Relative Liver to Body Wt.
0.057
Test Day
10
626397
335.7
17.422
0.052
10
626403
337.0
18.141
0.054
10
626406
333.8
17.180
0.051
10
626421
347.7
20.866
0.060
10
Animal Number 626402
626404
626409
626417
626418
Final
Body Wt. 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 Live r
to Body Wt. 0.062 0.077 0.068 0.079 0.064
Test Day
13 13 13 13 13
-32- CompanySaniffzaa. Does not comaTn TSCA C8I
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
H-2!3925 (TEST SUBSTANCE)
INDIVTDUAL FINAL BO:DY WEIGHTS (g) , LIVER WEIGHTS (g ) , AND
RELATIVE LI\rER TO BODY WEI GHT OF MALE RATS
Animal
Number 626405
626414
626419
626423
626423
Final
Body Wt.
417.7
402.5
395.6
427.7
396.6
GROUP IV
Liver Wt. 22.726 23.389 22.530 21.703 20.507
Relative Liver to Body Wt.
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 Wt. 430.1
440.0
508.3
509.7
513.6
GROUP V
Liver Wt.
18.450 19.086 20.697 21.089 20.960
Relative Liver to Body Wt.
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 Wt. 505.5
455.6
493.4
542.4
697.5
GROUP VI
Liver Wt. 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- CompanySanfffeeDdo. es notcontam TSCA CBt
H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2921 Supplement No. 1
APPENDIX B Individual Fluorine Levels
-34- Company Saniffzetf. Does not contain TSCA CBI
Terms: Active % Active
Mol Wt Active
Formulation Dose % F in Active
Mol Wt F
TERMS AND CALCULATIONS
Fluorine containing compound The % of formulation that is made up of fluorine containing components The molecular weight of the fluorine containing components (g/mole) The mg of formulation given per kg of animal body weight The % fluorine in the fluorine containing components of the formulation (weight basis) The molecular weight of fluorine g/mol
Compound Calculations:
Dose Active (mg/kg)
The mg of fluorine containing compound administered per kg of animal body weight.
= (% active/100) x Formulation Dose
Dose Active (mmole/kg)
The mmole of fluorine containing compound administered per kg of animal body weight
= dose [mg/kg] / Mol Wt Active [mg/mmol]
Dose F (mg/kg)
The mg Fluorine administered per kg of animal body weight = (% F in active/100) x Dose Active [mg/kg]
Dose F (mmol/kg)
The mmole of fluorine administered per kg of animal body weight = Dose P [mg/kg] / Mol Wt F [mg/mmol]
Molar Ratio (Active/F)
The moles of fluorine containing compound per mole of fluorine = Dose Active [mmol/kg] / Dose F [mmol/kg]
Formulation Dose Normalization Factor
The formulation dose that would be required to administer the amount of active needed to achieve the normalized dose
= (Normalized dose of Active [mmol/kg] / Dose Active [mmol/kg]) x Formulation Dose
-35- CwnpanySaniffzed. Does not contain TSCA CBt
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/kgDose
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)
/xmolar equivalents of active in liver
The ^molar [/xmol/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 CBt
FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS
Considerations: - The data used for kinetic analysis was from a limited screen and extrapolation should be done
cautiously. - Sample size is low - Analytical data used without validation - Steady-state not reached - Terminal phase may not be reached - Some compounds are mixtures of fluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - Each compound may have very different potency for producing toxicity
Assumptions: (May or may not be justified in all cases) - Liver fluorine concentrations are linear with respect to dose - Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of
individual components - Background Fluorine is 0.2 ppm - % F data is the % Fluorine of the active (Fluorine containing component(s) in the formulation) - Molecular weight is the molecular weight of the active component in the formulation
-37Company Sanitized. Does mH 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):
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
319.4
625330
10
355.4
625332
10
338.4
625341
10
305.0
625350
10
322.7
Group III
625321
13
625327
13
625342
13
625345
13
625348
13
255.9 275.3 277.9 260.4 275.6
Group VI
625326
94
625335
94
625346
94
625347
94
625351
94
79.8 89.6 73.8 62.5 82.1
ppm F in Liver
Minus Bkg 0.2 ppm
319.2 355.2 338.2 304.8 322.5
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
/{molar 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
1270.83 1367.25 1380.17 1293.19 1368.74
3933.85 4232.31 4272.31 4003.08 4236.92
395.61 444.32 365.79 309.63 407.04
1224.62 1375.38 1132.31 958.46 1260.00
-38CompanySanitized. Does not contain TSCA C5l
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):
426
Formulation Dose (mg/kg):
20
% Active (F Containing) in Formulation: 100
% F in Active:
69
Mol Wt. F (g/mol):
19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
20 0.047 42.6
Dose F (mmol/kg):
0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):_____13.8
Rat Number
Test Day Sample
ppmF
in Liver
Group II
627768
10
182.5
627774
10
109.0
627776
10
174.0
627782
10
147.5
627785
10
165.8
Group III
627758
13
627764
13
627787
13
627763
13
627788
13
61.5 40.5 33.3 51.3 45.2
Group VI
627778
94
1.9
627783
94
1.2
627762
94
1.9
627775
94
1.6
627769
94
1.8
ppm F in Liver
Minus Bkg 0.2 ppm
182.3 108.8 173.8 147.3 165.6
61.3 40.3 33.1 51.1 45.0
1.7
1.0
1.7
1.4
,
1.6
ppm F in L iver
Normalize!ito
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 of 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 CBt
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):
669.5
% F in Active:
55
Formulation Dose (mg/kg):
200
Mol Wt. F (g/mol):
19
% Active (F Containing) in Formulation: 29
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 Day Sample
ppm F in
Liver
Group II
626396
10
626397
10
626403
10
626406
10
626421
10
415.6 250.3 220.7 291.4 239.8
Group VI
626398
94
626401
94
626408
94
626410
94
626413
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 Lrver
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
/xmolar 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 SanfHzed. Does nol contain TSCA CR^