Document zQ2mLn949Z3Xv9M0nbYNdjg2R
AR226-3204
DuPont-2925 Supplement No. 1
TRADE SECRET
Study Title H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Supplement No. 1
Laboratory Project ID: DuPont-2925
AUTHOR: Carol Finlay, B.A.
STUDY COMPLETED ON: September 1, 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 Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
WORK REQUEST NUMBERj--------I--
JI^^^nT SERVICE CODE NUMBEI# --------I|
Sciences
Page 1 of 40 Company Sanitized. Does not contain TSCA CBB
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
^ ^ , J ^ 3 ^ L ^ f c 'Paul M. HhiderUter. PhJ>.
Postdoctoral Fellow
IZ-JPfc-TJ^I
Date
Reviewed tar:
Cj^^f. ff M<LdJLflj
77JudifliC.Stadler,TO.D..D.A.B.T.
v
Director
Py-^O^OO f
Date
Issued bTStmlyPirector:
/ & /LfrC ^t^ifL^_______
Cattl^iilay, BA. Q
Staff Scientist
^-Z -^Djg^^^b/
Date
-2-
Company Sanitized. Does not contain TSCA CBi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
TABLE OF CONTENTS
Page
CERTEFICATION.........................................................................................................................2
LIST OF TABLES .........................................................................................................................4
LISTOFmGURES.....................................................................................................^
LIST OF APPENDICES...............................................................................................................4 STUDY INFORMATION.............................................................................................................5 STUDY PERSONNEL...................................................................................................................? REASON FOR SUPPLEMENT NO. 1 ........................................................................................8
SUMMARY..........................................................................................................................
MATERIALS AND METHODS ...............................................................................................JO RESULTS AND DISCUSSION ..................................................................................................10
A. Liver Weights.....................................................................................................................l0
1. Test Substance........................................................................................................................................10 2. Positive Controls.................................................................................................................................... 10
B. Liver Fluorine Data............................................................................................................11
1. Positive Controls...........................--..............--.....................................................................................11 2. Test Substance........................................;............................................................................................... 11
CONCLUSIONS ..........................................................................................................................11
TABLES........................................................................................................................................12
FIGURES......................................................................................................................................16
APPENDICES.............................................................................................................................^
.3- Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
LIST OF TABLES
Page
1. MEAN BODY AND LIVER WEIGHTS............................................................................................................. 13
2. t^ANL^VERI^UORI^ffiIVELS..................................................................................................................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-24000 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
-4- Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION TEST SUBSTANCE:
Substance Tested: Synonyms/Codes] Haskell Number: 24000
Compositio:
Known Impurities: Not supplied by the sponsor
DuPont-2925 Supplement No. 1
Known Impurities: Unknown -5. Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION (Continued) POSITIVE CONTROL:
------z^ Substance Tested ^^^^H^l
Synonyms/Codes: H-24020
DuPont-2925 Supplement No. 1
^
Haskell Number: 24020
Sponsor:
E. I. du Pont de Nemours and Company Wilmington, Delaware 19898
U.S.A.
Study Initiated/Completed: April 30,1999 / (see report cover page)
In-Life Initiated/Completed: June 13,1999 / September 14, 1999
6-
Company Sanitized. Does not contain TSCA CBi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 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 P. Dinbokowitz
Laboratory Veterinarian: Wanda L. West, D.V.M., A.C.L.A.M.
.7-
Comp&ny Sanitized. Does not contain TSCA CBI
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-24000), 2 positive controls (H-24019 and H-24020),
and 2 negative controls (corn oil and corn oil/acetone).
- 8-
Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
SUMMARY
The objective of this study was to evaluate the potential for H-24000, 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 300 mg/kg/day of H-24000. 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 as described for H-24000.
The mean relative liver weight (liver/body weight) of rats dosed with the test substance, H-24000, was up to 36% 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 concentration of fluorine in the livers from rats dosed with the test substance, H-24000, was
648.16 juM equivalents on day 10 and 4.69 JU.Mequivalents on day 94. On day 10, mean ^M
equivalent concentrations of fluorine in the livers from rats dosed with the positive control materials were approximately 8-fold (H-24019) and 2-fold (H-24020) greater than the fluorine concentration in livers from rats treated with the test substance. By day 94, the concentrations were approximately 250x and 2x the fluorine concentration in rats treated with H-24000.
The pM. equivalents of fluorine in the livers from rats dosed 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-24000. However, fluorine levels in the liver were lower than levels in rats dosed with the positive control materials, H-24019 and H-24020. Liver weights were elevated in rats dosed with H-24000, at the end of the dosing period, but not at the end of the
recovery period.
-9-
Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 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 3-5; 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-24000, 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-24042, was up to 36% higher than the negative controls on day 10. On days 13,24, and 52 (recovery), the mean relative liver weights of rats dosed with the test substance were up to 11-37% higher than the negative
controls. The weights were similar by day 94.
2.
Positive Controls
The mean relative liver weights of rats dosed with the positive control H-24019 was 7% lower on day 10 than the mean relative liver weight of rats dosed with the test substance. At the end of the recovery period, the mean relative liver weight of rats dosed with H-24019 was 11% higher than the mean relative liver weight of rats dosed with the test substance. The mean relative liver weight of rats dosed with the positive control H-24020 was 28% 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 weights (% of body weight) of rats dosed with H-24019 were up to 27% higher during the dosing period than the negative controls. The mean relative liver weights of rats dosed with H-24019 were 13-39% higher than the negative controls on test days 13, 24, and
-10-
Conipany Sanitized. Does not contain TSCA CBi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
52 (recovery). At test day 94 (end of recovery), mean relative liver weights of rats dosed with H-24019 were 14 or 28% higher than controls.
The mean relative liver weights (% of body weight) of rats dosed with H-24020 were 43-75% higher during the dosing period than the mean relative liver weights of the negative controls. The mean relative liver weights were 30-63% higher than controls. On test days 52 and 94, the mean relative liver weights of the positive controls were similar to the negative controls.
Therefore, liver weights of rats dosed with the test substance, H-24000, were elevated at the end of the 10-day dosing period but not at the end of the recovery period.
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 /JiM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15
and 10.72 jiiM equivalents.
2.
Test Substance
Levels of total fluorine in livers from rats dosed with the test substance, H-24000, were lower than the levels in livers from rats dosed with the positive control materials. The total fluorine concentration in the liver from rats dosed with H-24000 was 648.16 jxM equivalents at day 10
and 4.69 (JIM equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 8x higher (day 10) and approximately 250x higher (end of study) than H-24000. For H-24020, the liver concentrations were approximately 2x higher (end of dosing) and 2x higher (end of study) than H-24000.
The /zM 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-24000. However, fluorine levels in the liver were lower than levels in rats dosed with the positive control materials, H-24019 and H-24020. Liver weights were elevated in rats dosed with H-24000, at the end of the dosing period, but not at the end of the
recovery period.
-11- Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
TABLES
-12- Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
Test Days 5 10 13 24 52 94
TABLE 1
MEAN BODY AND UVER 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
-13-
Company Sanitized. Dotes iioi contam "t'SCACBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925
Supplement No^l
Test Days 5 10 13 24 52 94
TABLE 2 (Continued)
MEAN BODY AND UVER WEIGHTS (g)
, H-24020 (POSITIVE CONTROL)
Absolute
Body Weight
I iver 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-24000 (TEST SUBSTANCE)
Absolute
Body Weight
liver Weight
236.6
12.090
279.6
16.710
304.3
19.183
378.7
18.146
512.8
21.158
608.6
22.816
Mean Relative Liver Weight(% of Body Weight)
0.051 0.060 0.063 0.048 0.041 0.037
-14- Company Sanitized. Does not contain "i3CA CBi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
TABUE2 MEAN UVER FLUORINE LEVELS
Negative Controls
Positive Controls
Test Substance
Corn
Test Corn Oil
OiLAcetone
H-24019
H-24020
H-24000
K)<02<S)3328.18 Days 94
(ppm)_____(ppm)______(ppm)
(ppm)_______(ppm)
(19.3)1'155.76 (29.2)486.32 (38.2)
0.6"______I8______77.56 (10.1)
1.68 (0.3)
3.72 (0.9)
a One value. Four of the values were below the level of detection (LOD).
b Standard deviation is in parentheses.
TABLE 3
MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Test Days
10
94
Positive Controls
Test Substance
H-24019
H-24020
H-24000
{JIM F Equivalents 5045.85 (296.91)2
ftM F Equivalents___/xM F Equivalents
1127.25 (211.37)
648.16 (50.9)
1190.15 (156.05)
10.72 (2.14)_______4.69 (1.1)
a Standard deviation is in parentheses.
-15- Company Sanitized. Does not contain TSCA CB1
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
FIGURES
-16- Company Sanitized. Does not contain T3CACB!
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 1 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND NEGATIVE CONTROLS
Test Day .17.
Company SimificedL Does not contain TS
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 2 CONPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
Test Day
-18-
Company Sanitized. Does not contain
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURES COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATI
FOR H-24000 AND NEGATIVE CONTROLS
Dy 10
Dnyl3
Dy94
Day 10
B Corn Oil (Negative Control) Corn Oil:Acetone (Negative Control) D H-24000 (Test Substance)
Dy94
.19'"o( contain
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 4 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
Day 10
Day 94
-20-
Company Sanitized. Does not contain
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
FIGURE 5 COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZED T
6000
5000
4000
'. 3000
a h
S 2000
1000
Blood / Liver
Blood / Liver
Blood / Liver
-21-
Company Sanitized. Does not contain TS
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
APPENDICES
-22- Co.panySan.f.ed.Ooesnotconta.nTSCACB
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
APPENDIX A Individual Final Body Weights, Liver Weights, and Relative Liver to Body Weight
-23- Company Sanitized. Does not contain TSCA CB{
H-24000: Biopersistciice Screening 10-Dose Oral Gavage,Study in Rats
DuPont-2925 Supplement No. 1
CORNr OIL (NEGATIVE CONTROL)
IMDI\riDUAL FINAL BOIDY WEIGHTS (g), LIVER WEIGHTS ((?), AMD RELATIVE LF/ER TO BODY WEIiGHT OP 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
.24-
Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistaace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDI\H:DUAL FINAL BODiY WEIGHTS (g),, LIVER WEIGHTS (gr), AND RELATIVE LIV]SR 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 Mumber 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
-25-
Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-DoseOral Gavage Study in Rats
DuPont-2925 Supplement No. 1
CORN Oil,: ACETONE (NEGA1riVE CONTROL) 1VSDT\7TDUAL FINAL BC>DY WEIGHTS (g),, LIVER WEIGHTS (g) , AND
RELATIVE LFVER TO BODY WEI GHT 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
-26-
Company Sanitized. Does not contain TSCA CB(
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
CORN OIL,: ACETONE (NEGA1:riVE C'ONTROL)
INDI\H:DUAL FINAL BOiDY WEIGHTS (g),, LIVE;R WEIGHTS (<?), AND RELATIVE LF/ER TO BODY WEI GHT 017 MALE RATS
Aniinal 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
Rela.tive 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 Wfc-
533.7
530.3
555.4
506.2
512.3
GROUP V
Liver Wt. 20.498 22.901 23.312 21.287 18.378
Rela tive 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
Rela fcive Liver to Body Wt. 0.033
0.034
0.030
0.032
0.035
Test Day
94 94 94
94
94
-27-
Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistenee Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
H-2-1019 (POSITIVE CONTROL)
IND:H/TDUAL FINAL BOIDY WEIGHTS (g), LIVER WEIGHTS (cr), AND
RELATIVE LI^/ER TO BODY WEI'GHT 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 Wfc. 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-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
H-2i1019 (POSITIVE CONTROL) INDI\fTDUAL FINAL BODY WEIGHTS (g),. LIVER WEIGHTS (g ) , AND
RELATIVE LI^/ER TO BODY WEI 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- Company Sanitized. Does not contain TSCA CBf
H-24000: Biopersistelace Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
H-2-1020 (POSITIVE iCONTROL)
IMDI\aDUAL FINAL BCIDY WEIGHTS (g), LIVER WEIGHTS ((?), AND RELATIVE LF/ER TO BODY WEHGHT OP 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-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
H-2.4020 (POSITIVE CONTROL)
INDIVI DUAL FINAL BC>DY WEIGHTS (g) ,, LIVER WEIGHTS (gr), AND RELATIVE LFi^ER TO BODY WEI GHT 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 TSCA CBi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
H-:24000 (TEST SUBSTANCE)
INDB^IDUAL FINAL BCIDY WEIGHTS (g), LIVER WEIGHTS (cr), AND RELATIVE LFiHER TO BODY WEI GHT OP MALE RATS
Animal
Number 627227
627229
627234
627237
627239
Final
Body Wt. 234.5
246.4
221.7
250.9
229.6
GROUP I
Liver Wt. 12.326 11.258 11.419 13.360
12.086
Relative Liver to Body Wt.
0.053
0.046
0.052
0.053
0.053
Test Day
5 5
5 5 5
Animal
Number 627213
627219
627231
627243
627244
Final
Body Wt. 301.3
262.2
295.1
262.6
276.6
GROUP II
Liver Wt. 18.118 16.329 17.325" 14.537 17.241
Relative Liver to Body Wfc.
0.060
0.062
0.059
0.055
0.062
Test Day
10 10 10
10 10
Animal
Number 627216
627235
627238
627240
627242
Final
Body Wt.
300.3
308.6
319.7
307.5
285.4
GROUP III
Liver Wt. 19.998
19.957 20.224 17.884 17.851
Relative Liver to Body Wt. 0.067 0.065
0.063 0.058 0.063
Test Day
13 13
13 13 13
-32-
CcBHgBaaGSyanitized. Doas ncsteaatain TSCA CBfi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
H-;24000 (TEST SUBSTANCE)
iinm7TDUAL FINAL BOiDY WEIGHTS (g), LIVER WEIGHTS (or), AMD
RELATIVE LP/ER TO BODY WEI'GHT OF MALE RATS
Animal Number 627220
627222
627223
627228
627232
Final
Body Wt. 354.3
379.6
382.0
407.4
370.2
GROUP IV
Liver Wt.
15.193 17.304 18.827 20.462 18.946
Relative Liver to Body Wt. 0.043
0.046
0.049
0.050
0.051
Test Day
24
24 24 24 24
Animal
Number 627212
627214
627221
627241
627245
Final
Body Wt. 520.5
482.8
447.8
595.7
517.2
GROUP V
Liver Wt. 19.870 18.189 17.604 26.836 23.292
Relative Liver to Body Wt. 0.038
0.038
0.039
0.045
0.045
Test Day
52
52 52 52 52
Animal Number 627218
627224
627230
627233
627236
Final
Body Wt. 594.4
594.6
510.1
679.3
664.8
GROUP VI
Liver Wt. 21.496 23.067 17.673 27.844 24.002
Relative Liver
to Body Wt. 0.036
0.039
0.035
0.041
0.036
Test Day
94 94 94 94 94
-33 Company Sanitized. Does not contain TSCA CQf
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
APPENDIX B Individual Fluorine Levels
.34- Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
Terms: Active % Active
TERMS AND CALCULATIONS
Mol Wt Active
Formulation Dose % F in Active
MolWtF
Compound Calculations:
Dose Active (mg/kg)
The mg of fluorine containing compound administered per kg of animal body weight.
= (% active/100) x Formulation Dose
Dose Active (mmole/kg)
The mmole of fluorine containing compound administered per kg of
animal body weight
= dose [mg/kg] / Mol Wt Active [mg/mmol]
Dose F (mg/kg)
The mg Fluorine administered per kg of animal body weight = (% F in active/100) x Dose Active [mg/kg]
Dose F (mmol/kg)
The mmole of fluorine administered per kg of animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol]
Molar Ratio (Active/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-
Coiapany Sanitized. Does not conSa;n T3CA CBi
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2925 Supplement No. 1
TERMS AND CALCULATIONS
Individual Animal Measurement:
ppm F in liver
The ppm fluoride measured in liver
Individual Animal Calculations:
ppm F in Liver minus Bkg 0.2 ppm
The ppm fluoride measured in liver minus the background fluoride measured in control animal liver. In this case the value was established at 0.2 ppm.
ppm F in Liver normalized to 0.1 mmol/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)
j^molar equivalents of
active in liver
The /xmolar [/zmol/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 CBi
_____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 - % P data is the % Fluorine of the active (Fluorine containing component(s) in the formulation) - Molecular weight is the molecular weight of the active component in the formulation
-37-
Company Sanitized. Does not contain TSCA CBf
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24019
DuPont-2925 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. P (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/P): 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 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
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
395.61 444.32 365.79 309.63 407.04
1224.62 1375.38 1132.31 958.46 1260.00
-38-
Conipaay SamSizetL Does noS contain TSCA CBf
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24020
DuPont-2925 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
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
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
1.7 1.0 1.7 1.4 1.6
ppm F in Liver Normalized to
O.I nunoles/kg Dose
388.30 231.74 370.19 313.75 352.73
3.62 2.13 3.62 2.98 3.41
jxmolar Equivalents of Active in
Liver
1321.01 788.41 1259.42 1067.39 1200.00
12.32 7.25 12.32 10.14 11.59
-39- Company Sanitized. Does not contain TSCA CBI
H-24000: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24000
DuPont-2925 Supplement No. 1
Given:
Mol Wt. Active (g/mole):
2500
% F in Active:
40
Formulation Dose (mg/kg):
1000
MolWt.F(g/mol):
19
% Active (F Containing) in Formulation: 25
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
250 0.100 1000.0
Dose F (mmol/kg):
5.263
Molar Ratio (Active/P): 0.019
Dose F (mg/kg):_____100
Rat Number
Test Day Sample
ppmF
in
Liver
Group II
627213
10
485.5
627219
10
511.3
627231
10
459.1
627243
10
535.1
627244
10
440.6
Group VI
627218
94
4.1
627224
94
3.5
627230
94
5.0
627233
94
3.1
627236
94
2.9
ppm F in Liver
Minus Bkg 0.2 ppm
485.3 511.1 458.9 534.9 440.4
3.9 3.3 4.8 2.9 2.7
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
^molar Equivalents of Active in
Liver
194.12 204.44 183.56 213.96 176.16
647.07 681.47 611.87 713.20 587.20
1.56 1.32 1.92 1.16 1.08
5.20 4.40 6.40 3.87 3.60
-40- Company Sanitized. Does not contain TSCA CBI