Document MGrGxyLjO9axbaQeGrJkQZm9L
AR226-3205
DuPont-2927 ______Supplement No. 1
TRADE SECRET Study Title
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Supplement No. 1 Laboratory Project ID: DuPont-2927
Author: Carol Finlay, B.A. Study C ompleted on: August 3,2000 Supplement No. 1 Completed on: December 12,2001 Performing Laboratory: E.I. du Pont de Nemours and Company
Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
lm&Work Request Number: 1mService Code Number:
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Company
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
C E R T IF IC A T IO N
We, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study.
Reviewed by:
' ''Paul M. Hinderliter, Ph.D. Postdoctoral Fellow
Reviewed by: { l * s d 6 e L 6 . / f Judith C. Siadfer, PhD.. D.A.B.T. [ J Director
Issued by Study Director:
C ( I JJb C iC M lJju , Carol ^inlay, B.A. Q
Staff Scientist
Date
0 # ' < G C '2 o d / Date
i l - A l U . --U O t Date
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rate
DuPont-2927 Supplement No. 1
TABLE OF CONTENTS
Page
C E R T IF IC A T IO N
2
LIST OF TABLES
4
LIST OF FIG U R ES................................. LIST OF APPRNDICES...................... .
........... ........................................... .
.......................................... ................... . 4
STUDY INFORMATION..
5
STUDY PERSONNEL
7
REASON FOR SUPPLEMENT NO. 1 ....................................... ................................................. 8
SUMMARY...................................................................................................................................... 9
MATERIALS AND METHODS1*..*.*..#...*.*...*...*.......................................................................
RESULTS AND DISCUSSION
.......................
A. Liver Weights
1. Test Substance.............................
2. Positive Controls.........................
B. Liver Fluorine D ata.................
1. Positive Controls.........................
2. Test Substance.............................
10
10
....................................10 ............................. ............................................................... 10
................................................. l i
" ......................................... h
.............
ii
C O N C L U S IO N S .. ........................
11
T A B L E S .........
a ....a ..................
at#..........a.........*.....*..**.*..*..*1J-2
FIG U RES.......
i..................
.... 16
A P P E N D IC E S ............
.....................
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 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-24018 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 D O SE....................................................................................................................................... 21
LIST OF APPENDICES
Page
A. INDIVIDUAL FINAL BODY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY W EIGHT........................................................................ ............................................................................................23
B. INDIVIDUAL FLUORINE LEVELS......................................................................................................................34
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
STUDY INFORMATION TEST SUBSTANCE:
Substance Tested: Svnonvms/Codes: H-24018
DuPont-2927 Supplement No. 1
Known Impurities:
POSITIVE CONTROL: SnVigtanp.fi Tested: Potassium perfluoroalkyl sulfonate
PFOS !
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
STUDY INFORMATION (Continued)
POSITIVE CONTROL: Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt
Svnonvms/Codes: H-24020
C-8 Perfluorooctanoate, ammonium salt APFO Ammonium perfluorooctanoate
Haskell Number: 24020
Sponsor: E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A.
Study Initiated/Comnleted: April 30,1999 / (see report cover page)
Tn-Life Initiated/Comnleted: June 13,1999 / September 14,1999
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Company
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
STUDY PERSONNEL
Study Director: Carol Finlay, B.A. Management: Judith C. Stadler, Ph.D., D.A.B.T.
Primary Technician: Richard P. Mathena
Liver Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D,, D.A.B.T.
Toxicology Report Preparation: Wanda F. Dinbokowitz
Laboratory Veterinarian: Wanda L. West, D.V.M., A.C.L.A.M.
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
REASON FOR SUPPLEMENT NO. 1
Th purpose o f this supplement is to present the results from analysis o f selected liver samples from rats treated with the test substance (H-24018), 2 positive controls (H-24019 and H-24020), and 2 negative controls (com oil and com oil/acetone).
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
SUMMARY
The objective o f this study was to evaluate the potential for H-24018, when administered by gavage, to be absorbed and to accumulate in a mammalian system. Six groups o f 5 male Crl:CD(SD)IGS BR rats each were given 1000 mg/kg/day o f H-24018. The test substance was administered to one group o f 5 rats for 5 consecutive days and to 5 groups for 10 days. On selected days (5,10, 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 o f total fluorine analyses in liver. Additionally, 2 negative controls, com oil and com oil/acetone (80:20), and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-24018.
The mean relative liver weight (liver/body weight) o f rats dosed with the test substance, H-24018, was up to approximately 27% higher than negative controls on day 10. On day 94, the weights were similar. On day 10, the mean relative liver weight o f rats dosed with the positive control H-24019 was up to 27% higher than the negative controls. By day 94, the mean relative liver weight was up to 28% higher than the negative controls. In rats given H-24020, the mean relative liver weight was up to 75% higher at day 10 than the negative controls. By day 94, the weights were similar.
The concentration o f fluorine in the livers from rats dosed with the test substance, H-24018, was 77.12 jxM equivalents on day 10 and 8.59 /xM equivalents on day 94. On day 10, the mean /xM equivalent concentrations o f fluorine in the livers from rats dosed with the positive control materials were approximately 65-fold (H-24019) and 15-fold (H-24020) greater than the fluorine concentration in livers from rats treated with the test substance. By day 94, the concentration for H-24019 was approximately 140x the fluorine concentration in rats treated with H-24018. The concentrations were similar for H-24018 and H-24020 at day 94.
The /xM equivalents o f fluorine in livers from rats dosed with the test substance, H-24018, were higher than levels in the blood.
Under the conditions o f this study, there was some absorption and retention o f fluorine in the liver from rats dosed with H-24018. Fluorine levels in the livers from rats dosed with the test substance were lower than levels in rats dosed with the positive control, H-24019. Fluorine levels in the livers from rats dosed with the test substance were lower on day 10 than levels in rats dosed with the positive control H-24020. On day 94, the fluorine levels were similar. Liver weights were elevated in rats dosed with the test substance, H-24018, at the end o f the dosing period, but not at the end o f the recovery period.
SCACB1 *>*" <'" <>" l' ' `
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 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 o f fluorine in the liver. See the original report for details o f the analysis.
RESULTS AND DISCUSSION
A. Liver W eights (Table 1, Figures 1-3, Appendix A)
As with body weights, comparison o f 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-24018, were older and heavier in weight than the com oil negative control and H-24020 positive control rats, but younger and fighter in weight than the com oil/acetone negative control and H-24019 positive control rats. Therefore, fiver weights relative to body weight are presented for comparison.
1. Test Substance
The mean relative fiver weight o f rats dosed with the test substance, H-24018, was up to approximately 27% higher than negative controls on day 10. On day 13, the weight was up to approximately 26% higher than the negative controls. The weights were similar throughout the remainder o f the recovery period.
2. Positive Controls
The mean relative fiver weight o f rats dosed with one o f the positive controls, H-24019, was similar on day 10 to the fiver weight o f rats dosed with the test substance, H-24018. On days 13, 24, and 52 (recovery), the mean relative fiver weights o f rats dosed with H-24019 were 10 or 20% higher than the fiver weights o f rats dosed with H-24018. By test day 94 (end o f recovery), the mean relative fiver weight o f rats dosed with H-24019 was 17% higher than the liver weight o f rats dosed with H-24018. The mean relative fiver weight o f rats dosed with H-24020, the other positive control, was 38% higher on day 10 than the fiver weight of rats dosed with H-24018. On days 13 and 24 (recovery), the mean relative fiver weights o f rats dosed with H-24020 were 29 and 22% higher than the fiver weights o f rats dosed with H-24018. On days 52 and 94, the weights were similar.
The mean relative fiver weight o f rats dosed with H-24019 was up to 27% higher at day 10 than the negative control groups. By day 94, the mean relative fiver weight of rats dosed with
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24019 was up to 28% higher than the negative controls. The mean relative liver weight o f rats dosed with H-24020 was up to 75% higher at day 10 than the negative controls. The mean relative liver weights o f rats dosed with H-24020 began to decrease during the recovery period and were similar by test day 94 to the negative controls.
Therefore, liver weights o f rats dosed with the test substance, H-24018, were elevated at the end o f the 10-day dosing period. The mean relative liver weight by the end o f recovery in rats dosed with the test substance was higher 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 and 3, Figures 3 through 5, Appendix B)
-
1. Positive Controls
The positive controls were H-24019 and H-24020. The concentrations o f fluorine in the livers on day 10 from rats dosed with the positive control materials were 5045.85 and 1127.25 iM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15
and 10.72 fiM equivalents.
2. Test Substance
.
Levels o f total fluorine in livers from rats dosed with the test substance, H-24018, were generally 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-24018 was 77.12 pM equivalents at day 10 and 8.59 /xM equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 65x higher (day 10) and approximately 140x higher (end o f study) than H-24018. For H-24020, the liver concentration was approximately 14x higher at the end o f dosing. The concentrations were similar by the end o f the study.
The /xM equivalents o f fluorine in the livers from rats dosed with the test substance, H-24018, were higher than levels in the blood.
CONCLUSIONS
Under the conditions o f this study, there was some absorption and retention o f fluorine in the liver from rats dosed with H-24018. Fluorine levels in the livers from rats dosed with the test substance were lower than levels in rats dosed with the positive control, H-24019. Fluorine levels in the livers from rats dosed with the test substance were lower on day 10 than levels in rats dosed with the positive control H-24020. On day 94, the fluorine levels were similar. Liver weights were elevated in rats dosed with the test substance, H-24018, at the end of the dosing period, but not at the end o f the recovery period.
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
TABLES
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 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
311.1 322.9 391.2 464.7 557.8
15.861 14.906 16.635 17.240 20.118
Mean Relative
Liver Weight(% o f 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(% o f Body Weight)
0.043 . 0.044
0.047 0.041 0.040 0.032
Test Days 5 10 13 24 52 94
H-24019 (POSITIVE CONTROL)
Absolute
Bodv Weight
Liver Weight
306.6 310.1 344.8 387.9 438.1 597.9
15.456 17.336 22.049 21.313 19.826 24.455
Mean Relative Liver Weight(% o f Body Weight)
0.050 0.056 0.064 0.055 0.045 0.041
c w m * * * * * ~ ~ Mniai" TSCA CB'
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 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(% o f Body Weight)
0.070 0.077 0.075 0.056 0.039 0.034
Test Days 5 10 13 24 52 94
H-24018 (TEST SUBSTANCE)
Absolute
Body Weight
Liver Weight
265.5
14.828
296.5
16.650
333.8
19.479
406.6
18.642
488.9
20.056
596.5
20.595
Mean Relative Liver Weight(% o f Body Weight)
0.056 0.056 0.058 0.046 0.041 0.035
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rate
DuPont-2927 Supplement No. 1
TABLE 2 MEAN LIVER FLUORINE LEVELS
Test Days
10 94
Negative Controls
Com
Com O il Oil:Acetone
(ppm)
(ppm)
<0.2 <0.5
0.6a l a
Positive Controls
Test Substance
H-24019 (PPm)
328.18 (19.3)b 77.56 (10.1)
H-24020
(PPm)_____ 155.76 (29.2)
1.68 (0.3)
H-24018 (ppm)
131.3 (30.1) 14.8 (7.9)
a One value. Four o f 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-24018
ixM F Equivalents ulM F Equivalents
uM F Equivalents
5045.85 (296.9l)a 1127.25 (211.37)
77.12 (17.7)
1190.15 (156.05)
10.72 (2.14)__________ 8-59 (4.7)
a Standard deviation is in parentheses.
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
FIGURES
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
0.060
FIGURE 1
COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE CONTROLS
0.050
0.040
>
Tn>OS> 0.030
ca<u
sos?'
0.020
0.010
0.000
5 10 13 24 52 Test Day
w
DuPont-2927 Supplement No. 1
94
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
0.080 0.070
FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS:
TEST SUBSTANCE AND POSITIVE CONTROLS
H H -24019 (Positive Control) H-24020 (Positive Control) H-24018 (Test Substance)
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0.000
Test Day -18-
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_________
_
DuPont-2927 Supplement No. 1
FIGURE 3
COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24018 AND NEGATIVE CONTROLS
Company Sanitized. Does
IS Com Oil (Negative Control) Com OihAcetone (Negative Control) H-24018 (Test Substance)
wo
>
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CD
1
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats______________
O
FIGURE 4 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
DuPont-2927 Supplement No. 1
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g.
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
FIGURE 5 COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE
ptM F Equivalents
Blood / Liver
Blood / Liver
Blood / Liver
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
APPENDICES
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
APPENDIX A Individual Final Body W eights, Liver W eights, and Relative Liver to Body W eight
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627789
627801 627805
627810
627818
Final Body Wt.
247.7
240.1 283.7
265.0
260.2
GROUP I
Liver Wt. 11.428 12.216 14.081 13.395 11.981
Relative Liver to Body Wt. 0.046
0.051
0.050
0.051
0.046
Test Day 5 5 5 5 5
Animal Number 627793
627797
627799
627800
627808
Final Body Wt.
283.5
324.2
294.7
331.3
321.8
GROUP II
Liver Wt. 15.292 15.803 14.285 16.344 17.580
Relative Liver to Body Wt. 0.054
0.049
0.048
0.049
0.055
Test Day 10 10 10 10 10
Animal Number 627796
627806
627807
627809 627816
Final Body Wt.
322.7
320.3 305.2
336.2
330.2
GROUP III
Liver Wt. 13.404 15.413 14.935 14.967 15.811
Relative Liver to Body Wt. 0.042 0.048
0.049
0.045
0.048
Test Day 13 13 13 13 13
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
CORN OIL (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627791
627794
627804
627812
627817
Final Body Wt.
365.6
416.7
378.1
388.4
407.3
GROUP IV
Liver Wt. 15.371 19.105 15.326 16.775 16.600
Relative Liver to Body Wt. 0.042
0.046
0.041
0.043
0.041
Test Day 24 24 24 24 24
Animal Number 627792
627798
627813
627814
627815
Final Body Wt.
496.8
517.3
416.8
386.9
505.9
GROUP V
Liver Wt. 16.359 19.293 15.234 14.358 20.958
Relative Liver to Body Wt. 0.033
0.037
0.037
0.037
0.041
Test Day 52 52 52 52 52
Animal Number 627795
627811
627819
627820
627821
Final Body Wt.
485.0
511.6
653.1 618.0
521.5
GROUP VI
Liver Wt. 16.649 17.405 21.828 26.766 17.943
Relative Liver to Body Wt. 0.034 0.034
0.033
0.043
0.034
Test Day 94 94 94 94 94
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H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
CORN OIL:ACETONE (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 625361
625363
625364
625380
625383
Final Body Wt.
349.7
286.7
312.0
305.5
313.1
GROUP I
Liver Wt. 16.800 11.236 .13.845 13.253 13.026
Relative Liver to Body Wt. 0.048
0.039
0.044
0.043
0.042
Test Day 5 5 5 5 5
Animal Number 625355
625362
625367
625368
625378
Final Body W t .
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-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
CORN OIL:ACETONE (NEGATIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 625354
625365
625371
625376
625381
Final Body Wt.
396.1
459.6
447.3
432.9
381.8
GROUP IV
Liver Wt. 16.383 18.313 17.859 19.168 14.135
Relative Liver to Body Wt. 0.041
0.040
0.040
0.044
0.037
Test Day 24 24 24 24 24
Animal Number 625356
625359
625360
625372
625374
Final Body Wt.
533.7
530.3 555.4
506.2
512.3
GROUP V
Liver Wt. 20.498 22.901 23.312 21.287 18.378
Relative Liver t o 'Body W t . 0.038
0.043 0.042
0.042
0-036
Test Day 52 52 52 52 52
Animal Number 625353
625369
625373
625375
625377
Final Body 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-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24019 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 625322
625323
625336
625343
625349
Final Body Wt.
329.0
298.0
318.1
294.0
293.7
GROUP I
Liver Wt. 16.409 14.830 16.545 13.809 15.685
Relative Liver to Body W t . 0.050
0.050
0.052
0.047
0.053
Test Day 5 5 5 5 5
Animal Number 625320
625330
625332
625341
625350
Final Body 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 W t .
371.2
355.3
324.4
336.9
336.4
GROUP III
Liver W t . 22.322 22.259 20.718 24.403 20.541
Relative Liver to Body W t . 0.060
0.063
0.064
0.072
0.061
Test Day 13 13 13 13 13
-28Company Sanitized. Does not contain TSCA CBS
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24019 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 625325
625328
625338
625340
625352
Final Body Wt.
381.3
423.9
339.9
419.0
375.3
GROUP rv
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-
Company Sanitized. Does not contain TSCACB1
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24020 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627756
627767
627771
627773
627777
Final Body Wt.
237.5
248.6
239.6
262.3
236.0
GROUP I
Liver Wt. 15.360 19.033 16.968 17.629 16.268
Relative Liver to Body Wt. 0.065
0.077
0.071
0.067
0.069
Test Day 5 5 5 5 5
Animal Number 627768
627774
627776
627782
627785
Final Body Wt.
293.5
338.2
254.4
268.2
267.9
GROUP II
Liver Wt. 22.536 27.332 18.821 20.172 20.857
Relative Liver to Body Wt. 0.077
0.081 0.074
0.075 '
0.078
Test Day 10 10 10 10 10
Animal Number 627758
627763
627764
627787
627788
Final Body Wt.
308.7
285.8
306.7
234.9
309.4
GROUP III
Liver Wt. 26.021 22.123 21.526 16.498 22.730
Relative Liver to Body Wt. 0.084
0.077
0.070
0.070
0.073
Test Day 13 13 13 13 13
-30Company Sanitized. Does not contain ISCACBS
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24020 (POSITIVE CONTROL)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627759
627761
627770
627779
627786
Final Body Wt.
380.1 359.4
379.8
406.2
326.1
GROUP IV
Liver Wt. 22.707 19.579 19.540 24.858 16.410
Relative Liver to Body Wt. 0.060
0.054
0.051
0.061
0.050
Test Day 24 24 24 24 24
Animal Number 627757
627760
627765 627772
627781
Final Body Wt.
436.7
456.5
455.6
513.5
421.0
GROUP V
Liver Wt. 16.899 18.240 16.505 20.225 16.695
Relative Liver to Body Wt. 0.039 0.040
0.036
0.039
0.040
Test Day 52 52 52 52 52
Animal Number 627762
627769
627775
627778
627783
Final Body Wt.
528.6
609.6
552.8
445.3
557.2
GROUP VI
Liver Wt. 17.732 19.324 17.296 15.628 21.044
Relative Liver to Body Wt. 0.034
0.032
0.031
0.035
0.038
Test Day 94 94 94 94 94
-31Company Sanitized. Does not contain ISCA CBi
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24018 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627252
627253
627260
627273
627274
Firlal Body Wt.
247.9
271.4
265.0
293.7
249.3
GROUP I
Liver Wt. 14.099 15.371 15.086 16.354 13.228
Relative Liver to Body Wt. 0.057
0.057
0.057
0.056
0.053
Test Day 5 5 5 5 5
Animal Number 627246
627258
627261
627268
627269
Final Body W t .
283.0
272.7
314.3
338.6 273.7
GROUP II
Liver Wt. 14.068 15.101 17.363 20.561 16.156
Relative Liver to Body W t . 0.050
0.055
0.055
0.061
0.059
Test Day 10 10 10 10 10
Animal Number 627249
627262
627265
627266
627277
Final Body W t .
357.2
322.7
312.7
309.1
367.3
GROUP III
Liver Wt. 22.366 19.749 17.171 17.390 20.717
Relative Liver to Body Wt. 0.063 0.061
0.055 0.056
0.056
Test Day 13 13 13 13 13
-32Company Sanitized. Does not contain TSCA CBi
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
H-24018 (TEST SUBSTANCE)
INDIVIDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LIVER TO BODY WEIGHT OF MALE RATS
Animal Number 627247
627250
627255
627257
627259a
Final Body Wt.
397.3
419.9 430.7
378.3
-
GROUP IV
Liver Wt. 17.316 21.643 20.139 15.468
-
Relative Liver to Body Wt. 0.044
0.052
0.047
0.041
-
Test Day 24 24 24 24 24
Animal Number 627256
627264
627271
627275
627278
Final Body Wt.
468.5
489.0
495.9
455.6
535.5
GROUP V
Liver Wt. 18.735 20.115 19.999 18.417 23.014
Relative Liver to Body Wt. 0.040
0.041
0.040
0.040
0.043
Test Day 52 52 52 52 52
Animal Number 627248
627251
627254
627263
627270
Final Body Wt.
591.0
651.5
628.7
531.5
579.6
GROUP VI
Liver Wt. 19.084 23.777 20.844 18.753 20.515
Relative Liver to Body Wt. 0.032
0.036
0.033
0.035
0.035
Test Day 94 94 94 94 94
a Rat found dead on test day 9
- 33-
Company Sanitized. Does not contain TSCA CBS
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
APPENDIX B Individual Fluorine Levels
- 34-
Contpany Sanitized. Does not contain TSCA CBt
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
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 o f the fluorine containing components (g/mole) The mg of formulation given per kg o f animal body weight The % fluorine in the fluorine containing components o f 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 o f animal body weight = (% F in active/100) x Dose Active [mg/kg]
Dose F (mmol/kg)
The mmole o f fluorine administered per kg of animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol]
Molar Ratio (Active/F) The moles o f fluorine containing compound per mole 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 o f active needed to achieve the normalized dose
= (Normalized dose of Active [mmol/kg] / Dose Active [mmol/kg]) x Formulation Dose
- 35Company Sanitized. Does not contain TSCA CBI
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 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 o f the actual active dose. This assumes linearity between administered dose and liver fluorine levels, but is needed because different doses o f active were used in the study.
= (0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in liver minus background)
/molar equivalents of active in liver
The /molar [/mol/L] concentration of fluorine containing compound in liver based on the ppm fluorine in liver normalized to 0.1 mmol/kg active dose. This assumes that all liver fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm -
1 mg/L = (Normalized ppm [mg/L] fluorine in liver / Mol Wt F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x
1000 /mol/mmol
- 36Company Sanitized. Does not contain TSCA CBi
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
DuPont-2927 Supplement No. 1
FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS
Considerations:
- The data used for kinetic'analysis was from a limited screen and extrapolation should be done cautiously.
- Sample size is low - Analytical data used without validation - Steady-state not reached - Terminal phase may not be reached - Some compounds are mixtures of fluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - Each compound may have very different potency for producing toxicity
Assumptions: (May or may not bejustified in all cases)
- Liver fluorine concentrations are linear with respect to dose
- Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of
individual components
.
- Background Fluorine is 0.2 ppm
- % F data is the % Fluorine of the active (Fluorine containing component(s) in the formulation)
- Molecular weight is the molecular weight o f the active component in the formulation
- 37Oompany Sanitized. Does not contain TSCAGBi
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24019
DuPont-2927 Supplement No. 1
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
497 10 100
% F in Active: Mol Wt. F (g/mol):
65 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
10 0.020 49.7
Dose F (mmol/kg):
0.342
Molar Ratio (Active/F): 0.059
Dose F (mg/kg):
6.5
Rat Number
Test ppm F Day in Sample Liver
Group II
625320
10 319.4
625330
10 355.4
625332
10 338.4
625341
10 305.0
625350
10 322.7
ppm F in Liver
Minus Bkg
0-2 PPm____
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
319.2 355.2 338.2 304.8 322.5
1586.42 1765.34 1680.85 1514.86 1602.83
{molar Equivalents of Active in
Liver
4910.77 5464.62 5203.08 4689.23 4961.54
Group VI 625326 625335 625346 625347 625351
94 94 94 94 94
79.8 89.6 73.8 62.5 82.1
79.6 89.4 73.6 62.3 81.9
395.61 444.32 365.79 309.63 407.04
1224.62 1375.38 1132.31 958.46 1260.00
- 38Company Sanitized. Does not contain TSCACBI
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H- 24020
DuPont-2927 Supplement No. 1
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
426 20 100
% F in Active: Mol Wt. F (g/mol):
69 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
20 0.047 42.6
Dose F (mmol/kg):
0.726
Molar Ratio (Active/F): 0.065
Dose F (mg/kg):
13.8
Rat Number
Test ppm F Day in Sample Liver
Group II 627768 627774 627776 627782 627785
10 10 10 10 10
182.5 109.0 174.0 147.5 165.8
ppm Fin Liver
Minus Bkg 0.2 ppm
182.3 108.8 173.8 147.3 165.6
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
pim o la r 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
Group VI 627778 627783 627762 627775 627769
94 94 94 94 94
1.9 1.2 1.9 1.6 1.8
1.7 1.0 1.7 1.4 1.6
3.62 12.32 2.13 7.25 3.62 12.32 2.98 10.14 3.41 11.59
Company Sanitized. Does not contain TSCA CB1
H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats
Data for H-24018
DuPont-2927 Supplement No. 1
Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation:
845 1000 34
% F in Active: Mol Wt. F (g/mol):
50 19
Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor:
340 0.402 248.5
Dose F (mmol/kg):
8.947
Molar Ratio (Active/F): 0.045
Dose F (mg/kg):
170
Rat Number
Test ppm F Day in Sample Liver
Group n 627246 627258 627261 627268 627269
10 126.4 10 130.6 10 169.4 10 86.6 10 143.5
ppm F in Liver
Minus Bkg 0.2 ppm
126.2 130.4 169.2 86.4 143.3
ppm F in Liver Normalized to 0.1 mmoles/kg
Dose
unolar Equivalents of Active in
Liver
31.36 32.41 42.05 21.47 . 35.61
74.24 76.71 99.53 50.82 84.29
Group VI 627248 627251 627254 627263 627270
94 94 94 94 94
10.7 8.9 27.6 17.4 9.4
10.5 8.7 27.4 17.2 9.2
2.61 6.18 2.16 5.12 6.81 16.12 4.27 10.12 2.29 5.41
- 40Company Sanitized. Does not contain TSCA CBS
