Document e5vjEv8qm2Jvp3vXxeOXZOwzg
AR226-2813
Study Title
Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
AUTHOR: Xing Han, Ph.D. STUDY COMPLETED ON: December 15, 2003
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
LABORATORY PROJECT ID: DuPont-13267
WORK REQUEST NUMBEj--R:--fU=|ff^1 SERVICE CODE NUMBER:|UHj
SPONSOR:
E.I. du Font de Nemours and Company Wilmington, Delaware 19898
U.S.A.
and
3M Company 3M Center Building St.Paul.MN 55144-1000
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GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
This study was conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards.
Study Director
J^^-------- Xing Han, Ph.D.
Research lexicologist E.I. du Pont de Nemours and Company
I^-D^-ZO^
Date
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
QUALITY ASSURANCE STATEMENT
Haskell Sample Number(s): 24921
Dates of Inspections: Protocol: June 18, 2003 Conduct: July 8,9,21,28,30, 2003
Records, Reports: October 27-30, 2003
Dates Findings Reported to: Study Director: June 18, 2003; November 3,11, 2003 Management: November 3,11, 2003
DuPont-13267
^Ported by:
/^^ J ^ W^o^nd_a^_J_te_lly________
Quality Assurance Auditor
f)-D^ate W
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
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CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Approved
by:
^'^2'>
----------/
^
6 ^ ^^%^J?^on/ f.^Ph.D.U
{i^^>^------
Principal Research lexicologist and Manager
/Z-' f^tc.-UcQ
--------Date--------
Approved by:; ^^PO&J^=^
Matthew S. BogdAiHyT
Research Manager
(Z-^,-zea3
Date
Issued by Study Director:
XingHan,PlLD. Research Tonicologist
^-D-^C-^0^
Date
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
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TABLE OF CONTENTS
Page
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.................................^ QUALITY ASSURANCE STATEMENT...................................................................................3
CERTIFICATION.......................................................................................................................^
LIST OF TABLES........................................................................................................................6 LIST OF FIGURES ......................................................................................................................6 LIST OF APPENDICES..............................................................................................................^ STUDY INFORMATION.............................................................................................................? STUDY PERSONNEL..................................................................................................................9
SUMMARY.................................................................................................................................. 10 A. Age Effect........................................................................................................................ 10 B. Sex Comparison................................................................................................................ 10
INTRODUCTION.......................................................................................................................11
MATERIALS AND METHODS................................................................................................ 11
A. Test Substance................................................................................................................. 11 B. Test System....................................................................................................................... 11 C. Animal Husbandry............................................................................................................ 12
1. Animal Housing .....................................................................................................................................12
2. Environmental Conditions......................................................................................................................12 3. Feed and Water.......................................................................................................................................12 4. Animal Health and Environmental Monitoring Program....................................................................... 12
D. In-Life Methods.....................................,..............................;............................................ 13
1. Animals.......................................................................................................................--........................13
2. Dose Preparation, Analysis, and Rates...................................................................................................13
3. Sacrifice and Blood Sampling................................................................................................................13
4. Experimental Procedure......................................................................................................................... 13
E.
Analytical
14
Methods...........................................................................................................
1. Verification of Dose Solution.................................................................................................................14
2. Extraction of PFOA from Plasma for LC/MS Analysis.........................................................................14
3.
14 Instrumentation.......................................................................................................................................
4. Chromatographic Methods.................................................--.........................--.....................................15
F. Statistical Analysis............................................................................................................ 15
RESULTS AND DISCUSSION.................................................................................................. 15
A. Dose Verification.............................................................................................................. 15
B. Plasma Concentration of PFOA........................................................................................ 16
1. Male Rats..................................................................................................................y...........................16
2. Female Rats...........................................................................................TT...............................................16
3.
Comparison
between
Male
and
Female
16 Rats...........................................................................................
CONCLUSIONS..........................................................................................................................16
RECORDS AND SAMPLE STORAGE ................................................................................... 16
Kliirdi\si/N\^EjS............................................................................................................................ 17 TABLES....................................................................................................................................... 18
FIGURES .....................................................................................................................................21
APPENDICES.............................................................................................................................25
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LIST OF TABLES
Page
1. Plasma concentration in rats 24 hours post-dose following oral gavage with
10
20
mg/kgPFOA......................................................................................................................
LIST OF FIGURES
Page
1. Plasma concentration in male rats 24 hours post-dose following oral gavage with 10 mg/kgPFOA......................................................................................................................22
2. Plasma concentration in female rats 24 hours post-dose following oral gavage with 10 mg/kgPFOA......................................................................................................................23
3. Plasma concentration in male and female rats 24 hours post-dose following oral gavage with 10 mg/kgPFOA.................................................................................................. 24
LIST OF APPENDICES
Page A. Dose Verification Data.............................................................................--............................27 B. Plasma Concentration Data..................................................................................................... 30
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STUDY INFORMATION
9th Collective Nomenclature: Octanoic acid, pentadecafluoro-, ammonium salt Synonyms/Codes: Ammonium perfluorooctanoate
C-8 Perfluorooctanoate, ammonium salt PFOA H-24921
Haskell Number: 24921 CAS Registry Numbed
Physical Characteristics: White solid
Stability:
The test substance appeared to be stable under the conditions of the study; no evidence of instability was
observed.
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Sponsor:
E.I. du Pont de Nemours and Company Wilmington, Delaware 19898
U.S.A.
and
3M Company 3M Center Building
St. Paul, MN 55144-1000
Experimental Start/Completion: June 30, 2003 / August 7, 2003 Study Initiated/Completed: June 27, 2003 / (see report cover page)
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________________DuPont-13267
STUDY PERSONNEL
Study Director: Xihg Han, Ph.D.
Primary Technicians: Management:
LaRue A. Manning, B.A.
Brian P. Shertz, A.A. Matthew S. Bogdanffy, Ph.D., D.A.B.T. Gary W. Jepson, Ph.D.
Toxicology Report Preparation: Lisa G. Burchfield, A.A. Management: Nancy S. Selzer, M.S.
Laboratory Veterinarian: Thomas W. Mayer, D.V.M., Diplomate A.C.L.A.M. Management: Janice L. Connell, M.S., B.A., C.I.H.
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SUMMARY
The relationship between age and plasma concentration of ammonium perfluorooctanoate (PFOA) in rats has been investigated in this study. Male and female rats between 4 and 8 weeks old were administered 10 ing/kg of PFOA by single oral gavage. The plasma concentration of PFOA at 24 hours post-dose was measured by LC/MS. The results showed that the plasma concentration of PFOA in rats was dependent on the age and sex of the rats.
A. Age Effect
In both male and female rats, the greatest age-related difference in PFOA plasma concentration was observed between 4 and 5 weeks of age. The 5-week old male rats had PFOA plasma concentrations that were 5.4-fold higher than the 4-week old male rats. In contrast, the PFOA plasma concentrations in the 5-week old female rats were 2.4-fold lower than in the 4-week old rats. PFOA levels in plasma were no longer significantly affected by age in rats of either sex that were older than 5 weeks (through study termination at 8 weeks).
B.
Sex Comparison
At 4 weeks of age, male rats had PFOA plasma concentrations that were 2.7-fold higher than in females. The PFOA plasma concentrations in 5- to 8-week old male rats were 34.7- to 65.1-fold higher than that in the females of the same age.
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____
INTRODUCTION
Ammonium perfluorooctanoate (PFOA) is a perfluorinated octanoic acid salt used as an industrial surfactant. Elimination of PFOA in rats is sex-dependent and hormone regulated. PFOA elimination is much faster in female rats than in male rats/0 down-regulated by testosterone in both female and castrated male rats/2'^ and up-regulated by estradiol in the male rats.^ However, these studies were conducted on sexually mature rats. In the current study, male and female post-weaning rats at different ages were dosed with PFOA, and the plasma concentrations of PFOA at 24 hours post-dosing were compared. This post-dosing time point was chosen because 24 hours after dosing, the sex difference in PFOA elimination in mature rats can be easily distinguished via the plasma concentration of PFOA/^ The objective of this study was te determine if immature male and female rats eliminate PFOA from blood at rates different than those observed in mature rats. This study was not designed to fully characterize the pharmacokinetics of PFOA in immature rats, and was not designed for use in determining definitive elimination rate constants.
MATERIALS AND METHODS
A.
Test Substance
PFOA (ammonium perfluorooctanoatqUlUUB^Kvas obtained from 3M (St. Paul, MN)
and assigned Haskell Laboratory Number H-24921 upon receipt. Available information on the purity, composition, contaminants, synonyms, hazards, and hazardous material classification(s)
0| f l H H I \ was provided by the venidor and documented in the study records and report.
CAS Registry Number:
Molecular Weight: Molecular Formula:
414.1
|
Sa--S--W----Sfie^SI
B.
Test System
Male and female Crl:CD(SD)IGSBR rats were obtained from Charles River Laboratories, Inc., Raleigh, North Carolina. The Sprague-Dawley rat was chosen for this study because of the
extensive experience with this strain and its suitability with respect to longevity, sensitivity, and low incidence of spontaneous diseases. Furthermore, the Sprague-Dawley rat has been used previously for toxicokinetic testing of PFOA and other fluorinated test materials/2'5'*^
At the time of dosing, rats were 4, 5, 6, 7, and 8 weeks of age and the weight variation did not exceed 20% of the mean weight by dose group and sex. Information on the cage labels
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included the Haskell animal number and an individual identification number assigned to each rat. The individual identification number was placed on the tail of each rat.
C. Animal Husbandry
1.
Animal Housing
Upon arrival at Haskell Laboratory, rats were removed from shipping cartons and housed in appropriate cages according to Standard Operating Procedures. Animals were maintained under quarantine for at least three days, had at least one recorded weight gain, and no abnormalities detected. After the quarantine period, rats were selected for study.
Throughout the dosing period with test substance, the rats were housed individually in appropriate cages according to the SOP.
2.
Environmental Conditions
Animal room(s) were maintained at a temperature of 18-26C (targeted to 22-24C) and a relative humidity of 30-70% (targeted to 40-60%). Animal rooms were artificially illuminated (fluorescent light) on an approximate 12 hour light/dark cycle. Unless judged by the study director or the laboratory veterinarian to have significantly affected the results of the study, the relative humidity and temperature ranges in the housing rooms was recorded but not included in
the final report.
3.
Feed and Water
All animals were provided tap water ad libitum and fed PMI Nutrition International, LLC Certified Rodent LabDiet 5002 ad libitum. Animals were fasted overnight for approximately 12 hours before dosing with test substance. Food was returned approximately two hours post-
dose.
4.
Animal Health and Environmental Monitoring Program
As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to ensure that contaminant levels are below those that would be expected to impact the scientific integrity of the study:
Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead,
and other contaminants.
Feed samples are analyzed for total bacterial, spore, and fungal counts.
Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers.
Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations of key contaminants, including
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specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study.
The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. Evaluation of these data did not indicate any conditions that affected the validity of the study.
D. In-Life Methods
1.
Animals
The experimental groups were as shown below:
Group
Weeks Age* Days
I
4
28
n
5
35
m
6
42
IV
7
49
v
8
56
Number of Rats
Male
Female
10
10
10
10
10
10
10
10
10
10
* Animals were 1 day from the target age at the time of dosing. The actual age at dosing is documented in the study records.
2.
Dose Preparation, Analysis, and Rates
The test substance was administered to each rat as a single oral gavage. This route was chosen because it is the route most commonly used for toxicity studies ofPFOA. PFOA was mixed with HPLC grade water to achieve the target dose concentrations. For all experiments, a single dose level was used equivalent to 10 mg PFOA/kg body weight, and the dose volume was approximately 4 mL/kg body weight. Dose solutions were prepared one day before the day of use and stored in a 4C refrigerator prior to use.
LC/MS methods that were used for dose verification are described in Section E, Analytical
Methods.
3.
Sacrifice and Blood Sampling
Rats were sacrificed by COz asphyxiation and exsanguinated by cardiae puncture.
4.
Experimental Procedure
Rats were administered PFOA as a single oral dose of 10 mg/kg. Twenty-four hours after PFOA administration, rats were sacrificed and whole blood samples were collected by cardiac puncture. Blood samples were all collected within 20 minutes of the desired sample time. Whole blood samples were placed into EDTA tubes and maintained on wet ice. Plasma was separated by centrifugation and frozen at <-10C until analysis.
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Kidneys and livers were collected at sacrifice and were flash frozen in liquid nitrogen and stored at <-60C for possible future biochemical analysis.
E. Analytical Methods
1.
Verification of Dose Solution
PFOA standards (1.0, 0.5, 0.2, 0.1, and 0.05 fig/mL) were prepared in HPLC-grade water. The dose solution was diluted by 10,000-fold prior to the analysis by LC/MS method.
2.
Extraction of PFOA from Plasma for LC/MS Analysis
Plasma samples were processed by protein precipitation (PPT) using Isolute Array protein precipitation columns (Jones Chromatography, Lakewood, CO). A 0.5 /tg/mL solution of perfluorononanoic acid (Aldrich Chemicals, Milwaukee, WI) in acetonitrile (ACN) was used as an internal standard for quantitation of PFOA. Plasma samples were thawed, and a 20 iiL aliquot of each sample was applied to the PPT array. The plasma samples were precipitated by adding appropriate dilution rate volumes of ACN/intemal standard solution to the PPT array.
Dilution rates, ranging from 1:4 (60 ^L of internal standard solution) to 1:50 (980 fiL of internal
standard solution), were utilized in order to capture the sample concentrations within the standard curve parameters. The array was slowly eluted under vacuum into a 96-well receiver
plate, centrifuged at -3000 rpm for 5 minutes, and extracts were analyzed by LC/MS.
3.
Instrumentation
System:
Detector: Mode: Source:
LM 1 resolution: HM 1 resolution
Ion energy 1: Entrance: Collision: Exit:
LM 2 resolution: HM 2 resolution:
Ion energy 2: Multiplier (V): Capillary (kV): Cone(V): Extractor (V): RF lens (V): Source temperature: Desolvation temperature: MS Method: Mode: Time: Chl:
Waters 2790 Liquid Chromatograph. equipped with quaternary pump, column heater, and autosampler Quattro Micro Mass Spectrometer
MRM
Negative Electrospray 10.0 10.0 1.0 5
2 5 14
14
2.0 650
2.0 15 0 0 130C 350C
MRM, 2 transitions
0-10.0 min
413.00 -> 369.00
Dwell:
0.25 sec
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
Ch2:
Collision energy:
463.00 -> 419.00
Dwell:
Collision energy:
15 eV
0.25 sec 15 eV
4.
Chromatographic Methods
Method: Column: Column temperature: Mobile phases:
Plasma concentration analysis Waters Xterra MS C18,2.1x30mm, 2.5 /(m Ambient A: 50 mM Ammonium Acetate B: Acetonitrile
Gradient:
Time (min)
%B
0.0
10
0:5
10
10.0
100
10.9
100
11.0
10
Row rate:
Stop time: Injection volume:
0.25 mL/min 11.0 min
5^L
DuPont-13267
F.
Statistical Analysis
Group data is represented by mean, standard deviation, standard error, and percent coefficient of variation. Statistical significance was assessed by the Student's t-test. Rejection of data is based on Chauvenet's criterion/^
RESULTS AND DISCUSSION
A.
Dose Verification
(Appendix A)
The concentrations of the test substance in the dose solutions were verified by LC/MS method as
described in Materials and Methods, Section E. Standards for dose verification and the dose solutions were prepared by different personnel. The determined meanJ'FOA concentrations of the dose solutions were all within 20% of the targeted concentration (2.5 mg/mL, Appendix A).
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
B.
Plasma Concentration of PFOA
(Table 1, Figures 1-3, Appendix B)
DuPont-13267
1.
Male Rats
The plasma concentrations of PFOA in the male rats are shown in Table 1. Their relationship with the age of the male rats is plotted in Figure 1. The PFOA plasma concentration was
7.32 1.01 ^g/mL (mean SD) in 4-weeks old male rats. In contrast, a higher PFOA
concentration (39.24 3.89 /^g/mL) was observed in the plasma of 5-week old male rats. The mean PFOA plasma concentrations in 6-,7-, and 8-week old male rats were in a range of 37.12 to 43.19 jug/mL, which, based on the results of unpaired t-test at 99% confidence interval, were not significantly different from the plasma concentration of PFOA in the 5-week old male rats.
2.
Female Rats
The plasma concentrations of PFOA in the female rats are shown in Table 1. Their relationship with the age of the female rats is plotted in Figure 2. The plasma concentration of PFOA in 4week old female rats was 2.68 0.64 iMg/mL, which was significantly higher (p<0.01) than the PFOA concentrations (mean ranged from 0.57 to 1.18 p-g/mL) in the elder females that were 5 to
8 weeks old.
3.
Comparison between Male and Female Rats
PFOA plasma concentrations in male and female rats are shown in Figure 3. PFOA plasma concentrations were higher in the males than in the females for all age groups. The most dramatic sex-dependent difference in PFOA concentration was observed for rats that were 5 weeks and older. The ratios of PFOA concentrations between male and female rats that were 5 to 8 weeks old were all greater than 34, whereas the ratio was only 2.7 for the 4-week old rats.
CONCLUSIONS
Age has a significant effect on the plasma concentrations of PFOA in both male and female rats following oral dosing with 10 mg/kg PFOA. The sex-related difference in PFOA plasma concentration is most pronounced for rats that are older than 5 weeks, with male rats having PFOA plasma levels more than 34-fold higher than female rats.
RECORDS AND SAl^IPLE STORAGE
Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware.
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ft
Ammonium Perfluorooctanoate: Age Effect on the
Plasma Concentration in Post-Weaning Rats Following Oral Gavage________________DuPont-13267 REFERENCES
1. Ohmori K., Kudo, H., Katayama, K., and Kawashima, Y. (2003). Comparison of the toxicokinetics between perfluorocarboxylic acids with different carbon chain length. Toxicology 184,135-140.
2. Vanden Heuvel, J.P., Davis, J.W., Sommers, R., and Peterson, R.E. (1992) Renal excretion ofperfluorooctanoic acid in male rats: inhibitory effect of testosterone. J. Biochem. Toxicol. 7,31-36.
3. Kudo, N., Suzuki, E., Katakura, M., Ohmori, K., Noshiro, R., and Kawashima, Y. (2001) Comparison of the elimination between perfluorinated fatty acids with different carbon chain length in rats. Chem. Biol. Interact. 134, 203-216.
4. Ylinen, M., Hanhijarvi, H., Jaakonaho, J., and Peura, P. (1989) Stimulation by oestradiol of the urinary excretion ofperfluorooctanoic acid in the male rat. Pharmacol. Toxicol. 65, 274277.
5. DuPont Haskell Labjaratory (2003), Perfluorooctanoic Acid: Toxicokinetics in the Rat. 6. Vanden Heuvel, J.P., Kuslikis, B.I., Van Rafelghem, M.J., and Peterson, R.E. (1991). Tissue
distribution, metabolism, and elimination ofperfluorooctanoic acid in male and female rats. J. Biochem. Toxicol. 6, 83-92, 7. Taylor, J.R. (1982). In An Introduction to Error Analysis. E.D. Commins, Editor, Oxford University Press.
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TABLES
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
TABLES
EXPLANATORY NOTES
ABBREVIATIONS:
SD
standard deviation
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Table 1:
Plasma concentration in rats 24 hours post-dose following oral gavage with 10 me/kg PFOA.
Group
Age (weeks)
Males
Mean
SD
Females
Mean
SD
I
4
7.32
1.01
n
5
39.24
3.89
m
6
43.19
3.79
rv
7
37.12
4.07
v
8
38.55
5.44
2.68 1.13 1.18 0.57 0.81*
0.64 0.46 0.52 0.29 0.27*
Data expressed as /xg/mL. * According to Chauvenet' s criterion,'7' the value of PFOA plasma concentration obtained from rat 511
(2.66 /xg/mL) was not included for the calculation of the mean and the SD.
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FIGURES
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
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Figure 1:
Plasma concentration in male rats 24 hours post-dose following oral gavage with 10 mg/kg PFOA.
Age (weeks)
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Figure 2:
Plasma concentration in female rats 24 hours post-dose following oral gavage with 10 mg/kg PFOA.
5 6 7 Age (weeks)
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%
Ammonium Perfluorooctanoate: Age.Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
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Figure 3:
Plasma concentration in male and female rats 24 hours post-dose following oral gavage with 10 mg/kg PFOA.
-Male -o- Female
5
6
Age(weeks)
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APPENDICES
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
APPENDICES
ABBREVIATIONS:
EXPLANATORY NOTES
LOQ SD SE % CV
limit of quantitation
standard deviation
standard error
percent coefficient of variation
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APPENDIX A Dose Verification Data
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Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
Number of
Replicates______Description______mg/mL
Dose Solution I - Group I
1
250 ng/mL dilution - 3A
2
250 ng/mL dilution - 3A
3
250 ng/mL dilution - 3A
4
250 ng/mL dilution - 3B
5
250 ng/mL dilution - 3B
6
250 ng/mL dilution - 3B
7
250 ng/mL dilution - 3C
8
250 ng/mL dilution - 3C
9
250 ng/mL dilution - 3C
2.64 3.01 2.70 2.83 2.60 2.84 2.80 2.75 2.78
Mean SD SE
%CV
Dose Solution II - Group n
1
250 ng/mL dilution - 3A
2
250 ng/mL dilution - 3A
3
250 ng/mL dilution - 3A
4
250 ng/mL dilution - 3B
5
250 ng/mL dilution - 3B
6
250 ng/mL dilution - 3B
7
250 ng/mL dilution - 3C
8
250 ng/mL dilution - 3C
9
250 ng/mL dilution - 3C
2.77 0.12 0.04 4.45
2.29 2.06 1.93 2.20 2.22 1.87 1.77 1.86 1.85
Mean SD SE
%CV
Dose Solution ffl - Group III
1
250 ng/mL dilution - 3A
2
250 ng/mL dilution - 3A
3
250 ng/mL dilution - 3A
4
250 ng/mL dilution - 3B
5
250 ng/mL dilution - 3B
6
250 ng/mL dilution - 3B
7
250 ng/mL dilution - 3C
8
250 ng/mL dilution - 3C
9
250 ng/mL dilution - 3C
2.01 0.19 0.06 9.47
2.69 2.65 2.62 2.59 2.82 2.55 2.60 2.73 2.76
Mean SD SE
%CV
2.67 0.09 0.03 3.32
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Number of
Replicates______Description______mg/mL
Dose Solution IV -
1 2 3 4 5 6 7 8 9
Group IV
250 ng/mL dilution - 3A 250 ng/mL dilution - 3A 250 ng/mL dilution-3A 250 ng/mL dilution - 3B 250 ng/mL dilution-3B 250 ng/mL dilution - 3B 250 ng/mL dilution - 3C 250 ng/mL dilution - 3C 250 ng/mL dilution - 3C
2.49 2.47 2.32 2.51 2.51 2.43 2.63 2.62 2.56
Mean SD SE
%CV
Dose Solution V 1 2 3 4 5 6 7 8 9
Group V 250 ng/mL dilution - 3A
250 ng/mL dilution-3A 250 ng/mL dilution-3A 250 ng/mL dilution - 3B 250 ng/mL dilution - 3B
250 ng/mL dilution - 3B 250 ng/mL dilution - 3C 250 ng/mL dilution - 3C 250 ng/mL dilution - 3C
2.50 0.09 0.03 3.74
2.28 2.19 2.11 2.18 1.78 2.07 1.85 2.20 2.21
Mean SD SE
%CV
2.10 0.17 0.06 8.13
Dose Solutions I-V
Mean SD SE
%CV
2.41 0.34 0.11 14.19
DuPont-13267
-29-
Company Sanitized. Does not contain TSCA CBI
Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
____DuPont-13267
__
APPENDIX B Plasma Concentration Data
-30-
Company SapEtized. Does not contain TSCA CBI
Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
Males
Animal Number
Group I 101 102 103 104 105 106 107 108 109 110
Mean SD SE
%CV
Group II
201 202 203 204 205
206 207 208 209 210
Mean SD SE
%CV
Group m 301 302 303 304 305 306 307 308 309 310
Mean SD SE
%CV
HgfmL
6.01 6.20 8.31 6.38 6.64 7.22 8.45 7.19 8.11 8.74
7.32 1.01 0.32 13.86
33.40 39.43 46.42 41.89 38.72 37.35 42.64 35.11 40.96 36.45
39.24 3.89 1.23 9.90
38.14 39.95 42.00 45.27 41.38 43.46 48.23 50.44 40.99 42.01
43.19 3.79 1.20 8.77
DuPont-13267
-31-
Company Sanit'zeri. Does not contain TSCAC38
Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
Males (continued)
Animal Number
Group IV
401 402 403 404 405 406 407 408 409 410
Mean SD SE
%CV
Group V 501 502 503 504 505 506 507 508 509 510
Mean SD SE
%CV
HS/mL
30.49 31.28 40.22 41.41 36.76 35.14 38.70 35.96 38.36 42.93
37.12 4.07 1.29 10.97
32.66 37.93 46.78 27.47 38.05 39.94 41.65 38.03 39.16 43.85
38.55 5.44 1.72 14.10
DuPont-13267
-32-
Company Sanitized. Does not contain TSCA CBI
Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
Females
. Animal Number
Group I 111 112 113 114 115 116 117 118 119 120
Mean SD SE
%CV
Group II
211 212 213 214 215 216 217 218 219 220
Mean SD SE
%CV
Group m 311 312 313 314 315 316 317 318 319 320
Mean SD SE
%CV
HS/mL
3.05 2.41 2.52 2.27 2.87 3.33 1.67 2.13 3.90 2.67
2.68 0.64 0.20 23.77
0.98 1.18 0.81 1.79 1.65 0.76 1.47 0.69 0.43 1.52
1.13 0.46 0.15 41.13
1.39 0.94 0.89 1.92 0.66 1.39 1.06 0.84 0.56 2.12
1.18 0.52 0.17 44.43
DuPont-13267
-33Company Sanitized. Does not contain TSCA C0(
Ammonium Perfluorooctanoate: Age Effect on the Plasma Concentration in Post-Weaning Rats Following Oral Gavage
Females (continued)
Animal Number
Group IV
411 412 413 414 415 416 417 418 419 420
Mean SD SE
%CV
Group V 511* 512 513 514 515 516 517 518 519 520
(te/mL
0.66 0.70 0.21 0.51 0.75 0.43 1.09 LOQ LOQ 0.23
0.57 0.29 0.09 51.00
2.66 0.87 0.63 0.95 1.36 0.67 0.64 0.89 0.50 LOQ
Mean SD SE
%CV
0.81 0.27 0.09 33.17
DuPont-13267
* According to Chauvenet's criterion/71 the value of PFOA plasma concentration obtained from rat 511 was not included for the calculation of the mean and the SD.
.34Company Sanitized. Does not contain TSCA CBl