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Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
(\ & T & -o 5 6 0
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
A nalytical Laboratory Report
ON THE
Determination of the Presence and Concentration
of Potassium Perfluorooctanesulfonate
(CAS Number: 2759-39-3)
in the Serum and Liver of Sprague-Dawley Rats Exposed to PFOS via Gavage
Laboratory Report No. <U2006> Requester Project No. <3M TOX 6295.9>
Study Dates
Study Initiation: 26 May 1998 Study Completion: At Signature
0C0038
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
3M Environmental Laboratoiy Report No. Tox-012
Laboratory Request Number (LRN>U2006
Table of Contents
Table o f Contents.........................................................................................................................................2
Study Personnel and Contributors.............................................................................................................. 3
Statement of Compliance.......................................................
4
Quality Assurance Statement......................................................................................................................6
In tro d u c tio n ..................................................................................................
7
Sample Receipt............................................................................................................................................ 7
Materials and Methods..............................................................................................................
8
Chemical Characterization................................................................................................................... 8
Method Sum m aries..............................................................................................................................9
Analytical Equipment.......................................................................................................................... 11
Protocol Deviations.............................................................................................................................12
Data Summary, Analyses, and Results.................................................................................................... 12 Summary of Quality Control Analyses Results..................................................................................12 Summary of Sample Results............................................................................................................. 13
Statistical M ethods......................................................................................................................
14
Data Quality Objectives and Data Integrity................................................................................................15
Statement of Conclusion............................................................................................................................15
R eferences................................................................................................................................................ 15
Attachment A: Report Signature Page
Attachment B: Table of Results of Analysis of Liver Samples from FO Male and Female Sprague-Dawley Rats Administered PFOS Daily by Gavage
Attachment C: Table of Results of Analysis of Sera Samples from FO Male and Female , Sprague-Dawley Rats Administered PFOS Daily by Gavage
Attachment D: Table of Results of Analysis of Liver Samples from F1 Male and Female Sprague-Dawley Rats Administered PFOS Daily by Gavage
Attachment E In-life Protocol and Analytical Study Protocol
Attachment F: Preparatory and Analytical Methods
Attachment G: Dose Analyses
000039
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Potassium Perfluorooctanesulfbnate CAS Number-2759-39-3
Study Personnel and Contributors
Analytical Chemistry Laboratories
3M Environmental Technology and Safety Services
3M Environmental Laboratory Fluorine Analytical Chemistry Team (FACT) 2-3E-09 935 Bush Avenue S t Paul, MN 55106
Kris Hansen, Ph.D., Analytical Study Director Lisa Clemen, Analytical Chemlst(s)
In-life Testing Laboratory
Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044
Raymond G. York, Ph.D., Study Director
Sponsor
3M Toxicology Services 3M Center Building 220-2E-02 S t Paul, MN 55144
Marvin Case, SponsorRepresentative
3M Environmental laboratory
Report No. Tox-012 Laboratory Request Number (LRN>U2006
000040
Page 3 of 16
Potassium Perfluorooctanesulfbnate CAS Number-2759-39-3
3M Environmental Laboratory
Report No. Tox-012 Laboratory Request Number (LRN)-U2006
Statement of Compliance
Study Title:
Analytical Laboratory Report on Presence and Concentration of Potassium Perfluorooctanesulfonate in Serum and Liver of Sprague-Dawley Rats Exposed to PFOS via Gavage
Study Identification Number . FACT Tox-012
This study was conducted in compliance with Food and Drug Administration Good Laboratory. Practice (GLP) Regulations for Nonclinical Laboratory Studies [Data Requirement(s): 21 CFR ( Part 58)], with the exceptions in the bulleted list below. In addition, the present study has been audited retrospectively by an independent quality assurance unit. Audit procedures and findings for audits performed at the 3M Environmental Laboratory and at participating contract laboratories are housed with documentation pertinent to this study in archives at the 3M laboratory and will be retained for at least 10 years. The analytical portion completed at the 3M Environmental Lab was performed in accordance with 3M Environmental Technology and Safety Services Standard Operating Procedures.
Exceptions to GLP compliance:
Storage containers for the reference substance, PFOS, were not labeled with name, CAS number, batch number, expiration date, or storage conditions.
Details o f the preparation, maximum storage time, and stability properties of the reference substance, PFOS, are unknown.
The identity, strength, purity, and composition defining the test or control article was not determined at the time of the study; however, analyses to detail the characterization were ongoing at the time of this analytical study.
Two separate study directors were assigned to the in vivo and the analytical portions o f this study.
From 15 September 1998 until 1 October 1998, the protocol was not sponsor approved and was during that time period not in compliance.
Deviations were not consistently approved by the study director as required by GLP regulations.
Data corrections were not always recorded as required by GLP regulations.
Data were not always attributed to the individual responsible for recording the data.
Records in electronic form do not meet the criteria set forth under 21 CFR(11).
A finalized sample log-in /tracking system was not in place at the time of the study; however, draft documents did exist and were used to log in and track samples.
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Potassium Perfluoroocfanesulfbnate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Not all raw data were verified by the study director.
No in-phase audits were conducted during the present study.
A t the time of the present study, personnel files did not always contain documentation o f equipment and method training; however, personnel files have been updated, and this noncompliance has been resolved.
Signature of Study Director
------- --- *
z ?' Oct 93
Signature o f Study Sponsor
n
\
&
Q M u a . /<? *
CC0042
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Potassium Perfluorooctanesulfonate CAS Number-2759-33-3
GLP Study Q uality Assurance Statement
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRNHJ2006
S tudy T itle :
Analytical Laboratory Report on Presence and Concentration of Potassium Perfluorooctanesulfonate in Serum and Liver of Sprague-Dawley Rats Exposed to PFOS via Gavage
S tudy ID Num ber:
FACT Tox-012
This study has been inspected by the 3M Environmental Laboratory Quality Assurance Unit (QAU) as indicated in the following table. The findings were reported to the study director and management
In s p e c t io n D a t e s
From
i
To
P hase
9 August 1999 27 September 1999 Final Report
Date Repo rted to
Management \ Study Director i
4 October 1999 4 October 1999
I
i I
I
IQ -l-qq
Date
o c0043
Page 6 of 16
Potassium Perfluorooctanesulfbnate CAS Number-2759-39-3
Introduction
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Potassium Perfluorooctanesulfonate
CAS Number 2795-39-3
Chemical Formula: CaF17S 0 3-K+
Molecular W eight 538
In the present study, groups of FO rats were administered 0.1, 0.4,1.6, or 3.2 mg PFOS kg/day in 0.5% Tween 80. (These doses correspond to concentrations of 0.02, 0.08, 0.32, and 0.64 mg/mL.) Groups of vehicle control F0 rats were administered only Tween 80. Male F0 animals were treated 42 days prior to mating and through the mating period; female F0 animals were administered PFOS daily 42 days prior to mating, through gestation, and up to 20 days following litter delivery. F1 male and female rats were exposed to the chemical in utero and during lactation. Following weaning at 21 days of age, selected F1 animals were treated during development and production of F2 animals. Various physical and biological parameters were monitored in F0, F1, and F2 animals.
In the analytical study reported here, liver and sera samples collected from the initial population of dosed animals (generation F0) and their offspring (generation F1) were analyzed for the presence of PFOS. (Analyses were performed to determine the presence of EtFOSE, PFOSA, POAA< PFOSEA, PFOSAA, and the monoester; however, these data were collected for informational purposes only, and were not reported.)
Liver samples were homogenized, and liver and sera samples were extracted by an ion-pairing extraction procedure. The extracts were quantitatively analyzed using high-pressure liquid chromatography/electrospray tandem mass spectrometry (HLPC/ESMSMS), and PFOS levels were evaluated against extracted standards. Analytical details are included in this report; further details are available in the study binder maintained by the 3M Fluor Analytical Chemistry Team (FACT).
Analyses assessing the toxicological effects of PFOS in the livers and sera of Sprague-Dawley rats were conducted in compliance with Good Laboratory Practice Regulations (21 CFR 58). Validated methods and standard operating procedures were followed during the preparation and analysis o f the samples associated with this study.
Sample Receipt
Tissue samples were received from Argus Research Laboratories sporadically, from August, 1998, through January, 1999. Samples received were packaged in dry ice. Specimens were registered with the 3M Environmental Laboratory and transferred to a freezer far storage. Sample receipt identification, storage, and chain o f custody protocols and data are located in the study folder for this report; the folder is located in the 3M archives.
Specimens analyzed at the 3M Environmental Laboratory will be maintained fo ra period of 10 years and will be stored at the laboratory at -20C 10C.
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Potassium Perfluorooctanesuifonate CAS Number-2759-39-3
3M Environmental Laboratory
Report No. Tox-012 Laboratory Request Number (LRN)-U2006
Materials and Methods
Chemical Characterization
Table 1 presents information regarding characterization of the test, control, and reference substances used in the analytical portion of this study.
Reference Substance Source Preparation Maximum Storage Time Storage Conditions '
Chemical Lot Number Physical Description and Identity Analyses Purity Stability Control Substance Source
Preparation Maximum Storage Time Storage Conditions Chemical Lot Number
Physical Description and Identity Analyses Purity Stability
A nalyses Perform ed a t th e 3M E n v ir o n m e n t a l La b o r a t o r y
Potassium perfluorooctanesuifonate 3M Specialty Chemical Division Details unknown Unknown In a sealed container at room temperature, exposed to light 193 White powder 99.28% Unknown Rat liver and serum matrices Rabbit liver--Argus Research Laboratories, Inc. Rabbit serum-- Sigma Chemical Company Unknown Unknown Frozen at -20C LiverF00012 Serum: 17H9306 Rabbit liver and serum Unknown Unknown
Table 1. Characterization and Treatment of the Control, Reference, and Test Materials in the Study of the Presence and Concentration of Potassium Perfluorooctanesuifonate in Serum and Liver of Sprague-Dawley Rats Exposed to PFOS via Gavage
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Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
3M Environmental Laboratory
Report No. Tox-012 Laboratory Request Number (LRN)-U2006
Analyses Perform ed a t th e 3M E n v ir o n m e n t a l L a b o r a t o r y
T est M aterial Source
Preparation
Maximum Storage Time Storage Conditions Chemical Lot Number Physical Description and Identity Analyses Purity Stability
Rat serum and liver
Sprague-Dawley rats exposed to PFOS via gavage during the in vivo portion o f 3M TOX 6295.9 at Argus Research Laboratories
Liver extraction per FACT M-1.0 (refer to Attachment F) Serum: extraction per FACT M-3.0 (refer to Attachment F)
1 week
Frozen at-20C
NA
Rat liver and serum
NA
NA
Table 1. C haracterization and Treatm ent o f the C ontrol, Reference, and Test M aterials in the S tudy o f the Presence and C oncentration o f Potassium P erfluorooctanesulfonate in Serum and Liver o f Sprague-Dawley Rats Exposed to PFOS via Gavage (continued)
Following analysis, remaining original specimens (test and control material) were stored frozen at -20C and will be maintained for a period of 10 years.
Method Summaries
Following is a brief description of the methods used during this analytical study by the 3M Environmental Laboratory. Detailed descriptions o f these methods are located in Attachment F.
Preparatory M ethods:
Method FACT-M-1.0: Extraction of Potassium Perfluorooctanesulfonate or Other Anionic Fluorochemical Surfactants from Liver for analysis Using HPLC/ESMS. In this method, an ion pairing reagent was added to the sample and the analyte ion pair was partitioned into MtBE. Four mLs of the MtBE extract was transferred to a centrifuge tube and put onto a nitrogen evaporator until dry. Each extract was reconstituted in 1.0 mL o f methanol, then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials.
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Potassium Perfluorooctanesulfnate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Method FACT-M-3.0: Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum or other Fluid for Analysis Using HPLC-Electrospray/Mass Spectrometry. Sera samples were extracted using an ion-pairing extraction procedure. In summary, an ion pairing reagent was added to the sample and the analyte ion pair was partitioned into methyl-ferf-butyl ether (MtBE). Four mL of the MtBE extract was removed and put onto a nitrogen evaporator until dry. Each extract was reconstituted in 1.0 mL of methanol and filtered through a 3-cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials.
Analytical Methods
Method FACT-M-2.0: Analysis of Fluorochemicals in Liver Extract Using HPLC-Eiectrospray/Mass Spectrometry The analysis was performed by monitoring a single ion characteristic of a particular fluorochemical, such as the perfluorooctanesulfonate (PFOS) anion, m/z = 499. Additionally, samples were analyzed using a tandem mass spectrometer to further verify the identity of a compound by detecting daughter ions of the selected parent ion.
Method FACT M-4.0: Analysis of Potassium Perfluorooctanesulfonate or other Fluorochemical in Serum or other Fluid Using HPLC-Electrospray/Mass Spectrometry The analysis was performed by monitoring a single ion characteristic of a particular fluorochemical, such as the perfluorooctanesulfonate (PFOS) anion, m/z= 499. Additionally, samples were analyzed using a tandem mass spectrometer to further verify the identify of a compound by detecting daughter ions of the parent ion.
Confirmatory dose analyses were conducted following the end of the study, and the results of these analyses are presented in Attachment G.
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Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
3M Environmental Laboratory
Report No. Tox-012 Laboratory Request Number (LRN)-U2006
Analytical Equipment
Following are typical analytical equipment settings for the procedures used in the present study. These settings vary somewhat during actual data collection. Exact settings during all phases of data collection are recorded and presented in the analysis section of the study binder for FACT Tox-12.
HPLC System: Hewlett-Packard Series 1100 Liquid Chromatograph
Column:
Keystone Betasil C18 Column 2X100 mm, 5pm particle size
Flow rate:
300 pDrain
Solvent A:
2.0 mM ammonium acetate
Solvent B:
Methanol
Solvent Gradient
40% to 90% B in 8.5 minutes Hold at 90% B for 3.0 minutes Return to 40% B in 1.0 minute Hold at 40% B for 1.0 minute
Injection Volume:
10 pL
Run Time:
13.5 minutes
Electrospray Tandem Mass Spectrometer: Micromass Quattro II API Mass Spectrometer
Mass Lynx 3.1 Software
Cone Voltage:
30--60V
Collision Gas Energy:
40 eV
Mode:
Electrospray Negative
Source Block Temperature: 115C
Desolvation Temperature: 250C
Electrode:
Z-spray
Primary lon/Daughter Ions: 499/80, 99, 130, 180, 230 amu
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Potassium Perfluorooctanesutfonate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRNJ-U2006
Protocol Deviations
There was one deviation to the protocol: The protocol called for the use of reference standard lot number 217; however, lot number 193 was used during the analytical portion of the study.
Data S ummary, A nalyses, and Results
Summary of Quality Control Analyses Results
Calibration Check Standards: A mid-level, extracted matrix calibration check was analyzed at least every 10 samples to monitor instrument drift Calibration check standards were compliant, and all calibration criteria were met (within 30%).
Blanks: Four blanks were extracted concurrently with each batch of samples. Two extraction blanks were prepared with water as a surrogate matrix. Two additional samples o f blank matrix were typically prepared with rabbit liver or sera and were used for extracted calibration curves. All blanks were nondetect for the compound(s) of interest
Duplicate matrix spike analyses including all target analytes were prepared and analyzed for one control animal. With a few exceptions, recoveries were within the acceptable range of 70130%.
Analysis o f two liver matrix spike samples resulted in 51 % and 61 % recoveries o f PFOS. These recoveries are below the lower boundary of the acceptable range; however, the second set of matrix spike samples were within the acceptable range. No action was taken to further characterize these matrix spikes.
Analyses of both sets of sera matrix spikes resulted in PFOS recoveries greatly above the high limit of the acceptable range. Although 1 mL of sera was used for the analysis of PFOS levels in the sera of test animals, only a small amount of serum (500 pL) was available for the matrix spike preparation; this limited quantity of matrix may have been a source of error. As the extraction is scaled-down to accommodate the available matrix, the resulting extract is particularly susceptible to loss of solvent due to evaporation. These evaporative losses, which would not effect the PFOS concentration (PFOS has a low vapor pressure) in the extract, could result in a greater-than-expected PFOS concentration in the extract When analyses of sera matrix spikes are performed with a full 1 mL of sera, high PFOS recoveries are unusual.
Matrix Spikes: Matrix spikes and matrix spike duplicates were analyzed every 40 samples, with a minimum of two per batch during analytical analyses. The results of these analyses are located in the archived study binder.
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Potassium Perfiuorooctanesulfonate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Summary of Sample Results
PFOS was detected in the livers of all control and exposed groups of FO animals and F1 animals. PFOS was detected in the sera samples collected from all of the control and dosed FO males and in sera samples collected from all dosed FO females. The PFOS levels found in the samples from control animals were quite low--generally 100-less than levels detected in the low-dose group. PFOS concentration in exposed animals increased with increasing dose group, generally In a dose-related manner.
PFOS levels in both sera and liver samples collected from female FO animals were much less than levels detected in samples collected from the corresponding group of FO males.
PFOS concentrations measured in pooled liver samples collected from the exposed groups of F1 animals shortly after birth were roughly equal to or lower than concentrations measured in the corresponding groups of FO females. No samples collected from FO females that received 3.2 mg PFOS per kg/day were submitted for analysis. Table 1 summarizes results of the quantitative analyses performed on the FO liver and sera samples examined.
G roup
D o se (mg/kg/day)
D ose C oncentratio n ,
(m g /m L)
Average C o ncentratio n of
P F O S in S erum (pc/iuiL)
A verage C o ncentratio n o f
P F O S in Liver (UG/G)
0.0 mg/kg/day
0.0
0.0
Female: 0.0307 Male: 0.0244
Female: 0.171
i
Male: 0.665
0.1 mg/kg/day
0.1
0.02
Female: 5.28
Female: 14.8
i
Male: 10.5
Male: 84.9
0.4 mg/kg/day
0.4
0.08
Female: 18.9
Female: 58.0
Male: 45.4
Male: 176
1.6 mg/kg/day
1.6
0.32
Female: 82.0
Female: 184
Male: 152
Male: 323
3.2 mg/kg/day
3.2
0.64
Female: NRa
Female: NRa
Samples were not received.
Male: 273
Male: 1360
Table 2. PFOS Concentrations in Serum and Liver Samples from FO Male and Female SpragueDawley Rats
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Potassium Perfluorooctanasuifonate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRNHJ2006
PFOS concentrations measured in pooled liver samples collected from the co ntro l and exposed groups of F1 animals shortly after birth were roughly the equal to or lower than concentrations measured in the corresponding groups of FO females. No samples collected from F1 males or females that received 3.2 mg/kg/day were submitted for analysis. PFOS was detected in the liver of all other exposed groups of F1 animals. Table 2 summarizes results of the quantitative analyses performed on the F1 sera samples examined.
G roup
D o se (mg/ kg/day)
0.0 mg/kg/day
I
| 0.0
0.1 mg/kg/day
0.1
0.4 mg/kg/day
0.4
1.6 mg/kg/day
1.6
3.2 mg/kg/day
3.2
Samples were not received.
D ose C o ncentration (mg/m L)
A v er a g e C oncentratio n of P F O S in Liver (IJG/G)
0.0 0.0511
0.020
6.19
0.080
57.6
0.320
70.4
0.640
NRa
l
Table 3. PFOS C oncentrations in Liver Sam ples from F1 Male and Female Sprague-Dawley Rats
Paper and electronic copies of raw data and study reports generated during the present studies will be kept for at least a period of time as established by regulation and by the 3M Standard Operating Procedures. Paper and electronic copies of reports, protocols, and methods generated by the 3M Environmental Technology and Safety Services laboratory will be retained with the study folder for at least a period of time as established by regulation and by the 3M Standard Operating Procedures.
Statistical Methods
For data generated by the 3M Environmental Laboratory, means and standard deviations were calculated using Microsoft Excel, and relative standard deviations were calculated manually.
Standard deviation is a measurement of analytical precision; thus it is used to gauge the precision o f the analyses. Relative standard deviation presents a measure of the magnitude of the standard deviation.
l nZx2 - ( Z c) 2
y n (n -l) Means are calculated by adding individual entities and dividing the resultant sum by the number of individual entities. Standard deviations were calculated using the equation:
Relative standard deviations were calculated by division of the standard deviation by the mean with subsequent multiplication of the quotient by 100.
C0 0 0 5 1
Page 14 of 16
Potassium Perfluorooctanesutfonate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Data Q uality O bjectives and Data Integrity
No circumstances existed during the present study that would have affected the quality or integrity of the data. The following data quality objectives (DQOs) were followed during the study:
No circumstances existed during the present study that would have affected the quality or Integrity of the data. The following data quality objectives (DQOs) were followed during the study:
Linearity-- The correlation coefficient (R2) of the standard curve was equal to or greater than 0.98 using 1/x weighting
Limits o f detection-- PFOS in serum=1.75 ppb Limits o f quantitation-- Equal to the lowest acceptable standard in the calibration curve Duplicate ffequency/acceptable precision-- <30% Spike ffequency/acceptable recoveries--70% to 130% Use o f confirmatory methods--no confirmatory methods were used Demonstration o f specificity--specificity was demonstrated by chromatographic retention time and
mass spectral daughter ion characterization
Statement of Conclusion
Statem ent of Conclusion:
Under the conditions o f the present studies, potassium perfluorooctanesulfbnate was observed in the livers and sera o f all groups of rats exposed to the chemical during the in vivo portion of the studies.
References
None
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Potassium Perfluorooctanesutfonate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRNJ-U2006
Attachments
Attachment A: Report signature page
Attachment 8: Table of results of analysis of liver samples from FO male and female SpragueDawley rats administered PFOS daily by gavage.
Attachment C: Table of results of analysis o f sera samples from FO male and female SpragueDawley rats administered PFOS daily by gavage.
Attachment D: Table of results of analysis of liver samples from F1 male and female SpragueDawley rats administered PFOS daily by gavage.
Attachment E: In-life protocol and study protocol
Attachment F: Preparatory and analytical methods
Attachment G: Dose Analyses
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Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
Attachment A Report S ignature Page
3M Environmental Laboratory Report No. Tox-012
laboratory Request Number (LRNHJ2C06
fi r-^ Nz------------------------------------------
Kristen J. Hansen, Ph.D., Study Director
Date
7 ~ Ss.. Marvin T. Case, D.V.M., Ph.D., Study Sponsor
Date
r o / y/ 9 ?
Date
54-
Page 17 of 16
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
Attachment B
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Dose Group
Sam ple Number
Method Blank Method Blank Matrix Blank Matrix Blank QC -- 100 ppb
0.0 mg/kg/day
0.1 mg/kg/day (0.02 mg/mL)
H20 Blank 1 H20 Blank 2 Rabbit Liver (Blank 1) Rabbit Liver (Blank 2) 8801F-MS 8801F-MSD 8801M-MS 8801M-MSD 8801F 8807F 8808F 8821F 8833F 8101M 8103M 8104M 8107M 8108M 8838F 8840F 8842F 8864F 8870F 8138M 8140M 8142M 8145M 8146M
Extracted W eigh t (g)
N/A N/A
PPOS C alculated C oncentration
(mG/G)
<MDL <MOL
Total Amount of
PFOS (Mg/g)
<MDL <MDL
Mean PFOS (Mg/g)
<MDL
R e la tiv e Standard Deviation
<MDL
N/A
N/A 1.0169 1.0169 1.0525 1.0525 1.0169 1.0384 1.0177 1.0439 1.0048 1.0525 1.0407 1.0568 1.0104 1.0321 1.0374 1.0168 1.0489 1.0608 1.0225 1.0617 1.0509 0.9961 1.036 0.9788
<MDL
<MDL 60 72 100 93 674 158 165 197 166 910 600 577 593 643 17251 14433 15557 13807 12784 80270 76629 92106 87938 ! 87451
<MDL
<MDL 50% 60% 87% 81% 0.674 0.158 0.165 0.197 0.166 0.910 0.600 0.577 0.593 0.643 17.3 144 15.6 13.8 12.8 80.3 76.6 92.1 87.9 [ 87.5
<MDL 55% 84% 0.171" 0.272
0.665
14.8
84.9
<MDL 18% 7% 10.1" 0.0172" 82.9 0.225
21.0 0.139
11.6 1.71
7.39 6.28
Table 4. Analyses o f Liver Samples from FO Sprague-Dawley Rats in the S tudy o f the Presence and C oncentration o f Potassium P erfluorooctanesulfonate in Serum and Live r o f Sprague-Dawley Rats Exposed to PFOS via Gavage
Proprietary and Confidential
0CG05
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
Attachment B (continued)
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Dose G roup
Sample Number
0.4 mg/kg/day
8890F
(0.08 mg/mL)
8893F 8895F
8902F
8905F
8172M
8174M
8175M
8176M
--------------------1 8181M
1.6 mg/kg/day
8919F
(0.32 mg/mL)
8921F
8926F
8934F
8937F
8209M
8213M
8219M
8221M
8225M
3.2 mg/kg/day
NR
(0.64 mg/mL)
NR
NR
NR
NR
8243M
8244M
8249M
8250M
8255M
Extracted W eig ht (g)
1.0346 1.0247 1.0154 1.0039 1.0426 1.0234 1.0067 1.0471 1.0335 1.0197 1.0756 1.0544 1.0219 1.022 1.654 1.0016 1.0211 1.0421 1.0153 1.0316 NR NR NR NR NR 1.0065 1.0397 1.0107 1.0522 0.9908
PPOS Calculated CONCENTRATION
(mG/G) 61853 62547 49543 64081 51922 137715 189256 183050 173288 198173 200313 212684 282001 183576 40958 374773 325087 279320 335349 301145 NR NR NR NR NR 1271156 1116251 2077414 1168784 1149853
T otal A mount o fPFOS
(pG/G) 61.9 62.5 49.5 64.1 51.9 138 189 183 173 198 200 213 282 184 41.0 375 325 279 335 301 NR NR NR NR NR 1271 1116 207 7 1169 1150
M ean PFOS (mg/g )
R elative S tandard D eviation
58.0
11.6 6.73
13.3 176 23.4
48.0 184 88.3
11.2 323 36.1
NR
;I_N_R_________ NR
1360
30.0 40.7
a Sample 8801F value was excluded from the calculations.
Table 4. A nalyses o f Liver Samples from F0 Sprague-Dawley Rats in the Study o f the Presence and Concentration o f Potassium Perfluorooctanesulfonate in Serum and Liver o f Sprague-Dawley6 Rats Exposed to PFOS via Gavage (continued)
Proprietary and Confidential
000056
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
Attachment C
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
D ose G roup
S am ple Number
Method Blank Method Blank Matrix Blank Matrix Blank QC -- 100 ppb
0.0 mg/kg/day
0.1 mg/kg/day (0.02 mg/mL)
0.4 mg/kg/day (0.08 mg/mL)
1.6 mg/kg/day (0.32 mg/mL)
H20 Blank 1 H20 Blank 2 Rabbit Liver (Blank 1)
Rabbit Liver (Blank 2) 8801F-MS 8801F-MSD 8801M-MS 8801M-MSD 8801F/M 8807F/M 8808F/M 8821F/M 8833F/M 8838F/M 8840F/M 8842F/M 8864F/M 8870F/M 8890F/M 8893F/M 8895F/M 8902F/M 8905F/M 8919F/M 8921 F/M 8926F/M 8934F/M 8937F/M
PFOS Calculated C oncentration
(ng/g)
<MDL <MDL
T otal A mountof
PFOS (pg'g)
<MDL
M ean PFOS (Mg/g)
<MDL
R e l a t iv e S tandard D eviation
(RSD)
<MDL
<MDL
<MDL
<MDL 60 72 100 93 369 51.5 53.2 52.7 46.9 5872 5149 7390 6745 5788 62050 61745 47738 63057 53492 78376 58893 91956 65023 57897
<MDL 50% 60% 84% 78% 0.369 0.0515 0.0532 0.0527 0.0469 5.87 5.15 7.39 6.74 5.79 62.1 61.7 47.7 63.1 53.5 78.4 58.9 92.0 65.0 57.9
<MDL 55% 81% 0 .0 5 1 1a 0.115
6.19
57.6
70.4
<MDL 18% 7% 5.58* 0.00285" 124 0.142
14.2 0.879
11.7 6.72
20.6 14.5
a Sample 8801F value not included in this calculation
Table 5. Analyses o f Liver Samples from F1 Sprague-Dawley Rats in the Study o f the Presence and Concentration o f Potassium Perfluorooctanesulfonate in Serum and Liver o f Sprague-Dawley Rats Exposed to PFOS via Gavage
Proprietary and Confidential
0C057
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
Attachment D
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Dose G roup Method Blank Method Blank Matrix Blank Matrix Blank QC -- 100 ppb
0.0 mg/kg/day
0.1 mg/kg/day (0.02 mg/mL)
0.4 mg/kg/day (0.08 mg/mL}
Sample N umber
H20 Blank 1 H20 Blank 2 Rabbit Liver (Blank 1) Rabbit Liver (Blank 2) 8801F-MS 8801F-MSD 8801M-MS 8801M-MSD 8801F 8807F 8808F 8821F 8833F 8101M 8103M 8104M 8107M 8108M 8838F 8840F 8842F 8864F 8870F 8138M 8140M 8142M 8145M 8146M 8890F 8893F 8895F 8902F 8905F 8172M 8174M 8175M 8176M 8181M
PPO S Reported (m g /m L )
<MDL <MDL <MDL <MDL 136% 133% 119% 121% 0.127 0.0229 0.0291 0.0445 0.0265 0.0306 0.0311 0.0177 0.0213 0.0214 5.14 4.93 5.24 5.23 5.89 11.8 9.54 9.65 11.0 10.4 20.9 18.6 18.4 17.4 19.1 40.7 54.1 41.6 47.4 43.3
M ean PFOS (Mg/mL)
<MDL <MDL 135% 120% 0.037* 0.0500
0.0244
5.28
10.5
18.9
45.4
R elative S tandard D eviation (R S D )
<MDL <MDL 1% 1% 30.9* 0.00950* 87.6 0.044
2.48 0.00605
6.78 0.358
9.02 0.946
6.89 1.30
12.1 5.49
Table 6. A nalyses o f Sera Samples from FO Sprague-Dawley Rats In the Study o f the Presence and C oncentration o f Potassium Perfluorooctanesulfonate in Serum and Liver o f Sprague-Dawley18Rats Exposed to PFOS via Gavage
Proprietary and Confidential
OC0058
Potassium Perfluorooctanesulfbnate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Dose G roup
Sample Number
PPOS Reported (m g /m L )
M ean PFOS (p g /m L )
R elative S tandard D eviation (R S D )
1.6mg/kg/day (0.32 mg/mL)
3.2 mg/kg/day (0.64 mg/mL)
8919F 8921F 8926F 8934F 8937F 8209M 8213M 8219M 8221M 8225M NR NR NR NR NR 8243M 8244M 8249M 8250M 8255M
75.3 79.2 113 74.4 68.4 144 151 148 165 153 NR NR NR NR NR 249 242 257 361 255
82.0 152 NR 273
21.4 17.5
5.20 7.91
NR NR
18.3 49.8
Sample 8801F value not included in this calculation
Table 6.
Analyses of Sera Samples from F0 Sprague-Dawley Rats in the Study of the Presence and Concentration of Potassium Perfluorooctanesulfonate in Serum and Liver of Sprague-Dawley0 Rats Exposed to PFOS via Gavage (continued)
Proprietary and Confidential
OC0059
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
3M Environmental Laboratory Report No. Tox-012
Laboratory Request Number (LRN)-U2006
Attachment E In-life Protocol and A nalytical Study Protocol
Proprietary and Confidential
000C60
