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FINAL REPORT
Epidemiology
Medical Department
3M Company
St. Paul, MN 55144
Date: March 15, 2002
Title: IdentificationofFluorochemicals in Human Sera. TIL Pediatric Participants in a Group A Streptococci Clinical Trial Investigation
Study StartDate: September 29, 2000
Protocol Number EPL0011
Principal Investigator: 3M Co-investigators:
Geary W. Olsen, D.V.M., Ph.D!
Jean M. Burris, MP.H., RN.!
James K. Lundberg, Ph.D? Kristen J. Hansen, Ph.D.
Jeffrey H. Mandel, M.D." Larry R. Zobel, M.D."
"
Study Sponsor:
Corporate Occupational Medicine, Medical Department, 3M Company, 220-3W-05, St. Paul, MN 55144
1. Medical Department, 3M Company, St. Paul, MN 55144. 2. Environmental Laboratory, 3M Company, St Paul, MN 55144
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ABSTRACT
aPraesloorinz
The purpose of this study was to better characterize the distribution of seven
fluorochemicals, including perfluorooctanesulfonate (PFOS, CsFinSO5) in 599 pediatric
samples obtained from a multi-centerclinical trial of group A streptococcal infections.
Serum samples were collected in 1994-1995 and frozen at -20 degrees Celsius. The
samples were void of personal identifiers. The only known demographic factors were:
age (2-12), gender and the state residence (n = 23 states and the District of Columbia).
Sera samples were extracted and quantitatively analyzed for seven fluorochemicals
using high-pressure liquid chromatography/electrospray tandem mass spectrometry. The.
seven fluorochemicals were perfluorooctanesulfonate (PFOS, CsFi7S03); N-ethyl
`perfluorooctanesulfonamidoacetate (PFOSAA, CaFirSO:N(CH:CHs)CH,CO0); N-
methyl perfluorooctanesulfonamidoacetate (M570, CsFi:SON(CH;)CH,CO0):
perfluorooctanesulfonamidoacetate (M556, CyFi7SON(CH)CH:C00): perfluorooctanesulfonylamide (PFOSA, CsFi7SO:NH,); perfluorooctanoate (PFOA, C:F1xCO0'; and perfluorohexanesulfonate (PHS, CoF13503).
Overall, the geometric mean measured concentration of PFOS was 37.5 ppb (95% CI
33.3:36.5). The measured PFOS concentrations ranged from 6.7 ppb to 515.0 ppb. Male children had a significantly (p < 01) higher geometric mean serum PFOS level compared to female children [male children geometric mean =40.1 ppb (95% CI 37.742.6) vs female geometric mean = 35.2 ppb (95% CI 33.3-37.2)). Bootstrap analysis was usedtocalculate a mean 95% tolerance limit of 88.5 ppb with an upper 95% confidence limit of 97.0 ppb. Additional geometric mean and tolerance limit data are reported for PFOA. PFHS. PFOSAA and MS70. A unique finding observed in these pediatric data
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ePevornia that was not observed in the adult or elderly data reported elsewhere, were the higher 95% tolerance limit mean concentrations for PFHS (64.5 ppb) and M570 (11.9 ppb) with upper 95% confidence limits of 80.6 ppb and 14.8 ppb, respectively. It is unlikely that these findings are a consequence of analytical systematic error between these three studies. There was a strong correlation between PFOS and both PFOA (r = 70) and PFHS (r= .66) with lower correlations with PFOSAA (r = 43) and M570 (r= 42). The `number of samples with measured concentrations of PFOSA and MSS6 less than the LLOQ prohibited meaningful statistical analyses for these compounds.
"The findings from this analysis of serum PFOS concentrations are consistent with serum PFOS levels of 645 American Red Cross blood donors and 238 elderly subjects from a longitudinal study of cognitive function. Along with other human data, the average serum PFOS concentration in non-occupational human populations may approximate 30 to 40 ppb with 95% of the population's serum PFOS concentrations below 100 ppb. Since serum PFOS concentration likely reflects cumulative human `exposure. this information will be useful for risk characterization. The higher mean 95% tolerance limits for PFHS and M570 suggest that some children may have had a greater exposure experience than adults and the elderly to products containing perfluorohexanesulfonyl fluoride (PHSF) and N-methyl perfluorooctanesulfonamidoethanol (N-MeFOSE) surface protectans.
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INTRODUCTION
ePage sr orst
In May, 2000 the 3M Company (3M) announced that it wouldvoluntarily cease
`manufacturing perfluorooctanesulfonyl- (POS, CsF1nSOSF) related materials after the
compound, perfluorooctanesulfonate (PFOS, CaFi7SO5), was found to be pervasive and
persistent in human populations, wildlife, marine mammals and piscivorous birds (3M
Company 2000; Hansen et al 2001; Giesy and Kannan 2001; Kannan et al 2001a; 2001b).
POSF, produced by an electrochemical fluorination process, is used as the basic
building block to create unique chemistries through the sulfonyl fluoride moiety using
conventional hydrocarbon reactions. For example, POS can be reacted with methyl or
ethyl amines to produce either N-ethyl or N-methyl perfluorooctanesulfonamide. At this
stage, these intermediates can be used to make amides, oxazolidinones, silanes,
carboxylates and alkoxylates as commercial products. Also, these intermediates can be
subsequently reacted with ethylene carbonate to form either N-ethyl or N-methyl
perfluorooctanesulfonamidoethanol which can be used to make adipates, phosphate
esters, fatty acid esters, urethane co-polymers and acrylates as commercialized products.
Depending upon the specific functional derivatizationor the degree of polymerization,
such POSF-based products may degrade or metabolize, to an undetermined degree, to
PFOS, a stable and persistent end-product that has the potential to bioaccumulate. While
not a major commercial product, PFOS itself has been used in some products, including
fire fighting foams
`The mechanisms and pathways leading to the presence of PFOS in human blood are
not well characterized but likely involve environmental exposure to PFOS or its precursor
`molecules and residual levels of PFOS or PFOS precursors in industrial and commercial
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3M Company
Pasas:
products. PFOS has been detected at low parts per billion (ppb) concentrations in the general population (Hansen et al 2001; 3M Company 2000) although the scope of these investigations has been limited. Using high pressure liquid chromatography/electrospray tandem mass spectrometry, Hansen et al (2001) detected an average PFOS concentration 0f 28.4 ppb (SD 13.6; range 6.7-81.5) in 65 commercial individual human sera samples. An analysis of pooled blood samples (n =3 to 6 pooled samples per location with 5 to 10 donors per pooled sample) from 18 blood banks in the United States resulted in a mean `measured PFOS serum concentration of 30 ppb with a range from 9 to 56 ppb (3M Company, 2000). Serum PFOS concentrations among production employees working in POSF-related processes were approximately 2 parts per million (ppm) depending on work activity (range 0.1 to 12 ppm) (Olsen et al 1999).
The purpose of this study was to better characterize the distribution of seven fluorochemicals, including PFOS and some of its precursors, using individual pediatric samples obtained from a multi-center clinical trial of groupA streptococcal infections. `The present study is the third formal assessment undertaken by the 3M Company to examine the distribution of PFOS in human sera. The previous two assessments examined serum fluorochemmical levels among American Red Cross adult blood donors (Olsen et al 2000a) and elderly participantsof a longitudinal cognitive function study in the Seattle (WA) area (Olsen et al 2000b).
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METHODS
Puc oirat2
Fluorochemicals The seven analytes detected and quantified in this study were: PFOS; N-ethyl
perfluorooctanesulfonamidoacetate (PFOSAA, CyF17SO;N(CH,CH3)CH,CO0): N-
methyl perfluorooctanesulfonamidoacetate (M570, CyF17SO;N(CHs)CH:COO: perfluorooctanesulfonamido acetate (M556, CaFi:SON(CHICH:CO0): `perfluorooctanesulfonylamide (PFOSA, C4F17SO:NH2); perfluorooctanoate (PFOA, C/F15CO0'; and perfluorohexanestlfonate (PFHS, CaF12SO5).
PFOSAA is an oxidation product of N-ethyl perfluorooctanesulfonamidoethanol (N-E(FOSE) and is a residual in N-E(FOSE-related chemistry which was primarily used in paper and packaging protectant applications. M570 is an oxidation product of N`methyl perfluorooctanesulfonamidoethanol (N-McFOSE) and is a residual of NMCFOSE-related chemistry which was used primarily in surface treatment applications (e.g., carpets, textiles). Therefore, POSAA and M570 can be considered markers of consumer-related exposure. Both PFOSAA and M570 can metabolize to M556 and PFOSA which, in tum can subsequently metabolize to PFOS. Unlike PFOSAA and M70, M556, PEOSA and PFOS are not specific to any one consumer application. Unlike the other analytes, PFOA and PFHS are not precursors, metabolites or residuals of PFOS. PFOA can be a residual by-product of the production of the POSF-related `manufacturing electrochemical fluorination process and was produced by 3M to be an emulsifier in a variety of industrial applications (e.g.. ammonium salt) (Olsen et al 2000). PFOA can also be an oxidation product or metabolite of the widely used telomer-based
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PageerTrota fluorochemicals manufactured by other companies. PFHS, the sulfonate form of perfluorohexane sulfonyl fluoride (PHSF) may be a residual by-product of POSF-related production. 3M produced the PHSF as a building block compound incorporated in fire fighting foams and specific post-market carpet treatment applications.
`Sample Collection `The sera analyzed in this study were collectaesd partof a large multi-center clinical
trial of 1,131 children, ages 2 to 12 years, who presented with signs and symptoms of acute-onset pharyngitis (Kaplan et al 1998). All 1,131 children had positive throat cultures for group A streptococci at an initial visit. The objective of the original research `was to determine age-specific geometric mean titers and upper limits of normal for antistreptolysin O and anti-deoxyribonuclease B. Sera for the clinical trial were obtained between January 1994 and March 1995. Sera were kept frozen at -20 degrees Celsius by the University of Minnesota Department of Pediatrics priotor the 3M request of an alloguot of 0.1 ml per sample for the present study (additional amounts were obtained for the reliability analysis - see below). Because of the uncertainty regarding the population istibution of PFOS, sample size was estimated by the use of tolerance limits (Natrella 1966). Provided below is the sampling distribution that was used. Percent sampled was the highest for the younger ages and included all samples four years of age and less.
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5PMiCgEoermsvpoiarnns!2y
Age Gr2oup 43 5 6 7 8 9 10 1
12 Toul
Toul N 27 51 81 122 116416 131 135 109 8817 1131
Sampled (%) 27 (100) 5811((110000)) 100 (82) 68003(575)) 44003(310)) 40 (37) 4400 ((4496)) 599 (53)
Fluorochemical Analysis Northwest Bioanalytical (Salt Lake City, Utah) analyzed the serum for the target
fluorochemicals using techniques similar to those described by Hansen et al (2001). Details of the specific analytical procedures are presented elsewhere (NWB 2002). Briefly, the analytical method consisted of a liguid:liquid extraction procedure followed by evaporation and reconstitution of the extract residue with 20 mM ammonium acetate in water:20 mM ammonium acetate in methanol (30:70, v/v). The samples were analyzed by high pressure liquid chromatography/tandem mass spectrometry. Quantitation of the target analytes in the serum samples was performed by comparing the chromatographic peak areas for each compound to those generated in a series of extracted. calibration standards prepared from control Chinese plasma. The samples were injected in a systematic order. Evaluation of quality control samples injected during each analytical run indicated that the reported quantitative results may have varied, on average, up t0 26 percent using human plasma calibration curves for all analytes except PFOSA which may have varied on average up to 43 percent.
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5M Company Paearst Also presented in this report is acalculated total organic fluorine (TOF) index.
`TOF was the percent of each of the seven fluorochemicals' molecular weight that was
attributed to organic fluorine [PFOS (64.7%); PHS (61.9%); PFOA (69.0%); PFOSAA
(55.3%); PFOSA (64.7%); MST0 (56.6%) and MSS6 (58.1%) multiplied by the ppb.
measured for each fluorochemical and then summed across all seven fluorochemicals.
Data Analysis Measures of central tendency applicable to log normally distributed data (median,
`geometric mean) were used for descriptive analyses. In those instances where a sample was measured below the lower limit of quantitation (LLOQ), the midpoint between zero and the LLOQ was used for calculation of the geometric mean. An assessment of this midpoint assumption and how it affected the calculation of the geometric mean was
In
performed using the 10 and 90TM percentile values between zero and the LLOQ for those
values LLOQ.
In order to minimize parametric assumptions in the estimation of extreme
percentiles of the population, the bootstrap method of Efron (1993) was used to generate
confidence intervals around the empirical percentiles for serum concentrations. Tn this
method. a large number of replicated estimatesofthe percentile are generated from full-
size samples of the original observations drawn with replacement, The distribution of the
deviations of replicates from the original-sample estimate mimics the underlying
sampling distribution for the estimate. Bias-corrected, accelerated percentiles were used
to minimize residual bias. The bias correction factor is derived by comparing empirical
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MPuec sl0oorns2 `percentiles to bootstrap percentiles and acceleration is accomplished by partial jackknifing.
An analysis of the reliability of the assay was conducted after the original samples were analyzed. The laboratory was blind to the identity of these samples as they related 10 the original values reported. Triplicate samples were analyzed for the highest one percent of the measured concentrations of PFOS, PFOA and PFHS. If there was insufficient serum sample left for analysis, the next highest sample was included for analysis. A 20 percent random sample of the next highest nine percent samples was also conducted but with only a single measurement. Finally, a five percent sample was randomly chosen of the remaining 90 percent of all samples. This five percent sample was also analyzed only once. Altogether, there were 62 samples reanalyzed representing sera from 44 unique children.
RESULTS The results for the reliability analysis for PFOS, PFOA, PFHS, PFOSAA and
M570 using the reanalyzed samples is displayed in Figure 1. There were no measured concentrations of PFOSA that were above the LLOQs. Only 12 of the 62 M556 concentration comparisons were above the LLOQ; thus, these graphs are not displayed. There were strong correlations for PFOS (r = 98), PFOA (r = .96) and PFHS (r = 93). Correlations were slightly less for PFOSAA(r = .69) and for M570 (r = 80). Both PFOSAA and M570 had many comparisons below the LLOQ as represented in the `graphs near the abscissa (0.0) on the identity (Iny = In x) line.
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reat
Provided in Table 1 is the distribution of the 599 children by age and gender. Altogether there were 299 males and 300 females. Presented in Table 2 is thedistribution by states (n = 23) and the District of Columbia. One subject (female) was not analyzed
due to an insufficient quantity of serum sample.
`The measured concentrations of PFOSA and M556 were: predominantly below the LLOQ. For PFOSA, there were no subjects with a concentration above the LLOQ, 457 subjects had concentrations <LLOQ (1.0 ppb), 82 subjects had concentrations <LLOQ (2.0 ppb) and 50 subjects had analyses below the LLOQ but failed to meet the
performance standards of the analytical method. As there was only 0.1 ml, on average, per sample, no subsequent analyses were conducted on these 50 samples for PFOSA. For
M556, 258 subjects had concentrations that ranged between 2.5 ppb and 9.9 ppb, 263
subjects had concentrations <LLOQ (2.5 ppb) and 77 subjects had concentrations < LLOQ (5.0 ppb). Assuming the midpoint between zero and the LLOQ, the geometric
`mean for M556 was 2.4 ppb (95% CI 2.2 - 2.5). Because PFOSA and M556 had `many
analyses <LLOQ, statistical analyses are not presented for these compounds. They were
included in the calculation of the TOF index using, for those PFOSA or M556 values
<LLOQ. the midpoint between zero and the LLOQ.
`The distributions of the five remaining fluorochemicals, PFOS, PFOA, PFHS,
PFOSAA and M570, are displayed in Figure 2 for the 598 children samples analyzed. Although the graphs are suggestive of log normal distributions, only the PFOS
distribution met such criteria based on the Shapiro-Wilk test. This lack of lognormality
may be due to the greater proportion of subjects with values <LLOQ for PFOA, PFHS,
PFOSAA and M570.
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PMueornss
`The range, interquartile range, number of samples < LLOQ, cumulative 90
percentile, median, geometric mean and 95% confidence interval of the geometric mean for PFOS, PFOA, PFHS, PFOSAA and M570 are provided in Table 3 for all children (N = 598), males only (N = 300) and females only (N = 298). Overall, the geometric mean concentration of PFOS was 37.5 ppb (95% CI 36.0-39.1). The POS values ranged from 6.7 ppb t0 515.0 ppb. Male children had a significantly (p < 01) higher geometric mean serum POS level compared to female children although the absolute difference was not substantial [male children geometric mean = 40.1 ppb (95% CI 37.7-42.6) vs female `geometric mean = 35.2 ppb (95% C1 33.3-37.2)]. Male children also had significantly higher geometric mean serum levels of PFOA and PFHS compared to female children. There were not gender-related geometric mean differences for PEOSAA and M570. The `geometric mean for the calculated TOF index was 38.9 ppb (95% CI 37.2407). The calculated TOF index range was 9.6 ppb to 803.7 ppb. Geometric means of male children (41.6 ppb, 95% CI 38.8-44.5) were significantly (p <.01) higher than female children (36.4 ppb. 95% C1 343-387).
Measures of central tendency for each of the ages, 2 to 12, are presented in Table 4. Provided in Figure 3 is a graphical distribution (natural log scale) of the five fluorochemicals by each age stratified by gender. The box covers the interquartile range of the natural log distribution. The circle within the box is the mean. The whiskers extend to the last observation within 1.5 times the interquartile range. The dots with lines. through them represent observations outside the 1.5 times interquartile range. Analyzed as a continuous variable in simple regression models. age was significantly (p < .05)
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Congr Puc 3ori2 negatively associated with PFOA and M570 in both males and females but not with PFOS, PFHS or PFOSAA. As discussed previously in the Methods, the geometric mean data were calculated under the assumption that, for individual serum fluorochemical values <LLOQ, the `midpoint between zero and the LLOQ was assigned. For PFOS, no subject had a value <LLOQ; thus this assumption did not affect is calculation of the geometric mean. However, many subjects had values less than the LLOQs for PFOA, PFHS, PFOSAA and MS70 (see Table 2). If these values were assumed to be 10% or 909% of this range between zero and the LLOQ, the respective range of the geometric means (95% confidence interval in parenthesis) became: PFOA 4.6 ppb (4.34.9) 0 5.0 ppb (4.8-5.2); PFHS 3.2 ppb (2.8-3.8)10 5.2 ppb (4.7-5.7); PFOSAA 2.5 ppb (2.2-2.7) 10.3.7 ppb (3.63.9 and M570 1.1 ppb (1.0-1.3) to 2.3 ppb (2.2-2.5). These geometric mean values were ot substantiallydifferent than those calculated using the midpoint between zero and the <LLOQ as presented in Table 2. Consequently, the midpoint between zero and the LLOQ was used for the analyses. Provided in Figure 4 is a graphical presentation of the fluorochemical data (natural log scale) by the 23 sates and the District of Columbia stratified by gender. Interpretation of the graphs is comparable to those discussed above for Figure 3. For PFOS, mean values were comparable for the various locations. Statistical analyses by state were problematic because of the limited sample size for any given age and gender combination. Scatter plots (log scale) between the five fluorochemicals are displayed in Figure 5. PFOS and PFOA were highly correlated (r = 70). PFOS had a lower correlation with
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Mesogny Pelorsz PFOSAA(r = 43) and M570 (r= 42). The correlation between PFOSAA and M570 was. less (r= 27). The remaining scatter plots display the correlations between PFOS and PFHS (r= 0.66) and PFOA and PFHS (r = 0.48). Both PFOSAA and M570, adjusted for age, gender and their interaction, were significant predictors of PFOS in a multivariable model (Table 5). Almost seventy percent of the variation of PFOS, however, was left unexplained. Adjusted for age, gender and their interaction, PFOA remained a significant predictor of PFOS (Table 6). A quadratic term was significant in the model `which examined the association between PFOS and PHS adjusted for age, gender and their interactions (Table 7). Presented in Table 8 are the results from bootstrap analyses conducted to provide tolerance limits. The tolerance limits represent the limit of each fluorochemical within which the stated proportion of the population is expected to be found. Presented are the
mean values of the five serum fluorochemicals and TOF for the 90%, 95TM and 99" percent
tolerance limits along with the upper limit (bound) from the 95% confidence interval.
For example, the mean of the 95% tolerance limit for PFOS was 88.5 ppb with an upper
95% percent confidence limitof97.0 ppb. At the lowest tolerance limit analyzed. (90%), the mean for PFOS was 70.6 ppb with an upper 95% confidence limit of 75.2 ppb. At the highest tolerance limit analyzed, the (99%). the mean was 140.6 ppb with an upper 95 percent confidence limitof 217.0 ppb. For other fluorochemicals analyzed, the mean of the 95% tolerance limit for PFOA was 10.1 ppb with an upper 95% confidence limit of 11.0 ppb. For PFHS, the mean of the 95% tolerance limit was 64.5 ppb with an upper 95% confidence limit of $0.6 ppb. The mean of the 95% tolerance limit for PFOSAA. was 10.4 ppb with an upper 95% confidence limit of 11.2 ppb. For M570, the mean was
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e Pasnar 11.9 ppb for a 95% tolerance limit with an upper 95% confidence limit of 14.8 ppb. Finally, for the calculated index of TOF, the mean was 112.1 ppb for the 95% tolerance limit with an upper 95% confidence limit of 125.0 ppb.
DISCUSSION As seen in Figure 6, the geometric mean measured concentrations for these
`pediatric samples is consistent with those reported for adult blood donors (Olsen et al 20008) and elderly participants of a longitudinal study of cognitive function (Olsen et al 20025). No substantial differences were observed for PFOS or PFOA between the three: study populations. Interpretation of the PFHS, PFOSAA and M70 is more problematic because the LLOQs varied slightly between studies and thus the assumption ofa `midpoint value may unduly influence a geometric mean calculation when comparing mean measured concentrations for the three studies.
Displayed in Figure 7 is another perspective regarding the differences in `measured fluorochemical concentration distributions between the pediatric, adult and elderly population data. I is clearly evident that the 95% tolerance limits for PFHS and, oa lesser extent MS70, were substanialy diferent in children than compared to the adult and the elderly populations whereas the mean concentrations of the 95% tolerance limits were similar for PFOS, PFOA and PFOSAA. These findings suggest a different exposure pattem for some children compared to the adult and the elderly populations. While residual PFHS related chemistry may have existed in POSF related materials, it was an intentional major ingredient only in fire fighting foam and an after market carpet protector, which was discontinued in 1999. One potential hypothesis to explain the
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FascElr6oatanz difference between adult and children sera PFHS levels could be the differential exposure to carpet known to exist between these two population groups. The mean 95 percent tolerance limit for MS70 was also greater in children than in the adults and the elderly. M570 can be a residual analyte of N-methyl perfluorooctanesulfonamidoacetate surface protectant which would include carpet and textile applications. An alternative hypothesis, which we suspect is much less likely, is that a segment of the pediatric population clears PFHS and M570 differently than adultsorthe elderly. There appeared to be similar comparisons between the three populations for the mean 95% tolerance limit for PFOSAA which may be a residual analyte associated with the N-EXFOSE paper and. packaging protectant products.
Previous measurements of human serum samples obtained in the United States have been comparable to what has been reported in the children, adult and elderly studies. The mean PFOS serum level was 30 ppb in 18 pooled blood banks, 44 ppb from a pooled commercial sampleof500 donors, 33 ppb from a different pooled commercial sample of 200 donors and 28 ppb in 65 commercial individual human sera samples (3M Company 2000; Hansen et al 2001). These findings were also comparable to a limited number of European samples which found mean serum PFOS concentrations at 17 ppb in 5 pooled samples from a Belgium blood bank, 53 ppb in 6pooled samples from the Netherlands, 37 ppb from 6 pooled blood samples from Germany and ranged between <LLOQ (3.2 ppb) to 85 ppb in 39 individual Swedes (3M Company, 2000). The mean calculated TOF index used in the present study was also consistent with the low ppb total organic fluorine measurements of general population samples that have been reported since the 1960's (Taves 1968; Taves et al 1976: Singer and Ophaug 1979: Belisle 1981).
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1 3M Company
:
As was also observed in the adult and elderly studies (Olsen ct al 2002a; 2002b),
we found a strong correlation between PFOS and PFOA in the children sera. Whereas
PFOS has been routinely measured in human populations, wildlife, marine mammals and
piscivorous birds (Giesy and Kannan 2001; Kannan et al 2001a; 2001b; Hansen et al
2001; 3M Company 2000), serum PFOA concentrations, to date, have been consistently
quantified (i.e., measured above the LLOQs) primarily in humans, This association is of
significant interest because PFOA cannot convert to PFOS (or vice versa). Whether this
association is due to the presence of PFOA as a by-product in POSF-elated production or
10 other non-related environmental exposures or consumer products from other
manufacturers (.g., higher chain telomers) remains to be answered. Another unanswered
question is whether perfluoroctanesulfonamides can metabolize in humans to PFOA. Any of these explanations coupled with the suspected long serum half-lives in humans for
_PFOS (8.7 years (SD =6.1)) and PFOA (4.4 years (SD =3.5)) as reported by Burris et
al (2002) could explain the strong correlation between PFOS and PFOA. It should also be
noted that the serum PHS half-life reported by Burrs et al (2002) was uninterpretable (-
2.27 years, SD = 23.1) but possibly indicative ofa long (years) serum half-life.
PFOS was also corelated with two fluorochermicals, PFOSAA and MSTO, known
10 be analytes from exposure to consumer products involving paper/packaging and
carpetftextile protectants, respectively. Overall, the data, to date, indicate that PFOS
bioaccumulation in animals may be primarily through environmental sources whereas
both environmental and consumer product exposures likely contribute to serum PFOS
concentrations in humans
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PMaege 18 ota2 As with any interpretation of data obtained from a study population, questions arise regarding its representativeness and the ability to generalize from the data collected. `We are confident that our sampling procedures allowed for an adequate representation of the original study database. We believe this population of children is not unique due to the high prevalence of group A streptococcal infections in children. The only other information available for analysis were the age, gender and residence (state)ofthe children. We are unaware of any database that can be considered generalizable to the diverse United States pediatric general population without measures of random and systematic bias incorporated in the data analysis Given the consistency of the data analyzed, to date, we hypothesize that the average serum PFOS concentrations in non-occupational adult populations likely ranges between 3010 40 ppb with 95% oaf population's serum PFOS below 100 ppb. Understanding these serum PFOS levels in human populations will be useful in risk characterization since serum PFOS likely reflects cumulative human exposure (3M Company 2000). Currently available data (unpublished reports to U.S. EPA:Docket No. FYI-0500-01378) suggest, to date, that the serum concentrations observed in humans are substantially less than those required 0 cause adverse effects in laboratory animals (3M Company 2000). The data in the present study regarding the higher mean 95% tolerance limits for PFHS and M570, compared to those found in the adults and the elderly, suggest that some children may have had a greater exposure experience to products containing PHSF and N-MeFOSE surface protectants.
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ACKNOWLEDGEMENTS
3PaMgeCEo1vm9ip0an0ns2y
`We wish to acknowledge many contributors to this 3M final report. Dr. Edward Kaplan (University of Minnesota Department of Pediatrics) provided the samples for analysis. Laboratory analysis of the seven fluorochemicals was provided by a dedicated team at Northwest Bioanalytical, Inc which included Ann Hoffman, Connie Sakashita, Patrick Bennett, Dr. Rodger Foltz, Suzanne Newman and Emily Yardimici. Dr. Tim Church (University of Minnesota) provided biostatitical assistance.
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3M CEovmtpoainty Page ot2
REFERENCES 3M Company (2000). SIDS Initial Assessment Report Perfluorooctane Sulfonic Acid and its Salts. St. Paul:3M Company, September 20, 2000. Belisle J (1981). Organic fluorine in human serum: natural versus industrial sources. Science 212:1509-1510. DBeurtreirsmiJnMa,tiLounnodfbesregruJmK,haOllfs-elinveGsWo,f sSeivmerpaslofnlCuo,rMocahnedmeiclaJlHs. (2S0t0.2P)a.ulIn:t3eMriCmoRmeppaonrty:, January 11,2002. Efron B, Tibshiarani RJ. An Introduction to the Bootstrap. In: Cox DR, Hinkley DV, Reid N, Rubin DB, Silverman BW, eds. Monographs on Statistics and Applied Probability. Vol 57 New York:Chapman H Hall. Giesy JP, Kannan K (2001). Global distribution of perfluorooctane sulfonate in wildlife. Environ Sci Technol 35(7): 1339-1342. qHuaannstietnatKiJv,e CchlaermaectnerLiAz,atEilolneoffsoonrgMaEn,icJfolhunosroonchHemOic(a2l0s01ian).bioCloogmipcoaulnmda-tsrpiceecsi.fiEcnviron Sci Technol 35:766-770. KE.anNnyamnaKn,MK,oGisiteesnyenJPJ,(B2e00c1kbm).enAKc,cuEmvualnastiTo,nGoofrzpeelrafnlyuoJrFo,ocHtaannseesnulKfJo,naJtoeneins mPaDr,inHeelle mammals. Environ Sci Technol 35(8):1593-1598. KPearnfnlaunoroKo,cFtarnaensSounlfJoCn,atBeowinefrimsah-naWtiWng,wHataenrsebnirKdJs,inJcolnuedsinPgD,baGlideesayglJePs(a2n00d1). albatrosses. Environ Sci Technol-35(15):3065-3070. KdeaopxlyarnibEoLn,ucRloetahseermBeltitCerDs:, JnoohrnmsaolnvaDlRue(s1f9o9r8)c.hilAdnrteinstargeepsto2lytsoin12OinantdheanUtnii-ted States. Pediatrics 101:86-88. Leibrecht BC, Hogan JM, Luz GA, Tobias KI (1976). Donor and nondonor motivations. Transfusion 16:182-189. Natrella MG (1966). Experimental Statistics.NewYork:Wile & Sons, pp 15. Northwest Bioanalytical (NWB, 2002). Quantitative determination of PFOS and related compounds in human serum by LC/MS/MS. November 30, 2001
000088
3FaMgCEeopnmtpoooannaty O`alnsdehnepGaWti,c BaunrdrilsipJiMd,clMinaincdalelchJeHm,isZtorbyeltesLtRs i(n1f9l9u9)o.rocSheermuimcapleprrfolduuocrotoicotnaenmepsluolyfeoensat.e JOEM 41:799-806. OIdlesnetnifGicWat,ioBnurorfifslJuoMr,ocLhuenmdnibcearlgs JinK,huHamnasnesnerKaJ.,1M.anAmdeerliJcHa,nZRoebedlCrLoRss(2b0l0o2o2d).donors. St. Paul:3M Company (unpublished report) IOdlesnetnifGicWat,iBounrorfifslJuMo,roLchuenmdibcearlgs iJnK,huHmanasnesnerKaJ.,IIM.aEnlddeerllJyHP,arZtoibceilpaLntRs o(f20t0h2e0)A.dult Changes in Thought Study, Seattle, Washington St. Paul:3M Company (unpublished report). Singer L, Phaug RH (1979). Concentrations of ionic, total, and bound fluoride in plasma. Clin Chem 25:523-525. Taves D (1968). Evidence that there are two forms of fluoride in human serum. Nature 217:1050-1051. TaanvdecshaDr,acGteuryizWat,ioBn.reyInW: Fi(l1l9e7r6R),. edOsr.gaBniioccfhleumoirsotcrayrbIonnvsolivnihnugmCaanrbpolna-sFmlau:oriPnreevBaolnendcse `WashingtonDC:American Chemical Society, pp 117-134.
000089
Table 1 Distribution of All Children (N = 599) by Age and Gender
Age 2 3 4 5 6 7 8 9 10
1
12 TOTAL
Male
Female
Total
18
9
27
25
26
51
36
as
81
0
40
80
40
40
80
30
30
60
30
30
60
20
20
Fy
20
20
I
20
"20
40
20
2
1)
299
300
599
Meme Puc 220032
000090
Table 2 Distribution of Children (N = 599) by Location, Gender and Mean Age
State
Males Females Total Mean Age
Alabama
10
2
2
83
Arizona
7
n
18
68
Califomia
25
2
a
60
Colorado
2
n
EY
66
DistrictofColumbia
iy
n
2
61
Florida
16
2%
2
64
Idaho
10
6
16
8.1
linois
1
1
2
50
Kansas
10
3
3
64
Kentucky
1
7
18
69
Massachusetts
2
2
45
63
Michigan
s
4
83
Missouri
4
6
10
78
North Carolina
15
18
3
67
Nebraska.
2
3
5
63
New Jersey
.
15
38
69
New Mexico
1"
14
2%
76
New York
7
3
30
63
Ohio
16
7
33
63
Oklahoma
10
18
ES
63
Pennsylvania,
6
3
9
59
Texas
20
2
41
60
Utah
10
16
2
68
Virginia
4
19
23
83
Totoa w l 61
000091
3 CEoPmLpOaOnIy Page Bors
.
Tm se eseofCn Toreyof erm rhTees foANChin 598 adby Gr saeown Geese ment meas <oeasms ono-es
aun onCumulative 90%
.
awwon awonn ae tcoouwnn
a08 :
u8s
u83
i
u73
aoekon00 ness pe
en
amenst an
2
amen uwmn "
aseo-mr ccuuem
coarse ctooownn 2
p= Rae we <Log
Cumiive 90% Median Goomesic Mean M9e5%nCL. Gevmenic:
61-1680 70-463
.
os 7
32 Ba-m2
ina
<LOQUASISE 35-03
<106) <200) 0 0 " as-a0
<L0QUA-IT00 <LOQUO-DS <LOQ(O-381
<L0Q-102
22-56
<L0QUO-37
ras
<1san
<100
<2
<26a
<20e)
2s
8
3
8
"
i)
3
1
33-45
aoa
16-20
------------
000093
nar
| or won mon am |
Table. Mesosfernisrl TendofnSceruym lorochenicals for All Children (4 598) by Age
oe
az
Range
88-20
aa
rem
Cumlive sot 2
Metin
a
Geomet Mean x6
sGsomca Mesn ata-301
0QUY-M2 25-62
2 " us 33-63
AOQUA-W0 a0QUe-1m
us 36 " 20-85
4OQU-10S 1-52
2s 2 2 24-45
<LOQUO-163 00a 51
100 2s 2s 16-35
ae Range
ae Comiaive0% Metin [-- oGesomm. Mesn
41-180
243-506 toa 03 us 0-118
<0QUD-IG
sa-67 os sa si ren
<LOQUA-1M0
aoeee-n2 $00 2 in
30-28
<LOQUG-ISE 21-54
0 3 at 25-38
<LOQUO-34 w00@0)-39
2 20 2 16-28
CU0094
aes |
He-2s0
aa
ne-s1
Cumtaive 90% ws
Medi
Geomeri Mem GseonmcaticMean
340
13 s18-32
aes Range ao Cumsine 50%
155-965 303-564
nt
| Median
7
| Geomeric Mem ws
GseommeetricMen 268-405
20-561 12-72 01
55
Cs si-o4
A0QIA-4I60 19-02 13
3s
as 24-60
0QQ9-US 3mm 2
ss
sa 49-59
40QUA-06 1-110 ns
a
a 23-58
ar
<L0QUO-BS 20-60
2s
39
34 29-40
LOR -450 0000-46
s0
22
2 17-28
OQUS-196 23-66 103
a1
"0 23-47
<LOQUIO)-38I w00@0-47
107
23
n 13-30
000095
J Age6
Bass 40qE9-m2 0QUA-NT dOQUO-RS 0-3
ao
00-503 aes
23-5 262 4000049
| ce es
we
ns
os
ss
|
woo
"
"
2
| Gomierem a0
"
Ml
2
osice em wzws ass 38-68
30-42
16-28
se Agel
WMO 400EN-MD 4OQUA-M2
a
J ssa
26-5
cm 2
510
_
wr
w
o
| Geometric Mean
4OQNO-9 15-44 o
40QU0-ITA 06-33 o "
000096
aes | Ree
| Cumulative 0% Matin Geometric Mean sGseocme.tic Mean
awed Range Q-e [---- Metin Geometic Mean GseommectricMen
02-1160
EN EY ws 573-468
05-120 1-507 a2 wa ws 6-487
21-164 0QUA-I00 A0QUE-2T
8
505
us
52
a
a8
54
a
39
49-60
33-69
3-47
A0Q@Y-16
38-63 0 sa I 43-56
AOQUA-USO
22-16 368
5 s6 36-86
<LOQUE-1L6
400@e-52 5 a
2 21-35
har
40QUO-178
0 1 19 15-25
<LOQUO)-64 4000-30
a" 1s 1s 12-20
000097
Maelo
hee
| Cumulative 90%
Metin Geometric Mean
| Gaesanet en
men Runge a-o Cumlaive 0%
Median
Geometric Mean
eGeosmeetric Men
102-989
0.1
19 w1 05-037
104-1060 254-489
@1
38
ns
252-397
4009-8)
72
a 6 ai-s2
40QUA-BT
353
2s 5 20-49
0QU9-90 26-54 0
38 :
36
30-43
LOQUA-TS4 40QUH-66
510
18
27
17-43
40QUO-27
87
36 52 25-41
mr
40QUO-0
57
19 18 14-24
AOQUO-18T A0QRO-6E
wr
a2
37
27-50
<LOQUO-IL <L0QUY-25
50
<L0Q 0)
12
09-16
000098
nFaetra Agerz
Range
14-1240 <LOQUY-146 <AOQUAH-3ST <LOQUO-13 <LOQ(LO)-310
BEE
27-93
26-49
| Cmulaiovoe
9
ss
08-101 22
15-53
<L0QUL0-26
2
a1
Median
0
38
39
37
3
Geometric Mean
28
3s
3s
30
14
| --C CG9ee5oo8m.veCterLiiceMMeeann 29-385 30-42 O 23-54 24-39 10-18
000099
PMageeS2onti2
Table 5
.
MultivariableRegression Model of PFOS" by
PFOSAA", M570", Age, GenanddTeherir Interaction
Coefficient
SE
tratio
Intercept
31
005
8.1
PFOSAA"
02
0.02
97
M570"
02
0.02
93
Age
002
0.006
25
Gender
004
005
038
Age x Gender
-0.003
0.006
06
N=598
"Natural log Adjusted * = 31
Gender: females = 1; males =0 ratio = coefficient/SE (standard error)
p value <.0001
<.0001 <.0001
o1 40 58
000100
Pueerrtna
.
Table 6
Multivariable Regression Model of PFOS" by
PFOA', Age, Gender and Their Interaction
------ Intercept
CoCeofeffiicciieent.
SSEE
22
007
PFOA"
08
003
Age
003
0.005
Gender
0.0008
0.04
Age x Gender
-0.004
0.005
-_--
N=598
"Nawral log
Adjusted r* =.53
=
Gender: females = 1; males = 0
tratio = coefficient/SE (standard error)
ttrraattioo 35 254 61 002 038
pavlaleue <.0001 <.0001 <.0001 98 41
000101
PoEesririad
Multivariable
Table Regression
7 Model
of
PFOS"
3
by
PFHS", Age, Gender and Their Interaction
Coefficient
SE
tratio
Intercept
32
004
74
PFHS"
0.1
0.03
46
(PFHS]*
Age
003
0006
51
001
0.005
22
Gender
003
0.04
071
Age x Gender
-0.008
0.005
16
N=598
"Natural log
-
Adjusted 7 = 47
Gender: females = 1; males = 0
tratio = coefficientSE (standard error)
p value <.0001
<.0001
<.0001
03 1 12
000102
`Tolerance
Limits
and
Their
Table Asscciated
Means
and
Upper
95
Percent
Confidence
Limits for Total
OSregraunimcFFlluuoorroicnheemIincdaelxs
and
Calculated
`Tolerance
Limit
Mean
`Upper95 Percent Confidence Limit
PFOS
90%
706
752
95%
88.5
97.0
9%
1406
217.0
PFOA
90%
84
20
95%
10.1
10
9%
166
202
PFHS
90%
339
387
95%
64.5
80.6
9%
156.3
4160
PFOSAA 90%
86
91
95%
104
1n2
9%
178
207
M570
90%
72
82
95%
1
148
9%
257
38.1
TOF
90%
7.8
Ls
95%
122
1250
99%
203.0
482.1
P3aMgeCEor35miopofoarn2ty .
000103
Figure 1. AnalysisofSplit Samples for Reliability Assessment for PFOS, PFOA, PFHS, PFOSAA and M570
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000104
sir pe
Pagessora2
ii-- "l|| iLl[I ooek ]| Figure 2. Pediatric Study Population Distribution of Measured Fluorochemical Concentrations
=f
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IEEE rr eee 1
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eifet irATasesraziarasieniidis
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or
000105
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fladissiiiai 1 Usisissins;
: gedssatiizg : Bititineags
: 7 * agesmiastbimasy
PY pestiasbinnasy
ETardiaiarelangas iFe IRRRRE.RAREoh
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ilPc7a]ghpoesdaigeholt.assCiose.
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Figure 5. Scatter Plots (log scale) of Fluorochemical Associations
--
Er
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Erag o
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|
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000108
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Figur6e. CoPmFpHaSr,isPoFnOoSfAGAeoamnedtMiSc7M0efaonrsthoanPdod9i5a%tiCco(lNid=e5n6c6)a,IAndourlvta(lsN =f6or4P5)FOS, PFOA,
[
I [eee -- and E-- der (N= 238) Studios
|
_.
|
z
Ea
Sw 332s Ex
`WChidren |
macs
|oEdory J
45
10
5
o pros
PFOA
PrHS
PrOSAA
ws70
000109
| = UTppoerFh3i5-Pg% oC7.odCnot(maNop=naSri9am9osf tfohr PMb0e0a,n 9Fy5O%RToPln EoRrSrcPLeiO nnsaan tMhET
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