Document rEG2QOojL9z7DLz5j1y71OBv

. : AR226_103 PagSe or03% FIENpAiLdemRiEolPogOyRT Medical Department 3M Company St. Paul. MN 55144 Date: February 25, 2002 Title: Identificationof Fluorochemicals in Human Sera. 1. American Red Cross Adult Blood Donors Study Start Date: September 29, 2000 Protocol Number EPL-0013 Principal Investigator: 3M Co-investigators: Geary W. Olsen, D.V.M., PhD. Jean M. Burris, MPH, RN." James K. Lundberg, Ph.D.* Kristen J. Hansen, Ph.D. Jeffrey H. Mandel, M.D." Larry R. Zobel, MD! Study Sponsor: C3oMrpCooramtpeanOcyc,up2a2t0i-o3nWa-l0M5e,diStc.iPnaeu,l,MeMdiNcal55D1e4p4artment, 1. Medical Department, 3M Company, St. Paul, MN 55144 2. Environmental Laboratory, 3M Company, StPaul, MN 55144 2 35 Ez2 no3 3]Sn ~ 1 000003 ABSTRACT "Through cooperation with six American Red Cross blood banks, 645 serum samples from adult donors (ages 20-69, equally represented of both sexes) were obtained for fluorochemical analyses. Blood bank locations were Los Angeles (CA), Portland (OR), Minneapolis-St. Paul (MN), Charlotte (NC), Hagerstown (MD) and Boston (MA). `Samples were void of personal identifiers. Age, gender and location were the only known demographic factors. Sera samples were extracted and quantitatively analyzed for seven fluorochemicals using high-pressure liquid chromatography/electrospray tandem mass spectrometry and evaluated versus an extracted curve from a human plasma matrix. The seven fluorochemicals were perfluorooctanesulfonate (PFOS, CsF17S05); N-ethyl perfluorooctanesulfonamidoacetate (PFOSAA, CsFi7SO;N(CH;CH3)CH,COO); N+ `methyl perfluorooctanesulfonamidoacetate (M570, CsFiSO:N(CH3)CH:CO0): `perfluorooctanesulfonamidoacetate (M556, CsF17SO:N(CH)CH:CO0-); perfluorooctanesulfonylamide (PFOSA, CsF17SO;NHy); perfluorooctanoate (PFOA, C+F13C00"); and perfluorohexanesulfonate (PFHS, CeF13505). Overall, the geometric mean measured concentration of PFOS was 34.9 ppb (95% CI 33.3-36.5). The measured PFOS concentration ranged from less than the lower limit of quantitation (LLOQ) of 4.1 ppb to 1656.0 ppb. The geometric mean for PFOS was significantly higher among males (37.8 ppb; 95% CI 35.540.3) than females (31.3 ppb; 95% CI30.0-34.3). No significant difference was observed with age. Charlotte (NC) had the highest geometric mean serum PFOS concentration (51.5 ppb) and Boston (MA) the lowest (29.5 ppb). Bootstrap analyses were used to calculate a 95% tolerance limit for od 000004 3M Comey [rer PFOSof 88.5 ppb with an upper 95% confidence limit of 100.0 ppb. Additional `geometric mean and tolerance limit data are reported for PFOA, PFHS, PFOSAA and M570. The geometric mean and 95% tolerance limits of these fluorochemicals were, on average, an order of magnitude (or more) lower than PFOS. PFOS and PFOA were highly correlated (r = .63). PFOS had lower correlations with PFOSAA (r= 42), PFHS (r= 38) and M570 ( = 20). The number of samples with measured PFOSA and M556 concentrations below the LLOQ prohibited meaningful statistical analyses for these compounds. `The findings from this analysis ofserumPFOS concentrations are consistent with those previously reported. The human data, to date, suggests the approximate average serum concentration in non-occupational adult populations may be 30 to 40 ppb with 95%ofa population's serum PFOS concentrations below 100 ppb. Since serum PFOS concentrations likely reflect cumulative human exposure, this information will be usefui for risk characterization. 000005 3 Pageedont36 INTRODUCTION In May, 2000 the 3M Company (3M) announced that it would voluntarily cease `manufacturing perfluorooctanesulfonyl- (POSF, CsFi7SO:P) related materials after the compound, perfluorooctanesulfonate (PFOS, CyF17SOy), 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 20012; 20010). POSE, produced by an electrochemical fluorination process, is used as the basic building block to create unique chemistries through the sulfony] fluoride moiety using conventional hydrocarbon reactions. For example, POSF 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 cither 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 derivatization or 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 POS or POS precursors in industrial and commercial o 600006 MPageetn36 `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 pressureliquid chromatography/electrospr; tandem mass spectrometry, Hansen et al (2001) detected an average PFOS concentratior: o 28.4 ppb (SD 13.6; range 6.7-8L.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 i(C donors per pooled sample) from 18 blood banks in the United States resulted in amean `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 partspermillion (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 in an adult population by analyzing sera samples obtained from donors at six American Red Cross blood banks. Am assessment of the serum fluorochemical distribution was performed in relation to three demographic atributes (age. gender and location) of the anonymous blood donors. METHODS Fluorochemicals The seven analytes detected and quantified in this study were: PFOS; N-cthyl perfluorooctanesulfonamidoacetate (PFOSAA, CsFirSO:N(CH:CH3)CH,COO: Nmethyl perfluorooctanesulfonamidoacetate (M570, CsFi7SO:N(CH3)CH:C00): perfluorooctanesulfonamido acetate (M556. CF1zSO:N(CHICH:COO): s 000007 SM Company Pees perfluorooctanesulfonylamide (PFOSA, CsFi7SO:NH.); perfluorooctanoate (PFOA, C3F13C00'); and perfluorohexanesulfonate(PFHS, CeFiSO5). PFOSAA is an oxidation product of N-ethyl perfluorooctanesulfonamidoethanol (N-EXFOSE) and is a residual in N-EtFOSE-related chemistry which was primarily used in paper and packaging protectant applications. MS70 is an oxidation product of N`methyl perfluorooctanesulfonamidoethanol (N-MeFOSE) and is a residual of NMeFOSE-related chemistry which was used primarily in surface treatment applications (e.8., carpets, exiles). Therefore, PEOSAA and M570 can be considered markers of consumer-related exposure. Both PFOSAA and MS70 can metabolize to M556 and 'PFOSA which, in tum can subsequently metabolize to PFOS. Unlike PFOSAA and MS70, M556, PFOSA and PFOS are not specific to any one consumer application. Unlike the other analytes, PFOA and PHS are not precursors, metabolites or residuals of PFOS. PFOA can be a residual by-productofthe production of the POSF-related `manufacturing electrochemical fluorination process and was produced by 3M to be an emulsifier in a variety of industrial applications (c.g., ammonium salt (Olsen etal 2000). PFOA can also be an oxidation product or metabolite of the widely used telomer-based fluorochemicals manufactured by other companies. PFHS, the sulfonate form of perfluorohexane sulfonyl fluoride (PHSF), is 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. 3 000008 IM Comey rere `Sample Collection Through cooperation with six AmericanRed Cross blood banks, 645 serum samples from adult donors (ages 20 - 69, equally represented of both sexes) were obtained for analysis. [Note: This final report supercedes the June 25, 2001 3M interim report whict: indicated 652 samples. Seven samples were not included in the final report because the subjects were determined to be ineligible by age (2 70). Additional separate 3M sponsored studies were designed to examine the distribution of serum samples among elderly adults and children.] The six American Red Cross blood banks represented : donors from the following areas: Los Angeles, CA; Portland, OR; Minneapolis-St. Paul. MN; Charlotte, NC; Hagerstown, MD and Boston, MA. Samples were void of personal identifiers. The only known demographic factors were age, gender and location. Each blood bank was requested to provide approximately 10 samples per 10 year age intervals (20-29, 30-39, 40-49, 50-59 and 60-69) for each sex. Fluorochemical Analysis Northwest Bioanalytical (Salt Lake City, Utah) analyzed the serum for the target fluorochemicals using techniques similar to those described by Hansen ct al (2001). Details of the specific analytical procedures are presented elsewhere (NWB 2002). Briefly, the analytical method consisted of a liquid-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 serum samples was performed by comparing the 7 000009 may Pasesol3e 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 to 26 percent using human plasma calibration curves for all analytes except PFOSA which may havevariedon average upto 43 percent. Also presented in thisreport is a calculated total organic fluorine (TOF) index. TOF was the percent of each of the seven fluorochemicals' molecular weight that was attributed to organic fluorine [POS (64.7%); PHS (61.9%); PFOA (69.0%); PFOSAA (55.3%); PFOSA (64.7%); M570 (56.6%) and MSS6 (58.1%)] multiplied by the ppb `measured for each fluorochemical and then summed across all seven fluorochemicals. Data Analysis Measuresof 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. performed using the 10% and 90 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. In this 8 000010 MPaseedotn36 `method,a large number of replicated estimates of the 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 distributionforthe estimate. Bias-corrected, accelerated percentiles were usec. to minimize residual bias. The bias correction factor is derived by comparing empirical percentiles to bootstrap percentiles and acceleration is accomplished by partial jackknifing. Twenty-four samples were split and analyzed to provide an estimoaftthee reliability of the analyses conducted. The analytical laboratory was blind to the identity of these split samples. These analyses were performed concurrently with all other analyses of the study to minimize experimental error. Five split samples were analyzed from Charlotte, Los Angeles, Hagerstown and Portland and four split samples from Boston. Inadvertent, no reliability analyses were performed on the Minnesota samples. RESULTS The results for the reliability analysis for PFOS, PFOA and PFHS are displayed Figure 1. Only six split samples for PEOSAA and seven split samples for M570 had values that were above the LLOQ. None of the PFOSA and M536 split samples were above the LLOQ. Therefore, only the reliability results for PFOS, PFOA and PFHS are displayed in Figure 1. There was a strong correlation between the split samples (r=.9 for each of these three fluorochemicals. It shouldbe noted that 13 of the split sample analyses for PFHS had the identical LLOQ (2.1 ppb). This is represented in the graph =. the single point near the abscissa (0,0) on the identity (In y = In x) line. 9 000011 FaseE1r00a03n6 Provided in Table 1 is the distribution of the donor subjects by 10 year age intervals, gender and location. Altogether there were 332 male donors and 313 female donors. As could be expected with the age stratification design used for sample collection, the study subjects' mean ages were comparable by gender: 44.6 years for males and 43.9 years for females. The measured concentrations ofPFOSA and M556 were predominantly below the LLOQ. For PFOSA, there was one subject with a value above (2.1 ppb) the LLOQ, 196 subjects <LLOQ (1.0 ppb) and 448 subjects <LLOQ (1.4 ppb). The different LLOQ values were determined on different analytical runs. For MS56, twelve subjects were above the LLOQ. Their M556 values ranged from 3.6 to 12.9 ppb. There were 145 subjects with M556 values <LLOQ (2.5 ppb) and 488 subjects with M56 values LLOQ (3:2 ppb). Because of the few subjects whose serum concentrations exceeded the LLOQ, statistical analyses are not presented for PFOSA and M36. Although PFOSA and M556 are not presented in the subsequent analyses, they were included in the calculation of the TOF index. For those measured concentrations of PFOSA and M556 < LLOQ, the midpoint between zero and the LLOQ was used in the calculation of the TOF index. The frequency distributions of the five remaining fluorochemicals, PFOS, PFOA, PFHS, PFOSAA and M570, are displayed in Figure 2. 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 log normality is due to the greater percentage of subjects with values <LLOQ for PFOA, PFHS, PFOSAA and M570. The range, interquartile range, number of samples < LLOQ. raw cumulative 90 percentile. median, geometric mean and 95% confidence interval of the geometric mean 10 000012 MPaEge 11 736 for PFOS, PFOA, PFHS, PFOSAA and M570 are provided in Table 2 for all subjects, males only and females only. Overall, the geometric mean levels of PFOS was 34.9 ppib (95% C133.3-36.5). The range of PFOS values was < LLOQ (4.3 ppb) to 1636.0 ppb. Male subjects had significantly (p <.05) higher geometric means for PFOS than female: subjects [male geometric mean = 37.8 ppb (95% C135.5-40.3) vs female geometric me:an =32.1 ppb (95% C1 30.0-34.3)]. Males also had significantly higher serum levels of PFOA and PFHS compared to females although the mean levels for both sexes were approximately one order of magnitude lower than that of PFOS. It should be noted that the overall geometric mean for the calculated TOF index was 31.7 ppb (95% C1 30.4 33.0) (data not shown). The calculated TOF index ranged from 5.7 ppb to 1083.2 ppb. Provided in Figure 3 is a graphical distribution (natural log scale) of the five fluorochemicals by 10 year age intervals 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. As shown in Figure 3, age was not an important predictor of adult serum fluorochemical concentrations. In those instances where there were many outliers (c.g. M570 concentrations in males aged 40-49 and 60-69), this was the resultof a large percentage of values <LLOQ that were within the 1.5 x interquartile range. As discussed previously in the Methods, the geometric mean data were calculate: under the assumption that for individual serum fluorochemical values <LLOQ the `midpoint between zero and the LLOQ was assigned. For PFOS, only one subject had 2. value <LLOQ: thus this assumption did not affect ts calculation of the geometric mean. 1" 000013 PasEeir2e00s3 `However, considerably more subjects had LLOQs for PFOA, PFHS, PFOSAA and M570 (see Table 2). If these values were assumedtobe 10% or 90% of this range between zero and the LLOQ the respective range of the geometric means (95% confidence interval in parenthesis) became: PFOA 4.0 ppb (3.74.1) to 4.8 ppb (4.6-5.0); PFHS 0.9 ppb (08-1.0) 102.5 ppb (2.4-2.6); PFOSAA 0.8 ppb (0.7-0.9) to 2.8 ppb (2.7-3.0) and M570 0.5 ppb (0.5-0.6)t0 1.9 ppb (1.8-2.0). These geometric mean values were not substantially different 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. Presented in Table 3are the range, interquartile range and medians for the six locations combined across age andgenderfor the five fluorochemicals. A graphical presentation of these data (natural log scale) is presented in Figure 4. Interpretation of the graphs is comparable to those discussed above for Figure 3. Provided in Table4 are the results from a bootstrap analysis which calculated mean serum fluorochemical values for each of the six locations adjusted for 10 year age intervals, gender and their interaction terms. The highest mean value for PFOS was Charlotte (39.0 ppb) with the lowestbeing Boston (29.0 ppb). Los Angeles, Minneapolis-St. Paul and Hagerstown had comparable mean PFOS levels of approximately 35.0 ppb with Portland slightly lower (32.8 ppb). The range of means for the other fluorochemical analytes was narrow and thus difficultto distinguish any substantial differences by location. Because PFOS is the primary contributor to the calculated TOF index, the bootstrap analysis findings for TOF mirrored those of PFOS. 1& 000014 30 Compa nore Scatter plots (log scale) between the five fluorochemicals are displayed in Figure: 5. PFOS and PFOA were highly correlated (r = 63). PFOS haad lower correlation wit PFOSAA (r = 42) and lower yet with M570 (r = 20). The correlation between PROS and M570 was weak (r = 12). The remaining scatter plots display the correlation between PFOS and PFHS (r= 0.38) and PFOA and PFHS (= 0.32). Both PFOSAA andl M570, adjusted for age, gender and their interaction, were significant predictors of PFOS in a multivariable model albeit PEOSAA was the strongerofthe two independent Variables (Table 5). Nevertheless, approximately eighty percent of the variation of PFOS was left unexplained. Age and gender were not significant predictors in models that examined the significant association between PFOS and PFOA (Table 6) or PFHS (Tabie: 7). None of the models in Tables through 7 had lack of fit F ratios that were statistically significant (p < 05). Presented in Table 8 are the results from bootstrap analyses conducted to provide `mean concentrations of several 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%, 95 and 99TM 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% confidence limit of 100.0 ppb. At the lowest tolerance limit analyzed (90%), the mean for PFOS was 70.7 ppb with an upper 95% confidence limit of 74.3 ppb. At the highest tolerance limit analyzed (99%), the mean was 157.3 pp with an upper 95% confidence limit of 207.0 ppb. For other fluorochemicals analyzed. the mean of the 95% tolerance limit for PFOA was 12.1 ppb with anupper 95% 12 000015 ) oPtie laot confidence limit of 13.6 ppb. ForPHS, the mean of the 95% tolerance limit was 9.5 `ppb with an upper 95% confidence limit of 10.8 ppb. The mean of the 95% tolerance limit for PEOSAAwas 7.6 ppb with an upper 95% confidence limit of 8.5 ppb. For M570, the mean was 5.0 ppb foar 95% tolerance limit with an upper 95% confidence: limit of 5.4 ppb. Finally, for the calculated indexof TOF, the mean was 75.1 ppb for the 95% tolerance limit with an upper 95% confidence limit of 80.9 ppb. DISCUSSION : The findings from this analysis of serum PFOS concentrations in 645 adult donors are consistent with previous human data. Previous measurements of human serum samples obtind in the United States showed mean PFOS concentrations of 30 ppb in 18 pooled blood banks, 44 ppb from a pooled commercial sample of 500 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 6 pooled samplesfromthe Netherlands and 37 ppb from 6 pooled blood Samples from Germany and 39 individual Swedes whose serum PFOS ranged between <LLOQ 32 ppb to 85 ppb (3M Company, 2000). The mean calculated TOF index of 31.7 `ppb 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 ct al 1976: Singer and Ophaug 1979; Belisle 1981). la 000016 Poe tet Unique to the present study was its large individual sample size which facilitatead the characterization of the serum fluorochemical distribution. This included the calculation of tolerance limits and their upper bounds. The highest serum PFOS measurement (confirmed by re-assay of the sample) was 1656.0 ppb. Because donor samples were anonymous, it is not possibletodetermine anything about this individual besides gender (male), age (67 years) and location (Portland). This PFOS sample approximated the average serum PFOS levels observed for POSF-related production workers (Olsen et al 1999). The next highest. donor level for PFOS was considerably lower at 329 ppb (also a male, age 62 from the Portland arca) and the subsequent next eight highest serum PFOS values (range 139 ppb to 226 ppb) were `measured in four females and four males representing Charlotte (n =4), Hagerstown (n = 2), Los Angeles: (n= 1) and Minneapolis-St. Paul (n = 1). Also distinctive to this effort was the study's capability to examine potentialassociations between PFOS and age (no association) anc. gender (males slightly higher than females). Our findings showed a strong correlation between PFOS and PFOA. Whereas PEOS has been routinely measured in humanpopulations, wildlife, marine mammals amid `piscivorous birds (Giesy and Kannan 2001; Kannan et al 20012; 2001b; Hansen et al 2001), serum PFOA concentrations, to date, have been consistently quantified (i.e. measured above the LLOQ) primarily in humans. This association is of significant interest because PFOA cannot convert directly from PFOS (or vice versa). Whether this: association is due to the presence of PFOA as a by-product in POSF-related production or 10 other non-related environmental exposures or consumer products from other `manufacturers (.g.. higher chain telomers) remains to be answered. Another Ie 000017 germs Page l6ots 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. PFOS was also correlated with two fluorochemicals, PFOSAA and M570, known to be analytes from exposure to consumer products involving paper/packaging and carpettextile protectants, respectively. Overall, the data, to date, indicate that PFOS bioaccumulation in animals may be primarily through environmental sources whereas both environmental and product exposures likely contribute to serum PFOS concentrations in humans (Giesy and Kannan 2001; Kannan et al 20012; 2001b). As with any interpretation of data obtained from a study population, questions arise regarding the representativeness and ability to generalize the data collected. Clearly, American Red Cross blood donors are a self-selected group from the United I" States population (Leibrecht et al 1976; Oswalt 1977; Bumett 1982: Allen and Butler 1993; Andaleeb and Basu 1995: Chiavette et al 2000). Motivations to donate have included altruism, desire for personal credit, low self-esteem, social pressure and testseeking behavior. Motivations notto donate have included fear, medical excuses, apathy and inconvenience. Donors tend to have a greater trust in institutions, more interest in their personal health and higher risk-taking behavior. Demographically, white males and older individuals have historically constituted a larger percentage of the donor pool. Donors also tend to be more educated, married and have more children than nondonors. In the present study, donors were not asked whether a sample of their blood donation couldbe used for fluorochemical assay nor was there any linkage between the sample 16 000018 SM Cormmmy Po 1te collected and personal identifiers. No information was obtained about past exposure: histories to POSF-related chemistries and materials (or the other fluorochemicals that were analyzed). Therefore, we believe the selection process of donors used in this stucy resulted in a reasonable representation of the overall blood donor population that was providing blood at the time when these donors were sampled. We are unaware of any. database that can be considered generalizable to the diverse United States general aduiz: population without measures of random and systematic bias incorporated in the data analysis. : Given the consistency of the data analyzed, o date, we hypothesize that the average serum PFOS concentrations in non-occupational adult populations likely rangess between 30 10.40 ppb with 95% of a population's serum PFOS concentrations below L090 ppb. Understanding these serum PFOS levels in human populations will be useful for 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 that the serum concentrations observed in humans are: substantially less than those required to cause adverse effects in laboratory animals (3M. Company 2000) 11 000019 -- rcaotns ACKNOWLEDGEMENTS We wish to acknowledge many contributors to this 3M final report, American Red Cross bCelnotordaldoBnloorocdolSleercvtiicoens)wawistchoroergdiionnaatledasusnidstearnctehefgruoimdtahnecefoolfloDwr.inJg:ohnDrMs.ilJloerhn(NAorrmtihtage. (Carolinas Region) Ross Herron and George Garratty (Southern Califomia Region), Z`AalhlreaghMeendiehsdiRzeagdieohnk)aasnhdi M(aParcyifOi'cNNeoilrlth(wNeeswt REenggiloannald)R,eJgoihonn)N.obLialbeottriat(oGrryeaatnearlysis of the seven fluorochemicals was provided by a dedicated team at Northwest Bioanalytical `which included Ann Hoffman, Connie Sakashita, Patrick Bennett, Dr. Rodger Foltz, Suzanne Newman, Toni Peacock and Emily Yardimici. Biostatistical assistance was provided by Dr. Tim Church of the University of Minnesota. Ig 000020 3PMagCeEo1rr9r2mo6ar0ns3 REFERENCES 3M Company (2000). SIDS Initial Assessment Report Perfluorooctane Sulfonic Acid and its Salts. 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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. Hansen KJ, Clemen LA, Ellefson ME, Johnson HO (2001a). Compound-specific quantitative characterization of organic fluorochemicals in biological matrices. Environ Sci Technol 35:766-770. Kannan K. Koistenen J, Beckmen K, Evans T, Gorzelany JF. Hansen KJ, Jones PD, Hele E. Nyman M, Giesy JP (2001b). Accumulation of perfluorooctane sulfonate in marine `mammals. Environ Sci Technol 35(8):1593-1598. Kannan K. Franson JC, Bowerman WW, Hansen KJ, Jones PD, Giesy JP (2001). Perfluorooctane Sulfonate in fish-eating water birds including bald eagles and albatrosses. Environ Sci Technol 35(15):3065-3070. 19 000021 3M CEarmipionng Page 200036 Leibrecht BC, Hogan JM, Luz GA, Tobias KI (1976).Donorand nondonor motivations. `Transfusions 16:182-189. Northwest Bioanalytical (NWB, 2002). 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Washington DC: American Chemical Society, pp 117-134. ao 000022 Tae1 na an Disuibution ofAmerican Red Cross Blood Donor Suits by Age, Gender and Location Lossion Lams Boston MSuherpls- Cale Pd Hgmom wwe 20:20 30:30 10-49 50-9)_ 0-60 Subic _20-29_ Wns nw @ 6 eww nor ww WW WW 0 0 0 mw 7 #0 0 0 0 0 0s 0 0 0 0 0 B 0 we wee wm % wes 30-3 40-09 0-59 wom oR ow wow 00 0 wm 0 0 0m 0 uw 000 ew a 0-6 Sue _ OTvaa 0 oe ws wom Ww 0 0 Ww 5 @% 9 sw 9 4s www 21 000023 .a m oa n MessfC TedeeyofSr Fle aArtinRedCossBlood oes N=64 by Geer , Movean Rae aa <L0Q ante) Cuma 50% Mean Gone Gsenocm Mem Matava) Rae aa <L0QNene Cumsive 50% etn Geman petmne Mem <LOQUN-IH0 auras <n or ass us na-us <LOQUN-3 1-66 se aan 2 w as was <LOGMN-63 <l0QED-M ison aren " is 0 1-20 <LOQUO-@1 <l0QED-34 <1s00n <asam 52 coon 0 19-21 <L0QUD-164 <0aup-22 <1owy <tsam a <woous 1 ois 0QUN-1660 <LOQUN-PD 231-07 360 <aiam ne wr En " ns a 155-003 6-53 <L0QUH-663 <100@n-3s <1s00 2100 u 2 a20n-24 <LOQUO-WI coo@n-13 pr <2s040) oan 0 18-21 <LOQUO- 164 <roaun-22 <1000 <rsaey 3s <woqus n 1214 000024 | Eemals 2313 m a a oo 20-458 u-62 <L00ED-28 <L0QEH-36 <LOQUE-22 <L0Q umber <i9e) <u <teay <toan <aen <aram <ass sae Cumiaive 00% 1 se s0 a 0 Medan a3 a" <tooan <tooes <toQis GeometicMean 2 a 1 2 [E ] Gseanecrlic Mean 200-33 39-45 15-18 20-23 12-14 . 22 000025 iade | Messe of Cnt Tendency ofceilabya Si AmericanReCros Blood Bk Locations | wos pron eens prosan ws Los nels Ronee 66-50 0Q@H-341; AOQRI-124 LOQEH-26 <LOQH- 164 ao 05-57 20-66 40QED-30 L0Q@H-42 400-22 | Comics 01 22 ss a as Metin 22 ws aooan si anos GomeicMen 04 a 1 2 1 Gsenocm Mean 0-410 36-42 11-21 24-30 12-16 bee | Range AQEN-2 15-09 <0QUA-126 LORE 67 <LOQUH-130 ao 208-31 4573 400@D-30 <LOQU6-29 LOQUH-24 Comicon a7 0 sa " as Metin 20s ss u aoe 1s Geometic Me 280 sa 1 1s 14 Gsenmcma Mean 254-310 s0-s8 16-22 Lois 12-16 000026 aeMp-lSt.sPa.ul ame d0QUN-20 40QUA-1S2 40QUA-129 40-106 aa Noem3 3251 a0000-25 40006-26 40QU0-23 Mein wr " " M i" ] Geometric Mean 31 as 1s 16 12 | Seni en Charlotte ao Range. Comers Nein Wie 193-1660 1053 ws awsoss <LOQ(21)-290 150-s48 <LOQ(14)-224 40Qee-42 <LOQ(16)-60.1 w ws 40UH-28 <LOQ(1.0)-108 " IN Ceti Men 000027 mfain | Portand a-a Cumisive50% Cem GeometricMean sGeosmecticMean 2-1 w4 20 no Bsa 28-47 40QEN-25 40QRH-38 A0QUH-29 6 5s 4 En uw doen aoees <ooan 16 16 25 1 22-40 Laos 22-29 12-15 Hagerstown Range Q-@ Comaive 90% Median Geometic Mean Gseommcetic Mean 76-260 4-481 os 387 3 s18-392 40QE@N-23 32-59 7% 4 "2 38-48 i <LOQEN-663 <0QRD-38 3 doen 2 17-24 LOQEH-22 <LOQEH-14 4 do0en 7 15-19 AOQUH-79 <LOQU-19 a 400s 12 11-14 I" 000028 ] Mean and 95% Confidence IneraTlastCeld ete fomBoosrap Analyses Fore Six Locations, AdjusiedforAg, Genandd Tehari Interaction Term | mos pro Men 9%Cl Men 9SGCI_ | Awe 050 Dads 46 4ads Boston MO 0-3 53 dass MSichml o ME 9-33 45 41-47 Chote M0 M2407 50 46-SI Pot RS 0S-M2 43 404d Movs M9 M835 4S 42-46 pins. Men OCI 09 1820 Ge Mean 08 6-20 22 20-24 03 17-20 19 03-21 rrosaa Men 95% 20 19-22 06 Mes 08 1320 2 09-24 20 20-3 09 18-20 ws Men 9c 03 A313 das 13 Ll 04 1300S 43 d2-l4 13 12-14 m Boes Tor Mem osucl %2 m0 2 ema 27 Zaem Ba 33-37 ms 27-304 M0 24-30 27 0000;29 nCeoni Multivariable Regression Table5 Model of PFOS" by Lo PFOSAA', M570", Age, Gender and Age x Gerider Interaction Coefficient SE Intercept 33 007 PFOSAA" 03 003 M570" 0.1 0.03 Age -0001 0001 Gender -02 007 Age x Gender 0.001 0.001 tratio p value 463 <.0001 17 <.0001 4.1 <.0001 08 40 24 0 09 35 N=645 "Natural log Adjusted #=022 - Gender: females =1; males = 0 ratio = oefficienVSE (sandard error) 22 000020 3M Compan, Multivariable RegressiToanbMloed6el of PFOS. "by PFOA'. Age, Gender and Age x Gender Interaction Coefficient SE tratio pvalue Intercept 27 007 364 <.0001 PFOA" Age 06 0.03 203 <.0001 -0.001 0.001 10 29 Gender 002 0.06 04 n Age xGender 0.001 0.001 10 a1 -------- ee -- ---- N=64s "Natural log Adjusted = 0.40 Gender: female=s1;male=s0 - tratio = coefficienvSE (standard error) 29 000021 smc Multivariable Table 7 Regression Model of PFOS . by PFHS . Age, Gender and Age x Gender Interaction : Coefficient SE tratio p value Intercept 34 0.07 455 <.0001 PFHS' 03 003 99 <.0001 Age 0.0003 0.002 02 8 Gender -006 007 08 4 Age x Gender 0.0002 0.002 02 87 N=645 "Natural log Adjusted * =0.15 Gender: females = 1; males = 0 tratio = coefficienySE 000032 30 P`eTrabcleent8.CToonlfeirdaenncceeLLiimmiittssafnodr STheeriurmAFslsuoocrioactheedmMiecaalns aannddUCpalpceurla9t5ed Total Organic Fluorine Index ToLliemraintce Mean UCpopnefird9e5ncePeLricmeintt PFOS 90% 707 3 95% 88.5 1000 99% 1573 2070 PFOA 90% 94 10.1 95% 12.1 136 99% 198 258 PFHS 90% 63 70 95% 95 108 99% 170 24 . PMugECeottmpaoafnniy PFOSAA 90% 95% 99% M570 90% 95% 99% TOF 90% 95% 9% 53 59 76 85 194 276 37 40 50 54 81 103 599 63.1 75.1 509 1373 1875 31 000033 Figure 1. Analysis of Split Samples for Reliability Assessment for PFOS, PFOA and PFHS : WV Pd veo3 Ce %. || | J } || || | 5 i . | il B] ! i k ; : A | | 1 maeSsrtsoonrsy wr 32 000034 Figure. Adu: Say Pogulion iscibuon of Messed Florochemical Concentrations - L Lb i | "Wrineaeess8221133280828000 = eee RTT ARNE i _-- 4 CL L Cd hi +3 FN mevee~secczmspzossseg i -- = kbl I 7 i i TgrgnrcereeiiEEEes id JE ETL 000035 ------ rete 33 eFing.B Bon adT Wao lofSerumFlsstemiclCobny egenstdoGen-ndser -- a0 =I E=2I22 TF : loosle fog == = sEm5 a= bt EzEsi bade 2 3 z = |ow IL a ee 2 I 2 = = Ta S20 He ah SEES `ageat last birthday I `ageat last birthday asada} skes - Haldia 3 l. azzszs -- ERE int) Lh _ ----- +a | [| meena g . 13 -= || `Ed eTs=ETa=s TWE E EE ------------ _-- -- Hesse Ul {Hee "1 ! --_-- ---- fa 7| EF TF oz a | FB. = L 2 i am TL oT I Biu 22 SE BW ee 0 we oe 2= 22=~. 2= 2I || T Eae E=== s== =EL 2029 Es 3039 04s S05 ss 3u 000036 _--is3sss; AEI=c3 Figure 4. Box and Whisker Plots of Serum Fluorochemical Concentrations by Age anc = z=:-= a ------ iE === o E22s 28: | HM EefE=E==s (| b- apB=TilL|nz|! dI a--a-====== | # == ETT IR EITE = =-- | er city =n { BITZm-gTEITa+%lll.|==E== 2===23 T= E oea= rmrge Em--T s-- iB :. rEr a z==o --=I = n Ie 000037 Figure 5. Scatter Plots (log scale) of Fluorochemical Associations --_-- --r Ho agg PE digas. | fLo | dn bDlolpeeed Lo| an 1 7 Hid Sent" Dip onal ---- I 1 I Hija bd TH poses || CE | CLvo | ] [8 whe Ta | DE nme ; CoopmEEE Lop |