Document 6BK72Ob1pr2peBvJg9ynajX4o

BACK TO MAIN SELECTED FLUOROCHEMICALS IN THE DECATUR, ALABAMA AREA Preparedfor: 3M St. Paul, Minnesota Prepared by: Entrix, Inc. John P. Giesy, PhD. John L. Newsted, Ph.D. East Lansing, Michigan Project No. 178401 June, 2001 1 BACK TO MAIN 1 EXECUTIVE SUMMARY A monitoring study was conducted to determine the concentrations of selected fluorochemicals in biotic and abiotic phases within the Tennessee River in the vicinity of the 3M Decatur, Alabama Facility. Surface water, sediments, clams and fish were collected from locations upstream and downstream of the 3M facility and analyzed for four fluorochemicals: perfluorooctanesulfonate (PFOS), perfluorooctanesulfonamide (FOSA), perfluorooctanoate (PFOA), and perfluorohexanesulfonate (PFHS). Surface waters and sediments were collected from five primary locations. These areas consisted of a location near the point of discharge of a combined industrial effluent (3M and a neighboring industrial facility) (Outfall), a stream that receives this combined effluent (Bakers Creek), a location about five miles downstream (Fox Creek), a location upstream of 3M Facility but below the City of Decatur Drinking Water Treatment Plant and adjacent to the Decatur Wastewater Treatment Plant, and a reference location upstream of Guntersville Dam. Clams were collected from the Guntersville location and the Fox Creek location. Fish were collected from the Outfall and from the Guntersville location. All samples were collected according to standard procedures and analyzed for fluorochemicals according to 3M Environmental Laboratory methods. Results from the monitoring study indicate temperature and conductivity differed from the reference location only in the immediate vicinity of the Outfall. Dissolved oxygen and pH did not differ from the reference location. Average surface water PFOS concentrations ranged from 0.009 to 151 pg/L for Guntersville and Outfall, respectively. Average PFOS sediment concentrations ranged from 0.180 to 5930 pg/kg (wet weight) for Guntersville and the Outfall, respectively. However, while fluorochemical concentrations in sediments and surface waters were greater in locations downstream of the Outfall, the downstream sample locations within the main stem of the Tennessee River (Fox Creek) were not statistically greater than those upstream (WWTP and Guntersville). Only samples collected from the Outfall and Bakers Creek locations were significantly greater than those in the reference location. From these results, it can be concluded that the effluent from the combined industrial Outfall does not significantly 2 BACK TO MAIN affect fluorochemical concentrations in sediments or surface waters in the main stem of the Tennessee River. Fluorochemicals were detected in all fish collected from both the Guntersville reference area and Outfall. The least fluorochemical concentrations were observed in fish collected from Guntersville while the greatest concentrations were observed in fish from the Outfall. For instance, at Guntersville, average whole body concentrations for PFOS, FOSA, PFOA, and PFHS were 59.1, 9.43, 11.7 and 7.50 gg/kg, (wet weight) respectively. For fish from the outfall, average whole body concentrations for PFOS, FOSA, PFOA and PFHS were 1332.0, 20.1, 106.4, and 86.9 gg/kg, (wet weight) respectively. While there were species specific differences in the accumulation of the selected fluorocarbons into fish collected from either Guntersville or Outfall locations, these differences were not statistically significant due to the small sample size and among-individual variability. Concentrations of FOSA and PFOA in clams collected from the Outfall were greater than concentrations observed in clams collected from the Guntersville reference site. However, PFOS and PFHS tissue concentrations in Outfall clams were not greater than concentrations in clams collected from Guntersville. In conclusion, fish downstream of the Outfall contained greater concentrations of fluorochemicals than did fish from the Guntersville area but the significance of these differences is difficult to assess due to the small sample size and variability. 3 BACK TO MAIN 2 INTRODUCTION Entrix, Inc. (ENTRIX) is providing a report on activities to assess the impact of effluents containing fluorochemicals from the combined industrial Outfall in Decatur, Alabama to the Tennessee River. Water, sediment, clams and fish were collected from upstream and downstream locations of the 3M Facility and were used to evaluate the magnitude of releases of target compounds from the Facility to the Tennessee River. The objectives of the research are to determine concentrations of selected fluorochemicals in abiotic and biotic phases of the Tennessee River system in the vicinity of the 3M Facility at Decatur, Alabama. Fluorochemicals selected for evaluation in this study included: perfluorooctanesulfonate (PFOS), perfluorooctanesulfonamide (FOSA), perfluorooctanoate (PFOA) and perfluorohexane sulfonate (PFHS) (Appendix A). 3 FIELD SAMPLING 3.1 Site Description Four primary locations were selected for the field investigation (Figure 1, Appendix B). These areas consisted of a location near the point of discharge of a combined industrial effluent (3M and a neighboring industrial facility) labeled Outfall, a stream that receives this combined effluent (Bakers Creek), a location about five miles downstream (Fox Creek), a location upstream of 3M Facility but below the City of Decatur Drinking Water Treatment Plant and adjacent to the Decatur Wastewater Treatment Plant, and a reference location upstream of Guntersville Dam. Specifically, samples were collected adjacent to the Outfall (includes Outfall and Bakers Creek) that corresponds to River miles 301-302. Samples were also collected downstream of the 3M Facility near the mouth of Fox Creek (River miles 296-297). Sediment and water were collected upstream adjacent to the Decatur Wastewater Treatment Plant (approximately river mile 303.5). Finally, fish, clams, sediment, and water were collected from a reference site above the Guntersville Dam (approximately river mile 370). To better evaluate the impact of the effluent from the Outfall, the Bakers Creek location was divided into two sub-locations, Bakers Creek1 included Stations 1, 2 and 3 that were collected near the Outfall and within Bakers 4 Creek. Stations 4, 5 and 6 were collected near the interface of the mouth of Bakers Creek and the Tennessee River and are identified as Bakers Creek-2. BACK TO MAIN 5 Z Bakers Creek Decatur Wastewater Treatment Plant Fox Creek Guntersville Reference Site Chattanooga Figure 1. Tennessee River and sampling locations evaluated during study. Flow Direction Guntersville Reservoir 6 BACK TO MAIN Fox Creek sampling locations were also divided into two sub-locations. Fox Creek-1 included Stations 7, 11, 12 and 26/27 that were collected from the mouth of Fox Creek. Fox Creek-2 included Stations 8, 9, and 10 that were collected on the northern side of the Tennessee River across from the mouth of Fox Creek. Locations and sample codes for samples collected from the Tennessee River are given (Table 1). 3.2 Field Sampling Logistics Sampling at selected sites on the Tennessee River was initiated on June 19, 2000 by ENTRIX. Collection of water and sediment samples continued until June 21, 2000. During the collection of sediments, clams were also collected. Fish collection was initiated on June 21 and continued through June 22, 2000. Water, sediment, clams and fish were collected as outlined in Entrix sampling protocols (Appendix C). 3.3 SAMPLE COLLECTION 3.3.1 Water Collection Water samples were collected using ENTRIX sampling protocols from a total of 26 locations (Table 1, Appendix B). At each location, water quality parameters were measured using YSI Model 63 and 95 meters. Six samples were collected from the areas surrounding the Outfall, the City of Decatur wastewater treatment facility and the upstream reference location. Seven water samples were collected from the area near Fox Creek. In addition to these 25 samples of surface water, water was collected from the combined industrial effluent stream. All sampling locations were documented using TRIMLBE PRO-XRS Global Positioning Satellite technology and these locations were recorded on topographic maps. Samples were properly labeled, stored on ice at 4oC, and delivered to the 3M Environmental Laboratory, St. Paul, MN. 3.3.2 Sediment Collection Sediment samples were collected from the same location, as were the water samples (Table 1, Appendix B). A total of 25 grab samples were collected from the Tennessee River. Six samples were collected from areas surrounding the Outfall, the City of Decatur's wastewater treatment facility, and the upstream reference location using either a Petite PONAR dredge. Seven samples were collected from the downstream area in the 7 BACK TO MAIN vicinity of Fox Creek. All sampling locations were documented using TRIMLBE PROXRS Global Positioning Satellite technology and these locations were recorded on topographic maps. Sediments were collected by use of a PONAR dredge following standard protocols. Sediments were placed into labeled 250 ml wide-mouth LDPE containers, stored on ice at 4oC, and shipped to the 3M Environmental Laboratory, St. Paul, MN. Table 1. Location and identification of sediment and water samples collected from the Tennessee River in Alabama Water Sample Sediment Sample Location Sublocation/Code SW-01 SED-01 Bakers Creek BCR1 SW-02 SED-02 Bakers Creek BCR1 SW-03 SED-03 Bakers Creek BCR1 SW-04 SED-04 Bakers Creek BCR2 SW-05 SED-05 Bakers Creek BCR2 SW-06 SED-06 Bakers Creek BCR2 SW-07 SED-07 Fox Creek FCR1 SW-08 SED-08 Fox Creek FCR2 SW-09 SED-09 Fox Creek FCR2 SW-10 SED-10 Fox Creek FCR2 SW-11 SED-11 Fox Creek FCR1 SW-12 SED-12 Fox Creek FCR1 SW-13 SED-13 Decatur WWTP WWTP SW-14 SED-14 Decatur WWTP WWTP SW-15 SED-15 Decatur WWTP WWTP SW-16 SED-16 Decatur WWTP WWTP SW-17 SED-17 Decatur WWTP WWTP SW-18 SED-18 Decatur WWTP WWTP SW-19 SED-19 Guntersville GTVL SW-20 SED-20 Guntersville GTVL SW-21 SED-21 Guntersville GTVL SW-22 SED-22 Guntersville GTVL SW-23 SED-23 Guntersville GTVL SW-24 SED-24 Guntersville GTVL SW-25 SED-25 Outfall OUTF SW-26 Fox Creek FCR1 SW-27 SED-27 Fox Creek FCR1 3.3.3 Clam Collection Samples of the Asian clam (Corbicula fluminea) were targeted for collection from a location downstream of the 3M Facility and from a location upstream of Guntersville 8 BACK TO MAIN Dam. Historically, various state and federal agencies have used clams in monitoring programs for organic and inorganic pollutants in aquatic systems, as a result, they were also included n this study. Furthermore, due to their ubiquitous distribution in the Tennessee River and their sessile behavior, they provide site-specific information concerning the potential accumulation of fluorochemicals into aquatic biota. Due to the habitat requirements of C. fluminea, clam samples were not co-located with water or sediment samples. Clams were collected using a Petit PONAR dredge. However, due to difficulties in locating clam beds as well as interference of substratum with the PONAR dredge, the collection of clams was limited to two composite samples from locations in the vicinity of sediment samples. One sample (approximately 25 clams/sample) was taken from a location downstream of the 3M Facility (SED-08, -09, -10) and one upstream sample was collected from the Guntersville location (SED-20, - 21, - 22, -23, 24). Clams from each location were placed into labeled plastic bags and shipped on ice to the MSU-ATL (Michigan State University-Aquatic Toxicology Laboratory), East Lansing, MI. 3.3.4 Fish Collection Fish were collected from two locations (Table 2). One location was in the vicinity of the Outfall and the other was located within the reference area upstream of Guntersville Dam. The fish samples were collected using both hook and line as well as gill nets. Gill nets (passive gear) were placed in the lower energy environments where concentrations of fish were generally greater. Due to the techniques employed for collection of fish, all fish caught in each of the sampling locations were retained for analysis. Collected fish were removed from the nets, measured (total length), weighed, and immediately wrapped in aluminum foil and stored in plastic bags. Samples were labeled placed on ice, and shipped to the MSU-ATL (East Lansing, MI) for processing. 4 CHEMICAL ANALYSIS Water and sediment samples were collected from the Tennessee River and analyzed by the Centre Analytical Laboratories, State College, PA (CLA, Report No. 023-0140; Appendix D). The analytical methods used for the water samples were validated by Centre. 9 BACK TO MAIN The methods were modified for the analysis of sediment samples but were not fully validated for this matrix. Fish livers and clam samples were extracted and processed at MSU-ATL. LC/MS/MS characterization of fluorochemicals was conducted at 3M Environmental Laboratory. Summaries of each protocol are given: Table 2. Sample identification and location of individual fish collected from the Tennessee River. Sample ID______ Location_____________________ Species LM1 Guntersville Largemouth bass Micropterus salmoides LM2 Outfall Largemouth bass Micropterus salmoides SHAD1 Outfall Skipjack Herring Alosa chrysochloris SHAD2 Outfall Skipjack Herring Alosa chrysochloris SHAD3 Outfall Skipjack Herring Alosa chrysochloris SHAD4 Outfall Skipjack Herring Alosa chrysochloris SHAD5 Outfall Skipjack Herring Alosa chrysochloris SHAD6 Outfall Skipjack Herring Alosa chrysochloris SHAD7 Outfall Skipjack Herring Alosa chrysochloris SHAD8 Outfall Skipjack Herring Alosa chrysochloris WP1 Outfall White Perch Morone americana WP2 Outfall White Perch Morone americana CAT1 Guntersville Catfish Ictalurus punctatus CAT2 Guntersville Catfish Ictalurus punctatus CAT3 Outfall Catfish Ictalurus punctatus CAT4 Outfall Catfish Ictalurus punctatus GAR1 Guntersville Gar Lepisosteus sp. GAR2 Guntersville Gar Lepisosteus sp. GAR3 Guntersville Gar Lepisosteus sp. GAR4 Guntersville Gar Lepisosteus sp. GAR5 Outfall Gar Lepisosteus sp. GAR6 Outfall Gar Lepisosteus sp. SB1 Guntersville Striped Bass Morone saxatilis SB2 Guntersville Striped Bass Morone saxatilis SB3 Guntersville Striped Bass Morone saxatilis SB4 Guntersville Striped Bass Morone saxatilis SB5 Guntersville Striped Bass Morone saxatilis SB6 Guntersville Striped Bass Morone saxatilis SB7 Guntersville Striped Bass Morone saxatilis SB8 Guntersville Striped Bass Morone saxatilis SB9 Guntersville Striped Bass Morone saxatilis SB10 Guntersville Striped Bass Morone saxatilis SB11 Guntersville Striped Bass Morone saxatilis SB12 Guntersville Striped Bass Morone saxatilis SB13 Guntersville Striped Bass Morone saxatilis SB14 Guntersville Striped Bass Morone saxatilis 10 BACK TO MAIN 4.1 Water Analysis Water samples were initially treated with 200 pl of 250 mg/L sodium thiosulfate to remove residual chlorine. Solid phase extraction was used to prepare the samples for LC/MS/MS analysis. A forty-milliliter portion of sample was transferred to a Ci8SPE cartridge and the cartridge was eluted with 5 ml of 40% methanol in water. The eluate was discarded and the SPE column was eluted with 5 ml of 100% methanol. The eluate was collected for analysis by LC/MS/MS. This treatment resulted in an eight-fold concentration of the samples. 4.2 Sediment Analysis For sediment samples, a 5 g portion was extracted into 5 ml methanol. The extracts were filtered and diluted to final volume of 40 ml with ASTM Type I water. The diluted extracts were then treated in the same manner as the water samples, beginning with the solid phase extraction. Sediment and water extracts were analyzed by use of a Hewlett-Packard HP1100 HPLC system coupled to a MicroMass Ultima MS/MS. Analysis was performed using selected reaction monitoring (SRM). HPLC conditions and MS/MS methods used for analysis and instrument parameters are given in the procedure described below. 4.3 Fish Liver Analysis Fluorochemical surfactants were extracted from fish livers using an ion pairing reagent and methyl-tert-butyl ether (MtBE). Details of the analytical procedure have been outlined in 3M Environmental Laboratory (St. Paul, MN) standard operating procedures of the analysis of fluorochemicals in tissues. The title of the SOP is: ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Liver for Analysis using HPLC-Electrospray/Mass Spectrometry". Briefly, fish were thawed and livers were removed, weighed and then homogenized in HPLC grade water. An aliquot of liver homogenates were spiked with a surrogate standard and then 0.5 M tetrabutyl ammonium hydrogen sulfate (TBA) and 0.25 M sodium carbonate/sodium bicarbonate was added to each sample. Five ml methyl-tert-butyl ether was added to each tube and the sample was capped and shaken for 20 min. The samples were then centrifuged for 25 min at 3500 rpm and 4.0 ml of the organic layer was 11 BACK TO MAIN transferred to a fresh tube. The extracts were evaporated under nitrogen gas to dryness then reconstituted in methanol. The methanol extracts were then passed through 0.2 pm nylon filters into glass autovials for chromatographic analyses. 4.4 Whole Body Fish Analysis Perfluorochemicals and other fluorochemical surfactants were extracted from whole fish tissues using an ion pairing reagent and methyl-tert-butyl ether (MtBE). Details of the analytical procedure have been outlined in the 3M Environmental Laboratory (St. Paul, MN) standard operating procedures for the analysis of fluorochemicals in tissues. Briefly, whole fish were thawed and then homogenized in a Hobart grinder. The homogenates were placed in I-ChemTM bottles and stored at -20 C until chemical analysis. Whole fish homogenates were weighed and then further homogenized in HPLC grade water. Aliquots of tissue homogenates were spiked with a surrogate standard and then 0.5 M tetrabutylammonium hydrogen sulfate (TBA) and 0.25 M sodium carbonate/sodium bicarbonate was added to each sample. Five ml methyl-tert-butyl ether was added to each tube and the sample was capped and shaken for 20 minutes. Following the shaken period, the samples were centrifuged for 25 minutes at 3500 rpm and 4.0 ml of the organic layer was transferred to a fresh tube. The extracts were evaporated under nitrogen gas to dryness then reconstituted in methanol. The methanol extracts were then passed through 0.2 pm nylon filters into glass autovials for chromatographic analysis. 4.5 Clam Tissue Analysis Clam tissues were extracted with acetonitrile followed by a solid phase column cleanup of the extract. Briefly, frozen clam tissues were thawed and the tissues were composited and homogenized in an Omni homogenizer. A 5 gram sample was weighed a 50 ml polypropylene tube and 15 ml acetonitrile was added. Samples were shaken for 30 min and then centrifuged for 30 min at 3500xg. The supernatant was passed through a column of Florisil and activated charcoal and the eluate was collected in 50 ml polypropylene tube. The column was eluted with 15 ml methanol and both eluates were 12 BACK TO MAIN combined. Two drops of octanol were added to the eluate and then the sample was reduced to near dryness under a flow of nitrogen gas. The sample was resuspended in 1 ml methanol and passed through a 0.2 pm nylon filter into autovials for chromatographic analysis. 5 QA/QC Standard operating procedures for sample collection and preparation were maintained during the entire project. Proper QA/QC samples, as required by 3M, were collected in the field (Appendix C). Two field matrix spikes and two field control samples were collected. Field matrix spikes solutions were prepared by collecting one liter of site water and spiking with 20 or 100 pl of a 10 ppm stock solution containing PFOS, FOSA, PFOA, and PFHS to yield 200 and 1000 ppt solutions, respectively. The same procedure was followed to spike one liter of distilled water to prepare the field spike control samples. All samples were shipped under ENTRIX chain of custody forms and a field book was used to document conditions and activities. A field camera was used to document all locations and to provide a visual record of the conditions of the site during the sampling events. In addition, ambient water quality parameters, such as temperature, conductivity, salinity, pH and dissolved oxygen were measured at each location during each sampling event. Since the method detection limit (MDL) and limit of quantification (LOQ) are analyte and matrix specific, method and matrix blanks along with matrix spikes were used to determine accuracy and precision of the extractions and final chemical determination. Water and sediment concentrations were not corrected for either matrix spike recoveries or for purity of the fluorochemical standards. Fish tissue fluorocarbon concentrations were reported as wet weight and were not converted to a dry weight basis. 6 STATISTICAL ANALYSES Statistical analyses were preformed with SAS (Version 8, SAS Institute, Cary NC, USA). General linear models (PROC GLM) were used to test for differences among locations for all endpoints measured in the study. If values of F tests indicated a significant difference, Tukey's HSD test for multiple comparisons was used to compare means of the different locations. 13 BACK TO MAIN 7 RESULTS 7.1 Water quality parameters Water quality parameters of river water from each location are reported (Table 3, Appendix E and F: figures 1a, 1b, 1c, and 1d). Dissolved oxygen and pH were variable within the study, however there were no significant differences observed between values measured in waters collected from the Outfall or Bakers Creeks and those from upstream or downstream locations. Conductivity and temperature measured at the Outfall and within Bakers Creek (BCR1) were significantly different from the upstream and downstream sample locations. However, there was no significant difference between that from Bakers Creek 2 samples and those collected from either the upstream and downstream sampling locations. This result indicates that the effect of the combined industrial effluent on water quality parameters was limited to a small area near the Outfall. Table 3. Water quality parameters for samples collected from the Tennessee River, in the Decatur Alabama area. Values are reported as means and standard deviations DO Conductivity Temperature pH Location (mg/L) (pmhos/cm) (oC) (su) Guntersville 7.6 192.8 29.0 8.2 (0.8) (5.4) (0.3) (0.4) WWTP 10.0 * 184.7 29.3 8.8 (1.3) (3.4) (0.3) (0.2) Outfall 6.5 4650 * 33.2 * 7.2 Bakers Creek-1 7.75 (14) 1843.3 * (1749.0) Bakers Creek-2 9.0 (0.6) 463.0 (108.9) Fox Creek-1 8.1 (0.4) 179.0 (2.9) Fox Creek-2 8.8 (13) 162.7 (8.1) * Significantly different from Guntersville Reservoir at a = 0.05 30.7 * (14) 29.6 (0.6) 27.8 (0.2) 27.8 (0.5) 7.7 (0.5) 8.0 (0.1) 8.2 (0.3) 8.2 (0.5) 14 BACK TO MAIN 7.2 Fluorochemicals in Surface waters Fluorochemicals were detected in all surface waters collected from the sample locations in the Tennessee River (Table 4) (Appendix F: figures 2a, 2b, 2c, and 2d). The least concentrations of all measured fluorochemicals were observed in samples taken at Guntersville while the greatest concentrations were observed at the Outfall. In increasing order, the concentrations of PFOS, FOSA, PFOA and PFHS in surface water was as follows: GTVL < WWTP < FCR1 < FCR2 < BCR2 < BCR1 < Outfall. Concentrations of surface water fluorochemicals at only the Outfall and Bakers Creek 1 locations were significantly greater than those measured at Guntersville Reservoir. Table 4. Mean concentration of fluorochemicals in surface waters of the Tennessee River in the Decatur, Alabama area. Concentrations means and standard deviations PFOS FOSA PFOA PFHS Location (pg/L) (Pg/L) (Pg/L) (Pg/L) Guntersville 0.009 0.004 0.0081 0.0031 (0.002) (0.002) WWTP 0.053 (0.013) 0.016 (0.025) 0.028 (0.006) 0.006 (0.006) Outfall 150.50* (7.778) 7.985* (0.148) 1900.00* (0.00) 42.70* (1.697) Bakers Creek-1 82.260* (56.672) 5.424* (3.074) 1023.60* (712.64) 22.374* (14.665) Bakers Creek-2 13.838 (2.351) 1.191 (0.244) 129.375 (31.892) 4.034 (0.575) Fox Creek-1 0.54 (0.05) 0.169 (0.022) 2.647 (0.614) 0.566 (0.102) Fox Creek-2 0.179 (0.110) 0.055 (0.033) 1.001 (0.441) 0.168 (0.114) * Significantly different from Guntersville Reservoir at a = 0.05 1 Limit of quantitation for all fluorochemicals water is 0.025 qg/L. All fluorochemical concentrations reported below the LOQ are estimates. Additional statistical analyses were conducted on fluorochemicals in surface waters to further examine the difference between sample locations (GTVL, WWTP, BCR2, FCR1, 15 and FCR2) located within the Tennessee River. Results from the first analysis showed that both the Outfall and BCR1 could be influencing the results of the statistical analyses by acting in a manner similar to that of an outlier. For instance, the concentration of PFOS at the Outfall and BCR1 were approximately 17,000-fold and 9,700-fold, greater, respectively, than the concentration observed at the Guntersville location. Removal of the Outfall and BCR1 samples from the analysis allowed for the evaluation of the ,hypot,hesi.s that there were no stat.isti.ca.lly. s.ign.ificant differences i.n f.luorochemBicAalCK TO MAIN concentrations within the main channel and near shore locations within the river. Results from the second analysis (Table 5) show that for water concentrations of PFOS and PFOA, only BCR2 was statistically greater than Guntersville Reservoir concentrations. Furthermore, while the concentrations of FOSA and PFHS at BCR2 and FCR1 were significantly greater than Guntersville, they were not significantly different from the furthest downstream location. Thus, at FCR2, the furthest downstream location, the concentration of targeted fluorochemicals was not significantly different from that observed at Guntersville Reservoir, which was selected as an upstream reference location. Table 5. Tukey's range test results for surface water fluorochemicals taken from within or near the main channel of the Tennessee Rivera Site PFOS FOSA PFOA PFHS BCR2 A A AA FCR1 B B BB FCR2 B B,C B C WWTP B C B C GTVL BCB a Sites with the same letter are not significantly different at a = 0.05 C From these results, it can be concluded that the Outfall does not significantly affect concentrations of fuorochemicals farther downstream than FCR2. In addition, there were no statistically significant differences in the concentrations of fluorochemicals 16 BACK TO MAIN within the main channel of the river immediately below Bakers Creek as compared to those measured at the reference location. 7.3 Sediment concentrations offluorochemicals Fluorocarbons were detected in all sediments collected from the study sites (wet weight: Appendix F-figures 3a, b, c, d and dry weight: Appendix F-figures 4a, b, c, d). 17 Table 6. Mean concentration of fluorochemicals in sediments taken from the Tennessee River in the Decatur Alabama areaa Wet weight sediment concentrations Dry weight sediment concentrations Location PFOS FOSA PFOA PFHS PFOS FOSA PFOA PFHS Guntersville (pg/kg) 0.18 (0.07) (Pg/kg) 0.08 (0.01) (Pg/kg) 0.08 (0.01) (Pg/kg) 0.08b (Pg/kg) 0.43 (0.20) (Pg/kg) 0.19 (0.04) (Pg/kg) 0.19 (0.04) (Pg/kg) 0.19 (0.04) WWTP 0.98 (0.42) 0.15 (0.06) 0.09 (0.02) 0.08 (0.01) 1.72 (0.83) 0.26 (0.11) 0.16 (0.04) 0.14 (0.04) Outfall 5930.00* (254.56) 1200.00* (42.43) 1855.00* (106.07) 135.00* (4.24) 12600.00* 2555.00* (565.69) (91.92) 3950.00* (226.27) 287.50* (9.19) Bakers Creek 1 1298.67* (751.89) 283.30* (176.21) 892.00* (345.80) 42.10* (22.05) 2275.67* (142.76) 503.17* (364.31) 1548.17* (774.63) 74.6* (48.57) Bakers Creek 2 192.00 (191.27) 54.09 (36.33) 237.61* (283.01) 11.46* (8.08) 272.64 (279.05) 75.91 (53.03) 338.43* (410.79) 16.18 (1191) Fox Creek 1 2.58 (137) 1.70 (109) 1.81 (0.77) 0.22 (0.11) 4.51 (2.88) 2.99 (2.21) 3.11 (168) Fox Creek 2 0.93 0.44 0.80 0.08b 2.02 0.95 1.73 (0.10) (0.05) (0.22) (0.22) (0.10) (0.46) * Significantly different form Guntersville Reservoir at a = 0.05 a Concentrations given as means with standard deviations inparenthesis bLimit of quantification of all fluorocarbons in sediment is 0.20 pg/kg wet weight. All values reported belowthe LOQ are estimates. 0.38 (0.23) 0.17b 18 BACK TO MAIN Least concentrations of fluorochemicals in sediments were observed at Guntersville Reservoir while the greatest concentrations were observed in sediments collected in the vicinity of the Outfall (Table 6). Concentrations of fluorochemicals in sediment decreased in the following order: GTVL < WWTP < FCR2 < FCR1< BCR2< BCR1< OUTF. For PFOS and FOSA, only concentrations in sediments at the Outfall and BCR1 locations were significantly greater than those collected from the reference location in Guntersville Reservoir. In addition, concentrations of PFOA and PFHS in sediments at the Outfall, BCR1 and BCR2 were significantly greater than concentrations in sediments from Guntersville Reservoir. However, the fluorochemicals in sediments at BCR2 were not significantly different from concentrations measured at WWTP, FCR1 or FCR2. When sediment concentrations of targeted fluorochemicals were evaluated on a dry weight basis, only concentrations at the Outfall and BCR1 were significantly different from those measured at Guntersville Reservoir. As was done with the surface water data, statistical analyses were conducted that examined only the Tennessee River channel sample locations (GTVL, WWTP, BCR2, FCR1, and FCR2). Results from the second set of analyses showed that only the BCR2 concentrations of targeted fluorochemicals were greater than those observed at Guntersville Reservoir reference location on both a wet and dry weight basis. Thus, while concentrations at the Outfall and within Bakers Creek were statistically greater than the reference location, concentrations in sediments downstream of the 3M Facility were not statistically greater than those measured at the upstream reference location. The results of both of these analyses indicated that that there is little accumulation of the target fluorochemicals in sediments downstream of the Outfall. 7.4 Fish Mean length, whole body weight and liver weight data for all fish species collected from the Tennessee River in the Decatur Alabama area are given (Table 7, Appendix G). 19 BACK TO MAIN Table 7. Mean length, weight, and liver weight for fish collected from the Tennessee River (Collected on 06/21/00-06/22/00)1. Sample ID Length Weight Liver wt Location (cm) (g) (g) LM1 Guntersville 26.5 195 1.32 Outfall 32 454 2.92 SHAD WP1 CAT GAR SB Outfall Outfall Guntersville Outfall Guntersville Outfall Guntersville 34 (7.6) 16.8 (0.36) 32.5 (2.12) 40.5 (12.0) 58.4 (5.25) 48.3 (5.3) 23.3 (2.17) 352 (160) 76 (7.1) 376 (112) 659.2 (531.2) 765 (170) 460.7 (170.4) 168 (55.6) 1.95 (0.985) 0.42 (0.04) 5.6 (0.18) 6.8 (3.8) 6.4 (4.3) 3.6 (1.9) 1.01 (0.34) values are means with standard deviations (in parentheses) Due to the small sample size and large amount of variability in the endpoints, there were no statistically significant differences between the sites for species common to both the Outfall and Guntersville locations. 7.4.1 Concentrations o fPFOS, FOSA, PFOA, AND PFHS infish livers. Fluorochemicals were detected in all livers removed from fish collected at both the Guntersville and Outfall locations (Table 8, Appendix H). The least liver concentrations for all measured fluorocarbons were observed in fish collected from the Guntersville location while the greatest concentrations were observed in Outfall fish. Average for PFOS, FOSA, PFOA, and PFHS liver concentrations for all fish collected from the Outfall were approximately 19-fold, 340-fold, 109-fold and 710-fold greater than those observed in Guntersville fish, respectively. 20 BACK TO MAIN Average liver concentrations for PFOS, FOSA, PFOA, and PFHS for all fish collected at the Guntersville site were 1036, 22.0, 22.1 and 3.79 gg/kg wet weight, respectively. Table8. Liver concentrations of selected fluorocarbons in fish collected from the Tennessee River in the Decatur, Alabama area. a PFOS FOSA PFOA PFHS Species Guntersville Nc (hg/kg) (hg/kg) (hg/kg) (hg/kg) Catfish 2 87.3 20.3 18.8b 3.75b (22.2 (5.0) Gar 4 249.3 20.95 18.8b 4.58 (185.9) (16.4) (1.65) Striped Bass 9 1330.0 (763) 23.19 (10.0) 24.7 (17.6) 3.75b Largemouth Bass 1 643 18.8b 18.8b 3.75b Outfall Catfish 2 Gar 2 Shad 5 White perch 2 Largemouth bass 1 10093.5 (5,777.8) 13892.0 (3073.1) 9195.2 (13252.2) 49285.0 (37657.7) 27143.0 14521.5 (2090.9) 5,089.0 (3,739.2) 4,420.0 (4047.3) 17403.0 (14679.5) 2456.0 252.9 (277.3) 1,934.5 (652.7) 96.9 (49.0) 1628.5 (1228.2) 3.75b 30.2 (28.6) 256.5 (89.9) 129.1 (145.3) 2069.5 (1941.0) 11.6 a Concentrations are reported as means and standard deviations. b Results were reported as the limit of quantitation: 18.8 gg/kg for PFOS, FOSA, PFOA and 3.75 gg/kg for PFHS. All other values below the LOQ are estimated values. cNumber of fish analyzed. Differences in liver fluorochemical concentrations were also observed between fish species collected at Guntersville. Liver concentrations for the selected fluorocarbons increased in the following manner: PFOS: Catfish < Gar < Largemouth bass < Striped bass FOSA: (Largemouth bass)< Catfish < Gar < Striped bass PFOA: (Catfish = Largemouth bass) < Gar < Striped bass PFHS: (Striped bass = Catfish < Largemouth bass) < Gar 21 BACK TO MAIN The fish in parentheses indicate liver fluorochemical concentration below the LOQ. Except for PFOS, there were no statistically significant differences in fluorochemical liver concentrations between fish species collected at Guntersville. For PFOS, there was a 160-fold difference between catfish that had the least concentration and striped bass with the greatest liver concentration. FOSA, PFOA, and PFHS liver concentrations in fish from Guntersville were all less than their respective LOQs. The significance of these results and a true understanding of the magnitude of the observed differences at this site are unknown at this time due to the variability in the data and small sample size. Average liver concentrations for PFOS, FOSA, PFOA, and PFHS for all fish collected from the Outfall location were 15,692, 7,195, 581, and 384 pg/kg wet weight, respectively. As was observed at the Guntersville site, differences in liver fluorochemical concentrations were also observed between fish species collected at the site. Liver concentrations of the selected fluorochemicals at Outfall site increased as follows: PFOS: FOSA: PFOA: PFHS: Shad < Catfish < Gar < Largemouth Bass < White Perch Largemouth Bass <Shad< Gar < Catfish < White Perch Largemouth Bass < Shad < Catfish < White Perch < Gar Largemouth Bass < Catfish < Shad < Gar < White Perch Except for PFOA, there were no statistically significant differences in species specific liver concentrations of the selected fluorocarbons for fish from the Outfall. For PFOS, FOSA, and PFHS there were 5.4, 7.1 and 178-fold differences between the least and greatest liver concentrations. However, due to the small sample size and large amount of variability observed in the data, these differences were not statistically significant. For PFOA liver concentrations, there was a 516-fold difference between the least and greatest concentration. However, only shad and gar liver concentrations were significantly different. Largemouth bass was not included in the analysis due to the fact that there was only a single value for this species. The magnitude and significance of these differences is unknown at this time due to the variability in the data and small sample size. 22 BACK TO MAIN 7.4.2 Wholefish concentrations o fPFOS, FOSA, PFOA, and PFHS Fluorochemicals were detected in all fish collected from both the Guntersville and Outfall locations (Table 9, Appendix I). Table 9. Whole body concentrations (wet weight) of selected fluorochemicals in fish collected from the Tennessee River in the Decatur Alabama area. a PFOS FOSA PFOA PFHS Species Guntersville Catfish Nc (hg/kg) 2 7.5b (hg/kg) 7.5b (hg/kg) 20.1b (hg/kg) 7.5b Gar Striped Bass 4 14 15.1 (15.2) 65.6 (23.2) 7.5b 10.4 (0.96) 20.1b 8.03b 7.5b 7.5b Largemouth Bass 1 230 8.76 8.03b 7.5b Outfall Catfish 2 Gar 2 Shad 8 White perch 2 1190 (170) 1863 (1679) ND ND ND 119.6 11.4 (140.6) (5.52) ND 154.5 35.5 (40.3) (25.0) ND 54.0 38.9 (48.2) (60.2) ND 278 405.5 (69.3) (129.4) Largemouth bass 1 ND 557 20.1b 7.5b a Concentrations are reported as means and standard deviations. b Results were reported as the limit of quantification:7.5 qg/kg for PFOS, FOSA, PFHS, 8.03 qg/kg for PFOA. cNumber of fish analyzed ND signifies that the compound is present but not quantified due to the data not meeting quality control criteria. The least fluorochemical concentrations were observed in fish collected from the Guntersville location while the greatest concentrations were observed in fish from near the Outfall. Mean PFOS, FOSA, PFOA and PFHS concentrations for all fish collected 23 BACK TO MAIN from the Outfall were approximately 23-fold, 2-fold, 9-fold and 12-fold greater than those measured in Guntersville fish, respectively. Mean whole body concentrations for PFOS, FOSA, PFOA and PFHS for all fish collected from the Guntersville site were 59.1, 9.43, 11.7 and 7.50 pg/kg wet weight, respectively. Species differences in whole body concentration at the site were also observed with concentrations increasing as follows: PFOS: FOSA: PFOA: PFHS: Catfish < Gar < Striped bass < Largemouth bass Catfish = Gar < Largemouth bass < Striped bass Largemouth bass = Striped bass < Catfish < Gar All species equal due to being at LOQ Due to the small sample size, there were no statistically significant differences between whole body concentrations for the species collected at this site. Furthermore, except for PFOS, there was less than a 3-fold difference between the least and greatest fluorochemical concentration in the species being evaluated in this study. For PFOS, there was a 30-fold difference between the catfish and the largemouth bass concentrations. Mean whole body concentration for PFOS, FOSA, PFOA and PFHS for fish caught at the Outfall were 1332.0, 20.1, 106.4, and 86. 9 pg/kg wet weight, respectively. As observed at the Guntersville location, differences in whole body fluorochemical concentrations were also observed between species caught on site. Whole body concentrations of fluorochemicals in Outfall site increased as follows: PFOS: FOSA: PFOA: PFHS: Gar < Catfish (Shad, White perch, Largemouth bass) Largemouth bass (Gar, Catfish, Shad, White perch) Largemouth bass < Shad < Catfish < White perch Largemouth bass < Catfish = Shad < White perch 24 BACK TO MAIN The species listed in the parentheses were not quantified during the analyses and could not be evaluated in this report. As with the Guntersville fish, small sample size precluded any statistical analysis of the data to evaluate differences between species at the Outfall location. Thus, while these data show that there are differences between locations and between fish in each location, the small sample size precludes an evaluation of the significance and magnitude of these differences. 7.5 Fluorochemicals in Clam Tissues Fluorochemical concentrations were also measured in clam tissues collected at both sites (Table 10, Appendix H). PFOS concentrations did not differ between the sites and ranged from 15.6 pg/kg to 14.1 pg/kg wet weight at Guntersville and the Outfall, respectively. In contrast, there was a 42.8-fold difference in FOSA clam tissue concentrations with 25.1 pg/kg and 1074 pg/kg, wet weight, being measured at Guntersville and Outfall locations, respectively. There were no significant differences in the concentration of PFOA and PFHS in clam tissue collected at each of the two sites. The results of the clam analyses reflect the areas from which the samples were collected. Due to substrate problems, Outfall clams were collected at the Fox Creek location. This location had surface water and sediment fluorochemical concentrations that were statistically similar to those observed at the Guntersville location. Thus, these results indicate that for at least clams, the environmental concentrations of fluorochemicals downstream of the Outfall are comparable to background levels in the Tennessee River. 25 BACK TO MAIN Table 10. Tissue concentrations of selected fluorochemicals in clams collected from the Tennessee River in the Decatur Alabama area. 1 PFOS FOSA PFOA PFHS Location (bg/kg) (bg/kg) (bg/kg) (bg/kg) Guntersville 15.62 25.1 4.382 0.9462 Outfall 14.12 1074 8.422 0.372 (13.7) (1358) (7.18) - r 1 C oncentrations presented as means and standard deviations. (0.53) Limits of quantitation: 18.8 qg/kg for PFOS, FOSA, PFOA, 3.75 qg/kg for PFHS. Concentrations reported below LOQs are estimated values 8 REFERENCES Hansen, K.J. and H.O. Johnson, 2000. Determination of perfluorooctane sufonate (PFOS), perfluorooctane sulfonylamide (FOSA), and perfluorooctanoate (PFOA) in water by liquid-solid extraction and high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). 3M Environmental Laboratory method, number ETS-8-154.0. 3M Environmental Laboratory, St. Paul, MN. LIMs Report numbers: E00-2361, E001958, E01-0520. 26 BACK TO MAIN 9 APPENDICES Appendix A. Structure and chemical characteristics of selected fluorochemicals monitored in water, sediment and biota of the Tennessee River in the Decatur Alabama area. Appendix B. Water and Sediment locations within the Tennessee River study areas Appendix C. Water, sediment, clams, and fish sampling protocols Appendix D. Centre Analytical Laboratories analytical report for the characterization of fluorochemicals in water and sediment. Appendix E. Water quality data collected at sample locations. Appendix F Topographic maps of sample locations in the Tennessee River study area. Maps include water quality data, water or sediment fluorochemical concentrations for sample locations. Figure 1. Water quality parameters Figure 2. Fluorochemicals in surface waters Figure 3. Fluorochemicals in sediments, wet weight Figure 4. Fluorochemicals in sediments, dry weight Appendix G. Physiological data for fish collected from the Tennessee River. Appendix H. Fluorochemicals concentrations in fish livers and clam tissues collected from the Tennessee River in the Decatur Alabama area. Appendix I. Whole body fluorochemical concentrations in fish collected from the Tennessee River in the Decatur Alabama area. 27 BACK TO MAIN APPENDIX A Selected fluorochemicals monitored in water, sediment, clams, and fish collected from the Tennessee River, Decatur Alabama area. Chemical Name : Molecular structure: Molecular ion: Perfluorooctanesulfonate C8F17SO3 499 O C g F ^ --OO PFOS Chemical Name: Molecular structure: Molecular ion: Chemical Name: Molecular structure: Molecular ion: Perfluorooctanesulfonamide C8F17SO2NH2 498 O CgFnS-- NH2 O FOSA Perfluorooctanoate C7 F1 5 COO" 413 O C7F 15CO- PFOA Chemical Name: Molecular structure: Molecular ion: Perfluorohexanesulfonate C6F13SO3399 O C6F13^OO PFHS BACK TO MAIN APPENDIX B Location and identification of water, sediment and clam samples collected from the Tennessee River, Decatur Alabama Water Samples Sediment Samples Clam Samples Locations Latitude/Longitude SW-01 SW-02 SW-03 SW-04 SW-05 SW-06 SW-06DUP SW-07 SW-08 SED -01 SED -02 SED-03 SED -04 SED-05 SED -06 SED -06DUP SED -07 SED-08 SW-09 SED-09 SW-10 SED-10 SW-11 SW-11FMS1 SW-11FMS2 SW-12 SW-12DUP SW-13 SW-14 SW-15 SW-16 SW-17 SW-18 SW-18DUP SW-19 SW-20 SED-11 SED-11FMS1 SED-12 SED-12DUP SED-13 SED-14 SED-15 SED-16 SED-17 SED-18 SED-18-DUP SED-19 SED -20 SW-21 SED -21 SW-22 SED -22 SW-23 SED-23 SW-24 SED -24 SW-25 SW-26 SW-FSCS1 SW-FSCS2 SW-FB01 SED-25 SED -27 Mussels Outfall1 Mussels Outfall1 Mussels Outfall1 Mussels Guntersville2 Mussels Guntersville2 Mussels Guntersville2 Mussels Guntersville2 Mussels Guntersville2 Bakers Creek Bakers Creek Bakers Creek Bakers Creek Bakers Creek Bakers Creek Bakers Creek Fox Creek Fox Creek Fox Creek Fox Creek Fox Creek Fox Creek Fox Creek Fox Creek Fox Creek Decatur (WWTP) Decatur (WWTP) Decatur (WWTP) Decatur (WWTP) Decatur (WWTP) Decatur (WWTP) Decatur (WWTP) Guntersville Guntersville Guntersville Guntersville Guntersville Guntersville 3M Outfall Fox Creek Fox Creek QA/QC QA/QC QA/QC N34o 38' 29.40260" N34o 38' 28.81649" N34o 38' 29.53395" N34o 38' 28.42481" N34o 38' 26.86800" N34o 40' 33.40841" W87o 0 2 ' 03.04910" W87o 0 1 ' 59.71998" W87o 0 1 ' 57.65374" W87o 0 1 ' 57.42383" W87o 0 1 ' 54.56769" W87o 0 2 ' 00.33515" N34o 40' 39.05480" W87o 06' 32.40907" N34o 41' 35.30707" W87o 06' 03.32079" 00 00 00 o "Jo "Jo N34o 41' 31.76767" 05' 57.66518" N34o 41' 28.78044" 05' 49.76882" N34o 40' 14.54885" 06' 33.16910" 00 "Jo N34o 40' 23.97538" 06' 08.31686" N34o 37' 31.83448" N34o 37' 29.73268" N34o 37' 32.67510" N34o 37' 35.16076" N34o 37' 31.81098" N34o 37' 37.18274" W87o 0 0 ' 00.38611" W87o 0 0 ' 02.75138" W87o 0 0 ' 05.30664" W87o 0 0 ' 08.46005" W87o 0 0 ' 08.96597" W87o 0 0 ' 14.65060" N34o 30' 20.88355" W8 6 o 1 0 ' 12.94233" N34o 30' 24.43653" W8 6 o 1 0 ' 48.37409" N34o 30' 23.28174" W8 6 o 1 0 ' 51.61750" N34o 30' 20.24378" W8 6 o 1 0 ' 54.99066" N34o 30' 16.66486" W8 6 o 1 0 ' 56.88539" N34o 30' 15.15118" W8 6 o 1 0 ' 54.36650" N34o 38' 29.33970" W87o 0 2 ' 06.28543" N34o 40' 23.29995" W87o 05' 43.29199" N34o 40' 13.80610" W87o 05' 44.03756" 1 Mussels collected from the vicinity of each sediment/water sampling location downstream of the 3M Outfall. 2 Mussels collected from the vicinity of each sediment sample. BACK TO MAIN APPENDIX C Water Sampling Standard Operating Procedure 3M Decatur Sampling Trip 1.0 PURPOSE The purpose o f this Standard Operating Procedure (SOP) is to provide details for the collection o f water using a MasterFlex peristaltic pump or some comparable unit. This SOP outlines the general procedures for collection o f water samples. Specific sampling details should be outlined in the site-specific work plan and quality assurance project plan for each individual project SCOPE Water samples collected under this SOP are to be taken with a MasterFlex tubing pump or some comparable unit. These samplers are fairly simple devices that consist o f a peristaltic pump and tubing appropriate for the specific project. The tubing fits within a groove within the pump head. Several rollers are designed within the pump head to role over the tube in a clockwise fashion. As the rollers move over the tube they create a vacuum that draws the water up through the tubing and dispels it on the other side. These units are optimal for surface water sampling in cases where it is important that only the selected tubing comes in contact with the sample. No portion of the pump comes in contact with the sample. A new portion o f pre-cleaned tubing can be used with each sample to minimize any risk o f contamination. EQUIPMENT Equipment and supplies needed may include, but are not limited to the following: One boat or sampling barge for the purpose of collecting in situ parameters and surface water samples Direct-read and properly calibrated water quality sampling instruments capable of recording temperature, DO, pH, conductivity, and salinity (YSI Models 63 and 95) MasterFlex tubing pump or some comparable unit Sample containers and coolers with wet ice for sample storage Field data sheets, COC forms, and related materials Items needed to document data collection activities and to label sample jars for shipment, including: camera, film, field notebook, pens, sample labels, custody seals Personal Protective Equipment PPE as required by the site Health and Safety Officer, Decontamination equipment Prior to the collection o f surface water samples, direct-read field meters will be deployed to measure the water quality parameters at each sample station. The direct read instruments will be calibrated each day. Suggested parameters targeted for measurement at each sampling station may include but are not limited to: Water depth (m) Temperature (C) Dissolved oxygen (mg/L) Conductivity (gmhos/cm) MS Word 97  Salinity (7M) pH (s.u.) In some instances, the collection o f additional water quality parameters may be warranted or required. Water quality meters to collect additional parameters will be added as needed. PROCEDURES 1. Obtain the sample location information from either the scope-of-work, work outline, or the Work Plan. This may be in the form o f an actual coordinate or a description o f the general sampling location. 2. Position the sampling barge or boat over the selected sample location. The field crew should always work from the most downstream location to the most upstream location unless otherwise directed. 3. Once you arrive at the proper location measure and record depth o f water and record sample location identification as identified on the sampling log. 4. Make sure that all data sheets correspond to the correct sampling location. 5. Measure the sampling location using the best available technology (e.g., global positioning satellite system) and record sample coordinates on the sampling log. 6. After warm-up and calibration o f the field meters, water quality parameters will be measured and recorded. The depth o f readings should reflect the depth at which water samples are collected and any other requirements o f the site specific work plan. 7. Water samples will then be collected using an appropriate pump with pre-cleaned tubing. 8. The specific type and size o f tubing for sampling should be obtained. The length o f this tubing depends on the depth o f sample to be collected. 9. Weave the tubing through the designated slot on the pump. Make sure that the peristaltic head is correctly situated. 10. Lower the end o f the tube into the water to the desired depth. Turn the pump on and wait for it to prime itself. 11. Run water through the tube until you have completed at least three volume additions o f the tubing (calculated by taking the radius o f the tubing and the length - TTr2l). 12. Once approximately three volume additions have run through the tubing, begin collecting your sample. 13. Sample containers will be sealed with the appropriate caps, labeled, and placed on wet ice in an insulated container or cooler. 14. Appropriate COC documentation will accompany the samples as required by the projectspecific QAPP. 15. After collection o f the sample, remove the tubing and discard. Obtain a new section of tubing and navigate to the next sampling location. Sediment and Soil Sampling Petite Ponar or Eckman Dredge Standard Operating Procedure For the 3M Decatur Sampling Trip 1.0 PURPOSE The purpose of this Standard Operating Procedure (SOP) is to provide details for the collection of surface sediments using either a Petite Ponar or Eckman Dredge. Procedures outlined below detail the general methods required to collect surface sediment samples using either o f these sampling devices. Although the details of sample collection will be influenced by site-specific conditions, certain aspects of sample collection can be standardized. These procedures give descriptions of equipment, field procedures, and documentation necessary to collect sediments using either of these methods. 2.0 SCOPE Petite Ponar The Petite Ponar is widely used in both fresh and salt waters for collecting sediments from soft to relatively hard bottoms. It is especially useful in sand, gravel, consolidated marl or clay bottoms where standard dredges will not work. The Ponar is a self-closing sampler that releases when the unit comes in contact with the sediment. The sample area with this unit is 15 x 15 cm and the unit weight is approximately 55 kg. Eckman Dredge The Eckman Dredge is designed to collect samples in soft, finely divided littoral bottoms. In systems that have high amounts o f debris or rocky bottoms, these units typically will not work well. The unit is typically lowered to the bottom via a rope and a messenger is sent down the line to release the spring-loaded doors. The doors are designed to close tight, encompassing a 15 x 15 cm area of sediment. This SOP will cover the equipment required to collect sediment samples using either of these sampling devices and the specific procedures that are followed. 3.0 EQUIPMENT Equipment and supplies needed may include, but are not limited to the following: Petite Ponar and Eckman Dredge Stable boat or platform barge Stainless steel mixing containers Stainless steel spoons for homogenizing sediments Sample containers (project specific) and coolers with wet ice for sample storage Field data sheets (water quality/chemistry, general data sheets) Chain-of-custody forms  Items needed to document data collection activities and to label sample containers for shipment, including: camera, film, field notebook, pens (write in the rain and Sharpie), sample labels, custody seals Personal protection equipment as required by the site Health and Safety Officer, including organic vapor meters (OVMs) Decontamination equipment specific for the project 4.0 SAMPLE COLLECTION PROCEDURE Surface sediment collection using the Ponar or Dredge samplers will involve the following procedures: 1. Obtain the sample location information from either the scope-of-work, work outline, or the Work Plan. This may be in the form of an actual coordinate or a description o f the general sampling location. 2. Position the sampling barge or boat over the selected sample location. When sampling sediments, the field crew should always work from the most downstream location to the most upstream location unless otherwise directed. 3. When sampling sediments in deep waters, attempt to set the barge in place with either a two to three point tie down (e.g., spud poles). This will provide some stability for the platform and ensure that the platform does not drift while sampling. 4. Once you arrive at the proper location measure and record depth of water and record sample location identification as identified on the sampling log. 5. Make sure that all data sheets correspond to the correct sampling location. 6. Measure the sampling location using the best available technology (e.g., global positioning satellite system) and record sample coordinates on the sampling log. 7. Obtain the decontaminated dredge or ponar (depends on the sediment/substrate type). 8. Make sure that the dredge or ponar are in the open position and slowly lower them over the side of the boat. This is to prevent the units from premature closure. Once the units are slightly submerged, release them and let them fall to the bottom of the system you are sampling in. The weight of the units should force them into the sediment bottom. 9. If using the Eckman Dredge, make sure that the unit is upright on the bottom (tug lightly on the line). Pull the line taut and release the messenger. Once the messenger releases the doors, pull the unit off the bottom and slowly through the water to the boat. 10. For the Ponar, the force of the unit hitting the sediment should cause the pin holding the doors open to release. Therefore when you pull the line holding the unit, the doors will close (collecting the sample). 11. Once the unit is in the boat, place all the contents into one of the decontaminated stainless steel mixing containers. Use a decontaminated, stainless steel spoon to homogenize the sediment. 12. Once homogenized, use the stainless steel spoon to place sufficient sample volumes into the proper sample containers. It is critical that the sample label for this sample be placed on the container. At a minimum, this label should contain the sample ID, date, time collected, initials of the persons collecting, and type of analysis for this sample. BACK TO MAIN 13. Collect the appropriate QA/QC samples as outlined in the QAPP attached to the project specific work plan. 14. Complete the chain-of-custody form. 15. Place all samples in the proper sample shipping/storage container, which should contain sufficient wet ice to keep the samples at a temperature of 4C until receipt by the laboratory. Slight modifications may be required in the field, depending upon the type of sample that is being collected. The field crew will always have a working knowledge o f the equipment prior to sampling, and will be familiar with the specific requirements o f the project. Any modifications or changes that are needed for sampling specific systems should be documented in field data sheets and reflected in the final report. MS Word 97 Fish Sampling Using Nets, Seines, Or Traps General Standard Operating Procedures 3M Decatur Sampling Trip 1.0 PURPOSE/SCOPE The purpose of this Standard Operating Procedure (SOP) is to define the general procedures to be followed for the collection o f fish within the Tennessee River and associated tributaries. This work is completed for partial fulfillment o f the work agreement with 3M. AH fish collections will be performed using the most practicable methods using nets, seines, trawls, traps, or a combination of these methods. Although the details of sample collection will be influenced by site-specific conditions, certain aspects o f sample collection can be standardized. These procedures give descriptions of equipment, field procedures, and documentation necessary to collect fish. All potential sampling techniques will be detailed below, however, not every technique may be used for this particular project. 2.0 EQUIPMENT Equipment to be used during the collection o f fish may include, but is not limited to the following: Sampling vessel (specified upon initial site verification visit) Gill net, seine, and/or trawl Standard minnow traps and bait Weights and buoys (or floats) Filet knives Fish measuring board Electronic scale Anatomical examination checklist Field guides and taxonomic keys Plastic buckets and/or steel wash tubs Sample containers Bubble wrap Ice (wet or dry) Insulated coolers Sample identification labels/tags Waterproof marking pens Plastic Ziploc bags 3.0 FISH COLLECTION The following general procedures will be followed for collecting fish samples from the appropriate sampling locations. Gill Nets Gill nets are most useful when deployed in lentic shallow water habitats where obstructions and debris are mostly absent. This net is especially useful in catching highly mobile fish species. Gill nets o f varying lengths and comprised o f various mesh sizes (i.e., 1.0 in, 1.5 in., 2.5 in. 3.0 in., 3.5 in., and 4.0 in.) are typically utilized. For this particular investigation, 50 ft. gill nets with 1.5 in. mesh size will be used. Nets are selected to consist o f mesh types appropriate to capture targeted fish. Each net is equipped with lead weights and floats designed to hold the net vertically in the water column (i.e., after deployment, the bottom of die net will be suspended at least one foot above the bottom to avoid contact with bottom debris). The nets will be anchored with appropriate weights, and buoy lines will be rigged taut within 1-2 feet with respect to the next predicted high tide following deployment. To comply with federal boating regulations for navigable waterways, buoys will not be set in navigable channels. This requirement may influence the actual location of the gill net. These deployment techniques will ensure reasonable positioning o f the net in the water column throughout the tidal cycle (if working in a tidally influenced system). If necessary, gill nets with alternating mesh sizes may be used during field activities. Gill nets will be deployed perpendicular to shore during the late afternoon - early evening hours and retrieved the following morning, as practicable. Generally, fish activity increases during the night, and the catch retrieved the following day will be more representative o f species movement within the area. During deployment, it may be determined that checking the gill nets more regularly is warranted. This will be decided based on best professional judgement o f the field crew. The following general procedures will be followed for collecting fish with gill nets. 1. Position the vessel (i.e., boat) where the gill net is to be set. 2. Attach floats and anchor weights to surface float lines and bottom lead lines, respectively. 3. Identify and cover with duct tape any cleats, exposed screws, and irregularities in deck rail of the vessel where the net might become entangled during deployment. 4. Deploy gill net perpendicular to shore/current from bow of vessel while vessel is in reverse. Note the tim e and location o f net deployment in field log-book. 5. Make sure that both ends o f the gill net are either tied in place or anchored securely. 6. Retrieve gill net after the desired time interval. Approach the net from the downwind end and slowly pull the net into the boat. 7. Stack the gill net into a cooler or wash tub in coils or figure eights, carefully removing fish as the net is pulled out o f the water. 8. Place fish captured in the net into a clean, labeled, holding container (e.g., insulated cooler). 9. Identify, measure, and/or process the fish as specified in the Work Plan. 10. If adequate numbers and types o f fish are not collected, replace the gill net for another sampling event. Trawls Trawls are bag-shaped nets that are dragged by boat through the water column or along bottom sediments to collect fish. The mesh size used will be dependent on the fish targeted for collection. Trawl nets will be deployed during daylight hours. The following general procedures will be followed for collecting fish with trawls. 1. Floats, rollers, and/or weights will be attached to the trawl as appropriate. 2. Lower the trawl into the water from the side o f the boat, making sure that the trawl is attached securely to the boat. Care should be taken to ensure that the trawl does not contact the boat propeller. 3. The trawl will be deployed and dragged for a time interval or distance appropriate for site conditions and the fish targeted for collection. 4. Note the time, distance, and location of trap deployment in the field log-book. If appropriate, use GPS to document the location of the trawl. 5. After the appropriate distance interval, retrieve the trawl. 6. Empty the trawl into a pre-cleaned holding container (e.g., insulated cooler). 7. Identify, measure, and/or process the fish as specified in the Work Plan. Seine Seines are nets that encircle fish with a long fence-like wall of netting. The top edge of the net is buoyed with floats and the bottom edge is weighted with weights. Seines are most useful in lentic shallow water habitats where the seine can encompass the entire vertical water column. Mesh sizes are typically small for capturing small, relatively immobile fish inhabiting shallow waters. Seines will be deployed during daylight hours, although sampling may also occur in late afternoon and early evening, as many shallow water species may come to the surface to feed at this time. The following general procedures will be followed for collecting fish with seines. 1. Attach floats or buoys along the top edge and weights to the bottom edge of the seine. 2. Two persons while wading from shore will operate the small two-stick seines used for sampling. Seines will be deployed during the late afternoon to early evening hours. 3. The seine will be deployed for a set distance perpendicular to the shoreline. Ensure that the trap is dragged along the bottom sediment to reduce fish escaping. 4. Note the time, distance, and location of seine deployment in the field log-book. 5. After the appropriate distance, retrieve the seine. 6. Empty the fish captured in the seine into a pre-cleaned holding container (e.g., insulated cooler). 7. Identify, measure, and/or process the fish as specified in the Work Plan. Baited Minnow Traps Baited minnow traps can be deployed to catch small bottom dwelling fish seeking food or shelter. This trap is made from reinforced aluminum mesh (1/4 in) where fish pass through a conical-shaped funnel to reach the bait. Baited minnow traps for collecting forage fish will be preferentially set during the day on incoming tides as possible based on the schedule of sampling activities. If sampling activities do not allow for deployment of baited minnow traps during the day, traps are deployed in the late afternoon or early evening hours and retrieved the following morning. The following general procedures are typically followed for collecting fish with minnow traps. 1. Place the bait into a mesh bag or on a hook attached to the center bow of the trap. Attach a float or buoy to end of minnow trap line. 2. Lower the trap into the water from the side of the boat, making sure that the trap is securely anchored and oriented on the river bottom, A buoy should be clearly visible on the surface of the water so that the minnow trap can be easily identified and retrieved. 3. Note the time and location of trap deployment in the field log-book, 4. After the appropriate time interval, retrieve the trap. 5. Empty each trap into a pre-cleaned holding container {e.g., insulated cooler) by slowly pulling the two ends o f the trap apart. 6. Identify, measure, and/or process the fish as specified in the Work Plan. Trotline A trotline is typically used in situations where bottom-dwelling fish are needed. A trotline is a long line that is anchored with weights on each end and individual baited hooks spaced throughout. The trotline can be placed in just about any water body and can collect a large volume and variety of fish. The following general procedures will be followed for collecting fish with a trotline. 1. Bait the hooks at set intervals along the length of the line. Attach a weight and a float or buoy to each end of the line. 2. Lower the line into the water from the side of the boat, making sure that the line is securely anchored. Allow the line to sink to the bottom. A buoy should be clearly visible on the surface of the water so that the line can be easily identified and retrieved. 3. Note the time and location of line deployment in the field log-book. 4. After the appropriate time interval, retrieve the line. 5. Start at one end of the line by pulling it out of the water and draping it into the boat. Place fish into a pre-cleaned holding container {e.g., insulated cooler). 6. Identify, measure, and/or process the fish as specified in the Work Plan. 4.0 SAMPLE HANDLING AND PRESERVATION Fish collected for identification or population surveys should be identified in the field and released. Fish collected for tissue analysis should be placed in plastic bags labeled by sampling station and sampling time, and placed on wet ice in an insulated cooler until further sample preparation is performed. Collected fish will be sacrificed using a filet knife or scalpel to sever the spinal cord just posterior to the brain. Alternative sacrificial methods will consist of placing the fish on wet ice. After sample processing, all fish are placed on wet ice and shipped to the laboratory. APPENDIX D PART I - REPORT Analytical Report Fluorochemical Characterization of Water and Sediment Samples MSU-Entrix (Tennessee River) FACT-GEN-037 (E00-1958) Centre Analytical Laboratory Report No. 023-014O (Revision 2) Revision Date 6/28/01 Testing Laboratory Centre Analytical Laboratory, Inc. 3048 Research Drive State College, PA 16801 3 M E nvironm ental Laboratory C ontact Kent R. Lindstrom Bldg. 2-3E-09 P.O. Box 33331 St. Paul, MN 55133-3331 Phone: (651) 778-5352 R equester Dale Bacon Ph.D. 3M Environmental Technology & Safety Services Bldg. 2-3E-09 P.O. Box 33331 St. Paul, MN 55133-3331 PAGE1OF6 1 Introduction Results are reported for the analysis of a series of water and sediment samples received by Centre Analytical Laboratories, Inc. (Centre) from the 3M Environmental Laboratory. The samples were collected from the Tennessee River and are part of 3M Project E00-1958. The Centre study number assigned to the project is 023-014. Specific fluorochemical characterization by liquid chromatography / tandem mass spectrometry (LC/MS/MS) was requested for all samples. A total of 66 samples were received for analysis. The samples were prepared and analyzed by LC/MS/MS for the following list of fluorochemicals: Table 1: Target Analysis Compound Name Perfluorooctane Sulfonate Perfluorooctane Sulfonylamide Perfluorooctanoate Perfluorohexane Sulfonate Acronym PFOS PFOSA POAA PFHS The analytical methods used for water samples were validated by Centre. The validation protocol and results are on file with Centre. The methods were modified for the sediment samples, however the procedures have not been fully validated for this matrix. Data presented here is the highest quality data available at this time. 2 Sam ple R eceipt The samples were submitted in individual plastic containers and were not preserved. Sixty-six individual sample containers were received. Samples were received on 7/26/00. The samples were collected between 6/19/00 and 6/22/00. Chain-of-custody information is presented in Attachment C. 3 H old ing Tim es The analytical met hod used was validated against a maximum holding time of 14 days. Samples were received after the 14-day holding time. However, it should be noted that field fortifications in water and other matrices have shown acceptable recoveries at 100 and 1000 ng/L for periods longer than 14 days. PAGE2 OF6 4 M ethods - A nalytical and P reparatory 4.1 LC/MS/MS 4.1.1 Sample Preparation for LC/MS/MS Analysis Water samples were initially treated with 200 uL of 250 mg/L sodium thiosulfate solution to remove residual chlorine. Solid phase extraction (SPE) was used to prepare the samples for LC/MS/MS analysis. A forty-milliliter portion of sample was transferred to a Q 8 SPE cartridge. The cartridge was first eluted with 5 mL of 40% methanol in water solution. The eluate was discarded and the SPE column was then eluted with 100% methanol. A 5 mL portion of methanol was collected for analysis by LC/MS/MS. This treatment resulted in an eight-fold concentration of the samples prior to analysis. For the sediment samples, a representative portion of sample (5 grams) was first extracted into 5 ml of methanol. The extracts were filtered and diluted to a final volume of 40 mL with Type I water. The diluted extracts were then treated in the same manner as the water samples, beginning with the solid phase extraction. 4.1.2 Sample Analysis by LC/MS/MS In HPLC, an aliquot of extract is injected and passed through a liquid-phase chromatographic column. Based on the affinity of the analyte for the stationary phase in the column relative to the liquid mobile phase, the analyte is retained for a characteristic amount of time. Following HPLC separation, ES/MS provides a rapid and accurate means for analyzing a wide range of organic compounds, including fluorochemicals. Electrospray is generally operated at relatively mild temperatures; molecules are ionized, fragmented, and detected. Ions characteristic of known fluorochemicals are observed and quantitated against standards. A Hewlett-Packard HP1100 HPLC system coupled to a Micromass Ultima MS/M S was used to analyze the sample extracts. Analysis was performed using selected reaction monitoring (SRM). Water samples were extracted on 8/2/00 and 8/3/00 and analyzed by MS/MS between 8/3/00 and 8/17/00. Sediment samples were extracted 8/4/00 and 8/10/00 and were analyzed by MS/MS between 8/8/00 and 8/17/00. The HPLC and MS/MS methods used for analysis and instrument parameters can be found in Attachments D and E. 5 A nalysis 5.1 Calibration A 7-point calibration curve was analyzed at the beginning and end of the analytical sequence for the compounds of interest. The calibration points were prepared at 0, 25, 50, 100, 250, 500, and 1000 ng/L (ppt). The instrument response versus the concentration was plotted for each point. Using linear regression with 1/x weighting, the slope, y-intercept and correlation coefficient (r) and coeffi2cient of determination (r2) were determined. A calibration curve is acceptable if r > 0 .9 8 5 (r2 > 0.970). Calibration standards are prepared using the same SPE procedure used for samples. PAGE3 OF6 Calibration check standards were analyzed periodically (every three to five sample injections) throughout the analysis sequence. Compliance is obtained if the standard analyte concentrations are within +/-20% of the actual value. For the results reported here, calibration criteria were met. 5.2 Blanks Extraction blanks were prepared and analyzed with every extraction batch of samples. The extraction blanks should not have any target analytes present at or above the concentration of the low-level calibration standard. For these samples, the extraction blanks were compliant. Instrument blanks in the form of clean methanol solvent were also analyzed after every highlevel calibration standard, and after known high-level samples. Again, the blanks should not have any target analytes present at or above the low-level calibration standard. For the samples presented here the instrument blanks are compliant. 5.3 Surrogates Surrogate spikes are not a component of the LC/MS/MS analytical method. 5.4 Matrix Spikes Matrix spikes were prepared for every field sample using all compounds of interest. Matrix spike recoveries are given in Attachment B. Field spikes were submitted with one sediment and one surface water sample. Field spike recoveries are also included in Attachment B. The sediment sample showed no recovery of the field spike, indicating that there was a problem with the sediment field spiking procedure. Field control samples spiked at 200 ppt and 1000 ppt were submitted. The field control samples showed recoveries between 70 and 130%. The results are also included in Attachment B. 5.5 Duplicates All field samples were analyzed in duplicate. Results are given along with the sample results in Attachment A. 5.6 Laboratory Control Samples For LC/MS/MS analyses, Milliq water was spiked with all compounds of interest at 25 and 250 ng/L during each extraction set. All recoveries for all compounds were between 70-130% in each LCS. Results are given along with the raw data in Attachments D and E. 5.7 Sample Related Comments There are no other sample related comments for this data set. 6 Data Sum m ary Please see Attachment A for a detailed listing of the analytical results. Surface water results are reported in parts per trillion (ppt) (ng/L). Sediment sample results are reported in parts per billion (ppb) (ug/Kg) on both an as-received and dry-weight basis. PAGE4OF6 7 Data/Sample Retention Samples are disposed of one month after the report is issued unless otherwise specified. All electronic data is archived on retrievable media and hard copy reports are stored in data folders maintained by Centre. 8 Attachm ents 8.1 Attachment A: Results 8.2 Attachment B: Matrix Spike Recoveries 8.3 Attachment C: Chain of Custody 8.4 Attachment D: LC/MS/MS Raw Analytical Data (Surface Water Samples) 8.5 Attachment E: LC/MS/MS Raw Analytical Data (Sediment Samples) 9 Signatures John M. Flaherty, Operations Manager Kevin J Lloyd, Vice President Other Lab Members Contributing to Data Karen Smith Date Date PAGE5 OF6 APPENDIX E-1 W ater quality param eters for the collected by Entrix, June 2000. Sam ple ID DO C onductivity (m g/L) (pm hos/cm ) SW -01 6 .0 4156 SW -02 7.2 2240 SW -03 8.7 505 SW -04 9.1 472 SW -05 9.4 540 SW -06 8 .6 386 SW -07 8 .0 176 SW -08 10.3 172 SW -09 8.4 158 SW -10 7.7 158 SW -11 7.8 178 SW -12 7.8 179 SW -13 8.9 187 SW -14 1 2 .6 188 SW -15 9.5 188 SW -16 9.6 180 SW -17 9.5 183 SW -18 9.9 182 SW -19 9.3 182 SW -20 7.3 195 SW -21 7.3 196 SW -22 7.1 194 SW -23 7.2 196 SW -24 7.3 194 SW -25 6.5 4650 SW -26 8 .6 183 Tennessee Salinity (o/oo) 1.9 1.0 0 .2 0 .2 0 .2 0 .2 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 2 .0 0.1 R iver and tributaries Tem perature (oC ) 32.5 31.3 29.6 29.5 30.0 29.1 28.0 28.4 27.6 27.5 28.0 27.6 29.1 29.7 29.4 29.4 29.1 28.9 29.6 28.7 28.9 28.8 28.9 29.2 33.2 27.6 pH ( s .u .) 7.2 7.4 8.1 8.1 8.1 7.9 7.8 8.7 8.1 7.7 8.4 8 .2 8.4 9.0 8.7 8 .8 8 .8 8 .8 8.9 8 .0 8 .0 8 .0 8 .0 8.1 7.2 8.5 APPENDIX E-2 W ater quality param eters for sam ples collected from the Tennessee R iver dow nstream o f the W heeler Dam . Sam ples w ere collected by 3M and Pace A n a ly tica l L a b o r a to ry . 1____________________________________________________ W ater D .O . C onductivity Tem perature pH S am ple (m gA )) (pm hos/cm ) (oC ) ( s .u .) W D D05 7.3 170 23.5 7.0 W D D06 7.5 170 23.0 7.1 W D D07 7.8 170 23.5 7.2 W D D08 8.5 170 23.5 7.4 W D D09 1 0 .2 160 24.0 7.7 W D D10 10.4 160 23.5 7.8 W D D11 11.5 160 24.0 8.1 W D D12 11.7 160 24.0 8.3 W D D13 10.5 160 23.5 8.1 W D D14 10.4 160 23.0 8 .2 W D D15 9.2 160 23.0 7.9 W TI 9.0 160 23.0 7.5 1 Sample locations as given in Appendix B. WDD is Wheeler Dam Downstream with WDD being furthest downstream. WTI is Water Treatment Intake near Wilson Dam. jum ping dialion failk i e plant SW-18 D.O. = 9.9 Cond. = 182 Sal. = 0.1 Temp. = 28.9 pH = 8.8 SW-16 D.O. = 9.6 Cond. = 180 Sal. = 0.1 Temp. = 29.4 pH = 8.8 SW-17 D.O. = 9.5 Cond. = 183 Sal. = 0.1 Temp. = 29.1 pH = 8.8 SW-14 D.O. = 12.6 Cond. = 188 Sal. = 0.1 Temp. = 29.7 pH = 9 A 'w >:r . In, , , -, l r- ry ;. . a ';rC_ L a%.. j k j'- 1e - SW-15 D.O. = 9.5 Cond. = 188 Sal. = 0.1 Temp. = 29.4 pH = 8.7 LEGEND: 5 Sampling Locations D.O. = Dissolved Oxygen (m g/L) Cond. = C onductivity (um hos/cm ) Sal. = S alin ity (o/oo) Tem p. = T em perature (C) pH = pH (s.u.) SW-13 D.O. = 8.9 Cond. = 187 Sal. = 0.1 Temp. = 29.1 pH = 8.4 SWAM MAN A N A 500 0 500 1000 Feet rj f C,i Vl Vj i 'A T --. m 'p 'Z -'- SSl = T o Jjrif ' _ v ". Figure 1A Decatur W aste W ater Treatm ent Plant Surface W ater Chemistry 3M Project DATE: 2/19/01 .Finley Island Cabin Site rea SW-06 C h e m 6^1 SW-01 D.O. = 6 Cond. = 4156 Sal. = 1.9 Temp. = 32.5 D.O. = 8.6 Cond. = 386 Sal. = 0.2 Temp. = 29.1 pH = 7.9 pH = 7.2 SW-03 D.O. = 8.7 Cond. = 505 Sal. = 0.2 Temp. = 29.6 pH = 8.1 : SW -25 - ; D.O. = 6.5 Cond. = 4650 Sal. = 2 Temp. = 33.2 1pH = 7.2 \S ~ X s J } \^ Y i\ SW-02 D.O. = 7.2 Cond. = 2240 Sal. = 1 Temp. = 31.3 pH = 7.4 : :.i < SW-05 D.O. = 9.4 Cond. = 540 Sal. = 0.2 Temp. = 30 pH = 8.1 SW-04 D.O. = 9.1 Cond. = 472 Sal. = 0.2 Temp. = 29.5 pH = 8.1 LEGEND: Sampling Locations D.O. = Dissolved Oxygen (m g/L) Cond. = C onductivity (um hos/cm ) Sal. = S alin ity (o/oo) Tem p. = T em perature (C) pH = pH (s.u.) IT>Purrifjir -I. 400 N A 0 400 800 Feet Figure 1B 3M Outfall Area Surface Water Chemistry 3M Project DATE: 2/19/01 x Mile'295 SW-08 D.O. = 10.3 Cond. = 172 Sal. = 0.1 Temp. = 28.4 pH = 8.7 SW-09 D.O. = 8.4 Cond. = 158 Sal. = 0.1 Temp. = 27.6 pH = 8.1 SW-10 D.O. = 7.7 Cond. = 158 Sal. = 0.1 Temp. = 27.5 pH = 7 .7 jr f f f 1t ; 1 .l \ ) - i- SW-07 D.O. = 8 Cond. = 176 x Sal. = 0.1 Temp. = 28 pH = 7.8 i\S i.r XT ~\ 1 IJ ) \ \ \ SW-11 r< 1 {( v ' si D.O. = 7.8 1 \N ( 'C ' Cond. = 178 - Sal. = 0.1 Temp. = 28 pH = 8.4 SW-12 D.O. = 7.8 Cond. = 179 Sal. = 0.1 Temp. = 27.6 pH = 8.2 SW-27 S/. WSfi 2 IL $ 5 30 SW-26 D.O. = 8.6 Cond. = 183 Sal. = 0.1 Temp. = 27.6 pH = 8.5 LEGEND: $ Sampling Locations D.O. = Dissolved Oxygen (m g/L) Cond. = C onductivity (um hos/cm ) Sal. = S alin ity (o/oo) Tem p. = T em perature (C) pH = pH (s.u.) 500 0 500 1000 Feet Figure 1C Fox Creek Area Surface Water Chemistry 3M Project DATE: 2/19/01 SW-19 D.O. = 9.3 Cond. = 182 Sal. = 0.1 Temp. = 29.6 pH = 8.9 SW-23 D.O. = 7.2 Cond. = 196 Sal. = 0.1 Temp. = 28.9 pH = 8 SW-21 D.O. = 7.3 Cond. = 196 Sal. = 0.1 Temp. = 28.9 pH = 8 SW-20 D.O. = 7.3 Cond. = 195 Sal. = 0.1 Temp. = 28.7 pH = 8 SW-24 D.O. = 7.3 Cond. = 194 Sal. = 0.1 Temp. = 29.2 pH = 8.1 SW-22 D.O. = 7.1 Cond. = 194 Sal. = 0.1 Temp. = 28.8 pH = 8 LEGEND: $ Sampling Locations D.O. Cond. Sal. Temp. pH Dissolved Oxygen (mg/L) Conductivity (umhos/cm) S alinity (o/oo) : T em p eratu re (C) = pH (s.u.) N 500 0 500 1000 Feet Figure 1D Upstream (Guntersville) Reference Location Surface Water Chemistry 3M Project DATE: 2/19/01 Pumping station * J H in i S * " 1,11 :iint S W -18 PFOS = 0.03305 FOSA = 0.00555 PFOA = 0.02025 PFHS =0.0025 S W -17 PFOS = 0.0548 FOSA = 0.00955 PFOA = 0.01445 PFHS =0.00435 S W -16 PFOS = 0.0532 FOSA = 0.0097 PFOA = 0.0289 PFHS =0.0045 S W -15 PFOS = 0.0547 FOSA = 0.00945 PFOA = 0.02795 PFHS =0.00345 SW -14 PFOS = 0.0655 FOSA = 0.012 PFOA = 0.0311 PFHS =0.0153 S W -13 ; H PFOS = 0.0627 P FOSA = 0.01265 ; ! PFOA = 0.03135 PFHS =0.00435 LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/L) FOSA = Perfluorooctanesulfonamide (ug/L) PFOA = Perfluorooctanoate (ug/L) PFHS = Perfluorohexanesulfonate (ug/L) SA3 500 0 500 1000 Feet Figure 2A Decatur W aste W ater Treatment Plant Concentrations in Surface W a ter 3M Project DATE: 2/15/01 - rT* S^Finley Island Cabin Site Area S W -06 SW-01 PFOS = 142.5 FOSA = 8.38 PFOA = 1750 PFHS = 37.05 PFOS = 11.9 FOSA = 1.003 PFOA = 104 PFHS =3.86 S u b statL O T -* S W -25 PFOS = 150.5 FOSA = 7.985 PFOA = 1900 PFHS =42.7 SW -02 PFOS = 67.2 FOSA = 4.62 PFOA = 750.5 PFHS =17.35 S W -03 PFOS = 12.6 FOSA = 1.14 PFOA = 121.5 PFHS =3.265 SW -04 S? 34 POT ? PFOS = 12.7 FOSA = 1.089 PFOA = 104.5 PFHS =3.73 1 S W -05 PFOS = 16.5 FOSA = 1.52 PFOA = 174.5 PFHS =4.58 LEGEND: f) Sam pling Locations PFOS = Perfluorooctanesulfonate (ug/L) FOSA = Perfluorooctanesulfonamide (ug/L) PFOA = Perfluorooctanoate (ug/L) PFHS = Perfluorohexanesulfonate (ug/L) 300 0 300 600 Feet Figure 2B 3M Outfall Area Concentrations in Surface W ater 3M Project DATE: 2/15/01 SW-08 PFOS = 0.317 FOSA = 0.09785 PFOA = 1.53 PFHS =0.3145 S W -09 PFOS = 0.0982 FOSA = 0.03575 PFOA = 0.5705 PFHS =0.0848 S W -10 PFOS = 0.1125 FOSA = 0.032075 PFOA = 0.9035 PFHS =0.10345 S W -07 PFOS = 0.4865 FOSA = 0.146 ^ PFOA = 2.135 PFHS =0.497 SW-11 PFOS = 0.531 FOSA = 0.146 PFOA = 1.985 PFHS =0.428 S W -12 PFOS = 0.5695 FOSA = 0.186 PFOA = 2.838 PFHS =0.6455 S W -27 V. WSR 28 L $ S W -26 PFOS = 0.5855 FOSA = 0.1845 PFOA = 3.46 PFHS =0.658 LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/L) FOSA = Perfluorooctanesulfonamide (ug/L) PFOA = Perfluorooctanoate (ug/L) PFHS = Perfluorohexanesulfonate (ug/L) N A 500 0 500 1000 Feet Figure 2C Fox Creek Area Concentrations in Surface Water 3M Project DATE: 2/15/01 S W -19 PFOS = 0.0081 FOSA = 0.0025 PFOA = 0.0075 PFHS =0.0025 SW-21 PFOS = 0.0089 FOSA = 0.0045 PFOA = 0.0075 PFHS =0.0025 S W -23 PFOS = 0.00905 FOSA = 0.00285 PFOA = 0.0075 PFHS =0.0025 S W -24 PFOS = 0.0063 FOSA = 0.0034 PFOA = 0.0075 PFHS =0.0025 S W -20 PFOS = 0.01175 FOSA = 0.008 PFOA = 0.0075 PFHS =0.0025 S W -22 PFOS = 0.0081 FOSA = 0.0036 PFOA = 0.0075 PFHS =0.0025 vtV .,.; " : - n-\ IB s LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/L) PFOSA = Perfluorooctanesulfonamide (ug/L) PFOA = Perfluorooctanoate (ug/L) PFHS = Perfluorohexanesulfonate (ug/L) 500 0 500 1000 Feet Figure 2D Upstream (Guntersville) Reference Location Concentrations in Surface Water 3M Project DATE: 2/15/01 dumping slatin I iv faim ei/vfje <hsp5sa1 p la n t S W -18 PFOS = 0.82 FOSA = 0.1375 PFOA = 0.096 PFHS = 0.08 S W -16 PFOS = 0.513 FOSA = 0.08 PFOA = 0.08 PFHS = 0.08 S W -17 PFOS = 0.963 FOSA = 0.1255 PFOA = 0.08 PFHS = 0.08 SW -14 PFOS = 0.468 FOSA = 0.0985 PFOA = 0.08 PFHS = 0.08 D s-sftjjr 2? 1 1{5==n] 1I K n 1 H 1t e s V 1M 11 i: 1. <4 l1l' l- T= fex 'V qI*---L 1*+1 . 'fe*.t 4- '' . LA E S W -15 PFOS = 1.415 FOSA = 0.2005 PFOA = 0.087 PFHS = 0.08 LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) S W -13 'CO H PFOS = 1.5 FOSA = 0.24 PFOA = 0.1205 PFHS = 0.099 500 0 500 1000 Feet Figure 3A Decatur W aste W ater Treatment Plant Sediment Concentrations (W et Weight) 3M Project DATE: 2/19/01 --Finley Island Cabin Site Area S W -06 C h e m si SW-01 PFOS = 15645 FOSA = 2935 PFOA = 536 PFHS = 38.05 PFOS = 74.15 FOSA = 21.2 PFOA = 42.2 PFHS = 6.275 S W -03 PFOS = 371 FOSA = 84.4 PFOA = 490 PFHS = 19.45 Sub 1f SW -04 II. .IL1 %Wa . >- SW -25 ,-p--JT1 ---- ---- l " ' ; Val'/j PFOS = 5930 . statar FOSA = 1200 n J PFOA = 1855 '' y fl >Sfi 34 POT 3 PFOS = 471.5 FOSA = 101.95 PFOA = 648.35 PFHS = 23.2 \ SiftPsF 135 SW -02 jr State SW-05 PFOS = 1965 FOSA = 472 PFOA = 1250 PFHS = 68.4 V PFOS = 77.7 FOSA = 55.75 1PFOA = 120 PFHS = 7.505 . -/Q --------------------------------- iW ( LEGEND: Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 300 0 300 600 Feet Figure 3B 3M Outfall Area Sediment Concentrations (W et Weight) 3M Project DATE: 2/19/01 SW-08 PFOS = 1.028 FOSA = 0.481 x PFOA = 0.725 ' PFHS = 0.08 S W -09 PFOS = 0.916 FOSA = 0.4575 PFOA = 1.075 PFHS = 0.08 S W -10 PFOS = 0.848 FOSA = 0.383 PFOA = 0.597 PFHS = 0.08 S W -07 PFOS = 0.941 FOSA = 0.5755 PFOA = 0.864 PFHS = 0.081 = SW-11 \ \ ; })/ ? i ovlWVL; ,, ' PFOS = 1.59 FOSA = 0.832 _ PFOA = 1.175 PFHS = 0.13 J v \r S W -12 PFOS = 3.16 FOSA = 2.285 PFOA = 2.325 PFHS = 0.285 'SW -26 S W -27 PFOS = 3.585 FOSA = 2.225 PFOA = 2.625 PFHS = 0.325 LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 500 0 500 1000 Feet Figure 3C Fox Creek Area Sediment Concentrations (W et Weight) 3M Project DATE: 2/19/01  m t S W -19 PFOS = 0.093 FOSA = 0.08 PFOA = 0.08 PFHS = 0.08 SW-21 PFOS FOSA PFOA PFHS 0 .1 3 8 5 0.08 0.08 0.08 S W -23 PFOS = 0.169 FOSA = 0.08 PFOA = 0.08 PFHS = 0.08 SW -24 PFOS FOSA PFOA PFHS 0 .2 6 1 5 0.0915 0.093 0.08 S W -20 PFOS FOSA PFOA PFHS 0 .2 6 7 5 0.087 0.08 0.08 S W -22 PFOS FOSA PFOA PFHS 0 .1 4 4 0.08 0.08 0.08 -fgt rv~* LEGEND: Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 500 0 500 1000 Feet Figure 3D Upstream (G untersville) Reference Location Sedim ent Concentrations (W et W eight) 3M Project DATE: 2/19/01 11jum ping slalin r ' " n. jLi-; *---ii. VI v r - ** v- + I r V 't M~1 !e"(isp^sal oL&nt S W -18 ^3 ;jSB'y " fc. | _ ^ * * , 111 I a h k ^ ; i t -*j, -- A . fcl PFOS = 1.35 FOSA =0.2265 PFOA = 0.1585 PFHS = 0.132 S W -17 PFOS = 2.6 FOSA =0.339 PFOA = 0.216 PFHS = 0.216 BACK TO MAIN S W -16 PFOS = 0.7455 FOSA =0.116 PFOA = 0.116 PFHS = 0.116 V.>, v - r"-,i * fc . x . tf 2t ^ j_ S W -15 va" - ' PFOS = 2.175 FOSA =0.3085 PFOA = 0.134 PFHS = 0.123 SW -14 V> . , PFOS = 0.6225 FOSA =0.131 PFOA = 0.106 PFHS = 0.106 S W -13 ! .I ^ ;! PFOS = 2.52 FOSA =0.4035 PFOA = 0.2025 PFHS = 0.166 LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 500 0 500 1000 Feet Figure 4A Decatur W aste W ater Treatm ent Plant Sediment Concentrations (Dry W eight) 3M Project DATE: 2/19/01 BACK TO MAIN ^Finley Island Cabin Site Area r" r r"4 1,1;. *a * 9 h> Chem tu ri 3 S W -01 PFOS = 2395 FOSA = 451 PFOA = 1435 PFH S = 5 8 .4 5 - N 3 -5 ^ . f . SW -06 PFOS = 101.95 FOSA =29.1 PFOA = 57.95 PFHS = 8.62 Subsjtaji jfi SW -25 PFOS = 12600 FOSA =2555 PFOA = 3950 PFHS = 287.5 r. hr litatief. Jr 8 P S W -02 . PFOS = 3865 FOSA =929 PFOA = 2460 PFHS = 135 SW -03 PFOS = 567 FOSA =129.5 PFOA = 749.5 PFHS = 30.35 \ <h SW -04 SR 34 J0 T ? PFOS = 680.5 FOSA =147 PFOA = 935.5 PFHS = 33.5 S W -05 PFOS = 105.3 FOSA =75.465 PFOA = 162.5 PFH S = 1 01.1.1335 \ . LEGEND: f) Sam pling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 300 300 600 Feet - J- Figure 4B 3M Outfall Area Sediment Concentrations (Dry W eight) 3M Project DATE: 2/19/01 S W -08 PFOS = 2.24 FOSA =1.05 PFOA = 1.585 PFHS = 0.174 BACK TO MAIN S W -09 PFOS = 0.195 FOSA =0.9755 PFOA = 2.29 PFHS = 0.171 S W -10 PFOS = 1.855 FOSA =0.838 PFOA = 1.305 PFHS = 0.175 S W -07 y r ijy PFOS = 1.27 FOSA =0.7755 PFOA = 1.165 PFHS = 0.1085 v " \\ v\ I J,j i AI-i-. 1 Vlk Jw I k VV ` ^V ' \\\\ JL c* 1 S V1'1 'I If V VJ v\ / \ I! SW -11 ; PFOS = 2.09 FOSA =1.095 PFOA = 1.545 PFHS = 0.171 v4 y S W -12 PFOS = 5.885 FOSA =4.25 PFOA = 4.33 PFHS = 0.5335 SW -26 $\ S W -27 PFOS = 6.46 FOSA =4.01 PFOA = 4.73 PFHS = 0.586 LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 500 0 500 1000 Feet Figure 4C Fox Creek Area Sediment Concentrations (Dry W eight) 3M Project DATE: 2/19/01 V J//C BACK TO MAIN S W -19 PFOS = 0.1245 FOSA =0.107 PFOA = 0.107 PFHS = 0.107 SW-21 PFOS = 0.3625 FOSA =0.209 PFOA = 0.209 PFHS = 0.209 S W -23 PFOS = 0.4415 FOSA =0.209 PFOA = 0.209 PFHS = 0.209 SW -24 PFOS = 0.59 FOSA =0.2065 PFOA = 0.21 PFHS = 0.181 S W -20 PFOS = 0.7013 FOSA =0.224 PFOA = 0.206 PFHS = 0.206 S W -22 PFOS = 0.385 FOSA =0.214 PFOA = 0.214 PFHS = 0.214 _________ Hfu,_l A * di-. 8M 9 PN LEGEND: $ Sampling Locations PFOS = Perfluorooctanesulfonate (ug/kg) FOSA = Perfluorooctanesulfonamide (ug/kg) PFOA = Perfluorooctanoate (ug/kg) PFHS = Perfluorohexanesulfonate (ug/kg) 500 0 500 1000 Feet Figure 4D Upstream (Guntersville) Reference Location Sediment Concentrations (Dry W eight) 3M Project DATE: 2/19/01 BACK TO MAIN APPENDIX G Length, weight, condition factor and liver som atic index for individual fish collected from the Tennessee River. S am ple L en g th W eight Liver w t ID LM1 LM2 SHAD1 SHAD2 SHAD3 SHAD4 SHAD5 SHAD6 SHAD7 SHAD8 WP1 WP2 CAT1 CAT2 CAT3 (cm) 26.5 32 42 35 36 42.5 37 34 25 21 17 16.5 31 34 32 (g) 195 454 567 340.2 396.9 510.3 396.9 354.4 154 96 81 71 296 454 283.5 K1 1.0478 1.3855 0.7653 0.7935 0.8507 0.6648 0.7836 0.9017 0.9856 1.0366 1.6487 1.5805 0.9936 1.1551 0.8652 (g) 1.32 2.92 0.87 na3 3.43 1.32 1.84 2.29 na na 0.44 0.39 5.77 5.52 4.17 L S I2 0.6769 0.6432 0.1534 na 0.8642 0.2587 0.4636 0.6462 na na 0.5432 0.5493 1.9493 1.2159 1.4709 CAT4 GAR1 49 1034.8 0.8796 64.5 963.9 0.3592 9.48 2.87 0.9161 0.2977 GAR2 61 850.5 0.3747 10.72 GAR3 54 623.7 0.3961 9.45 GAR4 54 623.7 0.3961 2.41 GAR5 44.5 340.2 0.3861 4.88 GAR6 52 581.2 0.4133 2.27 SB1 23 149 1.2246 0.59 SB2 24 161 1.1646 0.92 SB3 26.5 261 1.4025 1.42 SB4 21 114 1.2310 0.81 SB5 27 289 1.4683 1.29 SB6 22.5 130 1.1413 1.04 SB7 21 114 1.2310 1.18 SB8 19 103 1.5017 0.69 SB9 25 208 1.3312 0.83 SB10 23.5 148 1.1404 0.69 SB11 22 143 1.3430 0.76 SBT2 24 196 1.4178 1.78 SBT3 22.5 137 1.4178 0.81 SB14 24.5 193 1.2027 1.29 T 2 Condition factor (K) was calculated as K= (Wt x 100)/L33 Liver somatic index (LSI) was calculated as LSI= (Liver wt/total wt) x100 3 na is not applicable since the liver was not removed from sample 1.2604 1.5152 0.3864 1.4345 0.3906 0.3960 0.5714 0.5441 0.7105 0.4464 0.8000 1.0351 0.6699 0.3990 0.4662 0.5315 0.9082 0.4133 0.9416 Gen-042 Analytical Results: Fish Liver and Clam Tissues APPENDIX H 3M Environmental Technology and Services PO Box 33331 St. Paul, M N 55133-3331 61277 6442 BACK TO MAIN 3M November 30,2000 JohnNewsted Entrix 4295 Okeinos Road Suite 101 Okemos, MI 48864 Dear JohnNewsted: Enclosedaie 11raw datapackets for study GEN042. These datapackages include all data from extraction date 8/14/00 and die reextractionon 10/18/00 & 10/19/00. If youhave any questions or comments regarding the data transfer, please contact me at (651) 778-5568. Sincerely, Lisa A. Clemen Advanced Chemist enclosure Page 1 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN FACT-r^ -N042 fi r *i * iiI n i11 i i J 1 t i iSf i3 s SS 2 i1 s S S 8+ if HH U IJi31 II !S igasss SS1s Si*2IlIlHS 11 i l l i i II aoi i II i i i i i i i i i l i i 111 il Ii ii II II i i i i i i i i i i i i i i II! il! iiii Ilii im Ul i ii II ii S3si i l s 2S2g i s s i l i i i 1-- as 8SS -- -- --S22S88 - g-28 - " jjj n n m mjSSs a1SSSSiS SS3Si Sa SBSiS S i i d s f ! n ni l l ! II 1 11 i 1 Ibis ? * nS Is1 Issf l i tf i 1* all J 5i H i fl *1*11 Ji , f1i tii l l lll l i H sss ss ss Si SS SSSS s s s s s s s s ssssss sss sssss ssssss 1er HSill II jjjsy i l I! i i l p i i i 1^11i 1II I 1111! l m j !i m mSj". i l l 111! II i! i l 11oidiHlOlrtHHl i 11! i i i i i l i i i l ii iil sss 1 II I iillil 1 II I 1 ill ! f ss ss ss SS H! l i T ! !`i T " I H i; ii * m m * pais ii i i II ii 1 fl 11 Siil iS li iiSJ i i i l i i j i i| l i 1 l I .j II 1 II a 1 Page 2 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN FACT-*" ' ' 042 Page 3 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN Page 4 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN FACT ~~N-042 Page 5 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN F A C T -r" N-042 Page 6 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN FACTrev' :BN-042 I 1 Page 7 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN Page 8 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN FA CT "~N -042 Page 9 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN 13 8 FA C T N -042 ~Sl?4C*jdi Page 10 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN FA CT ^-042 Page 11 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN 11 I F A C T t-042 ~S 1749CMJd* 1 Page 12 of 13 Gen-042 Analytical Results: Fish Liver and Clam Tissues BACK TO MAIN Page 13 of 13 BACK TO MAIN FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab Appendix I A n a ly tic a l L a b o ra to ry R ep o rt Title Quantitative Analysis of Fluorochemicals in Environmental Samples Obtained from Entrix (Decatur - Tennessee River Samples) on 03/30/2001 D ata R equirem ent Not Applicable A uthor 3M Environmental Laboratory A n alytical P hase C om pletion D ate at signing P roject Identification 3M Study Number: FACT-GEN-055 3M Laboratory LIMS No. E00-0520 Page 1 of 9 Summary of Findings BACK TO MAIN FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab F IS H (WHOLEBODY) American Shad - PFOS and PFOSA were measured above the limit of quantitation in all of the samples (n = 8). PFOA and PFHS were measured above the limit of quantitation in 5 out of 8 and 4 out of 8 samples, respectively. Catfish - PFOS and PFOSA were measured above the limit of quantitation in 2 out of 4 samples. PFOA and PFHS were measured above the limit of quantitation in 1 out of 4 samples. GAR - PFOS was measured above the limit of quantitation in 3 out of 6 samples. PFOSA, PFOA, and PFHS were measured above the limit of quantitation in 2 out of 6 samples. Large-Mouth Bass - PFOS and PFOSA were measured above the limit of quantitation in both of the samples. PFOA and PFHS were not measured above the limit of quantitation in any of the samples. Striped Bass - PFOS and PFOSA were measured above the limit of quantitation in all of the samples (n = 14). PFOA and PFHS were not measured above the limit of quantitation in any of the samples. White Perch - PFOS and PFOSA were measured above the limit of quantitation in both of the samples (n = 2). PFOA and PFHS were not measured above the limit of quantitation in any of the samples. Refer to the attached spreadsheets for more information. Method Summaries Following is a brief description of the 3M Environmental Laboratory analytical methods used during this study. 3M Environmental Laboratory Pr e p a r a t o r y M eth o d s ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Liver for Analysis using HPLC-Electrospray/Mass Spectrometry". An ion-pairing reagent was added to the sample and the analyte ion pair was partitioned into MtBE. The extract was transferred to a centrifuge tube and put onto a nitrogen evaporator until dry. Each extract was reconstituted in 1.0 mL of methanol, and then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials. Page 2 of 9 BACK TO MAIN FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab A n a lytic a l M eth o d s ETS-8-7.0, "Analysis of Potassium P erflu o ro octan esu lfo nate o r o th er F lu o ro ch em icals in L iver E xtracts U sing H P L C -E le c tro s p ra y /M a s s S p e c tro m e try ". T h e analyses w e re perform ed by m onitoring a single product ion selected from a prim ary ion characteristic of a particular fluorochem ical using H P L C -E S /M S /M S . For e x a m p le , m o le c u la r ion 4 9 9 , s ele cte d a s th e prim ary ion fo r P F O S (C 8F 17S O 3-) analysis, w a s fra g m e n te d fu rth e r to pro d u ce ion 9 9 ( F S O 3-). T h e c h a ra cteristic product ion 9 9 w a s m onitored fo r quantitative analysis. A n a l y t ic a l Eq u ipm en t T h e following are representative of the actual settings used during the analytical phase of this study. Liquid Chromatograph: Hew lett-Packard S eries 1100 Liquid Chrom atograph system A nalytical colum n: Keystone B etasilTM C 18 2 x 5 0 m m (5 p m ) Column tem perature: Am bient Mobile phase com ponents: Com ponent A: 2m M am m onium acetate C o m p o n e n t B: m ethanol Flow rate: 3 00 pL/min Injection volum e: 10 pL Solvent Gradient: 16.0 minutes Time (minutes) 0.0 1.0 6.0 11.0 13.0 %B 10% 10% 90% 100% 10% Mass Spectrometer: M icrom ass A P I/M a s s S p e c tro m e te r Q uattro IITM Triple Q uadrupole system Softw are: M ass LynxTM 3.4 C one Voltage: 3 0 -6 0 V Collision G as Energy: 2 5 - 4 5 e V Mode: Electrospray Negative Source Block Tem perature: 150C 1 0C Electrode: Z-spray Analysis Type: Multiple Reaction Monitoring (M R M ) Page 3 of 9 BACK TO MAIN FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab Table 3. Target Ions Monitored in 3M Laboratory Analyses Target Analyte PFOS PFOSA PFOA PFHS THPFOS Prim ary Ion (AMU) 499 498 413 399 427 Product Ion (AMU) 80, 99 78 169 99 80 Data Quality Objectives and Data Integrity The following data quality objectives were indicated in the method performance section of ETS-8-7.0, Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry: Linearity: The coefficient of determination (r2) equal to or greater than 0.980. Limits of Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration curve. Acceptable Precision: Precision is better than 30% for the method. Acceptable Spike Recoveries: 70-130% Blanks: Matrix or surrogate matrix blanks must have a peak area < % the peak area of the lowest acceptable standard (LOQ). Data Summary, Analyses, and Results Data quality objectives for the analytical phase of this study were met with the exceptions noted in this report. Summary of Quality Control Analyses Results for PFOS, PFOSA, PFOA, and PFHS Linearity: The coefficient of determination (r2) of the standard curve was >0.990. Calibration Standards: Quantitation of the target analytes was based on a quadratic curve weighted 1/x of a single, opening or two bracketing unextracted curves for each group of samples. High or low points on the curve may have been deactivated to provide a better fit over the curve range most appropriate to the data. Low curve points with peak areas less than two times that of the extraction blanks were deactivated to disqualify a data range that may have been significantly affected by background levels of the analyte. Occasionally, a single mid-range curve point that was an obvious outlier may have been deactivated. Quantitation of each analyte Page 4 of 9 FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab was based on the response of one or more specific product ion(s) using the multiple reaction-monitoring mode of the instrument (see Appendix C, Analytical Methods). Calibration standards were prepared to run, undiluted, approximately within the linear range of the instrument (approximately 5-1000 ng/g). Limits of Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration curve (defined as a standard within 30% of the theoretical value), and is at least two times the analyte peak area detected in the surrogate matrix blanks. Because low levels of the target analytes are ubiquitous in the laboratory, it is imperative that these criteria for LOQ determination be observed. The approximate LOQ for each analyte in each matrix is listed on the results sum BAC table. Blanks: All blanks were below the limit of quantitation for the compounds of interest. The well-characterized matrix of rabbit liver was used as surrogate tissue for matrix blanks. Matrix/surrogate matrix blanks were < % the level of the LOQ. Precision, Instrumental: Instrumental precision was not specifically determined within the course of this study. In other, similar liver/tissue studies, instrumental precision has been determined to be better than 7% for all target analytes. Matrix Spikes: When possible, at least one set of matrix spikes was prepared for each type of tissue from each type of animal; typically spikes were prepared at two different levels for each type of fish. Matrix spikes were prepared, after an initial screening of the samples, at appropriate levels (usually a "high" and "low" level), between 10-3500 ng/g. Please see the attached summary table for results of the spike studies. Matrix spike recoveries were not always within the expected range (+/-30%), indicating that some of the matrices challenged the analytical method. Although some spike recovery results were within +/-50% of the expected values, and thus may still be considered to support a semi-quantitative interpretation of the data, a few matrix spikes were recovered outside of this range. For these tissues, the presented results can be considered to be qualitative only. To verify the status of the extraction and analytical systems, matrix spikes were also prepared in rabbit liver. Matrix spike studies conducted in rabbit were all acceptable (+/-30%) for those analytical runs used when reporting data for the fish homogenates and fish matrix spikes. Prior to the extraction and analysis of the fish tissues by 3M, samples of the same fish homogenates were extracted by Entrix personnel using an alternative extraction method. 3M conducted the analysis on Entrix-prepared extracts. Results of the matrix spike studies conducted on the Entrix-prepared samples are provided in an attached summary table for comparison. Demonstration of Specificity: Specificity for analyte identification was demonstrated by chromatographic retention time and mass spectral daughter ion characterization. Additional confirmatory tests were performed for PFOS. In the electrospray tandem mass spectrometry (ESMSMS) system, the 499 Da ^ 80 Da transition can provide a stronger signal than the 499 Da ^ 99 Da transition of the PFOS analysis. However, in the analysis of tissue samples collected from some species of animals, an unidentified interferent was present in the 499 Da ^ 80 Da transition. Although this interferent is rarely observed, to ensure complete selectivity, quantitation was based on the 499 Da 99 Da transition. Page 5 of 9 FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab To verify the identity of the detected analyte, in samples determined to contain PFOS at > 70 ng/g (liver or other tissue), at least 2 transitions were monitored and showed quantitative agreement to within +/-30%. Typically, 499>99 and 499>80 transitions were monitored. On the occasions that these two transitions differed by more than 30%, the sample was re-analyzed monitoring the 499>130 transition. This third transition showed quantitative agreement with the reported data from the 499>99 determination. Statem ent of Data Quality BACK TO MAIN It is not possible to verify true recovery of endogenous analyte from tissues withe radiolabeled reference material. The only measurement of accuracy available at this time, matrix spike studies, indicate that the data can be reported with a stated accuracy of either +/-30%, +/-50%, or qualitative only. Please see the values in the attached summary table regarding data quality. References ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Liver for Analysis using HPLC-Electrospray/Mass Spectrometry". 3M Environmental Laboratory, St. Paul, MN. ETS-8-7.0, "Analysis of Potassium Perfluorooctanesulfonate or other Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" . 3M Environmental Laboratory, St. Paul, MN. Kris Hansen, PAI Date Page 6 of 9 ETS-8-7.0 Excel 97 BACK TO MAIN FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab FACT-GEN-055 Study; Product Number(Test Substance); Matrix; Method/Revision: Analytical Equipment System Number; Instrument Software/Version: Date o f Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reduct ioivAnalyst: Sam ple D ata GEN055, Decatur - Tennesee River Samples received from Entrix NA Wholebody Fish Homogenates ETS-8-6.0 & ETS-8-7.0 Amelia 062498 Mass lynx 3.4 04/10/01 RWW/OK 04/16/01,05/01/01,05/02/01 MMH 04/18/01,05/03/01,05/04/01 MMH Box# 01-025 Fish W holebody G ronp Bose Sample # American Shad Shad-1 (04/24/01) Shad-2 FT Shad-3 FT Shad-4 Shad-5 FT Shad-6 Shad-7 (04/24/01) Shad-8 Catfish Cat-l (04/24/01) Cat-2 Cat-3 (04/24/01) Cat-4 GAR GAR-1 (04/24/01) GAR-2 GAR-3 GAR-4 GAR-5 GAR-6 (04/24/01) Large-m outh Bass LM-1 (04/24/01) LM-2 (04/24/01) Striped Bass SB-1 FT (04/24/01) SB-2 (04/24/01) SB-3 FT SB-4 SB-5 FT SB-6 SB-7 FT SB-8 SB-9 FT SB-10 FT SB-11 FT S B -12 SB-13 FT S B -14 W hite Perch WP-1 WP-2 FT (04/24/01) X Sample contained this analyte. LAC 05/16/01 qualitative only t/-50% +/-30% C o n cen tratio n of PFOS ng/g or % Ree X X X X X X X X <LOQ (0 00750 ug/g) < I jO Q (0.00750 ug/g) 1.31 1.07 0.0379 <LOQ (0.00750 ug/g) <LOQ <0 00750 ug/g) <LOQ (0 00750 ug/g) 0.676 3.05 0.230 X 0.0421 0.0315 0.0934 0.0601 0.0865 0.0561 0.0764 0.0368 0.0709 0.0580 0.112 0.0451 0.0834 0.0666 X X C o ncentration Concentration of PFOSA of PFOA ug/g or * / Ree ug/g o r % Ree X : 0.0691 : X : : : 0.0692 . X 0.0286 X <LOQ (0.0201 lig/g) X <LOQ (0.0201 ug/g) X <LO Q (0.0201 ug/g) : XX X 0.151 < LOQ (0.00750 Ug/g) <LOQ (0.0201 ug/g) < LOQ (0.00750 ug/g) <LOQ (0.0201 ug/g) X 0.219 X <LOQ (0.0201 ug/g) < LOQ (0.00750 ug/g) <LOQ (0.0201 ug/g) < LOQ <0.00750 ug/g) <LOQ (0.020! ug/g) < IjOQ <0.00750 ug/g) <LOQ (0.0201 ug/g) < LOQ (0.00750 ug/g) <LOQ (0.0201 ug/g) X 0.126 X 0.183 0.00876 < LOQ (0.00803 ug/g) 0.557 <LOQ (0.0201 ug/g) 0.0101 < LOQ (0.00803 ug/g) 0.0109 < LOQ (0.00803 ug/g) 0.00962 < LOQ (0.00803 ug/g) 0.0105 < LOQ (0.00803 ug/g) 0.00806 < LOQ (0.00803 ug/g) 0.0101 < LOQ (0.00803 ug/g) 0.0104 < lO Q (0.00803 ug/g) 0.0104 < LOQ (0 00803 ug/g) 0.0102 < LOQ (0.00803 ug/g) 0.0101 < LOQ (0 00803 ug/g) 0.0119 < LOQ (0.00803 ug/g) 0.0101 < LOQ (0.00803 ug/g) 0.0121 0.0106 < LOQ (0.00803 ug/g) < LOQ (0.00803 ug/g) X 0.229 X 0.327 P rO S = Perfluorooctanesulfonate PFO SA = Perfluorooc tanesulfonamide PFO A = Perfluoroocianoate PFH S = Peril uorohe xane sulfonate C oncentration of PFHS ng/g or % Ree <LOQ (0.00750 ug/g) < !O Q (0.00750 ug/g) 0.0697 0 .0 0 7 3 <LOQ (0.00750 ug/g) 0.0262 0.178 <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) 0.0153 <LOQ (0.00750 ug/g) <LOQ (.O7SO ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) 0.0178 .. .. 053I : ^ L <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LQQ (000750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LO Q (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LCKJ: (0.00750 tig/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g) <LQQ (0.00750 ug/g) 0.314 0.497 Date Entered/Analyst: Date Verified/Analyst: 05/03/01,05/04/01 KJR, 5/10/01 MMH/LAC 5/14/01 mmh 0 Results GEN-055~lrver-elec l 5/22/01 7:29 AM Page 7 of 9 FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab FACT-GEN-055 Sam ple Avg. PFO S Stated accuracy o f recoveries PFO S data Avg. PFO SA Stated accuracy recoveries o f PFO SA data Avg. PFO A recoveries Stated accuracy o f P F O A data Avg. PFH S Stated accuracy recoveries o f PFH S data American Shad Catfish Gar Large M outh Bass Striped Bass W hite Perch L ow Spike H igh Spike L ow Spike H igh Spike L ow Spike H igh Spike L ow Spike H igh Spike L ow Spike H igh Spike L ow Spike H igh Spike SE SE 97% 98% 112% 87% 92% SE 97% 74% NA 0% qualitative only qualitative only + /-3 0 % + /-3 0 % +/-5G % + /-5 0 % +/~50% qualitative only + /-5 0 % + /-3 0 % NA qualitative only X X * * * X * *x 34% 0% 39% 60% 0% 43% 55% 64% 61% 58% NA 28% qualitative only X qualitative only X qualitative only X qualitative only *x qualitative only X qualitative only X + /-5 0 % + /-5 0 % + /-5 0 % + /-5 0 % qualitative only qualitative only X 94% 84% 63% 4^ 21% 71% 89% 88% 69% 80% NA 81% SE = spiking error; spike prepared at inappropriate level, (<30% of endogenous) * high degree o f variability in spike recoeries X = qualitative data only; spike recovery studies do not support the accuracy o f the data to +/-50%. NA = Not applicable PFOS = Perfluorooctanesulfonate PFOSA = Perfluorooctanesulfonamide PFOA = Perfluorooctanoate PFHS = Perfluorohexanesulfonate + /-3 0 % X 113% qualitative only *X 113% +/*50% 107% + /-3 0 % 99% qualitative only X 67% ^0% 68% +/-3Q % 105% ^ /-3 0 /< # a 84% ^ -5 0 % 93% + /-3 0 % ^ . 93% qualitative only * NA + /-3 0 % 77% + /-3 0 % + /-3 0 % + /-3 0 % + /-3 0 % qualitative only X + /-5 0 % + /-3 0 % + /-3 0 % + /-3 0 % + /-3 0 % qualitative only + /-3 0 % ETS-8-7.0 Excel 97 QC Summary GEN-055-liver-elecl 5/22/01 7:29 AM 1 Page 8 of 9 FACT-GEN-055 E00-0520 Analytical Data Summary: 3M Environmental Lab FACT-GEN-055 Comparison of Matrix Spikes in PFOS GEN051 GEN055 Fish Entrix 3M American Shad 0% SE low spike SE A higA ke Catfish 24% 97% low spike W 98% high spike Gar 9% 112% low spike* 87% high spike* Large Mouth Bass 54% 92% low spike* SE high spike Striped Bass 8% 97% low spike* 74% high spike White Perch 104% NA low spike 0% high spike* SE = spiking error; spike prepared at inappropriate level, (<30% of endogenous) * high degree o f variability in spike recoeries NA = Not applicable ETS-8-7.0 and Entrix Method Excel 97 GEN055 GEN051 Comparison GEN-055-liver-elecl 5/22/01 7:29 AM Page 9 of 9