Supporting Information for:

Transcription

1 Supporting Information for: In-situ accumulation of HBCD, PBDEs and several alternative flame-retardants in the bivalve (Corbicula fluminea) and gastropod (Elimia proxima) Mark J. La Guardia * 1, Robert C. Hale 1, Ellen Harvey 1, T. Matteson Mainor 1 and Serena Ciparis 2 1 Department of Environmental & Aquatic Animal Health Virginia Institute of Marine Science College of William & Mary, Gloucester Point, Virginia Department of Entomology Virginia Polytechnic Institute and State University Blacksburg, Virginia, Contents: Quality Assurance - page S2 Instrument Methodology - page S3-S4 Table S1., Matrix (NaSO 4 and Crassostrea virginica) spiking results for GC-MS/SIM and UPLC-MS/MS analysis. Table S2., Selected BFR organic carbon-water (K oc ) and log transformed K oc and n-octanol-water (K ow ) partition coefficients. Figure S1., Log biota-sediment accumulation factors (log BSAF) for Corbicula fluminea and Elimia proxima indicating flame-retardant accumulation rates between species are not statistically difference (P = 0.092). Figure S2., Log bioaccumulation factors (log BAF w ) for Corbicula fluminea and Elimia proxima indicating flame-retardant accumulation rates between species are not statistically difference (P = 0.182). - page S5 - page S6 - page S7 - page S7 S1

2 Quality Assurance The extraction and analytical method used for the identification of PBDEs, TBB, TBPH, BTBPE, DBDPE and α-, β-, γ-hbcd used here has been previously reported for the analysis of these flame-retardants in sewage sludge (La Guardia et al., 2010 ES&T). To ensure that this method will also provide quality data when analyzing these flameretardants in diverse matrixes (i.e. sediments and mollusks), the flame-retardants of interest were included in a matrix spiking experiment of sodium sulfate (NaSO 4 ) and oyster tissue (Crassostrea virginica). Oysters were cleaned, shucked and homogenized prior to freeze-drying. Once dried, seven oyster aliquots (5-g dry weight (dw), each) were spiked with 15 PBDEs ranging from tri- to deca-brominated diphenyl ethers (listed in Table S1), TBB, TBPH, BTBPE, DBDPE and α-, β-, and γ-hbcd. Seven sodium sulfate (NaSO 4 ) samples (30-g dry-weight, each) were also prepared and spiked. All samples including a method blank (un-spiked NaSO 4 ) were extracted and analyzed by the method outlined in the manuscript. Results for the spiking experiment were calculated in the same matter as the samples in the manuscript. All spiking results were surrogate recovery corrected. Recoveries for the surrogate BDE-166 ranged from 94.2 to 110%, 13 C-α-HBCD ranged from 93.4 to 102.8% (Table S1). The method blank did not contain any analyte above detection limits (>1 ng g -1, dw). The mean recovery for the NaSO 4 spiked compounds (i.e. PBDEs, TBB, TBPH, BTBPE and DBDPE) analyzed by GC-MS/SIM ranged from 64.7 to 127%, with a %RSD of <10% for each compound. The recoveries for the spiked oysters ranged from 69.4 to 123%, with a %RSD of <10% for 17 of the 19 compounds. The %RSD for BDE-206 and TBPH were 15.1 and 16.6, respectively. The mean recovery for α-, β-, and γ-hbcd spiked NaSO 4 ranged from 91.8 to 97.2%. From spiked oysters recoveries of compounds ranged from 86.8 to 101.9%, with <10% RSDs for HBCDs in both matrixes. (Spiking recovery results, mean (n=7) and %RSD for each of the flame-retardants are listed by matrix in Table S1.) S2

3 Instrument Methodology GC-MS/SIM: Purified extracts were analyzed by GC (6890N, Agilent Tech., Palo Alto, CA) with MS detection (JMS-GC Mate II, JEOL, Peabody, MA.) Sample aliquots (1 µl) were introduced into the split/splitless injector, equipped with a glass liner (1 mm, ID), and separated on a 15-m DB-5HT (0.25 mm i.d., 0.1 µm, J&W Scientific, Agilent Tech.) column. The injector temperature was 300 o C and initial carrier gas (helium) head pressure was 50 psi. Four minutes after sample injection the split vent was opened and pressure reduced to 15.2 psi (flow 1.2 ml/min.). Column flow rate was kept constant (1.2 ml/min, temperature compensated) throughout the remaining portion of the analysis. Initial column oven temperature was 90 o C, held for 4 minutes, then increased to 150 o C at 30 o C/minute, then 10 o C/minute to 300 o C, and held for 20 minutes. It was then increased to 350 o C at 30 o C/minute and held at 350 o C for 5 minutes. Total run time was 47.7 minutes. The MS was operated in the electron capture negative ionization (ECNI) mode (methane reagent gas (99.99%)) using SIM. Ion source temperature was 200 o C and the GC transfer line was maintained at 300 o C. Quantitation ions for PBDEs, TBB, TBPH, BTBPE and DBDPE were m/z 79 ([ 79 Br] - ), 81([ 81 Br] - ), confirmation ions m/z 356, 463, 251 were monitored for TBB, TBPH, BTBPE, respectively and 486 and 488 m/z for decabromodiphenyl ether (BDE-209). A five-point calibration curve, r 2 > 0.995, was constructed from the analysis of calibration standards. PBDE standards were purchased from Wellington Laboratories, Inc., Ontario, Canada. TBB, TBPH and BTBPE were obtained from AccuStandards, New Haven, CT. DBDPE was purchased from TCI America, Portland, OR. UPLC-MS/MS: For ΣHBCD (α-, β-, and γ-hbcd) analysis purified extracts were analyzed using an Acquity UPLC (Waters, Milford, MA. USA) equipped with a Waters Acquity BEH (bridged ethyl hybrid) C18 column (150 mm, 2.1 mm i.d., 1.7 µm particle diameter), coupled with a 3200 Q-TRAP MS2 (Applied Biosystems/MDS Sciex; Toronto, Canada) triple quadrupole mass spectrometer. The UPLC mobile phase consisted of 75:25 water/methanol (A1) and 50:50 methanol/acetonitrile (B1). The initial S3

4 mobile phase composition was 80:20 A1/B1 at a flow rate of 200µL/minute. This was followed by a linear gradient to 100% B1 over the first 4-minutes of the run, and then held at 100% B1 for 6 minutes. Column regeneration was done using a 1-minute linear gradient back to 80:20 A1/B1, followed by a 5 minute hold at those conditions. The MS was operated in the electrospray ionization (ESI) negative ion mode, with the following parameters: curtain gas 30 psi (N 2 ), probe temperature 450 o C, nebulizer gas 62 psi (Zero air), auxiliary gas 20 psi (Zero air), interface heater on, collision gas medium (N 2 ), ion spray 1500 V, declustering potential 30 V, entrance potential 3.0 V, collision energy 38 V, and collision exit potential 1.0 V. For MS/MS acquisition, Q1 and Q3 were operated with unit resolution with a scan time of 1 s. Native isomers (α-, β-, and γ- HBCD) were identified by monitoring ion pairs produced from quadrupole scans (Q1/Q3): 640.9/78.9 and 640.9/80.9. For 13 C-α-HBCD ion pairs 652.1/78.9 and 652.1/80.9 and for d 18 - α-hbcd, 657.7/78.9 and 657.7/80.9 were monitored for identification. The quantitation ion was 78.9 m/z. A five-point calibration curve (r 2 > 0.995) for each compound was constructed from calibration standards purchased from AccuStandards, New Haven, CT. Reference: La Guardia, M. J.; Hale, R. C.; Harvey, E.; Chen, D. Flame-retardants and other organohalogens detected in sewage sludge by electron capture negative ion mass spectrometry. Environ. Sci. Technol. 2010, 44 (12): S4

5 Table S1., Matrix (NaSO 4 and Crassostrea virginica) spiking results for GC-MS/SIM and UPLC-MS/MS analysis. NaSO 4 Oyster (Crassostrea virginica) analytes spiked conc. ng/g, dry weight mean (n=7) % RSD* spiked conc. ng/g, dry weight mean (n=7) % RSD* GC-MS/SIM BDE % 3.89% % 8.36% BDE % 1.98% % 2.42% BDE % 2.34% % 3.14% BDE % 1.81% % 3.02% BDE % 3.11% % 4.03% BDE % 3.14% % 1.84% BDE % 1.07% % 7.90% BDE % 3.80% % 6.54% BDE % 3.81% % 6.56% BDE % 6.50% % 0.93% BDE % 4.79% % 4.84% BDE % 5.11% % 3.38% BDE % 5.52% % 4.17% BDE % 3.35% % 15.1% BDE % 9.45% % 9.96% BTBPE % 4.04% % 5.79% DBDPE % 3.58% % 2.40% TBB % 0.79% % 6.29% TBPH % 8.32% % 16.6% Surrogate, % rec. (BDE-166) % 3.61% % 3.33% UPLC-MS/MS α-hbcd % 2.86% % 3.06% β-hbcd % 4.08% % 2.21% γ-hbcd % 1.01% % 5.55% Surrogate, % rec. (C 13 -α-hbcd) % 4.04% % 2.84% *%RSD, % Relative standard deviation S5

6 Table S2., Selected BFR organic carbon-water (K oc ) and log transformed K oc and n-octanol-water (K ow ) partition coefficients. analyte K oc log K oc log K ow BDE-47 2,041,738 d 6.31 d 6.81 a (measured) BDE-85 3,090,295 d 6.49 d 7.37 a (measured) BDE-99 4,365,158 d 6.48 d 7.32 a (measured) BDE-100 3,019,952 d 6.45 d 7.24 a (measured) BDE-153 4,570,882 d 6.66 d 7.9 a (measured) BDE-154 4,365,158 d 6.64 d 7.82 a (measured) BDE-183 6,025,596 d 6.78 d 8.27 a (measured) BDE ,183,829 d 7.45 d b BDE ,183,829 d 7.45 d b BDE ,183,829 d 7.45 d b BDE ,183,829 d 7.45 d b BDE-203 8,317,638 d 6.92 d 8.71 b (measured) BDE ,703,180 d 7.73 d b BDE ,703,180 d 7.73 d b BDE ,703,180 d 7.73 d b BDE ,712,855 d 8.02 d b TBB 501,187 b 5.70 b 8.75 b TBPH 25,118,864 b 7.40 b b BTBPE 1,258,925 b 6.10 b 9.15 b α-hbcd 25,119 e 4.40 e 5.07 c β-hbcd 27,542 e 4.44 e 5.12 c γ-hbcd 56,234 e 4.75 e 5.47 c a Braekevelt, E.; Tittlemier, S. A.; Tomy, G. T. Direct measurement of octanol-water partition coefficients of some environmentally relevant brominated diphenyl ether congeners. Chemosphere 2003, 51, b EPISuite 4.1, U.S. EPA Office of Pollution Toxics and Syracuse Research Corporation (SRC), c Wu, J-P.; Guan, Y-T.; Zhang, Y.; Luo, X-J.; Zhi, H.; Chen, S-J.; Mai, B-X. Several current-use, non- PBDE brominated flame-retardants are highly bioaccumulative: Evidence from field determined bioaccumulation factors. Environ. Int. 2011, 37, d Chen, M-Y.; Yu, M.; Luo, X-J.; Chen, S-J.; Mai, B-X. The factors controlling the partitioning of polybrominated diphenyl ethers and polychlorinated biphenyls in the water-column of the Pearl River Estuary in South China. Marine Pollu. Bulletin 2011, 37, (equ. logk oc = logK ow ) e EPISuite 4.1, U.S. EPA Office of Pollution Toxics and Syracuse Research Corporation (SRC), (equ. log K oc = log K ow ) S6

7 ) ia im x r o p a lim (E F S A -1 B g -1.5 lo -2 y = x R² = logbasf (Corbicula fluminea) Figure S1., Log biota-sediment accumulation factors (log BSAF) for Corbicula fluminea and Elimia proxima indicating flame-retardant accumulation rates between species are not statistically difference (P = 0.092) a ) im x 7 r o p 6.5 ia 6 lim (E 5.5 w F A 5 B g lo y = x R² = logbafw (Corbicula fluminea) Figure S2., Log bioaccumulation factors (log BAF w ) for Corbicula fluminea and Elimia proxima indicating flame-retardant accumulation rates between species are not statistically difference (P = 0.182). S7