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Supplemental Data

Summary of NOAA SOP CCR-052: Analysis of TEH and PAH in seawater samples using C18-E SPE cartridges

Water samples (50-500mL) were collected from each CEWAFtest chamber, and replicatesamples were composited by treatment. Samples were acidified with 18% hydrochloric acid to a pH of 2 and then transferred into solvent rinsed volumetric cylinders. Extraction volumes were dependent on treatment strength and timepoint. In general, 50 mL was extracted for 100x and 50xCEWAFs and 100 mL was extracted for the 16.7x, 5.56x, 1.85x, 0.617x, 0.2x and control CEWAFs. QA/QC measures for each test included a blank and PAH spike (400ng). Some tests also included a sample duplicate and a TEH spike (5mg). All samples were spiked with PAH and TEH internal standards and mixed thoroughly. There were 18 deuterated PAH internal standards (d8-naphthalene, d10-1-methylnaphthalene, d8-acenaphthylene, d10-acenaphthene, d10-fluorene, d8-dibenzothiophene, d10-phenanthrene, d10-anthracene, d10-fluoranthene, d10-pyrene, d12-benz[a]anthracene, d12-chrysene, d12-benzo[b]fluoranthene, d12-benzo[k]fluoranthene, d12-benzo[e]pyrene, d12-benzo[a]pyrene, d12-perylene and d12-benzo[g,h,i]perylene [Cambridge Isotope Laboratories, Inc. Tweksbury, MA]) and 2 TEH internal standards (d26-dodecane and d42-eicosane [Cambridge Isotope Laboratories, Inc. Tweksbury, MA]).

Samples were extracted using Strata C18-E solid phase extraction (SPE) cartridges (1 g sorbent/6 mL column volume [PhenomenexTorrence, CA]). Prior to extraction cartridges were conditioned at gravity speed with two column volumes of dichloromethane, methanol and pH 2 DI water. Samples were then passed through the SPE cartridges followed by one column volume rinse of DI water. SPE cartridges were frozen for roughly 2-3 months prior to elution.

For elution, frozen SPE cartridges were thawed for 30 minutes and then placed on a vacuum manifold and dried for two hours to remove any residual water on the sorbent bed. Samples were then eluted with one column volume of hexane followed by two column volumes of dichloromethane. The hexane and dichloromethane fractions were combined, solvent exchanged once to hexane and concentrated under a stream of nitrogen in a water bath at 40°C to 200 μL. Extracts were transferred to 2 mL silanized amber auto sampler vials to a final volume of 0.5 mL. All samples were spiked with a recovery standard (d14-p-terphenyl, 50ng) and then analyzed for 50 PAH compounds (parent and alkylated PAHs; Suppl. Table 1) and TEH using gas chromatography mass spectrometry (GC/MS-Agilent 6890/5973N). PAH and TEH analysis was performed using two separate acquisition methods however the target analyte separation for both analyses was achieved on the same DB17-ms 60m x 0.25mm x 0.25μm column. PAH and TEH analyses were performed in selected ion monitoring (SIM) mode. The GC/MS was calibrated with standards ranging from 0.1-5000 ng/mL (PAHs) and 0.1-25 mg/mL (TEH).The TEH calibration curve was developed by diluting LSC oil to the desired calibration points. All calibration curves demonstrated a coefficient of determination value ≥ 0.99.

Supplemental Table 1: List of individual and alkylated PAHs that are included in Total PAH.

Individual and Alkylated PAHs in Total PAH
napthalene / C1-Naphthalenes
biphenyl / C2-Naphthalenes
acenapthene / C3-Naphthalenes
acenapthylene / C4-Naphthalenes
fluorene / C1-Fluorenes
dibenzofuran / C2-Fluorenes
dibenzothiophene / C3-Fluorenes
phenanthrene / C1-Dibenzothiophenes
anthracene / C2-Dibenzothiophenes
fluoranthene / C3-Dibenzothiophenes
pyrene / C4-Dibenzothiophenes
benz(a)anthracene / C1-Phenanthrenes/Anthracenes
benzo(b)naphtho(2,1-d)thiophene / C2-Phenanthrenes/Anthracenes
chrysene + triphenylene / C3-Phenanthrenes/Anthracenes
benzo(a)fluoranthene / C4-Phenanthrenes/Anthracenes
benzo(b)fluoranthene / C1-Fluoranthenes/Pyrenes
benzo(j)fluoranthene / C2-Fluoranthenes/Pyrenes
benzo(k)fluoranthene / C3-Fluoranthenes/Pyrenes
benzo(a)pyrene / C4-Fluoranthenes/Pyrenes
benzo(e)pyrene / C1-Chrysene/Benzanthracene
dibenzo(a,h)anthracene / C2-Chrysene/Benzanthracene
indeno(1,2,3-c,d)pyrene / C3-Chrysene/Benzanthracene
benzo(g,h,i)perylene / C4-Chrysene/Benzanthracene
C1-Naphthobenzothiophenes
C2-Naphthobenzothiophenes
C3-Naphthobenzothiophenes
C4-Naphthobenzothiophenes