The analytical standard mixture of PAH was purchased from Absolute Standards Inc. (cat #60026, Hamden, CT, USA). PAH are a class of bioactive compounds of concern for environmental exposures and are major components of complex petroleum substances; model PAH such as benzo[a]pyrene are often used to characterize the risks of mixtures [41,42]. Of the twenty-five PAH in the commercial mixture, twenty were the substances of interest in this study based on the availability of the quantitative analytical methods (see below), these are listed in Table S1. Analytical internal standards (ISTDs) naphthalene-d8 (cat #48715-U, CAS # 1146-65-2), acenaphthene-d10 (cat #48417, CAS #15067-26-2) and perylene-d12 (cat #48081, CAS #1520-96-3) were purchased from Supelco (Supelco, Bellefonte, PA, USA) and included in all GC-MS/MS analyses. The internal standard assigned to each PAH of interest for relative quantification are listed in Table S2. Dulbecco’s Modified Eagle Medium (DMEM, cat #11965092, ThermoFisher, Waltham, MA, USA), Fetal Bovine Serum (FBS, cat #97068-085, VWR, Radnor, PA, USA) and penicillin/streptomycin mixture (cat #10378016, ThermoFisher, Waltham, MA, USA) were used to prepare MAF and in cell-based experiments. Silicone micro-O-rings (inner diameter 0.056”, outer diameter 0.176”, width 0.06”) were from McMaster Carr (cat #9396K61; Elmhurst, IL, USA). DMSO extracts were prepared with cell culture-grade DMSO (cat #sc-202581B, CAS #67-68-5, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and cyclohexane (cat #C100307, Sigma-Aldrich, St. Louis, MO, USA). HPLC-grade methanol (MeOH, cat #34860), pentane (cat #34956), diethyl ether (Et2O, cat #309966), toluene (cat #650579), hexane (cat #650552) solvents were also purchased from Sigma-Aldrich, St. Louis, MO, USA. In vitro cytotoxicity assays were performed using CellTiter-Glo® 2.0 Luminescent Cell Viability Assay (cat #G9243, Promega, Madison, WI, USA) with EA.hy926 human endothelial cell line (cat #CRL-2922, ATCC, Manassas, VA, USA) and human HepG2 cell line (cat #HB-8065, ATCC, Manassas, VA, USA). Cells were cultured in 96-well black clear-bottom plates (cat #07-200-588; Corning, Kennebunk, ME, USA) and tetraoctylammonium bromide (TAB) was used as a positive control for the in vitro assays (cat #294136, CAS #14866-33-2, Sigma-Aldrich, St. Louis, MO, USA).