FIGURE 8.
Detection of oxoDHA-BME adducts and 14-HDoHE in BAL cells. Bronchoalveolar lavage cell pellets were incubated with 500 mm BME and extracted for LC-MS analysis. The loss of BME (78 atomic mass units) was monitored for oxoDHA-BME adducts (419.2 → 341.2) (A). Endogenous levels of 14-HDoHE (343.3 → 205.2) were detected in BAL cell pellets (B). Endogenous 14-HDoHE levels ranged from not detectable to 0.95 ng/106 cells in the clinical samples (C). Primary alveolar macrophages taken from bronchoscopies were plated at a confluency of 5 × 105 cells/well in a 24 well plate. Alveolar macrophages were incubated for 4 h with 2 μm 14-HDoHE ± 50 μm CAY10397, and 14-oxoDHA (D) and 14-oxoDPA (E) metabolites were quantified. **, p ≤ 0.01; ***, p ≤ 0.001.