FIGURE 8.
Selective reaction monitoring of M-45 generated by activated human neutrophils. Neutrophils (5 × 106/ml) in HBSS buffer (pH 7.4) were stimulated with 200 nm PMA (Complete) in the presence or absence of catalase (50 μg/ml), methionine (2 mm), or sodium azide (2 mm). The incubation was performed at 37 °C for 1 h. Supernatant was collected, and macromolecules were removed with a 3,000 Da cutoff spin filter. The filtered supernatant was analyzed by LC-MS/MS selective reaction monitoring of M-45 (m/z 568.1 → 550.1).