Figure 5.

Detection of free 3-NT from nitrated liver mitochondria by HPLC/ECD or HPLC/UV. Isolated liver mitochondria were nitrated by PN (50–500 µM). These samples were subjected to pronase digest before HPLC/ECD analysis (A) or HPLC/UV analysis (C). Representative chromatograms are shown for the HPLC/ECD quantification (B) or the HPLC/UV quantification (D). HPLC/ECD analysis was performed with 20 µL of sample at 27 °C with isocratic elution (1 mL/min, mobile phase: 26.3 mM sodium citrate and 10.9 mM sodium at pH 3.75 with 3.5 v/v% methanol; 3-NT eluted at 4.05 min). Data are presented as mean ± SD of n = 3–5 (A) and 3–5 (C) independent experiments. * indicates p < 0.05 versus Mito untreated control group.