Abstract
We described the first results of a quantitative ultra performance liquid chromatography–tandem mass spectrometry method for a novel antimicrobial peptide (phylloseptin, PSN-1). Chromatographic separation was accomplished on a Waters bridged ethyl hybrid (BEH) C18 (50mm×2.1mm, 1.7μm) column with acetonitrile–water (25:75, v/v) as isocratic mobile phase. Mass spectrometry detection was performed in the positive electrospray ionization mode and by monitoring of the transitions at m/z 679.6/120, 509.6/120 (PSN-1) and m/z 340.7/165 (Thymopentin, IS). Protein precipitation was investigated and the recovery was satisfactory (above 82%). The method was shown to be reproducible and reliable with intra-day precision below 5.3%, inter-day precision below 14.2%, and linear range from 0.02 to 2μg/mL with r>0.994. The method was successfully applied to a pharmacokinetic study of PSN-1 in rats after intravenous administration.
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