Rapid and Selective LC鈥揗S鈥揗S Method for the Determination of cyclo-trans-4-l-Hydroxyprolyl-l-serine (J
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A rapid and selective liquid chromatographic/tandem mass spectrometric method for the determination of JBP485 was developed and validated. Following protein precipitation, the analyte and internal standard (JBP923) were separated from human plasma using an isocratic mobile phase on an Elite Kromasil C18 column. An API 3200 tandem mass spectrometer equipped with a Turbo ionSpray ionization source was used as the detector and operated in the positive ion mode. Multiple reaction monitoring using the precursor to product ion combinations of m/z 201.2 → 86.2 and m/z 219.2 → 86.2 was performed to quantify JBP485 and JBP923, respectively. The method was linear in the concentration range of 0.10–50.00 μg mL−1 using 100 μL of plasma. The lower limit of quantification was 0.10 μg mL−1. The intra- and inter-day relative standard deviations over the entire concentration range were less than 6.65%. Accuracy determined at three concentrations (0.25, 4.00 and 25.00 μg mL−1 for JBP485) ranged from −0.78 to 2.74%in terms of relative error. Each plasma sample was chromatographed within 2.0 min. The method was successfully applied to characterize the pharmacokinetic profiles of JBP485 in rats after an intravenous injection of 6.25 mg kg−1 JBP485.

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