Rapid quantification of tiopronin in human plasma by high-performance liquid chromatography-mass spectrometry / 中国药科大学学报

ABSTRACT

Aim:To develop a rapid high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for the quantification of tiopronin in human plasma.Methods:Cysteine was chosen as antioxidant and firstly added into the whole blood firstly.After adding mycophenolic acid as internal standard (IS) and 1 mol/L HCl into the plasma,the samples were extracted with acetic ether and then determined by HPLC-MS.The chromatographic separation was performed on a Shim-pack VP-ODS C18 column (250 mm×2.0 mm,5 μm) using methanol-0.1% acetic acid (70∶30) as mobile phase with a flow rate of 0.2 mL/min.Results:The assay exhibited a linear range of tiopronin between 30～4 000 ng/mL.The precisions for intra- and inter-batch were all within 8.5%.The extraction recoveries were more than 70%.The total HPLC-MS analysis time was within 7.5 min per a run.The fully validated method was successfully applied to quantify tiopronin in human plasma for a bioavailability study.Conclusion:The assay proved to be accurate,sensitive,selective and convenient.The fully validated method can be applied to study the pharmacokinetics and bioavailability of tiopronin and tiopronin formulations in human.