High-performance liquid chromatography-electronspray ionization mass spectrometry for determination of tiopronin in human plasma.

A simple and sensitive HPLC/ESI-MS method for the determination of tiopronin in human plasma was described. Vitamin C and 2-mercaptoethanol (2-Me) were used as the reducer and the stabilitizer to release and stabilify tiopronin from a dimmer and mix forms with endogenous thiols in the treatment of plasma samples. The analytes were separated on a Johnson Spherigel analytical column packed with 5 microm C8 silica, using the formic acid aqueous solution (pH 4.5) including tris(hydroxymethyl) aminomethane (Tris) and 2-Me (0.5 and 1 mM, respectively) as a mobile phase. Cyclamate was used as the internal standard (I.S.) for the quantification of tiopronin. The correlation coefficient of the calibration curve were better than 0.998 in the range of 0.107-5.35 microg/ml in human plasma. The limit of quantification (LOQ) was 0.107 microg/ml (S/N 10:1, RSD 7.1%). The inter-day and intra-day accuracy was below 7.1 and 6.8%, respectively. As a preliminary application, this method has been successfully applied to the determination of tiopronin in the human plasma.