柱前衍生化高效液相色谱-质谱联用法测定人体内血浆中的硫普罗宁

Determination of tiopronin in human plasma by HPLC-MS with pre-column derivatization

目的：建立高效液相色谱-质谱联用法测定人血浆中硫普罗宁的浓度。方法：色谱条件——色谱柱为Waters SunFire^(TM)C_(18)(2.1 mm×20 mm,5μm),流动相为甲醇-0.05%冰乙酸水溶液(50∶50,V/V),流速为0.3 mL·min^(-1),柱温为室温；质谱条件——电喷雾离子化法(ESI)采集负离子,单离子方式检测硫普罗宁衍生化产物m/z 327.95和内标葛根黄豆苷元m/z 253.00。结果：方法的定量下限为0.1 mg·L^(-1),线性范围为0.1～4.0 mg·L^(-1)(r=0.996 2),提取回收率均大于(66±s 5)%,日内、日间精密度(RSD)均小于9.4%。结论：该法灵敏、专属,适用于硫普罗宁人体血浆药物浓度的测定。

AIM： To develop a high pressure liquid chromatography-mass spectrometry method for determination of tiopronin concentration in human plasma. METHODS： Chromatographic column was Waters SunFire^TM C18（2.1 mm× 20 mm,5 μm）. The mobile phase consisted of methanol - 0.05 % acetic acid（50 ： 50, V/V） with a flow rate of 0.3 mL·min^-1. MS detection in single ion monitoring mode （SIM） using electrospray ionization （ESI） as an interface was applied to determine negative ions at m/z 327.95 for TP-DNCB and m/z 253.00 for internal standard（IS,daidzein）. RESULTS： High specificity and a low quatitation limit （0.1 mg·L^-1） were achieved. Linearity was confirmed in the concentration range of 0.1- 4.0 mg·L^-1 with the correlation coefficient （r = 0.996 2）. The RSD values of intra-day and inter-day assay were less than 9.4 %. The extraction recovery rate of tiopronin was more than （66 ± s 5） %. CONCLUSION： HPLC-MS method is specific and applicable to the determination of tiopronin in human plasma.