柱前衍生-高效液相色谱荧光法测定大鼠血浆中硫普罗宁

建立了测定大鼠血浆中硫普罗宁含量的柱前衍生-高效液相色谱荧光法。血浆样品经二硫苏糖醇(DTT)还原后，再经N-(1-芘)马来酰亚胺(NPM)衍生化后进样测定。以0.2%乙酸(含0.015 mol·L^-1磷酸二氢钾)-乙腈(56∶44)为流动相，采用Kromasil C₁₈色谱柱分离，在λ_ex=340 nm，λ_em=375 nm下荧光检测。硫普罗宁线性范围为0.1～10.0 μg·mL^-1，定量下限为0.1 mg·L^-1，日内、日间精密度(RSD)分别为5.3%～10.8%和7.0%～10.8%，提取回收率在73.7%～79.7%。大鼠单剂量给予硫普罗宁后，药代动力学行为符合二室模型。该法准确可靠，专属性强，适用于硫普罗宁的药代动力学研究。

Determination of tiopronin in rat plasma by HPLC following fluorescent derivatization

A sensitive, rapid method for determining reduced tiopronin concentration in rat plasma has been developed by using a high-performance liquid chromatography (HPLC) technique in conjunction with the derivatizing agent N-(1-pyrenyl) maleimide (NPM). The analytes were separated on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) using 0.2% glacial acetic acid aqueous solution including 0.015 mol x L(-1) KH2PO4 and acetonitrile (56:44) as a mobile phase at a flow-rate of 0.8 mL x min(-1), and fluorescence detection wavelength were set at lamda(e x) = 340 nm and lamda(e m) = 375 nm, the column temperature was 30 degrees C. The calibration curve was found to be linear over a range of 0.1 - 10.0 microg x mL(-1), the limit of quantitation was 0. 1 mg x L(-1). The coefficients of the variation for the within-run and between-run precisions ranged from 5.3% to 10.8% and 7.0% to 10.8%, respectively. The percentage of absolute recovery ranged from 73.7% to 79.7%. The method was used to determine the concentration of tiopronin in rat plasma after a single intragastric administration of 25 mg x kg(-1) tiopronin to 6 healthy male Wistar rats. The pharmacokinetic process was fitted to a two-compartment model. The method has been successfully applied to the determination of tiopronin in rat plasma.