餐时血糖调节剂米格列奈钙的RP-HPLC法人肝微粒体药物浓度测定

Determination of mitiglinide in human hepatic microsomes by RP-HPLC

目的：建立人肝微粒体中米格列奈的高效液相色谱测定法。方法：采用Diamonsil—C18色谱柱（200mm×4．6mm，5μm）；流动相为乙腈-0．02mol·L^-1KH2PO4缓冲液（pH4．0）（43：57）；流速为1．0mL·min^-1；紫外检测波长为210nm；进样量为20mL。结果：米格列奈最低检测限为2μmol·L^-1；线性范围为5～1000μmol·L^-1（r=0．9999）；最低定量浓度为5μmol·L^-1（RSD〈5％，n=5）；低、中、高3种浓度日内、日间RSD（n=5）分别为1．9％，1．6％，1．4％和3．8％，4．0％，3．8％；绝对回收率平均为94．4％，相对回收率平均为101．2％。结论：本实验建立的人肝微粒体中米格列奈的高效液相测定方法操作简便、结果准确。

Objective：To establish a HPLC method to determine mitiglinide in human hepatic microsomes. Methods ： Mitiglinide was separated on a Diamonsil-C18 （200 mm × 4.6 mm,5 μm） column. The mobile phase consisted of acetonitrile-0.02 mol·L^-1 KH2PO4（adjusted to pH 4 with phosphoric acid） （43： 57） at a flow rate of 1 mL·min^-1 UV detection was set at 210 nm. Results：The regression was linear in the concentration range of 5 - 1 000 μmol·L^-1 （ r = 0. 999 9）. The limit of detection was 2 μmol·L^-1 and the lowest limit of quantification was 5 μmol·L^-1 Intra-day RSDs of mitiglinide in microsomal sample at three concentrations were 1.9% , 1.6% , 1.4% ; and inter-day RSDs were 3.8% ,4.0% ,3.8% （n = 5 ）. The mean absolute recovery was 94.4% and the mean relative recovery was 101. 2%. Conclusion： The HPLC assay was simple and accurate for the determination of mitiglinidc in human hepatic microsomes.