HPLCESIMS测定米格列奈钙的血浓度及其药动学研究.docVIP

HPLC-ESI-MS测定米格列奈钙的血药浓度及其药动学研究 摘要：目的建立人血浆中米格列奈钙片HPLC-MS测定法，研究米格列奈钙片在健康志愿者的药物动力学。方法： 以那格列奈为内标，血浆样品过Strata-X固相萃取柱，用60 ％的乙腈洗脱，20 uL注入Thermo C18色谱柱，柱温40 ℃。流动相组成为乙腈：0.1 ％甲酸溶液(60：40，V/V)，流速：1 mL/min，,质谱采用电喷雾电离源正源（ESI+），定量分析采用选择性离子监测（SIR）。30名健康志愿者（男女各半）随机分为三组，每组10人，分别单剂量口服米格列奈钙片5 mg（低剂量组）、10 mg（中剂量组）、20 mg（高剂量组），采集给药前和给药后血样,结果米格列奈在10-6054 ng/mL内线性关系良好，R=0.998,萃取回收率均％，方法回收率100.95 ％～103.88 ％，日内、日间RSD11 ％口服三种剂量所得MRTt1/2、Tmax相近体内吸收程度（Cmax、AUC0-8 ）与给药剂量有较好的线性关系结论HPLC-MS方法简单，准确度，可用于米格列奈钙片在人体内的药物动力学研究。关键词：米格列奈；；药物动力学: OBJECTIVE: To establish a high performance liquid chromatographic coupled with electrospray ionization mass spectrometry (HPLC- MS/ESI) method for the determination of Mitiglinide in human plasma; and apply to pharmacokinetic study. METHOD: Nateglinide was used as internal standard. The plasma samples were extracted by Strata-X solid-phase extraction. 60 ％ acetonitrile eluting. The fractions were injected into evaporated Thermo C18 column(250mm×4.6mm，5 μm), with the column temperature at 40 ℃. The mobile phase was composed of acetonitrile and 0.1 % formic acid (60:40, v/v), with flow rate at 1.0mL/min. Split ratio was 1∶4.The compound was ionized in the turbo electrospray ionization (ESI) ion source of the mass spectrometer and was detected in the selected ion recording (SIR) mode. 30 healthy volunteers (15 male and 15 female) were randomly divided into three groups, each group was administrated 5 mg, 10 mg, 20 mg, respectively, the blood samples was colleted before and after administrate mitiglinide and was frozen at –70 ℃ until analysis. RESUETS: The calibration curve was linear in the 10-6054ng/mL range for mitiglinide, R=0.9982. The extraction recovery was above 88 %. The methodological recovery ranged between 101.0 ％103.9 ％. The intra- and inter-day RSD were less than 11 %. There were no significant differences of MRT, t1/2 and Tmax among three dosages within the studied dose range. The absorption of miglinide data（Cmax、AUC0-8 ） show the linear relationship wit