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目的建立同时测定大鼠血浆中呱氨托美丁(MED-15)及其代谢产物MED-5与托美丁浓度的HPLC法。方法血浆样品用乙腈沉淀蛋白,采用HypersilODS C_(18)柱(250mm×4.6mm,5μm)进行分离,以甲醇-2%冰醋酸(60:40,V/V)为流动相,流速1.0mL·min~(-1);紫外检测波长313 nm。结果大鼠血浆中MED-15、MED-5和托美丁线性范围分别是0.050～5.6、0.042～5.4和0.039～5 mg·L~(-1)(r分别为0.995 1、0.995 7、0.999 0);定量下限分别为0.050、0.042和0.039mg·L~(-1);平均相对回收率分别是100.3%、99.4%和99.9%,平均绝对回收率分别88.9%、87.8%和88.7%;日内、日间精密度均<12%。血浆样品的稳定性符合要求。结论本研究所建立的大鼠血浆中MED-15、MED-5和托美丁浓度测定的HPLC方法灵敏、准确、精密、稳定,适用于大鼠体内MED-15的药动学研究。
OBJECTIVE To establish a HPLC method for simultaneous determination of the concentrations of MED-15 and its metabolites MED-5 and Tropimidine in rat plasma. Methods The plasma samples were precipitated with acetonitrile and separated on a Hypersil ODS C 18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-2% glacial acetic acid (60:40, V / V) min ~ (-1); UV detection wavelength 313 nm. Results The linear ranges of MED-15, MED-5 and tolmetin in rats were 0.050-5.6, 0.042-5.4 and 0.039-5 mg · L -1, respectively (r = 0.995, The average relative recoveries were 100.3%, 99.4% and 99.9%, respectively. The average absolute recoveries were 88.9%, 87.8% and 88.7%, respectively. The lower limit of quantitation was 0.050,0.042 and 0.039 mg · L -1, respectively. Days, day precision <12%. The stability of plasma samples meets the requirements. Conclusion The established HPLC method for the determination of MED-15, MED-5 and tolmetin in rat plasma is sensitive, accurate, precise and stable and suitable for the study of pharmacokinetics of MED-15 in rats.