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目的建立测定人血浆中克拉霉素浓度的RP-HPLC法。方法碱化后的血浆样品用正己烷-二氯甲烷-异丙醇(65:32:3)萃取、吹干富集,进样前用正己烷反萃取。采用Kromasil C18色谱柱(150 mm×4.6 mm,5μm),流动相为乙腈-50 mmol.L-1磷酸二氢铵(氨水调pH6.9)(46:54),流速1.0 m.lmin-1,柱温40℃,检测波长210 nm。结果克拉霉素血药浓度在0.156-4.98μg.ml-1范围内线性关系良好(r=0.9997),最低检测限为0.019μg.ml-1,日内RSD<2.0%,日间RSD<2.5%,平均提取回收率82.0%,平均方法回收率100.0%。结论所建方法快速、准确,适用于克拉霉素的临床药动学研究。
Objective To establish a RP-HPLC method for the determination of clarithromycin in human plasma. Methods Alkalized plasma samples were extracted with n-hexane-methylene chloride-isopropanol (65: 32: 3), dried and enriched. The sample was back-extracted with n-hexane before injection. Kromasil C18 column (150 mm × 4.6 mm, 5 μm) was used as the mobile phase. The mobile phase was acetonitrile-50 mmol·L -1 ammonium dihydrogen phosphate (ammonia water, pH6.9) (46:54), the flow rate was 1.0 m · lmin -1 , Column temperature 40 ℃, detection wavelength 210 nm. Results The linear range was 0.156-4.98μg.ml-1 (r = 0.9997), the lowest limit of detection was 0.019μg.ml-1, the intra-day RSD was less than 2.0% and the intraday RSD was less than 2.5% , The average extraction recovery rate of 82.0%, the average recovery rate of 100.0%. Conclusion The method is rapid and accurate and is suitable for the clinical pharmacokinetic study of clarithromycin.