反相高效液相色谱法同时测定大鼠血浆中紫杉醇硬脂酸酯及其活性代谢产物紫杉醇的含量 |
投稿时间:2017-12-18 修订日期:2018-02-28 点此下载全文 |
引用本文:程丹,陈建明.反相高效液相色谱法同时测定大鼠血浆中紫杉醇硬脂酸酯及其活性代谢产物紫杉醇的含量[J].药学实践杂志,2018,36(3):224~227,273 |
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中文摘要:目的 建立同时测定大鼠血浆中紫杉醇硬脂酸酯(PTX-SA)及其活性代谢产物紫杉醇(PTX)含量的RP-HPLC分析方法。方法 以地西泮作为内标,采用液-液萃取法,用甲基叔丁基醚萃取血浆中药物。以乙腈-甲醇-水作为流动相进行梯度洗脱,紫外检测波长227 nm,柱温30℃,流速1 ml/min,进样量20 μl。结果 大鼠血浆中PTX-SA和PTX在质量浓度分别为0.5~250和0.05~25 μg/ml的范围内均呈现良好的线性关系,相关系数r分别为0.997 6和0.997 9;两者的最低定量限(LOQ)分别为0.15和0.05 μg/ml;平均提取回收率均大于90%,RSD均小于5%;PTX-SA和PTX的日内与日间精密度均小于5%。结论 该方法简便快捷,准确度、精密度高,可用于药动学研究中PTX-SA和PTX的含量测定。 |
中文关键词:反相高效液相色谱 紫杉醇硬脂酸酯 紫杉醇 含量测定 |
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Simultaneous determination of paclitaxel-stearate (PTX-SA)and its bioactive metabolite paclitaxel (PTX)by RP-HPLC |
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Abstract:Objective To establish a reversed phase high performance liquid chromatography method which couldb be applied to determine the content of paclitaxel stearate(PTX-SA)and its bioactive metabolites paclitaxel(PTX)simultaneously.Methods Diazepam was chosen to be the internal standard and liquid-liquid extraction method was used for the preparation of sample solution with methyl tert-butyl ether. The two compounds were separated on Agilent Eclipse plus C18 column(4.6 mm×250 mm,5 μm) with gradient elution by using mobile phase of acetonitrile(A)-methanol(B)-water(C), the detection wavelength was 227 nm and the column temperature was 30℃. The flow rate was 1 ml/min and the injection volume was 20 μl.Results The standard curve exhibited good linearity within the concentration of PTX-SA from 0.5-250 μg/ml (r=0.997 6) and PTX from 0.05-25 μg/ml (r=0.997 9), meanwhile the LOQ of PTX-SA and PTX were 0.15 and 0.05 μg/ml, respectively. The recoveries were both larger than 90% with RSD less than 5%. Moreover, the RSD of intra-day and inter-day assays were both less than 5%. Conclusion The method was simple, effective, stable and accurate which could be suitable for the determination of PTX-SA and PTX in pharmacokinetic study. |
keywords:RP-HPLC PTX-SA PTX content determination |
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