反相高效液相色谱法测定人血浆中紫杉醇浓度

Determination of paclitaxel in human plasma by reversed-phase high performance liquid chromatography

目的建立反相高效液相色谱法测定人体血浆中紫杉醇的浓度。方法色谱柱为InertS ustain C18(250 mm×4.6 mm,5μm),以甲醇-乙腈-水(30∶30∶40)为流动相,流速为1.0 mL·min^(-1);紫外检测波长为230 nm,柱温为35℃。以地西泮为内标,以甲基叔丁基醚为提取剂,用液液萃取法处理血浆样品,考察该方法的专属性、精密度与回收率、稳定性和重复性。结果紫杉醇线性范围为2.75×10^(-1)~11.00μg·mL^(-1),回收率为96.28%~97.85%,萃取回收率为85.36%~90.83%,日内和日间的精密度均小于2.84%,血浆样品在-20℃下放置15天和反复冻融3次的稳定性良好。结论本方法处理样品的过程简捷、取样量少、重复性好且回收率高,可用于临床肿瘤患者血浆紫杉醇含量的测定。

Objective To establish a reversed-phase high performance liquid chromatography method to determine the paclitaxel concentration in human plasma. Methods The chromatographic column was InertS ustain C18（ 250 mm × 4. 6 mm,5 μm）. The mobile phase was consisted of methanol-acetonitrile-water（ 30 ∶ 30 ∶ 40） at the flow rate of 1. 0 mL ·min^-1. The ultraviolet detection wavelength was 230 nm and the column temperature was 35 ℃. The plasma samples were extracted by the method of liquid-liquid extraction with methyl tert-butyl ether,and diazepam was used as the internal standard. The specificity,precision,recovery,stability and reproducibility were analyzed. Results Calibration curves were linear over the concentration range of 2. 75 × 10^-1-11. 00 μg·mL^-1. The recovery rate was 96. 28% to 97. 85%,and the extraction recovery was 85. 36%-90. 83%. The intra-day and inter-day RSD were both less than 2. 84%. Paclitaxel in human plasma was stable after storage at-20 ℃ in 15 days or freeze-thaw for three times.Conclusion The method is simple,rapid and repeatable,with little quantity of sample and high recovery rate. It can be used to determine the paclitaxel concentration in the plasma of tumor patients.