固相萃取/高效液相色谱法对肝癌细胞中多西紫杉醇的测定

建立了固相萃取/高效液相色谱法测定肝癌细胞中多西紫杉醇浓度的方法。细胞样品经三氯乙酸沉淀蛋白,Bond-elut C18固相萃取柱提取,采用Agilent TC-C18(5μm,4.6 mm×150 mm)色谱柱分析,以乙腈-0.02 mol/L醋酸铵缓冲液(体积比50∶50,醋酸调至pH5.0)为流动相,流速为1 mL/min,检测波长为230nm,进样体积为20μL。多西紫杉醇的质量浓度在0.05~2.25 mg/L范围内线性关系良好,相关系数r=0.999 3,检出限为0.03 mg/L,提取回收率为93%~95%,其日内、日间精密度(n=5)不大于6.9%,且稳定性良好。方法简便、快速、灵敏度高、干扰小,可用于生物样品中低浓度多西紫杉醇的测定。

Solid Phase Extracted/HPLC Method for Determination of Docetaxel in Hepatocellular Cancer Cells

To establish a high performance liquid chromatographic(HPLC) method for determination of docetaxel in hepatocellular cancer cells.Docetaxel was extracted from hepatocellular cancer cells by a Bond-elut C_(18) solid phase extraction(SPE)column,and separated on a Agilent TC-C_(18)(5 μm,4.6 mm×150 mm) column by using mobile phase consisted of ammonium acetate buffer(0.02 mol/L,pH 5.0)- acetonitrile(50 : 50,by volume) with a flow rate of 1 mL/min at detection wavelength of 230 nm.The result indicated that the calibration of docetaxel was linear in the range of 0.05-2.25 mg/L with correlation coefficient of 0.999 3.The detection limit for docetaxel was 0.03 mg/L.The extraction recoveries of docetaxel form forin cells sample ranged from 93% to 95%.The within-day RSDs(n=5) and the between-day RSDs(n=5) were both not more than 6.9%.The method was convenient,accurate and sensitive,and could be applied in the determination of low level of docetaxel in biological samples.