Willey T A, Bekos E J, Gaver R C, Duncan G F, Tay L K, Beijnen J H, Farmen R H
Department of Metabolism and Pharmacokinetics, Bristol-Myers Squibb Company, Princeton, NJ 8543-4500.
J Chromatogr. 1993 Nov 24;621(2):231-8. doi: 10.1016/0378-4347(93)80100-i.
An isocratic high-performance liquid chromatographic method has been developed and validated for the quantitative determination of paclitaxel (Taxol), a novel antimitotic, anticancer agent, in human plasma. The analysis required 0.5 ml of plasma, and was accomplished by detection of the UV absorbance of paclitaxel at 227 nm following extraction and concentration. The method involved extraction of paclitaxel from plasma, buffered with 0.5 ml of 0.2 M ammonium acetate (pH 5.0), onto 1-ml cyano Bond Elut columns. The eluent was evaporated under nitrogen and low heat, and reconstituted with the mobile phase, acetonitrile-methanol-water (4:1:5, v/v/v) containing 0.01 M ammonium acetate (pH 5.0). The samples were chromatographed on a reversed-phase octyl 5 microns column. The retention time of paclitaxel was 10 min. The validated quantitation range of the method was 10-1000 ng/ml (0.012-1.17 microM) of paclitaxel in plasma. Standard curve correlation coefficients of 0.995 or greater were obtained during validation experiments and analysis of clinical study samples. The observed recovery for paclitaxel was 83%. Epitaxol, a biologically active stereoisomer, and baccatin III, a degradation product, were also chromatographically separated from taxol by this assay. The method was applied to samples from a clinical study of paclitaxel in cancer patients, providing a pharmacokinetic profiling of paclitaxel.
已开发并验证了一种等度高效液相色谱法，用于定量测定人血浆中的紫杉醇（泰素），这是一种新型抗有丝分裂抗癌药物。该分析需要0.5毫升血浆，通过在提取和浓缩后检测紫杉醇在227纳米处的紫外吸光度来完成。该方法包括用0.5毫升0.2 M醋酸铵（pH 5.0）缓冲的血浆将紫杉醇提取到1毫升氰基键合硅胶柱上。洗脱液在氮气和低热下蒸发，并用流动相乙腈 - 甲醇 - 水（4:1:5，v/v/v）复溶，该流动相含有0.01 M醋酸铵（pH 5.0）。样品在反相5微米辛基柱上进行色谱分析。紫杉醇的保留时间为10分钟。该方法经验证的定量范围为血浆中紫杉醇浓度为10 - 1000纳克/毫升（0.012 - 1.17微摩尔）。在验证实验和临床研究样品分析过程中，标准曲线相关系数达到0.995或更高。观察到的紫杉醇回收率为83%。该分析方法还能将具有生物活性的立体异构体表紫杉醇和降解产物巴卡亭III与紫杉醇进行色谱分离。该方法应用于癌症患者紫杉醇临床研究的样品，提供了紫杉醇的药代动力学特征分析。
