肿瘤细胞中长春新碱的高效液相色谱法测定

Determination of Vincristini Sulfas in Tumor Cells by High Performance Liquid Chromatography

长春新碱（ＶＣＲ）为重要和常用抗肿瘤药物之一。肿瘤细胞耐药性是导致化疗失败的主要原因。为了筛选耐药细胞的逆转剂，建立了测定肿瘤细胞内ＶＣＲ浓度的高效液相色谱法，色谱条件为：Ｚｏｒｂａｘ－ＯＤＳ反相柱２５ｃｍ×４．６ｍｍｉ．ｄ．，流动相：０．０２ｍｏｌ／ＬＫ２ＨＰＯ４（ｐＨ６．６）∶ＣＨ３ＯＨ（２０∶８０，Ｖ／Ｖ），流速：１．０ｍＬ／ｍｉｎ，检测波长：２６７ｎｍ。方法简单、快速、选择性好，在１０～２００ｍｇ／Ｌ范围内ＶＣＲ浓度－峰高呈良好的线性关系（ｒ＝０．９９９８），仪器灵敏度为４ｎｇ。所建立的方法适合于ＶＣＲ的血药浓度测定及ＶＣＲ在肿瘤细胞、组织中的代谢、分布、排泄、蓄积等研究。

Vincristini sulfas (VCR) is an important common antitumor drug. The resistance to anticancer drug is the main cause of the chemotherapy failure. To screen the drugs which can reverse VCR resistance for VCR resistant cell strain, an analytical method has been established for the determination of VCR concentration in tumor cells using high performance liquid chromatography (HPLC). The stainless steel column was 25cm× 4.6 mm i.d. packed with totally porous, spherical silica particles (5μm). A solution of methanol and 0.02mol/L dipotassium hydrogen phosphate (80∶20, V/V ) adjusted to pH 6.6 with H 3PO 4 was employed as the mobile phase. The flow rate was 1mL/min. Chromatography was performed with ultraviolet detector at 267nm. The method is simple, rapid and selective. Linear calibration curve for VCR was measured within the range of 10 to 200mg/L with correlation coefficient of 0.999 8. The lowest detection limit was 4mg/L tumor cells extract. The HPLC method described is suitable for clinical monitoring and pharmacokinetic study of VCR.