用梯度反相HPLC-DAD法分离鉴别异靛甲体外代谢产物  

SCREENING FOR IN VITRO METABOLITES OF MEISOINDIGO USING RP-HPLC-DAD WITH GRADIENT ELUTION

作　　者：彭莹[1] 王慕邹[1] 

机构地区：[1]中国医学科学院药物研究所,北京100050

出　　处：《药学学报》1990年第3期208-214,共7页Acta Pharmaceutica Sinica

摘　　要：本文报道梯度RP-HPLC-DAD法用于异靛甲未知体外代谢产物的分离鉴别。采用ODS柱,CH_3OH-H_2O(0.05%H_3PO_4)作流动相,线性梯度洗脱,代谢样品的分离过程用两个波长(270和380nm)同时检测,色谱峰光谱采集范围从220到440nm。经色谱图比较、色谱峰吸收光谱对照以及三维图鉴别,确认了异靛甲体外代谢样品中的10个代谢产物并计算了它们的相对含量。结果表明,本法分辨率高并有专属性较强的鉴定能力,只需一次进样,即可在线完成样品的分离鉴别。A method of screening for in vitro metabolites of meisoindigo using RP-HPLC-DAD (photodiode array detector) with gradient elution was reported. Meisoindigo, a new structural antitumor agent, was metabolized by rat liver microsomes in vitro. The extract of the metabolic sample was dissolved in MeOH-acetone (9: 1). An aliquot of the solution was injected onto a 5 μm YWGC18 (ODS) column (15 cm×4.0 mm I D)and separated with H_2O (0.05% H_3PO_4)—MeOH in linear gradient elution. The flow rate was 0.5 ml/min and the oven temperature was 40℃. Two simultaneous monitoring wavelengths (270 and 380 nm) were used in order to avoid detective losses of unknown metabolites. The spectral recording range of chromatographic peaks was set from 220 to 440 nm. After comparison of the chromatograms and the spectra of chromatographic peaks, ten metabolites were screened out which gave similar spectrum. Of them, the main chromatographic peaks (No. 7, 8) were confirmed as two monohydroxy metabolites by EI-MS. Their relative contents were calculated by area normalization method. The experimental results show that this method is characteristic of highly effective separation and specific detection. The whole analytical course can be completed in less than two hours. The effects of different elution models, detective parameters and components of mobile phase on the analytical results were discussed.

关 键 词：异靛甲代谢物 液相色谱 

分 类 号：R914.1[医药卫生—药物化学]