采用柱后衍生化高效液相色谱法分析治疗浓度的地高辛。

Analysis of digoxin at therapeutic concentrations using high-performance liquid chromatography with post-column derivatization.

作者信息

Kwong E, McErlane K M

出版信息

J Chromatogr. 1986 Sep 5;381(2):357-63. doi: 10.1016/s0378-4347(00)83601-4.

Abstract

A high-performance liquid chromatographic (HPLC) procedure has been developed for the analysis of digoxin in plasma at therapeutic concentrations. The assay method provides resolution of digoxin from its metabolites using a 15 cm X 4.6 mm HPLC column containing 3-micron octadecylsilane-bonded stationary phase. The effluent of the column is passed through a post-column reactor in which a fluorescent derivative is formed by the co-addition of hydrochloric acid and dehydroascorbic acid. Detection of the derivative is accomplished in a fluorometer with excitation at 336 nm and emission at 425 nm. The extraction efficiency for recovery of digoxin from plasma samples was 70% using chloroform-isopropanol (9:1) following a pre-wash with isooctane to remove endogenous substances. The calibration curve was linear (r = 0.9999) over the range 0.5-4 ng/ml digoxin in plasma using digitoxigenin as internal standard. The minimum detectable quantity of digoxin in plasma was 0.5 ng/ml at a signal-to-noise ratio of 4:1. Split-samples of digoxin control sera were assayed by the HPLC procedure and by the prescribed radioimmunoassay procedure. Excellent correlation was observed between the two methods (r = 0.999). No interference was noted when a selection of commonly co-prescribed drugs were evaluated for chromatographic co-elution or interference in detection with that of digoxin or the internal standard.

摘要

已开发出一种高效液相色谱（HPLC）方法，用于分析治疗浓度下血浆中的地高辛。该测定方法使用一根15 cm×4.6 mm、含有3微米十八烷基硅烷键合固定相的HPLC柱，可将地高辛与其代谢产物分离。柱流出物通过柱后反应器，在其中通过同时加入盐酸和脱氢抗坏血酸形成荧光衍生物。衍生物的检测在荧光计中完成，激发波长为336 nm，发射波长为425 nm。在先用异辛烷预洗以去除内源性物质后，使用氯仿 - 异丙醇（9:1）从血浆样品中回收地高辛的提取效率为70%。以洋地黄毒苷元作为内标，血浆中地高辛浓度在0.5 - 4 ng/ml范围内时，校准曲线呈线性（r = 0.9999）。在信噪比为4:1时，血浆中地高辛的最低检测量为0.5 ng/ml。对地高辛对照血清的分样采用HPLC方法和规定的放射免疫测定方法进行测定。两种方法之间观察到极好的相关性（r = 0.999）。在评估一系列常用的联合处方药物时，未发现色谱共洗脱或对与地高辛或内标物检测的干扰。