一种改进的测定体液中地尔硫及脱乙酰地尔硫的HPLC法

AN IMPROVEMENT OF THE DETERMINATION OFDILTIAZEM AND DESACETYLDILTIAZEM BYHPLC IN BIOLOGICAL FLUID

本文系统地研究和建立了用反相高效液相色谱法测定生物体液中的地尔硫及其活性代谢物脱乙酰地尔硫的方法。本方法中使用300mm×3.9mm(I.D)Waters不锈钢柱,μ-Bondapak C_(18)(10μm)作固定相,甲醇:缓冲液(0.8％TEMED,pH5.4)60:40(V/V)为流动相。流速为1.3ml/min。检测波长为238nm。以地尔硫(艹卓)的中间体为内标进行定量。本法对地尔硫(艹卓)及脱乙酰地尔硫(艹卓)的检测限均为2ng。血清中最低检测浓度均为5ng/ml。在20、200、1000ng/ml浓度范围内,两者的方法回收率接近100％。天间、天内精密度(CV)均小于5％。用本法对6例服用地尔硫(艹卓)控释片的心绞痛病人进行了临床监测,治疗血浓范围为cmin30～100 ng/ml。

A specific, sensitive and reproducible RP-HPLC method for determining of diltiazem and one of it's major active metobolites, desacetyldiltiazem, in plasma was established . A Waters HPLC instrument, equipped with a Waters Model 990 Photodiode Array Detector operated at 238 nm and a 300/3.9 mm (I. D.) stainless-steel column packed with the μ-Bondapak-018(10 μm) were selected. The Chroma-tography was carried out at temperature 25 + 0.5℃ with a mobile phase composed of methonol-TEMED (0.8%, pH6.4) = 60/40 (F/F) at the flow rate of 1.3ml/ min, The limit of detection of the method was 2ng, the minimal detectable concentration being 5ng/ml for both diltiazem and desaoetyldiltiazem in plasma sample. The recoveries of the method were 99.7% (CV3%), 100.5%(CV2%) and 100.7% (CV1.1%) for diltiazem and 101.9% (CV 2.7%),100.5%(CV 2.6%) and 101.6%(CV 2.5%) for desacetylditiazem, botn at the concentrations of 20ng/ml, 200 ng/ml and 1 000 ng/ml in plasma respectively. The CV% of intra-day and inter-day assays were less than 5% to both of them. The plsma concentrations of six angina patients were analysed after they had been treated only with the diltiazem hydroohloride tablets for 7 days.