HPLC-MAXPLOT UV检测法测定血清中丹那唑及其代谢产物

本文建立了同时测定人体血清中丹那唑(Ⅰ)及其主要代谢产物2-羟甲基妊娠素(Ⅱ)和其它代谢产物妊娠素(Ⅲ)的高效液相色谱法。方法用YWG C18柱并以甲醇—水(74:26)作为流动相,应用紫外最大吸收作图法(maxplot)检测技术,以UV 285,240nm分别测定Ⅰ,Ⅱ和Ⅲ。使原型药和代谢产物都达到较高检测灵敏度。同时对方法的专一性、准确性和精密度等方面进行了评价,并应用于临床监测。

ANALYSIS OF DANAZOL AND ITS MATABOLITES IN SERUM BY RP-HPLC WITH MAXPLOT UV DETECTOR

A sensitive, specific and rapid RP-HPLC method was developed for simultaneously determining a synthetic steroid drug danazol (Ⅰ) and its metabolites 2-hydroxymethylethisterone (Ⅱ) and ethisterone (Ⅲ). A column packed with YWG-C18, 10 μm and mobile phase of methanol-water (74:26, V/V) were used to separate C Ⅱ, Ⅲ and chloromadinone (I. S.). In order to get more satisfactory sensitivity of the four compounds, the detection wavelengths were set at 285 nm and 240 nm with maxplotechnique.he chromatographic peaks of Ⅰ and Ⅱ in serum were confirmed by UV spectra and mass spectra. The results showed that the limits of detection using 1 ml serum were 10 ng/ml for Ⅰ, 25.38 ng/ml for Ⅱ and 11.15 ng/ml for Ⅲ. Standard curves were constructed with 1 ml serum contaning Ⅰ (9.855～438.00 ng/ml), Ⅱ (50.75～812.00 ng/ml) and Ⅲ (14.99～428.40 ng/ml). The recoveries of Ⅰ, Ⅱ and Ⅲ from 1 ml serum were 99.04%, 63.17% and 96.08% respectively.The method was used to profile the serum levels of Ⅰ and Ⅱ in female with endometritis receiving 200 mg of danazol b. i. d. The result showed that the concentration of Ⅱ was 5～9 times highter than that of Ⅰ.