高效液相色谱法定量分析硫醇化合物的研究

Quantitative analysis of thiol compounds by high performance liquid chromatography

利用高效液相色谱(high performance liquid chromatography, HPLC)建立了一种柱前衍生化试剂丙烯酸2′-甲氧基苯酯定量分析硫醇化合物的方法.丙烯酸2′-甲氧基苯酯与半胱氨酸、高半胱氨酸、谷胱甘肽进行衍生化反应所得产物的色谱分离条件为:Cicaeagent Mightysil RP-18 GP (150 mm×4.6 mm, 3μm)色谱柱,水-乙腈为流动相(含体积分数为0.1%的甲酸),流速为0.3 mL/min,进样量为3μL.测定结果显示,硫醇化合物的线性范围为0.05~1.0 mmol/L,最低检测限为0.01 mmol/L(S/N=3).该结果表明使用丙烯酸2′-甲氧基苯酯柱前衍生化法可有效检测出L-半胱氨酸胶囊中硫醇化合物的含量,因此该方法对检测硫醇化合物含量具有良好的参考价值.

A pre-column derivatization reagent(2'-methoxyphenyl acrylate)was developed for the quantitative analysis of thiol compounds by high performance liquid chromatography.The chromatographic separation conditions of 2'-methoxyphenyl acrylate with cysteine,homocysteine and glutathione derivatives are as follows:Cicaeagent Mightysil RP-18GP(150mm×4.6mm,3μm)column,water-acetonitrile is the mobile phase(containing 0.1%formic acid by volume),with a flow rate of 0.3mL/min and an injection volume of 3μL.The results showed that the linear range of thiol compounds was 0.05-1.0mmol/L and the lowest detection limit was 0.01mmol/L(S/N=3),and the content of thiol compounds in L-cysteine capsules could be effectively detected.Therefore,the proposed method exhibit good reference value for the detection of thiol compound content.