高效液相色谱法测定血浆同型半胱氨酸

Determination of homocysteine in plasma by high performance liquid chromatography

目的 建立测定血浆同型半胱氨酸的高效液相色谱 (HPLC)方法。方法 血浆同型半胱氨酸经巯基乙醇还原后用碘乙酸封闭巯基 ,再与邻苯二甲醛 (OPA)反应完成衍生化 ;以磺基同型半胱氨酸为内标 ,经C18反相柱进行色谱分离 ,荧光检测器测定结果。结果 测定线性范围达15 0 μmol/L ,最低检测限为 1.0 μmol/L ,日内不精密度 3.7%～ 4.2 % ;日间不精密度 3.8%～ 4.3% ;方法回收率 93%～ 10 2 %。结论 本方法可用于临床实验室测定血浆同型半胱氨酸水平。

Objective A reverse phase high performance liquid chromatography (RP HPLC) method was established for determination of homocysteine in plasma with the use of pre column derivatization and fluorometric detection.Method Homocysteine was firstly reduced by β mercaptoethanol(MSH) and blocked the free thiol groups using iodoacetic acid. It was then reacted with o phthaldialdehyde (OPA) for derivatization and followed by HPLC and fluorescence detection. Results The linearity of this method was up to 150 μmol/L; the detection limit was 1.0 μmol/L; the within day imprecision was 3.7%～4.2%; the between day imprecision was 3.8%～ 4.3%; the recovery was 93%～102%.Conclusion This method could be used for the determination of plasma homocysteine in clinical laboratories.