同型半胱氨酸的高效液相色谱分析

ANALYSIS OF HOMOCYSTEINE BY HPLC

目的：建立一种用反相高效液相色谱测定同型半胱氨酸的新方法。方法：反应条件研究采用分子荧光光谱法，同型半胱氨酸测定采用反相高效液相色谱梯度洗脱法。结果：使用邻苯二甲醛作荧光衍生试剂，在温度为50℃，pH8．4，反应4min时，衍生物相对荧光强度最大。在3×10-7～1．6×10-6mol／L浓度范围内．荧光强度与浓度之间呈线性关系，检测限为2．44×10－8mol／L。结论：同型半胱氨酸经过氧化氢处理后，荧光强度可提高5倍。

Objective: To establish a method to determine homocysteine by reversed phase high perfor-mance liquid chromatography (RP-HPLC). Methods: The reaction conditions were studied by molecular fluorescence spectrophotometry. Homocysteine was determined by RP-HPLC. Results: Ophthaladehyde was used as fluorescent derivative reagent. When the reaction took place at 50℃ and pH8. 4 for 4 min,the fluorescent intensity reached the highest. In the concentration of 3 ×10-7~1. 6×10-6 mol/L, F% = 4.85 ×10;C (mol/L) -1. 91, r = 0. 99, CV = 2. 47 %,detection limit = 2. 44 ×10-8 mol/L. Conclusion: Treat-ed with H2O2, the fluorescent intensity will increase by 5 times.