精制左旋多巴中L-多巴含量及主要杂质成分的HPLC检测

Detection of L-dopa and related impurity in refined L-dopa from seeds of Stizolobium cochinchinensis(Lour.) by high performance liquid chromatography

L-多巴作(L-dopa)为治疗帕金森氏病的主要药物,可以作为食品添加剂开发针对特殊人群的功能性产品。从食药两用植物猫豆种子中纯化的精制L-多巴中通常会含有少量L-酪氨酸等杂质,从而影响精制品作为药物使用。本文建立了一种高效液相色谱方法,可以有效的检测从猫豆中提取的L-多巴主要杂质成分L-酪氨酸(L-tyr)和L-苯丙氨酸(L-phe)的含量。在紫外检测下(λ=260 nm),液相条件为:色谱柱:Dikma Spursil C18柱(250 mm×4.6 mm,5μm);流动相:2%乙酸溶液-甲醇(90∶10);流速:0.8 m L/min;检测波长:260 nm;柱温:室温;能够检测L-多巴和L-苯丙氨酸;在荧光检测下(λex=280 nm,λem=314 nm),液相条件为:色谱柱:Dikma Spursil C18柱(250 mm×4.6mm,5μm);流动相:0.8%甲酸-乙腈(98∶2);进样量:10μL;柱温:室温;能够检测L-多巴与L-酪氨酸。

L-dopa, a major drug for the treatment of Parkinson＇ s disease in clinical, can also be used as a food ingredient in functional products for special human subjects. The refined L-dopa from the seeds of Stizolobium cochinchinensis （L our） Tang et Wang, widely planted in Guangxi for its high economical values, usually contains trace amounts of impurities related L-dopa, e.g. L-tyr, L-phe, etc. The impurities would limit the refined L-dopa to be applied in clinical. In this study, a set of high performance liquid chromatography （HPLC） method was established to discriminate L-dopa and its related impurities efficiently. To discriminate L-dopa from L-phe, it was carried out on a Dikma Spursil C18 column （250 mm ×4.6 ram, 5μm） , with 2% acetic acid solution-methanol （90： 10） as the mobile phase at a flow rate of 0.8 mL/min, the column was kept at room temperature and the detection wavelength was 260 nm; To discriminate L-dopa from L-tyr, it was carried out on the same column, with 0.8% formic acid-acetonitrile （98： 2） as mobile phase at a flow rate of 0.8 mL/min, the column was kept at room temperature and the detection was undertaken with a fluorescence detector, with λex = 280 nm as the detection wavelength while λem = 314 nm as the reference wavelength.