高效液相色谱法测定复方氨基酸胶囊(8 -11)中氨基酸的含量

以1-(5- 氟 -2,4- 二硝基 - 苯基)-4- 甲基哌嗪(PPZ)为衍生化试剂,建立了一种测定氨基酸含量的柱前衍生化的高效液相色谱法(HPLC).色谱条件:采用Cica - reagent Mightysil RP - 18 GP(3 μm, 150 mm×4.6 mm)色谱柱,流动相为甲醇和水的混合液(甲醇和水的体积比为1:1且混合液中含体积分数为0.2%的甲酸),流速为0.3 mL/min, 紫外检测波长为354 nm, 进样体积为2 μL.衍生化条件:温度为40 ℃,反应时间为3 h, 氨基酸与衍生化试剂的摩尔比为1:10.对复方氨基酸胶囊(8 -11)中的8种氨基酸进行测定显示,在0.5 ～1 000 μmol/L浓度范围内工作曲线呈良好线性关系,且最低检测限为0.5 ～ 5.0 μmol/L, 加标平均回收率为90.0%～104.3%, 相对标准偏差为0.57%～2.29%.由此表明该方法具有良好的灵敏度、重复性和准确度,可用于测定药物中氨基酸的含量.

Determination of amino acids in compound amino acid capsule(8 -11)by high performance liquid chromatography

A pre - column derivatization method for the determination of amino acids by high performance liquid chromatography(HPLC)was developed using 1 -(5 - fluoro - 2,4 - dinitro - phenyl)- 4 - methylpiperazine(PPZ)as derivatization reagent.Chromatographic conditions: the chromatographic column of Cica - reagent Mightysil RP - 18 GP(3 μm, 150 mm×4.6 mm)was used, with mobile phase methanol and water volume ratio 1:1(0.2% formic acid by volume), flow rate of 0.3 mL/min, UV detection wavelength of 354 nm, and injection volume of 2 μL.The derivatization conditions was 40 ℃, the reaction time was 3 h, and the molar ratio of amino acids to derivatization reagents was 1:10.The analytical determination of the eight amino acids in the compound amino acid capsules(8 - 11)showed good linearity in the range of 0.5-1 000 μmol/L, and the minimum detection limits was 0.5-5.0 μmol/L.The average spiked recoveries was 90.0%-104.3%, and the relative standard deviations was 0.57% -2.29%.This method showed that the method has good sensitivity, reproducibility and accuracy and can be used for the determination of amino acids in drugs.