百令胶囊中17种氨基酸的柱前衍生化RP-HPLC法含量测定

目的: 测定与分析百令胶囊中17种氨基酸的含量。 方法: 样品以6 mol·L^-1盐酸于150 ℃水解1 h,以异硫氰酸苯酯(PITC)为衍生化试剂衍生后进行HPLC分析。采用Phenomenex Gemini C₁₈(250 mm×4.6 mm,5 μm)色谱柱,柱温为43 ℃;流动相A为0.1 mol·L^-1醋酸-醋酸钠缓冲液(pH 6.5)-乙腈(93∶ 7),流动相B为乙腈-水(4∶ 1),梯度洗脱,流速为1.0 mL·min^-1;检测波长为254 nm。 结果: 17种氨基酸浓度在1.02~43.32 μg·mL^-1范围内呈良好的线性关系(r=0.9996~0.9997);平均回收率除缬氨酸、甲硫氨酸为89.3%和84.8%外,其余15种氨基酸在94.5%~106.7%之间;RSD除苏氨酸、缬氨酸、甲硫氨酸为3.2%,3.6%,4.6%外,其余均小于3.0%。 结论: 本方法灵敏、准确,具有良好的重复性和稳定性,可用于百令胶囊中氨基酸的检测。

Objective: To determine and analyze 17 amino acids in Bailing capsules. Methods: The sample was hydrolyzed for 1 hour at 150 ℃, and derived with phenyl isothiocyanate(PITC).HPLC was performed on a Phenomenex C₁₈(250 mm×4.6 mm,5 μm)column with gradient elution of 0.1 mol·L^-1 acetic acid-sodium acetate buffer solution(pH 6.5)-acetonitrile(93∶ 7)(A)and acetonitrile-water(4∶ 1)(B)at the flow rate of 1.0 mL·min^-1,detected at 254 nm. Results: 17 amino acids had good linearity in the ranges of 1.02-43.32 μg·mL^-1(r=0.9996-0.9997).The average recoveries were 94.5%-106.7%,except Val 89.3%,Met 84.8%,and RSD were less than 3.0%,except Thr 3.2%,Val 3.6% and Met 4.6%. Conclusion: The method is sensitive and accurate with high repeatability and stability, which is helpful for the determination of amino acids in Bailing capsules.