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目的建立白茯苓中游离氨基酸的定量方法,同时测定白茯苓中4种游离氨基酸的含量。方法以6-氨基喹啉基-N-羟基琥珀酰亚氨基-甲酸酯(6-quinolyl-N-hydroxy-succinimidyl-carboxylate,AQC)作为衍生试剂,建立超高效液相色谱(ultra performance liquid chromatography,UPLC)柱前衍生法,测定3个省24批白茯苓中游离苏氨酸、丙氨酸、赖氨酸及酪氨酸的含量,采用Simca-p+11软件对不同批次样品氨基酸的含量进行聚类分析。结果 24个批次的白茯苓中均含有丙氨酸,含量范围为16.97~69.92μg·g~(-1);苏氨酸、酪氨酸和赖氨酸在有些批次的样品中未检出。经偏最小二乘法(partial least squares discrimination analysis,PLS-DA)分析,安徽和云南省收集的样本可以较好地聚类,湖北收集的各样品中氨基酸含量差异较大。结论 AQC柱前衍生化方法操作简便,快速,灵敏度高,可用于茯苓中游离氨基酸的测定。
Objective To establish a method for the quantitative determination of free amino acids in Poria cocos and determine the contents of four kinds of free amino acids in Poria cocos. Methods Using 6-quinolyl-N-hydroxy-succinimidyl-carboxylate (AQC) as derivatization reagent, ultra performance liquid chromatography , UPLC) pre-column derivatization method was used to determine the content of free threonine, alanine, lysine and tyrosine in 24 batches of Poria cocos from three provinces. Simca-p + 11 software was used to analyze the amino acid Content of cluster analysis. Results 24 batches of Poria cocos with alanine contents ranged from 16.97 to 69.92 μg · g -1; Threonine, Tyrosine and Lysine were not detected in some batches of samples Out By partial least squares discrimination analysis (PLS-DA), the samples collected in Anhui and Yunnan provinces clustered well, and the contents of amino acids in Hubei samples were significantly different. Conclusion AQC pre-column derivatization method is simple, rapid and sensitive. It can be used for the determination of free amino acids in Poria.