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16.97~69.92 μg·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。
Abstract
OBJECTIVE To establish the quantification method for free amino acids including threonine, alanine, lysine and tyrosine in the inner part of Poria cocos. METHODS AQC (6-quinolyl-N-hydroxy-succinimidyl-carboxylate) was used as deriving reagent, and pre-column derivatization combined with ultra-high performance chromatography was applied to quantify threonine, alanine, lysine and tyrosine in 24 batches of the inner part of Poria cocos from Yunnan, Hubei and Anhui provinces. The experimental data were analyzed by partial least squares discriminant analysis (PLS-DA) using Simca-p+11 software. RESULTS Alanine existed in every batch of sample, and its content ranged from 16.97-69.92 μg·g-1 in samples collected from different origins. Threonine, lysine and tyrosine were not found in some samples. The PLS-DA result showed that the samples from Yunnan and Anhui provinces clustered well. While the amino acid contents in the samples from Hubei province varied greatly. CONCLUSION AQC combined with pre-column derivatization is a convenient, rapid and sensitive method to quantify the free amino acids in Poria.
关键词
白茯苓 /
游离氨基酸 /
柱前衍生化 /
超高效液相色谱法 /
6-氨基喹啉基-N-羟基琥珀酰亚氨基-甲酸酯衍生化
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Key words
the inner part of Poria cocos /
free amino acids /
pre-column derivatization /
UPLC /
AQC derivatization
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中图分类号：
R284
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参考文献
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脚注
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基金
863课题药食同源重要功能因子提取,分离及规模化生物合成关键技术资助项目(2014AA0222201);十二五重大新药创制茯苓三萜类有效部分抗肿瘤或药性研究资助项目(2012ZX09103201-001);中医药物业专项道地药材国家重点实验室(培育)-茯苓等8味中药材近红外快速检测技术研究资助项目(GZ20140101)
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