基于AQC衍生和液质联用的茶叶游离氨基酸分析  

Analysis of free amino acids in tea based on AQC derivation with liquid chromatography-mass spectrometry

作　　者：陈思肜 赵峰[1,2] 王淑燕 金珊 周鹏[3] 危赛明 叶乃兴 CHEN Si-rong;ZHAO Feng;WANG Shu-yan;JIN Shan;ZHOU Peng;WEI Sai-ming;YE Nai-xing(College of Horticulture,Fujian Agriculture and Forestry University,Key Laboratory of Tea Science at Universities in Fujian,Fuzhou 350002,China;College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Inspection and Research Institute for Product Quality,Fuzhou 350002,China;Fujian Tea Import&Export Company Limited,Fuzhou 350014,China)

机构地区：[1]福建农林大学园艺学院/茶学福建省高校重点实验室,福州350002 [2]福建中医药大学药学院,福州350122 [3]福建省产品质量检验研究院,福州350002 [4]福建茶叶进出口有限责任公司,福州350014

出　　处：《南方农业学报》2019年第10期2278-2285,共8页Journal of Southern Agriculture

基　　金：国家自然科学基金项目（31270735）;福建农林大学科技创新专项（CXZX2016117）;国家茶叶质量安全工程技术研究中心开放项目（2018NTQS0301）;茶学福建省高校重点实验室开放项目（KLTS2018001）

摘　　要：【目的】开发一种能快速、准确、可靠地对各茶叶中蛋白质氨基酸和低含量非蛋白质氨基酸进行测定的方法,为分析茶叶氨基酸组分提供新途径。【方法】质控样品经提取和氨基喹啉-N-羟基丁二酰氨基甲酸酯(AQC)衍生后,通过三重四极杆液质联用仪进行测定,优化质谱条件,评估35种氨基酸的线性范围、检出限、定量限、重现性和回收率,并应用该方法对市售的六大茶类(红茶、绿茶、白茶、黄茶、乌龙茶和黑茶)共50批次样本的游离氨基酸含量进行测定。【结果】采用AQC衍生—液相色谱串联三重四极杆质谱法可在20 min内完成35种氨基酸的定性定量检测,线性回归良好(R>0.9900);定量限在2.1×10-6~1.8×10^-2 mg/L,检出限在6.3×10^-7~5.5×10^-3 mg/L;日内重现性≤4.6%,日间重现性≤5.4%;加标回收率81.0%~99.9%,相对标准偏差(RSD)≤9.5%。对六大茶类游离氨基酸的测定结果显示,除绿茶和黄茶的氨基酸构成相似外,其他茶类的氨基酸构成均呈现出各自的规律性特征,氨基酸的定量结果可通过3个主成分(累积方差为66.5%)实现茶类区分。其中,蛋氨酸、苯丙氨酸、肌氨酸、茶氨酸、瓜氨酸、3-甲基-L-组氨酸、精氨酸、色氨酸、组氨酸和1-甲基-L-组氨酸的变量权重值(VIP)大于1.0,对茶类判别贡献较大。【结论】AQC衍生结合液相色谱质谱联用是一种快速、可靠、有效的分析方法,能对茶叶中的蛋白质氨基酸和低含量非蛋白质氨基酸进行高灵敏度定量。【Objective】To develop a rapid,accurate and reliable method for the determination of protein amino acids and low content non protein amino acids in tea.【Method】The quality control samples(QC)were extracted and derived from 6-aminoquinolyl-n-hydroxysuccinyl carbamate(AQC),then determined by triple quadrupole liquid chromatographymass spectrometry,optimized the mass spectrum conditions,evaluated the linear range,detection limit,quantitative limit,reproducibility and recovery rate of 35 amino acids.The content of free amino acids in 50 batches of six kinds of tea(black tea,green tea,white tea,yellow tea,oolong tea and black tea)was determined.【Result】Using AQC derivative liquid chromatography tandem triple quadrupole mass spectrometry,35 kinds of amino acids were detected qualitatively and quantitatively in 20 minutes,with good linear regression(R>0.9900);quantitative limit was 2.1×10^-6-1.8×10^-2 mg/L,detection limit was 6.3×10^-7-5.5×10^-3 mg/L;daily reproducibility was less than or equal to 4.6%,daytime reproducibility was less than or equal to 5.4%;recovery rate was 81.0%-99.9%,relative standard deviation(RSD)was≤9.5%.The results showed that the amino acid composition of other tea groups showed their own regularity except green tea and yellow tea.The quantitative results of amino acids could be distinguished by three main components(cumulative variance was 66.5%).Among them,the variable weight value(VIP)of methionine,phenylalanine,sarcosine,theanine,citrulline,3-methyl-l-histidine,arginine,tryptophan,histidine and 1-methyl-l-histidine is greater than 1.0,which contributes a lot to tea discrimination.【Conclusion】AQC derivatization combined with liquid chromatography-mass spectrometry is a rapid,reliable and effective method for the quantitative analysis of protein amino acids and low content non protein amino acids in tea.

关 键 词：茶叶 游离氨基酸 AQC衍生 三重四极杆液质联用仪 

分 类 号：S571.1[农业科学—茶叶生产加工]