高效液相色谱法分析干旱胁迫下烟草中游离氨基酸的含量变化  

Analysis of Contents Change of Free Amino Acids in Tobacco Under Drought Stress by High Performance Liquid Chromatography

作　　者：刘欣 黄茹[2] 许永 李勇[3] 李晶 蒋佳芮 邢佳鑫 宋春满 杨光宇 高茜 黄海涛 LIU Xin;HUANG Ru;XU Yong;LI Yong;LI Jing;JIANG Jiarui;XING Jiaxin;SONG Chunman;YANG Guangyu;GAO Qian;HUANG Haitao(Technology Center of China Tobacco Yunnan Industrial Co.,Ltd.,Kunming 650106;School of Urban and Environment,Yunnan University of Finance and Economics,Kunming 650221;Institute of Agricultural Resources and Environment,Jiangsu Academy of Agricultural Sciences,Nanjing 210014)

机构地区：[1]云南中烟工业有限责任公司技术中心,云南昆明650106 [2]云南财经大学城市与环境学院,云南昆明650221 [3]江苏省农业科学院农业资源与环境研究所,江苏南京210014

出　　处：《分析科学学报》2023年第2期219-224,共6页Journal of Analytical Science

基　　金：中国烟草总公司科技重大专项项目(110202101004(JY-04));云南中烟工业有限责任公司科技项目(2021JC08);云南省烟草化学重点实验室开放课题(No.2021539200340239)。

摘　　要：建立了柱前衍生-高效液相色谱高灵敏、快速测定新鲜烟叶中21种游离氨基酸的新方法。新鲜烟叶样品经0.1 mol/L的盐酸超声提取30 min后,以6-氨基喹啉基-N-羟基琥珀酰亚胺基甲酸酯(AQC)作为衍生试剂,反向色谱柱分离,二极管阵列检测器测定。在最佳实验条件下,21种氨基酸在9 min内实现完全分离,在低、中、高3个浓度水平下的加标回收率在84.7%~104%之间,相对标准偏差(RSD)为1.8%~5.1%,定量限在0.02~0.10 mg/g之间。基于该方法,探讨了干旱胁迫下游离氨基酸含量的变化趋势。研究表明,在干旱胁迫下,总游离氨基酸、脯氨酸、天冬酰胺等含量明显升高,且大部分氨基酸在旺长期时变化最大,对明晰游离氨基酸参与烟草抗逆的调控机制提供了数据基础和技术支撑。A highly sensitive and rapid method for the determination of 21 free amino acids in fresh tobacco leaves was developed based on pre-column derivatization coupled with high performance liquid chromatography.The samples were extracted with 0.1 mol/L hydrochloric acid solution for 30 min by ultrasonic extraction,and AQC(6-quinolyl-N-hydroxy-succinimidyl-carboxylate)was used as a deriving reagent.The extracts were performed with AQC pre-column derivatization,reversed phase-column separation and detected by a diode array detector.Under the optimal conditions,the 21 kinds of amino acids were separated within 9 min.The recoveries were 84.7%-104%with relative standard deviations of 1.8%-5.1%.The limits of quantification were 0.02-0.10 mg/g.Based on this method,the contents change of free amino acids under drought stress was discussed.Results showed that the contents of total free amino acids,proline and asparagine increased significantly under drought stress,and most of the amino acids changed significantly at the prosperously growing stage,which provided the data basis and technical support for understanding the regulation mechanism of free amino acids involved in tobacco stress resistance.

关 键 词：氨基酸 柱前衍生 高效液相色谱 干旱胁迫 烟草 

分 类 号：O657.72[理学—分析化学]