AccQ-Tag-HPLC法测定烟叶生长过程中游离氨基酸的变化  

Detection of Free Amino Acids in Tobacco Leaves during Growth Process by Using AccQ-Tag-HPLC Method

作　　者：李方楼[1] 姬小明[1] 魏跃伟[1] 杨永锋[1] 刘国顺[1] 

机构地区：[1]河南农业大学国家烟草栽培生理生化研究基地,河南郑州450002

出　　处：《安徽农业科学》2011年第27期17010-17012,共3页Journal of Anhui Agricultural Sciences

基　　金：国家烟草专卖局科技攻关项目(110200401003)

摘　　要：[目的]在AccQ-Tag分析包的基础上建立一种适合烟草中游离氨基酸测定的方法。[方法]采用WATERS公司的AccQ-Tag试剂包作为构建方法的基本条件,通过优化烟草中游离氨基酸的前处理和色谱条件,以6-氨基喹啉-N-羟基琥珀酰亚胺基甲酸酯(AQC)为衍生试剂与各氨基酸柱前定量衍生后,再用AccQ-TagTM柱梯度洗脱,建立了烟叶中游离氨基酸的AccQ-Tag-HPLC测试方法。[结果]用该方法测定烟叶杀青样品中的游离氨基酸,能够分离检测烟叶中的16种游离氨基酸,其回收率均在82.8%～105.3%,标准偏差均在2.5%以内,具有较好的重现性和回收率。烤烟杀青样中游离氨基酸以脯氨酸、天门冬氨酸、丝氨酸和丙氨酸含量较高。[结论]AccQ-Tag-HPLC法是一种实用的烟草中游离氨基酸的分析方法,适于烟叶中游离氨基酸从旺长期到成熟期的不同时段含量变化的测试。[Objective] The study aimed to establish a method for the detection of free amino acids in tobacco by using AccQ-Tag-HPLC system.[Method] With the AccQ-Tag reagent bag from WATERS company as the basic condition of building method,through the optimization of pretreatment and chromatographic conditions for the free amino acid in the tobacco,the 6-amino decoquinate-N-hydroxyl succinyl imide formic ether（AQC） was taken as the derivative reagent to make for the column quantitative derivatization with each amino acid and then to elute by using AccQ-TagTM column gradient,thus,a AccQ-Tag-HPLC determining method for the detection of free amino acids in tobacco was established.[Result] When the method was used to determine the free amino acids in the kill out samples of tobacco,16 kinds of free amino acids in tobacco leaves were separated and detected by the method,with the recovery of 82.8%-105.3% and the standard deviation values of below 2.5%,showing a better reproducibility and recovery.The contents of proline,aspartate,serine and alanine in free amino acids in the kill out samples of the flue-cured tobacco were higher.[Conclusion] AccQ-Tag-HPLC method was a kind of practical method for the analysis of the free amino acid in the tobacco,which was suitable for the determination on the content variation of free amino acid in the tobacco in the different periods from the vigorous growing stage to maturity.

关 键 词：烟草 游离氨基酸 AccQ-Tag高效液相色谱法 

分 类 号：O657.72[理学—分析化学]