反相高效液相色谱法测定烟叶中的游离氨基酸

Determination of Free Amino Acids in Tobacco Leaves by Reversed-Phase High Performance Liquid Chromatography

摘要: 先用乙醇提取烟叶中的游离氨基酸,提取液经阳离子交换树脂纯化后,再用邻苯二甲醛(OPA)、氯甲酸芴甲酯(FMCF)联合柱前衍生化和反相高效液相色谱法测定了烟叶中17种带伯氨基和/或仲氨基的游离氨基酸。方法的重复性为:天冬氨酸(Asp)、谷氨酸(Glu)和组氨酸(His)的相对标准偏差(RSD)分别为8.0%、8.6%和9.0%,其它氨基酸的RSD均在3%~7%范围内;回收率,除苏氨酸(Thr)的为68.0%较低外,其它氨基酸的在81.0%~110.5%范围内,平均为93.9%。用该方法测定了不同等级烟叶中的游离氨基酸。

Abstract: The free amino acids in tobacco leaves were extracted with ethanol, and purified by a cationic ion-exchange resin column, then derived with o-phthalaldehyde(OPA) and 9-fluorenylmethylchloroformate(FMCF) in precolumn, and separated by reversed-phase high-performance liquid chromatography.Seventeen kinds of free amino acids were determined.The relative standard deviations of these amino acids were in the range of 3.0% to 7.0%, except that those of aspartic acid, glutamic acid and histidine were 8.0%, 8.6% and 9.0% respectively.The recoveries of these amino acids were from 81.0% to 110.5% with the exception of 68.0% for threonine, the mean recovery was 93.9%.The contents of free amino acids in tobacco leaves of different grades were determined by this method.