反相高效液相色谱法在测定白肋烟烟叶游离氨基酸含量中的应用

Application of Reversed-Phase High Performance Liquid Chromatography in Determination of Free Amino Acids in Burley Tobacco

应用反相高效液相色谱技术建立了白肋烟烟叶游离氨基酸的测定方法.试验前处理首先采用超声波萃取，而后用邻苯二甲醛/3-巯基丙酸作为衍生剂进行柱前衍生，衍生化后进行液相色谱分析.结果表明，该方法的回收率为95.3%-100.7%，RSD为2.32%-9.24%（n=6）.利用该方法对湖北恩施中二级的白肋烟烟叶中的游离氨基酸进行了测定，有17种氨基酸被测出，其中天氡氨酸的含量最高，约占总量的65%，胱氨酸含量最低，约占总量的0.2%.

A reversed-phase high performance liquid chromatographic method was developed for determining free amino acids in hurley tobacco. Pre-treatment of the test was done in a two-step procedure using ultrasonic wave in the first step to extract amino acids and using OPA and 3-mercaptopropionic acid as the pre-column derivatising reagents. Amino acids were derivatized and analyzed by LC. The results showed that the average recoveries of the method ranged from 95.3% to 100.7% with the relative standard deviation of 2.32% - 9.24%. Burley leaves in Enshi of Hubei were determined using the method. The results showed that seventeen kinds of free amino aeids were determined, of which asp is the highest,taking up 65% of the total,and Cys-2 is the lowest.