不同水解方法对植物叶片结合氨基酸测定影响分析  

Effects of different hydrolysis methods on the determination of bound amino acids in leaf of plant

作　　者：苏艳秋 陈伦兴 张雨晨 闵双楠 SU Yanqiu;CHEN Lunxing;ZHANG Yuchen;MIN Shuangnan(College of Life Science,Sichuan Normal University,Chengdu 610101,China;Key Laboratory of Land Resources Evaluation and Monitoring in Southwest,Ministry Education of China,Chengdu 610101,China;College of Life Science,Sichuan Agriculture University,Yaan 625000,China)

机构地区：[1]四川师范大学生命科学学院,四川成都610101 [2]四川师范大学西南土地资源评价与监测教育部重点实验室,四川成都610101 [3]四川农业大学生命科学学院,四川雅安625000

出　　处：《中国测试》2023年第11期102-109,共8页China Measurement & Test

基　　金：国家自然科学基金项目(32102759);四川师范大学实验技术与管理项目(SYJS2020030)。

摘　　要：为建立植物叶片结合氨基酸(bound amino acid,BAA)适宜检测方法,试验采用全自动氨基酸分析仪对4种植物叶片BAA进行测定:zuixinZG二极管,钠体系色谱柱(125 mmHR),柱后衍生光度法,570 mm/440 nm,单点校正。研究常规盐酸水解法、苯酚+盐酸水解法、β-巯基乙醇(β-mercaptoethanol,BME)+盐酸水解法、过甲酸氧化法对植物叶片17种BAA及其总量的影响。结果表明:过甲酸氧化法适合用于水稻和小麦叶片含硫氨基酸测定;BME+盐酸水解法适合用于4种植物叶片BAA测定。采用BME+盐酸水解26 h,植物叶片中多数极性正电和非极性氨基酸含量达到最大值;添加0.80%BME,植物叶片中极性正电氨基酸、负电氨基酸、中性氨基酸中的酪氨酸、胱氨酸及大部分非极性氨基酸含量达到最大值。该方法操作简单、重复性强、稳定性高,适合用于植物叶片BAA的检测。In order to establish a suitable hydrolysis method for bound amino acid content(BAA)in plant leaf,an amino acid analyzer was used for the determination of BAA in four plant leaves.The chromatography analysis was achieved using zuixinZG diode,sodium chromatographic column(125 mmHR),post-column derivation method.The detection wavelength was 570 nm and 440 nm and the test method was single point calibration method.The effect of different hydrolysis methods,including conventional hydrochloric acid,phenol+hydrochloric acid,β-mercaptoethanol(BME)+hydrochloric acid,performic acid+hydrochloric acid,on 17 amino acids and total amount of BAA in plant leaves was investigated.The results showed that the hydrolysis method of performic acid was suitable for the determination of sulfur-containing amino acids in rice and wheat leaves,and the hydrolysis method of BME+hydrochloric acid was suitable for the determination of BAA in leaves of four plants.After hydrolysis with BME+hydrochloric acid for 26 h,the contents of most polar positive and non-polar amino acids reached the maximum.When 0.80%BME was added,the contents of polar positive,negative amino acids,tyrosine and cystine in neutral amino acids,and most non-polar amino acids reached the maximum.The method is simple operation,reproducible and stable,and suitable for the determining of BAA in plant leaves.

关 键 词：氨基酸 植物叶片 盐酸水解 β-巯基乙醇 

分 类 号：Q331[生物学—遗传学]