色谱

色谱 ›› 2010, Vol. 28 ›› Issue (12): 1154-1157.DOI: 10.3724/SP.J.1123.2010.01154

• 研究论文 • 上一篇    下一篇

柱前衍生化-超高效液相色谱法快速测定酱油中的18种氨基酸

陈丽梅1,2, 尚艳芬2, 赵孟彬2, 刘虎威1*   

  1. 1. 北京大学化学与分子工程学院, 北京 100871; 2. 北京锦绣大地农业股份有限公司, 北京 100049
  • 收稿日期:2010-09-17 修回日期:2010-10-12 出版日期:2010-12-28 发布日期:2010-08-28
  • 通讯作者: 刘虎威,博士,教授. E-mail: hwliu@pku.edu.cn.
  • 基金资助:

    北京中关村科技园区海淀园博士后工作专项资助项目.

Rapid determination of 18 amino acids in soy sauce by ultra-performance liquid chromatography with pre-column derivatization

CHEN Limei1,2, SHANG Yanfen2, ZHAO Mengbin2, LIU Huwei1*   

  1. 1. College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China; 2. Beijing Glorious Land Agricultural Company Limited, Beijing 100049, China
  • Received:2010-09-17 Revised:2010-10-12 Online:2010-12-28 Published:2010-08-28
  • Contact:

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摘要： 建立了一种6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)柱前衍生,超高效液相色谱(UPLC)对酱油中18种氨基酸进行快速分离检测的方法。采用BEH C18色谱柱分离,在260 nm波长下检测,以乙酸铵-乙酸-乙腈-水和乙腈-乙酸为流动相,将流动相梯度和流速梯度相结合,在12 min内实现了18种氨基酸衍生物的分离。方法的线性回归系数(r2)均大于0.999,检出限为0.032～0.12 mg/L,日间相对标准偏差(RSD)为0.72%～4.05%,在酱油中18种氨基酸的加标回收率为90.2%～103.7%。该方法前处理过程简单,分离时间短,是检测酱油中氨基酸的有效手段,可用于酱油的质量评定。

关键词: 6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯, 氨基酸, 超高效液相色谱, 酱油, 柱前衍生

Abstract: A rapid ultra-performance liquid chromatographic (UPLC) method was developed for the separation and determination of 18 amino acids in soy sauce by using 6-aminoquinolyl-N-hydroxyl-succinimidyl-carbamate (AQC) as pre-column derivatization reagent. The 18 amino acids were separated within 12 min using a BEH C18 column, ultraviolet （UV） detection at 260 nm, ammonium acetate-acetic acid-acetonitrile-water and acetonitrile-acetic acid as the mobile phases with combined gradient elution and gradient flow-rate. A linear relationship between the UV absorbance and the concentration of each amino acid was obtained with the correlation coefficient (r2) above 0.999. The detection limits were ranged from 0.032 mg/L to 0.12 mg/L for different amino acids, and the overall relative standard deviations from 0.72% to 4.05%. The recoveries of 18 analytes in a spiked soy sauce were from 90.2% to 103.7%. With simple pretreatment of the samples and shorter analysis time, the proposed method can be applied to determine amino acids in soy sauce.

Key words: 6-aminoquinolyl-N-hydroxy-succinimidyl-carbamate （AQC）, amino acids, soy sauce, ultra-performance liquid chromatography (UPLC), pre-column derivatization