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

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

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260nm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12min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0.032～0.12mg/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。

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.