食品科学2018，Vol.39Issue(4)：165-170,6.DOI:10.7506/spkx1002-6630-201804025
AccQ-Tag柱前衍生反相高效液相色谱法测定谷物及食糜中氨基酸含量
Determination of Amino Acid Contents in Cereals and Ileal Digesta by Reversed-Phase High Performance Liquid Chromatography with AccQ-Tag Pre-column Derivatization Method
摘要
Abstract
The contents of amino acids in different cereals and ileal digesta were determined by reversed-phase high performance liquid chromatography (RP-HPLC) according to the AccQ-Tag method. The ratio of material to acidolysis agent was optimized in this paper. Quantification was performed using an internal standard method after pre-column derivatization with 6-aminoquinolinyl-N-hydroxysuccinimidate (AQC). The analysis procedure was carried out using a Waters 2695 HPLC system fitted with a Waters AccQ-Tag Nova-PakTMC18column (3.9 mm × 150 mm, 4 μm) and a Waters 2475 fluorescence detector (Waters Corp., USA) at an excitation wavelength of 250 nm and emission wavelength of 395 nm at 37 ℃. Sodium acetate (pH 4.95), acetonitrile and water were used respectively as mobile phases A, B and C for gradient elution and the injection volume was 10 μL. The results showed that the optimum ratio of material to acidolysis agent was 1:100 (mg/μL). The calibration curves of 17 amino acids were linear within the range of 25–500 μmol/L (12.5–250 μmol/L for cystine) with correlation coefficients (r) of 0.999 6–0.999 9. The average recoveries were in the range of 97.56%–103.92% with relative standard deviation (RSD) of 0.31%–2.75% (n = 3). This method is rapid, simple and accurate for the determination of amino acids in cereals and ileal digesta.关键词
反相高效液相色谱法/AccQ-Tag柱前衍生/谷物/食糜/氨基酸Key words
reversed-phase high performance liquid chromatography (RP-HPLC)/AccQ-Tag pre-column derivatization/cereals/ileal digesta/amino acids分类
轻工纺织引用本文复制引用
韩粉丽,韩飞,李爱科,陈曦,宋歌,范柳萍,余治权..AccQ-Tag柱前衍生反相高效液相色谱法测定谷物及食糜中氨基酸含量[J].食品科学,2018,39(4):165-170,6.基金项目
公益性行业(粮食)科研专项(201513003-8) （粮食）
