摘要
用AccQ-Tag法测定不同谷物及食糜中氨基酸的含量,并优化原料与酸解剂的比例。采用内标法定量,6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯为柱前衍生剂,用反相高效液相色谱仪,2475荧光检测器(激发波长250 nm、发射波长395 nm),Waters AccQ-Tag Nova-Pak^(TM)C_(18)柱(3.9 mm×150 mm,4μm),柱温37℃,以醋酸钠(p H 4.95)缓冲液为流动相A,纯乙腈为流动相B,水为流动相C进行梯度洗脱,进样量为10μL。结果发现,5 mg原料的最优酸解剂为500μL;17种氨基酸线性回归方程在25~500μmol/L范围内线性良好(胱氨酸(Cys)浓度在12.5~250μmol/L之间),相关系数r为0.999 6~0.999 9,测定的平均回收率为97.56%~103.92%(n=3),相对标准偏差为0.31%~2.75%(n=3)。本法快速简便,能够适用于谷物及食糜中氨基酸含量的测定。
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-Pak-（TM）C_（18） column（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（p H 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.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第4期165-170,共6页
Food Science
基金
公益性行业(粮食)科研专项(201513003-8)
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
reversed-phase high performance liquid chromatography （RP-HPLC）
AccQ-Tag pre-column derivatization
cereals
ileal digesta
amino acids
