微波水解衍生高效液相色谱法测定饲料中的氨基酸

建立了一种微波水解衍生高效液相色谱同时测定饲料中17种氨基酸含量的方法。采用2,4-二硝基氯苯作为柱前衍生试剂,利用C_(18)色谱柱分离。二极管阵列检测器进行检测,检测波长为360 nm,并对微波水解时间及温度进行优化。17种氨基酸在2.5~50 mg/L范围内呈良好线性关系,相关系数为0.990 7~0.999 9;相对标准偏差为0.88%~4.2%;加标回收率为90.6%~107.2%;检出限为0.15~2.37 mg/L。研究结果表明,150℃下微波水解16 min的结果与传统加热水解(110℃,24 h)的效果基本相同。该方法分析时间较短,灵敏度较高,可用于饲料中氨基酸含量的检测。

Determination of Amino Acids in Feed by High Performance Liquid Chromatography with Microwave Hydrolysis Derivatization

A high performance liquid chromatographic (HPLC) method with microwave hydrolysis derivation was developed for the simultaneous determination of 17 amino acids in feed.The method was based on pre-column derivatization with 2,4-dinitro-chlorobenzene.The compounds were separated on a C18 column.And the conditions of microwave hydrolysis were optimized.The detection was performed using a diode array detector at 360 nm.The calibration curves of 17 amino acids all exhibited good linearities in the range of 2.5-50 mg/L.The correlation coefficients(r2) for the determination varied between 0.990 7 and 0.999 9 with the relative standard deviations (RSDs) of 0.88％ to 4.2％,and the recoveries of 17 amino acids were in the range of 90.6％-107.2％ with the limits of detection of 0.15-2.37 mg/L.The results showed that the results of microwave hydrolysis in 16 min at 150 ℃ are basically the same as those of traditional heating hydrolysis(110 ℃,24 h).With the advantages of short analysis time,high sensitivity,the method could be used for the determination of amino acids in feed.