2,4-二硝基氟苯柱前衍生HPLC检测树莓中游离氨基酸  

High Performance Liquid Chromatographic Analysis of Free Amino Acids in Raspberry Using Pre-Column Derivatization with 2,4-Dinitrofluorobenzene

作　　者：赵英莲[1] 牟德华[1] 李艳[1,2] 

机构地区：[1]河北科技大学生物科学与工程学院,河北石家庄050018 [2]河北省发酵工程技术研究中心,河北石家庄050018

出　　处：《食品科学》2015年第6期178-182,共5页Food Science

基　　金：河北省科技支撑计划项目(11230604D-5-2)

摘　　要：目的:建立一种简单、快速、可同时检测22种氨基酸的色谱方法,并利用此方法对树莓中的游离氨基酸进行检测和分析,为树莓的开发提供科学依据。方法:以2,4-二硝基氟苯为柱前衍生试剂,Hypersil ODS2色谱柱,流动相A为40 mmol/L p H 6.4乙酸钠溶液,流动相B为体积分数50%乙腈-水溶液,梯度洗脱,反相高效液相色谱,二极管阵列检测器,检测波长360 nm。结果:22种氨基酸线性方程的相关系数范围为0.993 1~1.000 0,相对标准偏差在0.95%~4.98%之间,加标回收率为97.10%~103.39%,最低检出限为0.39~2.87μg/m L,定量限为1.29~9.47μg/m L。利用所建方法检测出树莓中含有14种游离氨基酸,其中6种人体必需氨基酸,占总氨基酸含量的29.36%。结论:所建方法操作简便、重复性好,树莓氨基酸含量丰富,有开发价值。A simple and rapid reversed-phase high-performance liquid chromatographic（RP-HPLC） method for the simultaneous analysis of 22 free amino acids was established and applied to analyze free amino acids in raspberries.The method was based on pre-column derivatization with 2,4-dinitrofluorobenzene（DNFB）.The compounds were separated on a Hypersil ODS2 column.The mobile phase used was a mixture of phase A containing 40 mmol/L sodium acetate buffer（pH6.4） and phase B containing 50%acetonitrile-water through the gradient elution.Detection was performed using a diode array detector at 360 nm.The method showed good linearity with coefficients（R2） of determination varying between 0.993 1and 1.000 0 and precision（relative standard deviation,RSD） ranging from 0.95%to 4.98%,and recovery rates were in the range between 97.10%and 103.39%for 22 amino acids.The limits of detection and quantification were 0.39-2.87μg/mL and 1.29-9.47μg/mL,respectively.This method proved to be simple and reproducible and was applied to detect 14 free amino acids in raspberries,including 6 essential amino acids（EEAs）.The contents of EEAs accounted for 29.36%of the total amino acids.The amino acid content of raspberries is abundant,indicating that this kind of berry has the potential to be developed into functional foods.

关 键 词：树莓 游离氨基酸 高效液相色谱法 柱前衍生 2 4-二硝基氟苯 

分 类 号：O657.72[理学—分析化学] TS255.7[理学—化学]