食品科学

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2,4-二硝基氟苯柱前衍生HPLC检测树莓中游离氨基酸

赵英莲1,牟德华1,李 艳1,2,*   

  1. 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
  • 出版日期:2015-03-25 发布日期:2015-03-17
  • 通讯作者: 李 艳
  • 基金资助:

    河北省科技支撑计划项目(11230604D-5-2)

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

ZHAO Yinglian1, MOU Dehua1, LI Yan1,2,*   

  1. 1. College of Biological Science and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, China;
    2. R & D Center for Fermentation Engineering of Hebei Province, Shijiazhuang 050018, China
  • Online:2015-03-25 Published:2015-03-17
  • Contact: LI Yan

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 pH 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0.39~2.87 μg/mL,定量限为1.29~9.47 μg/mL。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。

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

Abstract:

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 (pH
6.4) 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 1
and 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?2.87 μg/mL
and 1.29?9.47 μg/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.

Key words: raspberry, free amino acids, high performance liquid chromatography (HPLC), pre-column derivatization, 2,4-dinitrofluorobenzene

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