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中国农学通报

中国农学通报 ›› 2014, Vol. 30 ›› Issue (9): 279-284.doi: 10.11924/j.issn.1000-6850.2013-1455

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HPLC法测定发芽糙米中γ-氨基丁酸中不同衍生方式的比较研究

程威威 吴跃 林亲录 周婷 丁玉琴 王晓映   

  • 收稿日期:2013-05-23 修回日期:2013-06-27 出版日期:2014-03-25 发布日期:2014-03-25
  • 基金资助:
    “十二五” 国家科技支撑计划课题 “商品米品质提升与高效利用关键技术研究与示范” (2012BAD34B0202)。

The Comparative Study of Different Derivative Methods in the Determination of γ-Aminobutyric Acid in Germinated Brown Rice Acid by HPLC

  • Received:2013-05-23 Revised:2013-06-27 Online:2014-03-25 Published:2014-03-25

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摘要： 针对HPLC法测定发芽糙米中γ-氨基丁酸(GABA)所用衍生剂缺乏系统的比较分析，研究了3种常见衍生剂及其色谱方法比较。以邻苯二甲醛(OPA)、丹磺酰氯(Dansyl-Cl)、6-氨基喹啉基-N-羟基琥珀酰亚氨基氨基甲酸酯(AQC)3种物质作为柱前衍生剂，色谱柱为intertsil ODS-C18柱（前两者使用）、Waters AccQ Tag氨基酸分析柱，均为梯度洗脱，紫外吸收波长分别为332 nm、386 nm、250 nm，线性方程分别为Y=9.0×106X+2062.5(R2=0.9989)、Y=7.0×106X+11337(R2=0.9982)、Y=7.1×106X+10025(R2=0.9999)，检出限分别为0.005 mg/mL、0.01 mg/mL、0.001 mg/mL，同一样品中测得GABA峰面积的RSD分别为1.16%、1.98%、0.94%，加标回收率分别为98.5%~106.9%、95.8%~110.6%、99.4%~103.4%，同一样品中测得GABA含量分别为11.24、10.67、13.47 mg/100 g干重。从上述结果得知，HPLC法测定GABA的3种衍生剂中，测定结果最准确的是AQC，其次是OPA和Dansyl-Cl。AQC作为一种新型衍生剂，在γ-氨基丁酸乃至氨基酸分析中前景广阔。

关键词: 马铃薯, 马铃薯, 旱作, 覆膜方式, 产量, 土壤水分

Abstract: The derivative agent using in the determination ofγ-aminobutyric acid by HPLC lacked systematic comparative analysis. So this paper studied the comparison of three kinds of common derivative agent and chromatographic process. A gradient elution was used for the separation and quantification of GABA precolumn derivatization with OPA, Dansyl-Cl, AQC. The column were intertsil ODS-C18 (by the first two), waters AccQ·Tag for free amino acids. GABA was determined with UV detector at 332 nm, 386 nm, 250 nm, and linear equations were Y=9.0×106X+2062.5(R2=0.9989), Y=7.0×106X+11337(R2=0.9982), Y=7.1×106X+10025 (R2=0.9999), and detection limits were 0.005 mg/mL, 0.01 mg/mL, 0.001 mg/mL, and RSD of peak area of GABA measured in the same sample was 1.16%, 1.98%, 0.94%, and the recoveries were 98.5%-106.9%, 95.8%-110.6%, 99.4%-103.4%, GABA content measured in the same sample was 11.24, 10.67, 13.47 mg/ 100 g dry weight, respectively. We learned from the above results, that in three kinds of derivative agent determining GABA by HPLC, the most accurate one is AQC, followed by OPA and Dansyl-Cl, AQC, as a new derivative agent, has the broad prospect inγ-aminobutyric acid and even amino acid analysis.