Crops ›› 2016, Vol. 32 ›› Issue (1): 129-134.doi: 10.16035/j.issn.1001-7283.2016.01.024

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Method Optimization of Measuring γ-Aminobutyric Acid Contents in Rice by HPLC Coupled with Dabsyl Chloride Precolumn Derivatization

Shao Jinliang,Yang Dongshun,Fan Jianlin,Liu Xingyong,Du Lijuan,Wang Li,Wang Luxiang,Liu Hongcheng   

  1. Institute of Agriculture Quality Standards & Testing Technique,Yunnan Academy of Agricultural Sciences,Kunming 650223,Yunnan,China
  • Received:2015-10-22 Revised:2015-12-01 Online:2016-02-15 Published:2018-08-25
  • Contact: Luxiang Wang,Hongcheng Liu

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Abstract:

A method of measuring γ-aminobutyric acid (GABA) content in rice was established by high performance liquid chromatography coupled with dabsyl chloride pre-column derivatization. Using 80% ethanol as extracting solvent, and then 1.0mL extracted fluid pre-column derivatization with 0.20mL sodium bicarbonate, 0.40mL dabsyl chloride at 70℃ and 20min, chromatographic analysis was performed on C18 column eluted with an acetonitrile-sodium acetate trihydrate and ultraviolet detector at 436nm. The results showed that the fortified recovery of GABA was more than 87.60% and the standard deviation was less than 9.07%. A good linear range from 1.0mg/L to100.0mg/L concentration of GABA with correlation of 0.9995 (n=3) was obtained. GABA content in 14 rice and brown rice (6.38-108.7mg/kg) were determined by the method, and the content of GABA in Hunan rice was the highest.

Key words: DABS-Cl, Pre-column derivatization, HPLC, GABA, Rice

Fig.1

Effect of sodium bicarbonate solution volume on GABA peak area"

Fig.2

Effect of DABS-Cl derivatives solution volume on GABA peak area"

Fig.3

Effect of derivatives temperature on GABA peak area"

Fig.4

Effect of derivatives time on GABA peak area"

Fig.5

The stabilities of the derivatization products of GABA"

Fig.6

Elution ability of different mobile phase on GABA"

Fig.7

Effect of different flow speed on GABA retention time"

Fig.8

Maximum absorption wavelength of GABA"

Table 1

Accuracy and recoveries of standard addition of GABA"

添加水平（mg/kg）
Adding level		 精米Rice 添加水平（mg/kg）
Adding level		 糙米Brown rice
回收率（%）Recovery rate RSD（%） 回收率(%)Recovery rate RSD(%)
5 87.60 9.07 25 96.56 6.32
10 94.13 2.32 50 106.10 2.81
25 104.50 3.32 100 92.54 1.83

Table 2

The analysis of GABA contents in samples mg/kg"

样品来源Samples from GABA
精米Rice 糙米
Brown rice
中国云南（红河）Yunnan(Honghe)of China 6.38 34.5
中国云南（玉溪）Yunnan(Yuxi) of China 19.40 50.6
中国四川Sichuan of China 20.90 58.2
中国江苏Jiangsu of China 22.30 62.1
中国安徽Anhui of China 23.40 71.5
中国湖南Hunan of China 25.40 108.7
泰国Thailand 24.50 96.9
[1] 贾琰, 赵宏伟, 王敬国 , 等. 逆境胁迫下作物中-氨基丁酸代谢及作用的研究进展.作物杂志, 2014(5):9-15.
[2] 吴琴燕, 马圣洲, 张文文 , 等. 柱前衍生高效液相色谱法检测红茶中γ-氨基丁酸含量. 江苏农业学报, 2014,30(6):1534-1536.
[3] 郭超, 王昌禄, 陈勉华 , 等. HPLC法测定发酵乳中γ-氨基丁酸条件的优化. 现代食品科技, 2014,30(1):222-226,86.
[4] 程威威, 吴跃, 林亲录 , 等. 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. 中国农学通报, 2014,30(9):279-284.
[5] 刘三才 . 我国青稞的质量与品质研究进展.作物杂志, 2014(4):1-5.
[6] 郑鸿雁, 赵炜彤, 昌妍希 . 响应面法优化假丝酵母Y6产γ-氨基丁酸发酵工艺. 食品科学, 2015,36(9):130-135.
doi: 10.7506/spkx1002-6630-201509024
[7] 杜娟, 杨涛, 曾亚文 , 等. 分光光度法检测不同地区和类型稻种功能性成分的初步研究. 云南大学学报(自然科学版), 2011,33(2):232-237.
[8] 赵国群, 王维 . 比色法快速测定γ-氨基丁酸含量. 食品研究与开发, 2015,36(10):85-89.
[9] 张珺, 何义雁, 吴卫国 . 贝特洛显色法快速测定红米中γ-氨基丁酸的研究. 粮食与油脂, 2014,27(10):40-42.
[10] 李鹏飞, 王燕, 陶蓓蓓 , 等. 稳定同位素iTRAQ标记/高效液相色谱–串联质谱法同时定量分析人体中42种氨基酸及典型病例. 分析化学, 2012,40(5):762-767.
doi: 10.3724/SP.J.1096.2012.10917
[11] 苗雨田, 杨悠悠, 王浩 , 等. 全自动氨基酸分析仪法测定不同年份黄酒中游离氨基酸的含量. 食品安全质量检测学报, 2015,6(4):1154-1161.
[12] 顾宇翔, 葛宇, 巢强国 . OPA衍生-HPLC测定豆制品和发酵糙米中的γ-氨基丁酸. 食品工业, 2013,34(9):218-220.
[13] 陈佳, 魏锋, 程显隆 , 等. 柱前衍生化HPLC法测定不同基原珍珠粉及贝壳粉中7种氨基酸的含量. 中药材, 2015,38(4):693-696.
[14] 王欣美, 王柯 . 太子参氨基酸类成分的柱前衍生-HPLC指纹图谱分析. 中国卫生检验杂志, 2014,24(4):508-511.
[15] 周婷, 程威威, 林亲录 , 等. 丹磺酰氯柱前衍生发芽糙米中γ-氨基丁酸的HPLC分析. 食品科技, 2014,39(1):282-286.
[16] 赵英莲, 牟德华, 李艳 . 2,4-二硝基氟苯柱前衍生HPLC检测树莓中游离氨基酸. 食品科学, 2015,36(6):178-182.
doi: 10.7506/spkx1002-6630-201506033
[17] 洪健, 叶立斌, 励建荣 , 等. 2,4-二硝基氟苯柱前衍生RT-HPLC法测定γ-氨基丁酸. 中国粮油学报, 2011,26(4):104-106,111.
[18] 张俊杰, 赵春霞, 赵燕妮 , 等. 基于柱前衍生-超高效液相色谱-质谱联用技术的植物提取液中氨基化合物代谢谱分析. 色谱, 2015,33(6):613-621.
doi: 10.3724/SP.J.1123.2015.01028
[19] 胡家伟, 高榕, 曹敏杰 , 等. 水产食品中组胺的丹磺酰氯柱前衍生反相高效液相色谱测定方法的建立及应用. 食品科学, 2014,35(8):283-288.
doi: 10.7506/spkx1002-6630-201408057
[20] 董伟峰, 李宪臻, 林维宣 . 丹磺酰氯作为生物胺柱前衍生试剂衍生化条件的研究. 大连轻工学院学报, 2005,24(2):115-118.
[21] 赵怀鑫, 孙志伟, 夏莲 , 等. 新型荧光衍生试剂用于尿样中痕量游离雌二醇和雌三醇的反相高效液相色谱-荧光检测和质谱定性. 色谱, 2009,27(2):164-168.
[22] 曾琪, 张缙, 徐敦明 , 等. 柱前衍生–高效液相色谱–荧光检测法测定罐头食品中的尿素. 色谱, 2015,33(1):80-83.
doi: 10.3724/SP.J.1123.2014.08024
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