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
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