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中国精品科技期刊2020
段智红, 黄永梅, 吕应年, 梁力中, 黄燕霞, 叶华, 叶盛权. 2,4-二硝基氟苯柱前衍生HPLC-UV法测定海稻米中的γ-氨基丁酸含量[J]. 食品工业科技, 2019, 40(2): 257-261,270. DOI: 10.13386/j.issn1002-0306.2019.02.044
引用本文: 段智红, 黄永梅, 吕应年, 梁力中, 黄燕霞, 叶华, 叶盛权. 2,4-二硝基氟苯柱前衍生HPLC-UV法测定海稻米中的γ-氨基丁酸含量[J]. 食品工业科技, 2019, 40(2): 257-261,270. DOI: 10.13386/j.issn1002-0306.2019.02.044
DUAN Zhi-hong, HUANG Yong-mei, LV Ying-nian, LIANG Li-zhong, HUANG Yan-xia, YE Hua, YE Sheng-quan. Determination of γ-aminobutyric Acid in Sea Rice by HPLC-UV Method with 2,4-Dinitrofluorobenzene Pre-column Derivatization[J]. Science and Technology of Food Industry, 2019, 40(2): 257-261,270. DOI: 10.13386/j.issn1002-0306.2019.02.044
Citation: DUAN Zhi-hong, HUANG Yong-mei, LV Ying-nian, LIANG Li-zhong, HUANG Yan-xia, YE Hua, YE Sheng-quan. Determination of γ-aminobutyric Acid in Sea Rice by HPLC-UV Method with 2,4-Dinitrofluorobenzene Pre-column Derivatization[J]. Science and Technology of Food Industry, 2019, 40(2): 257-261,270. DOI: 10.13386/j.issn1002-0306.2019.02.044

2,4-二硝基氟苯柱前衍生HPLC-UV法测定海稻米中的γ-氨基丁酸含量

Determination of γ-aminobutyric Acid in Sea Rice by HPLC-UV Method with 2,4-Dinitrofluorobenzene Pre-column Derivatization

  • 摘要: 为建立海水稻活性成分γ-氨基丁酸(GABA)的高效液相色谱-紫外检测(HPLC-UV)方法。以2,4-二硝基氟苯(FDNB)为衍生化试剂,采用高效液相色谱法测定了精白米、玉米、海稻米、发芽海稻米24 h、发芽海稻米48 h、发芽海稻米72 h的γ-氨基丁酸(GABA)含量,研究了FDNB的投量,醋酸铵溶液-乙腈作为流动相以及样品的pH对GABA衍生物保留时间的影响,并考察了该方法的精密度、重复性和稳定性及加标回收率。结果表明,发芽海稻米24 h中GABA的含量最高,为34.35 μg/mg,玉米、未发芽海稻米、发芽海稻米48 h、发芽海稻米72 h中GABA的含量分别为为3.74、3.63、17.03、9.54 μg/mg,而精白米中GABA含量几乎为零;FDNB投量为0.3 mL时,可确保反应完全;用0.5%醋酸铵水溶液:乙腈(V:V)=85:15作流动相克服了泵压不稳定的现象;用硼酸盐缓冲液对衍生化后的样品进行稀释(1、5、10、15、20、40倍)后,样品的pH在9.0左右,GABA衍生物的保留时间几乎不漂移,在15.8 min左右;该方法精密度高、重复性和稳定性好,相对标准偏差值分别是0.08%、0.66%和0.60%;样品在24 h内稳定,回收率在99.29%~101.35%。结论 本实验方法简便、结果可靠,可用于高含量GABA功能性食品的质量控制。

     

    Abstract: 2, 4-dinitrofluorobenzene (FDNB) was used as the derivatization reagent in this study to establish a method for the determination of γ-Aminobutyric acid (GABA) of polished rice, corn, sea rice, germinated sea rice for 24 h, germinated sea rice for 48 h, germinated sea rice for 72 h. The effect on the retention time of GABA derivatives of FDNB volume, ammonium acetate solution-acetonitrile as mobile phase and the pH value of the sample were studied using an HPLC-UV analyzer. The precision, repeatability and stability of the method and the recovery rate of the standard were also investigated, the result was that the content of GABA in the germinated sea rice was the highest which was 34.35 μg/mg, and the content of GABA in corn, ungerminated sea rice, germinated sea rice for 48 h and germinated sea for 72 h were 3.74, 3.63, 17.03 and 9.54 μg/mg, respectively. while the GABA content in polished rice was almost zero;When the FDNB dosage was up to 0.3 mL, the reaction is guaranteed to be complete;Using 0.5% ammonium acetate aqueous solution:acetonitrile (V:V)=85:15 as the mobile phase to overcome the instability of pump pressure;After diluted 1, 5, 10, 15, 20 and 40 time by borate buffer the pH value of samples was aroud 9.0 which ensure that the retention time of GABA derivatives hardly drifts. The precision, repeatability and stability of the method was high and the relative standard deviation values were 0.08%, 0.66% and 0.60%, respectively. The sample was stable within 24 h and the recovery rate was 99.29%~101.35%. The method was simple and reliable, it could be used for quality control of high content GABA functional foods.germination (p<0.05). Sea rice is a plant resource with high GABA content, and the time of germination was shown to had a significant effect on the content of GABA. The sea rice with high content of GABA may be potentially applied in functional food. The simple and reliable method can be used for the quality control of functional food with high GABA content.

     

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