2,4-二硝基氟苯柱前衍生法测定植物中谷氨酸脱羧酶的活力  

Determination of Glutamate Decarboxylase Activity in Plant by Pre-column 30 Derivatization with 2,4-Dinitrophenylhydrazine

作　　者：吕莹果[1] 张晖[1] 孟祥勇[1] 王立[1] 郭晓娜[1] 陶冠军[1] 

机构地区：[1]江南大学食品学院,无锡214122

出　　处：《分析化学》2009年第3期347-350,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.20671044);"新世纪优秀人才支持计划"项目(No.NCET-06-0488);江南大学食品学院青年博士科研创新开放基金资助

摘　　要：建立了2,4-二硝基氟苯(FDNB)柱前衍生反相高效液相色谱紫外检测法测定植物中谷氨酸脱羧酶(GAD)活力的方法。利用GAD催化L-谷氨酸脱羧转变成γ-氨基丁酸(CABA)的性质,用FDNB柱前衍生GABA,HPLC Lichrospher C18色谱柱分离,在360nm下检测衍生物,从而得到生成GABA的含量,及不同植物组织中的GAD活力。本方法的线性范围0.2～5g/LGABA(r=0.9954),精密度RSD为0.21%、回收率高,且衍生化产物在一个月内都能保持稳定。用本方法测定得出稻米和茶叶中均有一定的GAD活力。在稻米组织中,GAD主要分布在米胚芽中,米糠也具有相对较高的GAD活性,为开发富含GABA的糙米食品提供了依据。A validated, selective, and sensitive chromatographic method for the determination of glutamate decarboxylase （GAD） activity in plant tissues by determining γ-aminobutyric acid （GABA） has been developed using a single （gradient） system. The plant tissues were reacted with substrate L-glutamic acid at 40 %; for 2 h before test. Determination was carried out by pre-column reaction of the samples with 2,4-dinitrophe- nylhydrazine （FDNB） and separating the corresponding derivatives with a reversed phase HPLC on a 5 micro Lichrospher C18 column and UV detection（360 nm）. A linear quantitative response curve was generated over a concentration range of 0.2 - 5 g/L GABA with a correlation coefficient of 0. 9954. The precision of this method is high （RSD =0.21% ）. This method also possesses high recovery rate （ 〉99% ） and good stability. The exprimental results showed that the rice germ and rice bran had relatively high GAD activity, and the GAD activity of brown rice was as good as tea leaves. The method is specific for the determination of GAD activity inplant, and can be adopted in the determination of GABA content in food or bio-systems.

关 键 词：柱前衍生化 高效液相色谱 2 4-二硝基氟苯 谷氨酸脱羧酶 Γ-氨基丁酸 

分 类 号：Q946[生物学—植物学]