高效液相色谱法测定桑叶γ-氨基丁酸和谷氨酸
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摘要
研究一种快速、精准可同时检测桑叶中γ-氨基丁酸和谷氨酸含量的色谱方法。通过研究料液比、提取温度、时间对提取率的影响,确立最佳提取条件为:80℃条件下,料液比5∶1(mg/mL),分两次浸提,每次5 min。以2,4-二硝基氟苯为柱前衍生试剂,Agilent TC-18色谱柱,以乙酸钠缓冲溶液-乙睛(50%)为流动相,梯度洗脱。此方法在0～0.05 mg/mL范围内线性关系良好,γ-氨基丁酸和谷氨酸测定结果相对标准偏差均<2%,平均加标回收率分别为88.024%、114.64%,所建立的方法可以用于桑叶中γ-氨基丁酸和谷氨酸含量的检测。为γ-氨基丁酸桑叶茶开发和利用提供理论与技术支持。
A rapid and accurate chromatographic method for simultaneous determination of γ-aminobutyric acid and glutamic acid in mulberry leaves was developed.By studying the effect of material to liquid ratio,extraction temperature and time on the extraction rate,the optimum extraction conditions were as follows:under the condition of 80 ℃,the ratio of material to liquid was 5 ∶ 1(mg/mL),and it was extracted two times,and each time was 5 min.2,4-two nitrobenzene was used as the pre column derivatization reagent,Agilent TC-18 column,and sodium acetate buffer solution-acetonitrile(50 %)as mobile phase,gradient elution.This method had a good linear relationship in the range of 0-0.05 mg/mL.The relative standard deviation value of γ-aminobutyric acid and glutamic acid was < 2 %,and the average recovery rate was 88.024 % and 114.64 %respectively.The method could be used for the detection of the content of γ-aminobutyric acid and glutamic acid in mulberry leaves.This study provides theoretical and technical support for the development and utilization of mulberry leaf tea rich in γ-aminobutyric acid.
A rapid and accurate chromatographic method for simultaneous determination of γ-aminobutyric acid and glutamic acid in mulberry leaves was developed.By studying the effect of material to liquid ratio,extraction temperature and time on the extraction rate,the optimum extraction conditions were as follows:under the condition of 80 ℃,the ratio of material to liquid was 5 ∶ 1(mg/mL),and it was extracted two times,and each time was 5 min.2,4-two nitrobenzene was used as the pre column derivatization reagent,Agilent TC-18 column,and sodium acetate buffer solution-acetonitrile(50 %)as mobile phase,gradient elution.This method had a good linear relationship in the range of 0-0.05 mg/mL.The relative standard deviation value of γ-aminobutyric acid and glutamic acid was < 2 %,and the average recovery rate was 88.024 % and 114.64 %respectively.The method could be used for the detection of the content of γ-aminobutyric acid and glutamic acid in mulberry leaves.This study provides theoretical and technical support for the development and utilization of mulberry leaf tea rich in γ-aminobutyric acid.
引文
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[2]邵文韵.提高茶叶γ-氨基丁酸含量的方法优化及其在速溶茶产品开发中的应用[D].杭州:浙江大学,2014
[3]李东,孙家义.2,4-二硝基氟苯柱前衍生高效液相色谱法测定18种氨基酸[J].化学分析计量,2004,13(1):18-20
[4]黄柳舒,沈莲清,王向阳.改良比色法测桑叶中γ-氨基丁酸含量及其热稳定性研究[J].食品科技,2010,35(8):328-331,335
[5]夏玉玲,徐立,杨翠凤.桑叶中γ-氨基丁酸含量的测定[J].中国农学通报,2009,25(20):209-212
[6]冀宪领,盖英萍.桑叶中γ-氨基丁酸含量的测定及其影响因素的研究[J].蚕业科学,2007,33(2):176-180
[7]Shozaburo Kitaoka,Yoshihisa Nakano.Colorimetric Determination ofω-Amino Acids[J].The Journal of Biochemistry,1959,66(1):87-94
[8]张晖,徐永,姚惠源.纸层析法定量测定米胚芽中的γ-氨基丁酸[J].无锡轻工大学学报(食品与生物技术),2004,23(2):101-103
[9]赵宏飞,宋伟,张柏林.食品中三种γ-氨基丁酸检测方法比较[J].中国乳品工业,2008,36(11):51-55
[10]唐涛.氨基酸柱前衍生化HPLC方法发展及应用[D].南京:南京理工大学,2006
[11]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[J].食品科学,2015,36(6):178-182