亲水作用色谱-串联质谱法测定茶叶中γ-氨基丁酸和茶氨酸
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摘要
粉碎后的茶叶样品用水在90℃水浴中提取10min(重复提取3次),提取液合并后,用水定容至50mL,移取1.00mL过0.22μm滤膜,采用亲水作用色谱-串联质谱法测定滤液中γ-氨基丁酸和茶氨酸的含量。以Waters Atlantis HILIC氨基色谱柱为固定相,以不同体积比的水和乙腈(9+1)溶液(含2mmol·L~(-1)乙酸铵和体积分数为0.1%的甲酸)的混合液为流动相进行梯度洗脱,串联质谱分析中采用电喷雾正离子模式和多反应监测模式。γ-氨基丁酸和茶氨酸的质量浓度均在5.00～500.00μg·L~(-1)内与其对应的峰面积呈线性关系,检出限(3S/N)依次为1.00,0.50μg·L~(-1)。方法用于茶叶样品的分析,加标回收率为97.8%～107%,测定值的相对标准偏差(n=9)为4.9%～7.1%。
The pulverized tea sample was extracted three times for 10 min with water in 90 ℃ water bath.Volume of the combined extract was made up to 50 mL with water.1.00 mL of the above solution was filtered on0.22μm filtering membrane.Hydrophilic interaction liquid chromatography with tandem mass spectrometry was applied to the determination ofγ-aminobutyric acid and theanine in the filtrate.Waters Atlantis HILIC NH_2 chromatographic column was used as stationary phase,and the mixture of water and acetonitrile(9+1)solution(containing 2 mmol·L~(-1) ammonium acetate and formic acid with 0.1% volume fraction)mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.Linear relationships between values of peak area and mass concentration ofγ-aminobutyric acid and theanine were kept in the same range of 5.00-500.00μg·L~(-1),with detection limits(3 S/N)of 1.00,0.50μg·L~(-1),respectively.The proposed method was applied to the analysis of tea samples,giving values of recovery found by standard addition method and RSDs(n=9)of determined values in the ranges of 97.8%~(-1)07%and 4.9%-7.1%,respectively.
The pulverized tea sample was extracted three times for 10 min with water in 90 ℃ water bath.Volume of the combined extract was made up to 50 mL with water.1.00 mL of the above solution was filtered on0.22μm filtering membrane.Hydrophilic interaction liquid chromatography with tandem mass spectrometry was applied to the determination ofγ-aminobutyric acid and theanine in the filtrate.Waters Atlantis HILIC NH_2 chromatographic column was used as stationary phase,and the mixture of water and acetonitrile(9+1)solution(containing 2 mmol·L~(-1) ammonium acetate and formic acid with 0.1% volume fraction)mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.Linear relationships between values of peak area and mass concentration ofγ-aminobutyric acid and theanine were kept in the same range of 5.00-500.00μg·L~(-1),with detection limits(3 S/N)of 1.00,0.50μg·L~(-1),respectively.The proposed method was applied to the analysis of tea samples,giving values of recovery found by standard addition method and RSDs(n=9)of determined values in the ranges of 97.8%~(-1)07%and 4.9%-7.1%,respectively.
引文
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[14]CARVALHO E,PUNYASIRI P A N,SOMASIRIH P P S,et al.Quantification of gamma-aminobutyric acid in Sri Lankan tea by means of ultra performance tandem mass spectrometry[J].Natural Product Communications,2014,9(4):525-528.
[15]赵贝贝,王丰琳,王风云,等.一种评价固定相表面硅羟基残余的新方法[J].分析科学学报,2008,24(5):517-521.
[16]黄亚辉,曾贞,郑红发,等.GABA茶中γ-氨基丁酸的TLC测定及提纯研究[J].氨基酸和生物资源,2008,30(3):11-15.
[2]师大亮,余继忠,郭敏明,等.γ-氨基丁酸茶的研究进展[J].杭州农业与科技,2009(5):23-26.
[3]TSUSHIDA T,MURAI T,OMORI M,et al.Production of a new type tea containing a high level of.GAMMA.-aminobutyric acid[J].Nippon Nogeikagaku Kaishi,1987,61(7):817-822.
[4]王芳,郑德勇,杨江帆.γ-氨基丁酸茶的研究进展[J].武夷学院学报,2009,28(5):39-43.
[5]LIN S D,MAU J L,HSU C A.Bioactive components and antioxidant properties ofγ-aminobutyric acid(GABA)tea leaves[J].LWT-Food Science and Technology,2012,46(1):64-70.
[6]ABDOU A M,HIGASHIGUCHI S,HORIE K,et al.Relaxation and immunity enhancement effects of gamma-aminobutyric acid(GABA)administration in humans[J].Biofactors,2006,26(3):201-208.
[7]ZHAO W,LI Y,MA W,et al.A study on quality components and sleep-promoting effects of GABAblack tea[J].Food and Function,2015,6(10):3393-3398.
[8]赵振军,包彬,邹万志.冲泡条件对茶树花茶茶汤内主要滋味成分的影响[J].湖北农业科学,2014,53(5):1096-1098.
[9]刘阳,陈根生,许勇泉,等.冲泡过程中西湖龙井茶黄酮苷类浸出特性及滋味贡献分析[J].茶叶科学,2015,35(3):217-224.
[10]TU Y F,YANG X,ZHANG S,et al.Determiantion of theanine and gamma-aminobutyric acid in tea by high performance-liquid chromatography with precolumn derivatization[J].Chinese Journal of Chromatography,2012,30(2):184-189.
[11]INOUE K,MIYAZAKI Y,UNNO K,et al.Stable isotope dilution HILIC-MS/MS method for accurate quantification of glutamic acid,glutamine,pyroglutamic acid,GABA and theanine in mouse brain tissues[J].Biomedical Chromatography,2016,30(1):55-61.
[12]HORIE H,KOHATA K.Analysis of tea components by high-performance liquid chromatography and high-performance capillary electrophoresis[J].Journal of Chromatography A,2000,881(1/2):425-438.
[13]王虹,康海宁,陈军辉,等.HPLC-APCI-MS法测定茶叶中的茶氨酸[J].茶叶科学,2008,28(6):20-26.
[14]CARVALHO E,PUNYASIRI P A N,SOMASIRIH P P S,et al.Quantification of gamma-aminobutyric acid in Sri Lankan tea by means of ultra performance tandem mass spectrometry[J].Natural Product Communications,2014,9(4):525-528.
[15]赵贝贝,王丰琳,王风云,等.一种评价固定相表面硅羟基残余的新方法[J].分析科学学报,2008,24(5):517-521.
[16]黄亚辉,曾贞,郑红发,等.GABA茶中γ-氨基丁酸的TLC测定及提纯研究[J].氨基酸和生物资源,2008,30(3):11-15.