Determination of Amino Acids in Tea by High-Performance Liquid Chromatography

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  • 高速液体クロマトグラフィーによる茶のアミノ酸類の定量
  • コウソク エキタイ クロマトグラフィー ニ ヨル チャ ノ アミノサンルイ ノ

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A method for the determination of amino acids in tea was investigated by high-performance liquid chromatography with a fluorescence detection system using o-phthalaldehyde and 2-mercaptoethanol.<BR>An assay solution was prepared as follows:100mg of pulverized tea added norvaline (0.1mg) as the internal standard were extracted with hot water (80°C) for 30 min and the exract was made up to 100ml with water.<BR>After the extract was filtered through a 0.45μm membrane filter, the filtrate was mixed each time with o-phthalaldehyde solution. The mixture (10μl) was injected into the HPLC immediately. Separation of amino acids derivatives were achieved on a column (4.6mmφ×250mm) packed with ODS-120A (10μm, Toyo Soda), with operating under the following conditions.<BR>Solvent A is the mixture of 1000 ml of 12%V/V ethanol and 3.5ml of citric acid buffer (pH 6.0) and solvent B is that of 1000ml of 50%V/V ethanol and 3.5ml of citric acid buffer (pH 6.0). Gradient solution was used at a solvent flow rate of 1 ml/min from A to B within 60 min.<BR>This method will ensure an accuracy with 1.10-6.26% of coefficient of varience except methionine. and 94.7-103.0% of recovery.

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