Determination of Homogentisic Acid in Bamboo Shoots by Column Chromatography on Sephadex G-10 and Reversed-Phase High-Performance Liquid Chromatography

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  • セファデックスG-10カラムクロマトグラフィー及び高速液体クロマトグラフィーによるタケノコのホモゲンチジン酸の測定法
  • セファデックス G-10 カラム クロマトグラフィー オヨビ コウソク エキタ

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A method for the separation and quantification of homogentisic acid (HGA) in bamboo shoots has been developed by the use of Sephadex G-10 column chromatography and high-performance liquid chromatography (HPLC). For the first purification step, the HGA in bamboo shoots was extracted with methanol: water (70:30, v/ v) and then fractionated into 5ml-fractions from Sephadex G-10 column with 0.05 M phosphate buffer (pH=8.0). The final purification was performed on HPLC (strong cation resin column), eluted with 0.2% phosphoric acid (pH=2.0) at the flow rate of 0.7ml/min. The column temperature was maintained at 55°C and the column effluent was monitered by uv spectrometry at 210nm. The identification of HGA was performed on gas chromatography-mass spectroscopy.<br>1. From the result of the elution pattern of authentic HGA, it was found that HGA was eluted in fractions 16-26.<br>2. Partially purified extract of HGA from the Sephadex column was separated and quantificated on HPLC. The peak of HGA from HPLC was well separated with the retention time of 13min.<br>3. It was found that the overall recovery of HGA was 92.4% and also that HGA was fairly unstable above 100°C.<br>4. By the determination of HGA in the three sections of bamboo shoots by Sephadex G-10 column and HPLC, it was found that the HGA content was the highest in the top section and also that the content decreased from the top to the bottom section.

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