Separation analysis of some food constituents and purification of enzyme in food by liquid chromatography
2004
Ohtsuki, K. (Kyoto Prefectural Univ. (Japan)) | Matsuo, A. | Obara, H. | Miyata, M. | Muranishi, T. | Sato, K. | Nakamura, Y.
Liquid chromatography and HPLC were successfully used for the determination of some food constituents and the purification of an enzyme in food. Free and total amino acids, water extracts and 6N-HC1 vapor hydrolysises in foods were analyzed in an ion-exchange HPLCamino acid analyzer with lithium citrate buffers and post-column fluorimetric detection with ophthalaldehyde - N-acetyl-L-Cysteine reagent. Tryptophan content in green tea, Mat-Cha, was also analyzed in an HPLC with fluorimetric detector, exci. 280nm, emi. 350nm, after barium hydroxide hydrolysis in a Teflon test tube and a 45 ml screw-capped vial in vacuo at 110 deg C. For the purification of ginger amino-peptidase, ion-exchange chromatography on Toyopearl SuperQ 650S and hydrophobic interaction chromatography on Octyl-Sepharose CL-4B were shown to be effective to speed up the purification. For the determination of the phytic acid in roasted brown rice flour and brown rice bread, an HPLC of Toyogel DEAE-5PW with a post-column detection system of Wade-reagent was very useful after 0.65 N HC1 treatment of the foods at 25 deg C for 2 hr. Analyses of cathechins and caffeine in various green-tea extracts were carried out on a single run of reverse phase HPLC and UV-detection at 280 nm and this method was convenient to evaluate the qualities among the various teas by comparing of their chromatograms. Liquid chromatography and HPLC were thus shown to be useful and simple for the analyses of the food constituents.
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