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Simultaneous Measurement of the Galacturonate and Neutral Sugar Contents of Pectic Substances by an Enzymic-HPLC
S Matsuhashi1, S Inoue1, C Hatanaka1
1a Department of Applied Biochemistry, Faculty of Applied Biological Science , Hiroshima University , Higashi Hiroshima 724 , Japan.
Abstract:
An enzymic-HPLC method was successfully applied to the simultaneous analysis of the galacturonate and neutral sugar contents of pectic substances. A mixture of seven neutral sugars that are present in oridnary pectins was eluted as one peak on a Shodex SUGAR SH1821 column, using 0.001 N sulfuric acid as the eluent; the peak was completely separated from that of galacturonate. No difference in the peak-area ratios of individual neutral sugars to glycerol (internal standard) was found among the seven; the relationship between the peak response and concentration was strictly linear throughout the entire concentration range studied. Seventeen pectic samples, including pectins, pectates, and rhamnogalacturonan fragments, were completely prehydrolyzed by Driselase, an industrial enzyme product from Irpex lacteus, and then analyzed for their constituent sugar contents. The enzymic-HPLC method was simple, accurate, and particularly useful for routine pectin analyses.
