PURPOSE: To enable the simultaneous quantification of six sweetening components of leaf with high sensitivity in one column, by using a high-speed liquid chromatography having a moving phase prepared by adding a small quantity of salt to a mixture solvent of acetonitrile/water and a fixed phase of amino-base- combined type silica.
CONSTITUTION: The moving phase of a high-speed liquid chromatography is prepared by a method wherein a phosphate such as potassium dihydrogen phosphate, sulphate or the like is added to the mixture liquid of acetonitrile/water in the volume ratio of 75/25 to 90/10 so that the added salt accounts for 1W 5millimol/l. For the fixed phase thereof, amino-base chemical combination type silica (PNH2-10/S2504 prepared by Shimazu Seisakusho) is used. Components extracted from stevia leaf are quantified by the high-speed liquid chromatography thus prepared. By this method, it becomes possible to separate and quantify precisely the rebaudeoside B and steviol bioside from other stevioside, rebaudeoside A and C, and zurcoside A which can not be separated and quantified by conventional methods, together with the other four components.
ISHIDA YASUO
IWAMURA JIYUNICHI
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