Method for simultaneously determining content of 2-keto-L-gulonic acid, content of vitamin C and content of 2-keto-L-gulonic acid methyl ester

Abstract

The invention provides a method for method for simultaneously determining content of 2-keto-L-gulonic acid, content of vitamin C and content of 2-keto-L-gulonic acid methyl ester, comprising the following steps of: dissolving and diluting a reference substance with mobile phase to prepare a reference substance solution, wherein the mobile phase is composed of organic solvent and buffer solution, the organic solvent is selected from methyl alcohol, acetonitrile or tetrahydrofuran, and the buffer solution is selected from phosphoric acid, phosphate, acetic acid or acetate buffer solution; dissolving and diluting a sample with the mobile phase to prepare a sample solution; injecting same amount of the reference substance solution and the sample solution into a chromatographic column, and performing chromatographic separation and chromatographic detection by a high performance liquid chromatography to generate liquid chromatogram maps of the reference substance solution and the sample solution, wherein the chromatographic column is selected from a C-8 column, a C-18 column or an amino column, and wavelength for detection is 210 nm; and obtaining response signals of the determined constituents in the liquid chromatogram maps, and respectively calculating the contents of the determined constituents in the sample by an external standard method. The method provided by the invention can simultaneously determine the contents of the three matters with faster speed.

Description

technical field [0001] The invention relates to the technical field of chemical analysis, in particular to a method for simultaneously measuring the contents of 2-keto-L-gulonic acid, vitamin C and 2-keto-L-gulonic acid methyl ester. Background technique [0002] Usually, the vitamin C esterification conversion process includes the following steps: an esterification reaction step, 2-keto-L-gulonic acid (also known as: 2-keto-L-gulonic acid) is catalyzed with concentrated sulfuric acid in methanol Esterification generates 2-keto-L-gulonic acid methyl ester; conversion reaction step, using NaHCO 3 Convert 2-keto-L-gulonic acid methyl ester to vitamin C sodium salt; acidification reaction step, add H to vitamin C sodium salt 2 SO 4 , acidified into vitamin C. [0003] In the above process stages, the possible products are: vitamin C, 2-keto-L-gulonic acid methyl ester. The esterification reaction is a reversible reaction, while the esterification, conversion, and acidificat...

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"And, this document only discloses the method of using high performance liquid chromatography to determine two components (gulonic acid and guronic acid methyl ester) in the production process of vitamin C

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