Multi-Functional Separation Mode-Ion Chromatography Using L-Pyroglutamic Acid Eluent for Simultaneous Determination of Sugars, Organic Acids, and Ethanol during Multiple Parallel Fermentation of Rice Wine

Abstract

An effective evaluation method for understanding fermentation is required to support the implementation of newly developed yeast strains in brewing. Our group developed a multi-functional separation mode-ion chromatography (MFS-IC) with conductivity and refractive index detectors to separate key species (glucose, ethanol, and organic acids: pyruvate, citrate, L-malate, and succinate) with ethyl α-D-glucoside and glycerol to understand the fermentation behavior in brewing. Optimal chromatographic resolutions for determining eight different key species in rice wine by the MFS-IC were obtained in 45 min on low-molecular size-exclusion chromatographic separation columns modified with a strong cation-exchanger as the stationary phase with an eluent of L-pyroglutamic acid. Understanding the behavior of analytes in rice wine samples during rice wine fermentation enables the monitoring of the progress of saccharification, glycolysis, ethanol fermentation, and the tricarboxylic acid (TCA) cycle. These results are expected to contribute to understanding fermentation characteristics and implementing newly developed yeast strains in the fermentation field.