The Application Of Ion Chromatography Analysing Water Soluble Metabolites In Bacteria Broth

There are some indispensable and water soluble substances in bacteria broth, suchas inorganic ions, organic acids, nucleotides, amino acids, carbohydrates and so on. Toimprove strains, analyze gene functions, optimize cell systems and design fermentationexperiment, researchers aim to quantitatively understand the metabolism of bacteriaand determine metabolic flux date. Therefore, the precise and efficient determinationthese metabolisms is of great significant.In this study, ion chromatagraphy coupled with single column was employed todetermine water soluble metabolites in bacteria broth. Multiple valves and pumpssystems were established successfully aimed to resolve the interference of amino acidsand carbohydrates. The valve switching and seperation conditions were optimized. Theprecise quantify of amino acids and carbohydrates were obtained. The method laids agood foundation for the simultaneous determination of multicomponents in cytosolcomplex samples.The content of this research mainly includes three parts:1. Conventional single column ion chromatographies were adopted to determineinorganic anions and ions, organic acids, carbohydrates, alcohols and amino acidsrespectively.2. The automatic switching ion chromatography systems coupled with multiplevalves and pumps were established. The systems with double valves and pumps, threevalves and pumps, double valves and three pumps were established.3. A three valves-three pumps system and a dual valves-three pumps system were used to simultaneously determine amino acids and carbohydrates in broth. The choiceof trap column, flow conditions, valve-switching time and flow conditions of twoanalitical columns were investigated. The optimized method was evaluated with amean coefficient of>0.99and repeatability of0.5-4.6%for eight replicates. Themethod was successfully applied in the analysis of amino acids and carbohydrates inaseptic media and in extracellular culture media of three phenotypes of Clostridiumthermocellum. The results of actual sample analysis is satisfactory.