离子排斥色谱法同时测定2,3-丁二醇发酵液中的葡萄糖和木糖及各种代谢产物

Simultaneous Determination of Glucose,Xylose and Metabolites in 2,3-Butanediol Fermentation Broth by Ion-Exclusion Liquid Chromatography

针对2,3-丁二醇发酵体系,以高交联度磺化苯乙烯-二乙烯基苯共聚物多孔微球为固定相的AminexHPX-87H离子色谱柱为分析柱,利用示差折光检测器对该体系中的6种代谢产物(2,3-丁二醇、3-羟基丁酮、乙醇、乙酸、甲酸和乳酸)以及2种残留底物葡萄糖和木糖进行了分离。在65℃柱温下,以5 mmol/L H2SO4作为流动相,发酵液样品中底物葡萄糖和木糖及2,3-丁二醇等各种代谢产物的线性相关系数均在0.9990～0.9999之间,回收率为98.8%～103.2%,精密度为0.3%～2.1%。在最佳色谱条件下,能够同时测定2,3-丁二醇发酵体系中的残留底物和各种代谢产物的含量,适合于实时定量监测2,3-丁二醇发酵过程。

An ion-exclusion liquid chromatographic method on a sulfonated polystyrene divinyl benzene column（Aminex HPX-87H） with a refractive index detector was used for the deternination of the co-substrates and metabolites in 2,3-butanediol fermentation broth. The mobile phase was 5 mmol/L sulfuric acid with a flow rate of 0.5 mL/min and the column temperature was 65 ~C. Under the optimal conditions, the co-substrates （ glucose and xylose） and 6 metabolites （2,3-butanediol, acetoin, ethanol, acetic acid, lactic acid and formic acid） were completely separated and simultaneously determined, All the correlation coefficients were between 0. 9990 and 0. 9999 with the recovery ranged from 98.8% to 103.2%. And the relative standard deviation was 0.23% to 2.1%. The method has been used to determine the co-substrates and metabolites in 2,3-butanediol fermentation broth.