高效液相色谱法测定克鲁维酵母菊芋发酵液中的乙醇，糖和有机酸类代谢成分

Simultaneous Determination of Ethanol,Sugar and Organic Acids in Kluveromyces marxious Broth by High Performance Liquid Chromatography-Ultraviolet/Refractive Index Detector

建立了高效液相色谱法同时分析菊芋发酵液中的乙醇和有机酸的方法。采用HPLC有机酸分析柱,流动相为0.01 mol/L H2SO4,流速为0.5 mL/min,以紫外和示差折光检测器作为双通道检测手段,同时对克鲁维酵母菊芋发酵液中的柠檬酸、α-酮戊二酸、葡萄糖、丙酮酸、果糖、琥珀酸、乳酸、甘油、乙酸、乙醇进行了定量分析,本方法的回收率为95.8%～109.6%;RSD为0.33%～4.0%,结果表明,本方法简单、快速、准确,适用于监控克鲁维酵母发酵产物并指导整个发酵过程条件的优化。

A high performance liquid chromatographic method was developed for the simultaneous determination of ethanol, sugar and organic acids in Kluveromyces marxinus broth by UV and RI （Refractive index） detector. Aminex HPX-87H organic acids analysis column was used for separation. The mobile phase was composed of sulfuric acid and water, and the concentration of sulfuric acid was 0.01 mol/L（pH 1.4）. The flow rate was 0.5 mL/min. The broth（containing glucose, fructose, ethanol, acetic acid, glycerol, and lactic acid） was separated by the dual-channel detector within 25 min. The method showed high average recoveries for the samples ranged from 95.8% to 109.6% and good precision（RSD） ranged from 0.3% to 4.0%. The results show that ethanol, sugar and organic acids can be determined rapidly and effectively, and is capable of being used for real-time analysis of metabolic components in fermentation broth.