离子排斥色谱法分析米根霉富马酸发酵液中的代谢产物  

Determination of Metabolites from Fumaric Acid Fermentation Broth of Rhizopus oryzae by Ion-exclusion Liquid Chromatography

作　　者：付永前[1] 高振[1] 李霜[1,2] 徐晴[1] 张昆[1] 黄和[1,2] 

机构地区：[1]南京工业大学制药与生命科学学院,江苏南京210009 [2]南京工业大学材料化学工程国家重点实验室,江苏南京210009

出　　处：《食品与发酵工业》2009年第1期131-134,共4页Food and Fermentation Industries

基　　金：国家高科技研究发展计划("863"计划)专题课题(No.2006AA02Z240);国家自然科学基金(No.20576054;No.20706031);国家重点基础研究发展计划("973"计划)(No.2007CB707805)

摘　　要：提出了利用离子排斥色谱法同时分析米根霉富马酸发酵液中主要代谢产物与葡萄糖的方法。在BIO-RAD Aminex HPX-87H色谱柱上,65℃柱温下,以5mmol/L H2SO4溶液作为流动相,流速0.8mL/min,采用示差折光检测器对8种有机酸(草酸、柠檬酸、α-酮戊二酸、L-苹果酸、丁二酸、L-乳酸、富马酸和乙酸)、葡萄糖及乙醇进行了成功的分离,确定了各物质工作曲线的回归方程、线性范围、相关系数和检出限。并应用于米根霉产富马酸发酵液的产品分析,对经预处理后的富马酸发酵液直接进样分离定量,其中的富马酸、-酮戊二酸、葡萄糖、L-乳酸和乙醇完全分离定量,确定了测定发酵液中有机酸、葡萄糖和乙醇的相对标准偏差在0.43%～4.69%,平均回收率在98.41%～104.33%。经多次实验证明,该方法是测定富马酸发酵液中各种主要代谢产物与葡萄糖的准确、有效的定量测定方法。An ion-exclusion chromatography method for simultaneous determination of mainly Metabolites and glucose in fumaric acid fermentation broth of Rhizopus Oryzae was developed. Aminex HPX --87H （300mm〉（7.8mm,9μm） was used and the mobile phase was 0. 005 mol/L sulfic acid solution with flow rate of 0.8 mL/min at 65℃column temperature. Eight organic acids （including oxalic, citric, α-ketoglutaric, L- malic, succinic, L-lactic, fumaric and acetic acid）, glucose, and ethanol were separated by refractive index detector （RID）. It was applied to the quantification of fumaric acid fermentation of Rhizopus oryzae ME- F12. The solution of samples pretreated would be determined directly. The fumaric acid, α-ketoglutaric acid, glucose, L-latic acid and ethanol were completely separated and determined. The regression equation, the linear range, the correlation coefficient and the detection limit were determined. The relative standard deviation （RSD, n=3） was in the range of 0. 432%-4. 691%, and the average recovery was in the range of 98. 41% -104.33% in samples. This method was proved an effective way to determine the mainly Metabolites and glucose in fumaric acid fermentation broth of Rhizopus oryzae.

关 键 词：离子排斥色谱 示差折光检测器 米根霉 富马酸 

分 类 号：TQ922[轻工技术与工程—发酵工程]