采用离子排斥色谱法分析发酵液中的琥珀酸等代谢产物

采用离子排斥液相色谱法对琥珀酸及发酵液中常见的代谢产物进行了分析。以高交联度磺化苯乙烯-二乙烯基苯共聚物多孔微球作为固定相的Aminex HPX-87H离子色谱柱为分析柱,利用示差折光检测器对9种有机酸(琥珀酸、柠檬酸、α-酮戊二酸、苹果酸、丙酮酸、乳酸、乙酸、甲酸和丙酸)、葡萄糖及乙醇进行了较成功的分离,并应用于琥珀酸放线杆菌发酵液的产品分析。通过色谱条件优化,选定了55℃柱温和10 mmol/LH2SO4作为流动相。发酵液样品中有机酸、葡萄糖和乙醇的平均回收率在96.0%-104.8%,精密度偏差在0.4%-3.5%范围内。

Determination of Succinic Acid and other Metabolites from Fermentation Broth by Ion-exclusion Liquid Chromatography

A method using ion-exclusion liquid chromatography was described for the simultaneous determination of succinic acid and other metabolites commonly found as products of fermentation.Nine organic acids(including citric,α-ketoglutaric,malic,pyruvic,succinic,lactic,formic,acetic and propionic acid),glucose,and ethanol were separated by a sulfonated polystyrene divinyl benzene column(Aminex HPX-87H) with a refractive index detector.It is found that the optimal conditions for separation were 55 ℃ column temperature and 10 mmol/L sulfuric acid,which were applied to the quantification of succinic acids and other metabolites of Actinobacillus succinogenes CGMCC 1593.The results showed that recoveries were within the range of 96.0%～104.8%,and the precision CV ranging from 0.4% to 3.5% in samples.