离子排斥色谱法分析发酵液中甘油酸等代谢产物  

Determination of glyceric acid and other metabolites in fermentation broth by ion-exclusion liquid chromatography

作　　者：方亚坤[1] 靳魁奇[1] 刘宇鹏[1] 

机构地区：[1]河南大学生命科学学院生物工程研究所,河南开封475004

出　　处：《食品与发酵工业》2015年第7期171-174,共4页Food and Fermentation Industries

基　　金：河南省科技厅攻关项目(No.132102210387)

摘　　要：建立了一个利用示差折光检测器检测发酵液中甘油酸等代谢产物的方法。利用磺化的苯乙烯-二乙烯共聚物作为固定相的Aminex HPX-87H离子色谱柱为分析柱，利用示差折光检测器对发酵液中的产物甘油酸、副产物二羟基丙酮和残余底物甘油进行了成功的分离检测。经过色谱条件优化后选择了柱温60℃，含有20％乙腈的5mmol／L的H2SO4为流动相。实验室条件下分离检测效果良好。甘油酸、二羟基丙酮和甘油的浓度线性范围和相关系数分别为：甘油酸为0．2～1．0g／L（R^2=0．999）；二羟基丙酮为0．2～1．0g／L（R^2=0．994）；甘油为0．2～1．0g／L（R^2=0．992），平均回收率分别为（99．52±1．30）％，（97．61±1．33）％，（98．52±1．58）％。该方法简便、灵敏、准确度高。A method using a refractive index detector to detect glyceric acid and other metabolites in the fermentation broth was described. Glyceric acid, glycerol and dihydroxyacetone were successfully separated by a sulfonated polystyrene divinyl benzene column （Aminex HPX-87H） with a refractive index detector. It is found that the optimal conditions were as follows： column temperature was 60 ℃ , and 5 mmol/L sulfuric acid containing 20% acetonitrile was used as mobile phase. These conditions were applied to the quantification. The linear range for glyceric acid, glycerol and dihydroxyacetone were 0. 2 - 1 g/L and the correlation coefficient of them were 0. 999, 0. 992, and 0. 994, respectively. Recoveries of glyceric acid, glycerol and dihydroxyacetone added to culture were （99. 52 ± 1.30） % , （97.61 ± 1.33）%, and（98.52 ± 1.58）%, respectively. The method was accurate, rapid and suitable for routine analysis.

关 键 词：高效液相色谱 甘油酸 二羟基丙酮 示差折光检测器 

分 类 号：TQ920.1[轻工技术与工程—发酵工程] O657.7[理学—分析化学]