反相HPLC同时测定产琥珀酸放线杆菌发酵液中的有机酸与葡萄糖

Simultaneous determination of organic acid and glucose in succinic acid fermentation broth of actinobacillus succinogenes by reversed-phase high performance liquid chromatography (RP-HPLC)

提出了一种利用高效液相色谱法同时分析产琥珀酸放线杆菌发酵液中葡萄糖和有机酸的方法。在EclipseXDB C8(150mm×Φ4.6mm,5μm)色谱柱上,以5mmol LH2SO4溶液(pH2.5)作流动相,流速为1mL min,采用示差折光检测器,一次进样可同时定性及定量分析待测试样中的有机酸及葡萄糖,每一样品的分析时间不超过9min。确定了测定各物质的工作曲线的回归方程、线性范围、相关系数和检出限。测定发酵液中葡萄糖、琥珀酸和副产物乳酸的相对标准偏差在0.41%～0.89%范围内,平均回收率在99.6%～100.8%之间。

A RP-HPLC method for simultaneous determination of organic acid and glucose in succinic acid fermentation broth of Actinobacillus succinogenes was developed. Eclipse XDB-C_8 column(4.6 mm i.d.×150 mm, 5 μm) was used and the mobile phase was 0.005 (mol/L) sulfic acid solution (pH 2.5) with flow rate of 1.0 mL/min. Organic acid and glucose in samples were determined by refractive index detector (RID) within 9 min. The regression equation, the linear range, the correlation coefficient and the detection limit were determined. The relative standard deviation (RSD, n =5) was in the range of 0.41%～0.89%, and the average recovery was in the range of 99.6%～100.8%.