高效液相色谱法同时测定生物质乳酸发酵液中有机酸及糖类化合物

Simultaneous determination of organic acids and saccharides in lactic acid fermentation broth from biomass using high performance liquid chromatography

建立了高效液相色谱(HPLC)同时测定生物质乳酸发酵液中有机酸及糖类的分析方法。使用Bio-RadAminex HPX-87H色谱柱,以5 mmol/L的H2SO4为流动相,在柱温55℃,流速0.6 mL/min条件下,采用示差折光检测器进行检测。结果表明,该方法可在17 min内实现发酵液中各种有机酸和糖类化合物等的完全分离与定量,6种有机酸和3种糖类化合物在0.15～5.19 g/L范围内的线性关系良好,回归方程的线性相关系数在0.999 8以上。将该法用于米根霉发酵液的检测,两个水平的加标回收率为96.91%～103.11%,相对标准偏差(n=6)为0.81%～4.61%。该法适用于微生物发酵液中多种有机酸和糖类的快速、高效分离和定量测定。

A high performance liquid chromatographic method for the simultaneous determina- tion of organic acids and saccharides in lactic acid fermentation broth from biomass was developed. A Bio-Rad Aminex HPX-87H cohumn was used at 55 ℃. The mobile phase was 5 mmol/L sulfuric acid solution at a flow rate of 0.6 mL/min. The samples were detected by a refractive index detector （RID）. The results showed that six organic acids and three saccharides in fer- mentation broth were completely separated and determined in 17 rain. The linear correlation coefficients were above 0. 999 8 in the range of 0. 15 - 5. 19 g/L. Under the optimized condi- tions, the recoveries of the organic acids and saccharides in Rhizopus oryzae fermentation broth at two spiked levels were in the range of 96.91% - 103. 11% with the relative standard deviations （RSDs, n=6） of 0.81% -4.61%. This method is fast and accurate for the quantitative analysis of the organic acids and saccharides in microbial fermentation broths.