摘要
【目的】建立分离5种有机酸标准品的反相高效液相色谱法,通过此方法分析凝结芽孢杆菌发酵液中有机酸代谢产物。【方法】选择优化好的液相色谱条件:色谱柱为Venusil ASB C18(5μm,250 mm×4.6 mm),流动相为乙腈0.02mol/L磷酸二氢钾缓冲溶液(磷酸水溶液调pH 2.5),流速1.0mL/min,柱温28℃,检测波长为210nm。【结果】此方法能够快速、准确分离5种混合酸标准液;检测出4种凝结芽孢杆菌发酵液中的有机酸,分别是L-乳酸、乙酸、丙酸、甲酸;相对标准偏差为1.2%~2.9%,回收率87.5%~107.5%,各种有机酸的线性相关系数r范围在0.999 1~0.999 6之间。【结论】建立一种快速分离5种有机酸的方法,并检测出凝结芽孢杆菌发酵液有机酸的组成。
[Objective]To establish a reversed phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous separation of five organic acid standards, The organic acid metabolites of the fermentation broth of Bacillus coagulans were analyzed by this method. [Methods]The optimized HPLC conditions is as follows: Venusil ASB C18 column (5μm, 250ram lows)was used and mobile phase contained acetonitrile-0.02 mol/L potassium dihydrogen phosphate buffer solution (pH 2.5 adjusted with phosphoric acid solution) . The flow rate was 1.0 mL/min and column temperature was 28℃. Detection wavelength was at 210 nm. [Results]This method could rapidly and accurately separate in five kinds of mixed standards. Meanwhile, four kinds of organic acids is include L-lactic acid, acetic acid, propionic acid and formic acid in the fermentation broth of Bacillus. The spiked recoveries were between 87.5% and 107.5% with the relative standard deviations (RSDs) of 1.2% -2.9%. The linear correlation coefficients were from 0.9991 to 0.9996 for all organic acids. [Conclusion] A method for rapid separation of five organic acids was established, and the types of organic acids in the fermentation broth of Bacillus coagulans could be determined.
出处
《北京农学院学报》
2017年第4期80-84,112,共6页
Journal of Beijing University of Agriculture
基金
2016北京市农委项目(PXM2016_014207_000057)
2016本科生科研训练项目