色谱

色谱 ›› 2012, Vol. 30 ›› Issue (01): 62-66.DOI: 10.3724/SP.J.1123.2011.09033

• 研究论文 • 上一篇    下一篇

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

马瑞, 欧阳嘉*, 李鑫, 连之娜, 蔡聪   

  1. 南京林业大学化学工程学院, 江苏 南京 210037
  • 收稿日期:2011-09-26 修回日期:2011-11-01 出版日期:2012-01-28 发布日期:2012-03-01
  • 通讯作者: 欧阳嘉,教授,主要从事生物质能源开发利用和酶工程研究. Tel: (025)85427587,
  • 基金资助:

    国家自然科学基金项目(31070513)、江苏省高校自然科学研究重大项目(10KJA22019)和江苏高校优势学科建设工程项目

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

MA Rui, OUYANG Jia*, LI Xin, LIAN Zhina, CAI Cong   

  1. College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China
  • Received:2011-09-26 Revised:2011-11-01 Online:2012-01-28 Published:2012-03-01

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1010

被引次数

67

摘要： 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-Rad Aminex 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0.9998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。

关键词: 发酵液, 高效液相色谱, 米根霉, 乳酸, 糖类化合物, 有机酸

Abstract: A high performance liquid chromatographic method for the simultaneous determination of organic acids and saccharides in lactic acid fermentation broth from biomass was developed. A Bio-Rad Aminex HPX-87H column 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 fermentation broth were completely separated and determined in 17 min. The linear correlation coefficients were above 0.9998 in the range of 0.15~5.19 g/L. Under the optimized conditions, 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.

Key words: fermentation broth, lactic acids, organic acids, Rhizopus oryzae, saccharides, high performance liquid chromatography (HPLC)