色谱

色谱 ›› 2015, Vol. 33 ›› Issue (8): 805-808.DOI: 10.3724/SP.J.1123.2015.04038

• 特别策划:第20届全国色谱学术报告会专栏 • 上一篇    下一篇

高效液相色谱法测定菌群降解纤维素产物中的糖、有机酸和醇

姜艳1,2, 范桂芳1,2, 杜然1,2, 李佩佩1,2, 姜立1,2, 赵刚2, 齐泮仑3, 李十中1,2   

  1. 1. 清华大学核能与新能源技术研究院, 北京 100084;
    2. 北京市生物燃料工程技术研究中心, 北京 100084;
    3. 中国石油天然气股份有限公司石油化工研究院, 北京 100000
  • 收稿日期:2015-04-22 出版日期:2015-08-08 发布日期:2015-07-27
  • 通讯作者: 李十中
  • 基金资助:

    国家高技术研究发展计划("863"计划)项目(2012AA101805);国家国际科技合作专项项目(2012DFG61720,2012DFG61700).

Determination of sugars, organic acids and alcohols in microbial consortium fermentation broth from cellulose using high performance liquid chromatography

JIANG Yan1,2, FAN Guifang1,2, DU Ran1,2, LI Peipei1,2, JIANG Li1,2, ZHAO Gang2, QI Panlun3, LI Shizhong1,2   

  1. 1. Institute of Nuclear and New Energy Technology, Tsinghua University, Beijing 100084, China;
    2. Beijing Engineering Research Center of Biofuels, Beijing 100084, China;
    3. Research Institute of Petroleum Processing of PetroChina Company Limited (CNPC), Beijing 100000, China
  • Received:2015-04-22 Online:2015-08-08 Published:2015-07-27

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摘要：

采用硫酸除钙和调节pH的样品预处理方法,利用高效液相色谱对菌群降解纤维素发酵液中的糖、有机酸和醇3类物质进行分析,待测组分与培养基成分能够得到有效分离。从菌群降解纤维素发酵液中检出纤维二糖、葡萄糖、乙醇、丁醇、甘油、乙酸与丁酸并进行了定量分析,7种组分的检出限范围为0.10~2.00 mg/L,线性相关系数均大于0.9996,线性范围为0.020~1.000 g/L,回收率为85.41%~115.60%,相对标准偏差(RSD)为0.22%~4.62%(n=6)。该方法准确可靠,可实现对菌群降解纤维素发酵液中糖、醇和有机酸的同时准确测定。

关键词: 醇, 高效液相色谱, 菌群, 糖, 同步生物加工, 纤维素降解, 有机酸

Abstract:

A high performance liquid chromatographic method was established for the determination of metabolites (sugars, organic acids and alcohols) in microbial consortium fermentation broth from cellulose. Sulfate was first added in the samples to precipitate calcium ions in microbial consortium culture medium and lower the pH of the solution to avoid the dissociation of organic acids, then the filtrates were effectively separated using high performance liquid chromatography. Cellobiose, glucose, ethanol, butanol, glycerol, acetic acid and butyric acid were quantitatively analyzed. The detection limits were in the range of 0.10-2.00 mg/L. The linear correlation coefficients were greater than 0.9996 in the range of 0.020 to 1.000 g/L. The recoveries were in the range of 85.41%-115.60% with the relative standard deviations of 0.22%-4.62% (n=6). This method is accurate for the quantitative analysis of the alcohols, organic acids and saccharides in microbial consortium fermentation broth from cellulose.

Key words: alcohols, cellulose degradation, consolidated bioprocessing (CBP), high performance liquid chromatography (HPLC), microbial consortium, organic acids, sugars

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