色谱 ›› 2017, Vol. 35 ›› Issue (3): 264-268.DOI: 10.3724/SP.J.1123.2016.10010

• 研究快报 • 上一篇    下一篇

毛细管离子色谱-串联质谱法测定热纤梭杆菌发酵液中的低相对分子质量有机酸

赵海杰1,2, 梁文辉2, 李震3, 史庆昊1,2, 建方方1, 法芸2,4, 刘会洲2   

  1. 1. 青岛科技大学化学与分子工程学院, 山东 青岛 266042;
    2. 中国科学院青岛生物能源与过程研究所公共实验室, 山东 青岛 266101;
    3. 青岛伊科思技术工程有限公司, 山东 青岛 266045;
    4. 中国科学院大学, 北京 100049
  • 收稿日期:2016-10-08 出版日期:2017-03-08 发布日期:2013-07-16
  • 通讯作者: 法芸,Tel:(0532)80662692,E-mail:fayun@qibebt.ac.cn;刘会洲,E-mail:liuhz@qibebt.ac.cn(刘会洲).
  • 基金资助:

    国家自然科学基金项目(21405166);科技部重大专项(2012YQ090229).

Capillary ion chromatography-mass spectrometry for determination of low relative molecular mass organic acids in culture media of Clostridium thermocellum

ZHAO Haijie1,2, LIANG Wenhui2, LI Zhen3, SHI Qinghao1,2, JIAN Fangfang1, FA Yun2,4, LIU Huizhou2   

  1. 1. College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China;
    2. Key Laboratory, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China;
    3. Qingdao YIKESI Technology Engineering Company Limited, Qingdao 266045, China;
    4. University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2016-10-08 Online:2017-03-08 Published:2013-07-16
  • Supported by:

    National Natural Science Foundation of China (No. 21405166); Major Projects of the Ministry of Science and Technology (No. 2012YQ090229).

摘要:

建立了一种测定热纤梭杆菌发酵液中的低相对分子质量有机酸的毛细管离子色谱-串联质谱方法。探索优化了色谱和质谱的检测条件,在最佳分析条件下同时检测8种有机酸。离子色谱以KOH水溶液作为流动相进行梯度洗脱;用IonSwift MAX-100毛细管柱进行分离;在喷雾电压为3.0 kV、喷雾气压强为2000 kPa、成源温度为275 ℃的条件下,选择离子监测(SIM)模式下运行质谱。结果表明,柠檬酸和异柠檬酸两种同分异构体能够得到很好的分离,8种有机酸在一定浓度范围内具有良好的线性关系,相对标准偏差为1.45%~5.99%,相关系数为0.9696~0.9986,平均加标回收率为89.0%~110.0%,8种有机酸的检出限为0.01~0.50mg/L。该方法进样量少,灵敏度高,重现性好,能够满足实际样品的检测要求,可用于嗜热厌氧菌发酵液中低分子量有机酸的测定。

关键词: 毛细管离子色谱, 热纤梭杆菌, 有机酸, 质谱

Abstract:

A capillary ion chromatography-mass spectrometry method was proposed to determine low relative molecular mass organic acids in culture media of Clostridium thermocellum. The conditions of analysis were optimized and eight organic acids were detected simultaneously. The target compounds were separated on an IonSwift MAX-100 capillary column with potassium hydroxide solution as mobile phase by gradient elution, and detected under selected ion monitoring (SIM) mode. The mass spectrometry conditions were: spray voltage 3.0 kV, sheath gas 2000 kPa, capillary temperature 275℃. The relative standard deviations ranged from 1.45% to 5.99%. The coefficients of determination ranged from 0.9696 to 0.9986 and the mean recoveries were 89.0%-110.0%. The limits of detection of the eight organic acids were 0.01-0.50mg/L. This method was used to detect low relative molecular mass organic acids in culture media of Clostridium thermocellum with high sensitivity and good reproducibility.

Key words: Clostridium thermocellum, capillary ion chromatography, mass spectrometry (MS), organic acids

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