色谱

色谱 ›› 2014, Vol. 32 ›› Issue (12): 1356-1361.DOI: 10.3724/SP.J.1123.2014.09016

• 研究论文 • 上一篇    下一篇

逐级提取-高效液相色谱法快速测定植物组织中8种有机酸

黄天志1,2,3, 王世杰1,2, 刘秀明1,2, 刘虹1, 吴沿友1,2, 罗绪强4   

  1. 1. 中国科学院地球化学研究所, 环境地球化学国家重点实验室, 贵州 贵阳 550002;
    2. 中国科学院普定喀斯特生态系统观测研究站, 贵州 安顺 562100;
    3. 中国科学院大学, 北京 100049;
    4. 贵州师范学院, 贵州 贵阳 550018
  • 收稿日期:2014-09-10 修回日期:2014-10-10 出版日期:2014-12-08 发布日期:2014-11-28
  • 通讯作者: 王世杰,E-mail:wangshijie@vip.skleg.cn.
  • 基金资助:

    国家重大科学研究计划项目(2013CB956700);中国科学院战略性先导科技专项(XDA05070400);国家自然科学基金项目(31100187).

Rapid determination of eight organic acids in plant tissue by sequential extraction and high performance liquid chromatography

HUANG Tianzhi1,2,3, WANG Shijie1,2, LIU Xiuming1,2, LIU Hong1, WU Yanyou1,2, LUO Xuqiang4   

  1. 1. State Key Laboratory of Environmental Geochemistry, Institute of Geochemistry, Chinese Academy of Sciences, Guiyang 550002, China;
    2. Puding Karst Ecosystem Research Station, Chinese Academy of Sciences, Anshun 562100, China;
    3. University of Chinese Academy of Sciences, Beijing 100049, China;
    4. Guizhou Normal College, Guiyang 550018, China
  • Received:2014-09-10 Revised:2014-10-10 Online:2014-12-08 Published:2014-11-28

PDF

385

被引次数

7

摘要：

针对植物组织中草酸存在的不同形态,建立了水和稀盐酸作为提取介质的逐级提取方法,获得了水溶态和酸溶态草酸及乙醇酸、乙醛酸、酒石酸、苹果酸、乙酸、柠檬酸、琥珀酸等有机酸。采用Hypersil ODS (200 mm×4.6 mm, 5 μm)色谱柱,以5 mmol/L磷酸二氢钾水溶液(pH 2.8)作为流动相,在进样量5 μL、检测波长210 nm、柱温30 ℃的条件下,通过分时段控制流速实现了8种有机酸的快速分离,同时去除了盐酸对酸溶态草酸测定的干扰。本方法精确灵敏、回收率高、重复性好,可应用于实际样品的测定分析。

关键词: 高效液相色谱, 有机酸, 植物, 逐级提取

Abstract:

A sequential extraction method was developed to determine different forms of oxalate and seven oxalate-metabolism-related organic acids (glyoxylic acid, tartaric acid, glycolic acid, malic acid, acetic acid, citric acid, succinic acid) in plant tissue. The ultra-pure water was used as the extraction medium to obtain water-soluble oxalic acid and the other seven water-soluble organic acids. After the extraction of the water-soluble organic acids, the residues were extracted by dilute hydrochloric acid successively to get the acid-soluble oxalate which entered the liquid phase. A Hypersil ODS column was used with 5 mmol/L potassium dihydrogen phosphate buffer solution (pH 2.8) as the mobile phase. The diode array detector was set at 210 nm and the column temperature at 30 ℃ with the injection volume of 5 μL. The flow rate was controlled at different times which allowed a good and rapid separation of the organic acids and hydrochloric acid. Under these conditions, the linear ranges of the method were 1-2000 mg/L for oxalic acid, 25-2000 mg/L for acetic acid, and 10-2000 mg/L for glyoxylic acid, tartaric acid, glycolic acid, malic acid, citric acid and succinic acid, with the correlation coefficients of the eight organic acids ≥ 0.9996. The average recoveries of the eight organic acids in leaves and roots were 93.5%-104.4% and 85.3%-105.4% with RSDs of 0.15%-2.43% and 0.31%-2.9% (n=7), respectively. The limits of detection ranged from 1 to 10 ng (S/N=3). The results indicated that the method is accurate, rapid and reproducible for the determination of organic acids in plant samples.

Key words: high performance liquid chromatography (HPLC), organic acids, plant, sequential extraction

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