色谱

色谱 ›› 2015, Vol. 33 ›› Issue (3): 309-313.DOI: 10.3724/SP.J.1123.2014.10010

• 技术与应用 • 上一篇    下一篇

高效液相色谱-串联质谱法直接定量分析植物酵素中多种氨基酸成分

郑重1, 孙琦3, 石永伟2, 曲佳乐2, 宋凤瑞1, 刘志强1   

  1. 1. 中国科学院长春应用化学研究所国家质谱中心(长春)&吉林省中药化学与质谱重点研究室, 吉林 长春 130022;
    2. 吉林敖东大高酵素有限公司, 吉林 延边 133700;
    3. 吉林敖东药业集团股份有限公司, 吉林 延边 133700
  • 收稿日期:2014-10-20 修回日期:2015-01-06 出版日期:2015-03-08 发布日期:2015-02-12
  • 通讯作者: 刘志强
  • 基金资助:

    吉林省重点科技发展计划项目(20120901);吉林省与中国科学院科技合作高技术产业化专项资金项目(2013SYHZ0008).

Direct quantitative analysis of amino acids in fermented beverage of plant extract using high performance liquid chromatography-tandem mass spectrometry

ZHENG Zhong1, SUN Qi3, SHI Yongwei2, QU Jiale2, SONG Fengrui1, LIU Zhiqiang1   

  1. 1. National Center for Mass Spectrometry in Changchun & Jilin Province Key Laboratory of Chinese Medicine Chemistry and Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
    2. Jilin Aodong-Ohtaka Enzyme Co., Ltd., Yanbian 133700, China;
    3. Jilin Aodong Medicine Industry Croup Co., Ltd., Yanbian 133700, China
  • Received:2014-10-20 Revised:2015-01-06 Online:2015-03-08 Published:2015-02-12

PDF

382

被引次数

14

摘要：

利用高效液相色谱-串联质谱(LC-MS/MS)方法直接分析植物酵素中多种未衍生化的氨基酸。样品用甲醇稀释5倍,超声提取30 min,离心10 min(速度为10000 r/min),取上清液分析。采用色谱柱Venusil ASB C18(250 mm×4.6 mm, 5 μm)分离,以甲醇-乙酸-水混合溶液为流动相体系进行梯度洗脱,流速为0.5 mL/min,质谱喷雾电压为3 kV,离子源温度为150 ℃,去溶剂温度350 ℃,去溶剂气体流速为800 L/h。碰撞气氩气的流速保持压力为0.17 Pa。以被测物的提取离子流图峰面积进行定量,该分析方法的线性范围为0.5~200 μmol/L(r2>0.99),回收率为86%~110%,可对植物酵素中16种氨基酸成分进行定量分析。该方法操作简便快速、准确可靠,还适用于食品、药品及天然产物中多种氨基酸成分的定量分析。

关键词: 高效液相色谱-串联质谱, 未衍生化氨基酸, 植物酵素

Abstract:

A method was established for underivatized amino acid determination in fermented beverage of plant extract. Samples were diluted with methanol for five times, extracted by ultrasonic vibration for 30 min, and high-speed centrifuged for 15 min at 10000 r/min. The supernatant was separated and detected by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The chromatographic column was Venusil ASB C18 (250 mm×4.6 mm, 5 μm). The elution was performed at a flow rate of 0.5 mL/min using the mobile phases of methanol-acetic acid-water mixture. The MS detector was set as follows: ion source voltage 3 kV, ion source temperature 150 ℃, solvent temperature 350 ℃, gas flow rate 800 L/h. The collision gas was argon with a pressure of 0.17 Pa. The quantitation analysis was carried out with peak area in extracted ion chromatograms. Good linearities were acquired in the range of 0.5-200 μmol/L (r2>0.99) for the amino acids. The recoveries were between 86% and 110%. There were 16 amino acids in the fermented beverage of plant extract quantitatively analyzed. The method is simple, rapid, accurate and reliable in quantitative analysis of amino acid samples in the fields of pharmaceutical, food and natural products.

Key words: fermented beverage of plant extract, high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS), underivatized amino acid

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