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园艺学报 ›› 2016, Vol. 43 ›› Issue (12): 2481-2490.doi: 10.16420/j.issn.0513-353x.2016-0479

• 新方法 • 上一篇    下一篇

羽衣甘蓝和白菜型油菜硫代葡萄糖苷超高效液相色谱—飞行时间质谱分析方法

Glucosinolate Analysis in Kale and Yellow Sarson Based on UHPLC–Triple–TOF–MS   

  1. ZHENG Shu-ning1,ZHANG Yan-guo1,GUO Ning2,LIU Zhi-yuan1,and XU Dong-hui1,*
  • 出版日期:2016-12-25 发布日期:2016-12-25

Glucosinolate Analysis in Kale and Yellow Sarson Based on UHPLC–Triple–TOF–MS

郑姝宁1,张延国1,郭 宁2,刘志远1,徐东辉1,*   

  1. (1Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Risk Assessment Laboratory for Vegetable Product Quality and Safety of Ministry of Agriculture(Beijing),Key Laboratory of Horticultural Crop Biology and Germplasm Creation of Ministry of Agriculture,Beijing 100081,China;2Beijing Vegetable Research Center,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China)
  • Online:2016-12-25 Published:2016-12-25

摘要:

利用超高效液相色谱—飞行时间质谱(UHPLC–Triple–TOF–MS)技术建立了羽衣甘蓝等蔬菜中硫代葡萄糖苷(Glucosinolate,简称硫苷)的分析方法。蔬菜样品采用70% 甲醇提取,采用UPLC BEH C18反相色谱柱,乙腈—水(均含0.1%甲酸)流动相系统,10 min快速梯度洗脱;在电喷雾质谱负离子模式下采用飞行时间质谱全扫描—信息关联采集—子离子扫描(TOF–MS scan–IDA–Product ion scan)复合扫描模式,1次进样分析可同时获得硫苷的一级质谱和二级质谱信息;基于TOF–MS的高分辨率性能,得到硫苷分子、离子和碎片离子的精确质荷比,大大提高硫苷定性分析的准确性。将建立的分析方法应用于白菜型油菜Yellow Sarson的种子和羽衣甘蓝的叶片组织,分别鉴定出24和15种硫苷,表明本方法具有灵敏度高、准确度高和高通量等优点,适合于十字花科蔬菜中硫苷的分析。

关键词: 羽衣甘蓝, 白菜型油菜, 硫代葡萄糖苷, 超高效液相色谱—飞行时间质谱, 黄酮醇, 生物合成, 代谢调控, 分子机制, 基因工程

Abstract:

A method for analysis of glucosinolates in cruciferous vegetable was developed by using UHPLC–Triple–TOF–MS. Vegetable samples were extracted using 70% methanol and injected after concentration. UPLC BEH C18 column was selected with acetonitrile–water(both 0.1% formic acid)as mobile phase. Chromatographic separation was achieved under gradient elution in 10 min. In ESI negative ion mode,TOF–MS scan–IDA–Product ion scan was performed to acquire both MS and MS/MS information from one injection. Based on high resolution TOF–MS,accurate masses of molecular ions and fragment ions were obtained for high accuracy-identification. This method was applied to the seeds of Yellow Sarson(Brassica rapa ssp. trillocularis)and the leaves of kale(Brassica oleracea). Finally,24 and 15 glucosinolates were analyzed in the seeds and leaves,respectively. The results demonstrated that this method has distinct advantages of high sensitivity,good accuracy and high throughput,which is suitable for analysis of glucosinolates in cruciferous vegetables.

Key words: kale, Yellow Sarson, glucosinolate, UHPLC–Triple–TOF–MS, Flavonol, biosynthesis, regulation, molecular mechanism, genetic engineering

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