色谱

色谱 ›› 2015, Vol. 33 ›› Issue (10): 1046-1050.DOI: 10.3724/SP.J.1123.2015.04039

• 研究论文 • 上一篇    下一篇

高效液相色谱-高分辨质谱法鉴定水稻稻曲病菌毒素

卞英芳, 于莎莎, 牟仁祥, 曹赵云, 孙伟华, 杨欢, 林晓燕, 陈铭学   

  1. 中国水稻研究所, 农业部稻米及制品质量监督检验测试中心, 农业部稻米产品 质量安全风险评估实验室, 浙江 杭州 310006
  • 收稿日期:2015-04-22 出版日期:2015-10-08 发布日期:2012-05-09
  • 通讯作者: 陈铭学
  • 基金资助:

    中央级公益性科研院所基本科研业务费专项(2014RG006-3);国家农产品质量安全风险评估专项(GJFP201500701);浙江省科技专项重大农业项目(2014C02002-1).

Identification of ustiloxins in false smut balls of rice based on high performance liquid chromatography-high resolution mass spectrometry

BIAN Yingfang, YU Shasha, MOU Renxiang, CAO Zhaoyun, SUN Weihua, YANG Huan, LIN Xiaoyan, CHEN Mingxue   

  1. Rice Product Quality Inspection and Supervision Center, Ministry of Agriculture, Laboratory of Quality and Safety Risk Assessment for Rice, Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China
  • Received:2015-04-22 Online:2015-10-08 Published:2012-05-09

PDF

273

被引次数

14

摘要：

建立了高效液相色谱-线性离子阱高分辨质谱(HPLC-LTQ/Orbitrap MS)鉴定稻曲球中5种稻曲病菌毒素的方法。样品匀浆后以水为提取剂超声提取10 min,混合阳离子交换柱PCX净化,采用Xselect HSS T3色谱柱(150 mm×2.1 mm, 3.5 μm)分离,以水(含0.1%(v/v)甲酸)和甲醇为流动相,梯度洗脱。在m/z 200~1000范围内进行一级质谱全扫描,结合准分子离子峰的精确质量数和同位素相对丰度进行质谱分析。结果表明,鉴定到5种稻曲病菌毒素,质量准确度小于1×10-6(1 ppm),同位素相对丰度偏差绝对值≤3.3%,二级质谱碎片离子与理论裂解一致,回收率为90%~105%。该方法简便,灵敏度高,定性准确,为稻曲病菌产毒能力等相关研究提供了技术手段。

关键词: 稻曲病菌毒素, 高分辨质谱, 高效液相色谱, 鉴定, 水稻

Abstract:

A sensitive method was developed for the simultaneous identification of five ustiloxins in the false smut balls of rice by high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry (HPLC-LTQ/Orbitrap MS). The samples were extracted with deionized water under ultrasonic condition for 10 min, then purified by a strong cation exchange column (PCX). The ustiloxins were separated on an Xselect HSS T3 column (150 mm×2.1 mm, 3.5 μm) by using 0.1%(v/v) formic acid water solution and methanol as mobile phases with gradient elution at a flow rate of 0.3 mL/min. The full scan range was m/z 200-1000. The confirmatory analysis of the target compounds was carried out by the accurate mass of quasi-molecular ion, isotope abundance ratio and qualitative fragments. The results showed that the five ustiloxins (A, B, C, D and F) were identified from the false smut balls with mass accuracy less than 1×10-6 (1 ppm) and the absolute values of the deviation of isotope abundance ratio were not more than 3.3%. The product ions were consistent with the theoretical fragment mode. The recoveries were 90% to 105%. This method is accurate and sensitive for the simultaneous identification of the five ustiloxins, which can provide technical means for the research of the ability in toxin producing by Ustilaginodea virens.

Key words: high performance liquid chromatography (HPLC), high resolution mass spectrometry (HRMS), identification, rice, ustiloxins

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