高效液相色谱-高分辨质谱法鉴定水稻稻曲病菌毒素  

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

作　　者：卞英芳 于莎莎[1] 牟仁祥[1] 曹赵云[1] 孙伟华[1] 杨欢[1] 林晓燕[1] 陈铭学[1] 

机构地区：[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

出　　处：《色谱》2015年第10期1046-1050,共5页Chinese Journal of Chromatography

基　　金：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)

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（150mm×2.1 mm,3.5μm）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 200~1 000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。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 / z200-1 000. 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.

关 键 词：高效液相色谱 高分辨质谱 稻曲病菌毒素 水稻 鉴定 

分 类 号：O658[理学—分析化学]