HPLC-MS/MS法测定烟叶中7种植物内源激素

Determination of seven endogenous phytohormones in tobacco by high performance liquid chromatography-tandem mass spectrometry

为获得烟草激素的质量分数数据,并分析其在抗性、生长发育等过程中的作用,采用高效液相色谱-三重四极杆质谱(HPLC-MS/MS)仪建立了同时分析烟叶中吲哚乙酸(IAA)、吲哚丁酸(IBA)、水杨酸(SA)、赤霉素(GA)、茉莉酸(JA)、脱落酸(ABA)和油菜素内酯(BR)等7种植物激素质量分数的方法。采用多离子反应检测模式,获得7种植物激素的特征碎片离子峰,并测定了烟草花叶病毒(TMV)感染的烟叶样品。结果表明:(1)7种植物激素在线性范围内的相关系数(r)为0.998 6～0.999 5,方法的检出限为0.07～3.00μg/L,日内日间相对标准偏差(RSD)为0.44%～9.76%,回收率为77.82%～117.63%;(2)烟草花叶病毒感染极显著(p<0.01)地增加了烟叶中SA、IBA、JA、ABA的质量分数,显著(p<0.05)增加了烟叶中IAA的质量分数,同时显著(p<0.05)降低了烟叶中GA的质量分数;TMV病毒感染对BR的影响不大。该方法具有良好的灵敏度、精密度和回收率,可用于植物相关激素的同时分析。

In order to determine the contents of endogenous phytohormones in tobacco and analyze the roles of phytohormones in the resistance, growth and development of tobacco,a method for simultaneously determining the contents of 7 endogenous phytohormones, including 3-indoleacetic acid （IAA）, 3-indolebutytic acid （IBA）, salicylic acid （SA）, gibberellin A3 （GA）, jasmonic acid （JA）, abscisic acid （ABA） and brassinosteroids （BR） in tobacco was developed by high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）. The TMV （tobacco mosaic virus）-infected tobacco samples were determined by obtaining the characteristic fragment ion peaks of the 7 endogenous phytohormones under multiple reaction monitoring mode. The results showed that：1） The correlation coefficients （r） of the 7 phytohormones were from 0.998 6 to 0.999 5 in the linear ranges. The limits of quantification （LOQ） for all analytes were between 0.07 and 3.00 μg/L, and the intra-and inter-day relative standard deviations （RSDs） were 0.44%-9.76% for all analytes. The recoveries were in the range of 77.82%-117.63%. 2） The contents of SA,IBA,JA,ABA in TMV-infected tobacco were significantly higher （p〈0.01） than the control sample, while which was significantly lower （p〈0.05） for GA. However the content of BR was basically unchanged. With good sensitivity, precision and recovery, this method is suitable for determining various endogenous phytohormones.