超高效液相色谱-高分辨质谱测定10种植物内源激素

Determination of Ten Plant Hormones by Ultra-high Performance Liquid Chromatography-High Resolution Mass Spectrometry

建立一种利用超高效液相色谱-飞行时间-质谱(UPLC-TOF-MS)快速高效地对植物激素进行定性定量测定的方法。对植物中常见的异戊烯腺嘌呤、玉米素、玉米素核苷、独脚金内酯、茉莉酸甲酯、脱落酸、赤霉酸、吲哚乙酸、茉莉酸和水杨酸10种内源激素,利用一级质谱全扫描、二级质谱及多反应检测模式对其进行测定,确定其检测限和定量限;采用不同的溶剂(异丙醇、丙酮、甲醇、乙腈)提取水稻、小麦、烟草、番茄等植物样品,利用固相萃取小柱从植物基质中回收激素标品,测定其回收率和精密度。结果表明,10种激素在质谱上均有良好的响应,浓度和响应值的线性相关性均达到0.99以上;固相萃取小柱对200、50 ng/mL激素的回收率均达到90%以上,对10 ng/mL激素的回收率在80%左右;乙腈+1%甲酸的提取效果最好;采用80%甲醇复溶可以更有效地回收激素。本方法简单、快速、重现性好,可用于同时测定多种植物内源激素。

This study aims to establish a rapid and efficient method for the qualitative and quantitative determination of plant hormones by UPLC-TOF-MS.Ten plant hormones,including isopentene adenine,zeatin,ribosylzeatin,strigolactone,methyl jasmonate,abscisic acid,gibberellic acid,indole-3-acetic acid,jasmonic acid and salicylic acid,were selected for determination in the mode of MS scan,MS/MS and multi-reaction monitoring.And the limits of detection and quantitation were determined.Solid phase extraction(SPE)column was used to clean-up hormones from plant matrix.The recovery and repeatability were determined.Different solvents(isopropanol,acetone,methanol,acetonitrile)were used to extract plant samples(rice,wheat,tobacco,tomato,etc).The results showed that the ten hormones had good response on mass spectrometry,and the linear coefficients between concentration and response value were above 0.99.The recovery rates of 200 and 50 ng/mL hormones by solid phase extraction column were over 90%,while the recovery rates of 10 ng/mL hormones were about 80%.There were significant differences in the extraction efficiency of several different solvents,among which acetonitrile added with 1%formic acid was the best.Hormones could be recovered more efficiently by dissolution in 80%methanol.The method is simple,rapid with good reproducibility,which is suitable for simultaneous determination of several endogenous hormones in plant samples.