Simple, Rapid, and Simultaneous Assay of Multiple Carboxyl Containing Phytohormones in Wounded Tomatoes by UPLC-MS/MS Using Single SPE Purification and Isotope Dilution
Simple, Rapid, and Simultaneous Assay of Multiple Carboxyl Containing Phytohormones in Wounded Tomatoes by UPLC-MS/MS Using Single SPE Purification and Isotope Dilution
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使用单 SPE 纯化和同位素稀释，通过 UPLC-MS/MS 简单、快速、同时测定受伤番茄中的多种含羧基植物激素
DOI：
10.2116/analsci.28.1081
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发表时间：
2012-11-01
影响因子：
1.6
通讯作者：
Yan, Cunyu
中科院分区：
文献类型：
--
作者：
Fu, Jihong;Chu, Jinfang;Yan, Cunyu
采用超高效液相色谱-三重四极杆质谱（UPLC-MS/MS）-负离子电喷雾离子源，建立了一种高效、简便的同位素稀释法，可同时测定200 mg新鲜番茄组织中4种含羧基植物激素。这四种植物激素是吲哚-3-乙酸（IAA）、脱落酸（阿坝）、茉莉酸（JA）和水杨酸（SA）。在粗提后，仅用Oasis MAX固相萃取（SPE）一步纯化即可富集目标植物激素。此外，还直接分离了JA的两种内源异构体（-）-JA和（+）-7-iso-JA。验证的方法已被应用于监测野生型和转基因35 S：：prosystemin（35 S：：PS）番茄品系的局部和系统叶片中JA，SA，IAA和阿坝的变化。同时，证实了在35 S：：PS中过表达的前系统素放大了JA爆发。分析了茉莉酸、水杨酸、阿坝和生长素的时空变化。
An efficient simplified isotope dilution method was developed to determine four carboxyl containing phytohormones simultaneously in 200 mg of fresh tomato tissues using ultra high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) with negative electrospray ionization. The four phytohormones are indole-3-acetic acid (IAA), abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA). Only one purification step of Oasis MAX solid phase extraction (SPE) was employed to enrich target phytohormones after crude extraction. In addition, two endogenous isomers of JA, (-)-JA and (+)-7-iso-JA, were separated directly. The validated method has been applied to monitor changes of JA, SA, IAA, and ABA in both local and systemic leaves of wild-type and transgenic 35S::prosystemin (35S::PS) tomato lines. Meanwhile, the JA burst amplified by the overexpressed prosystemin in 35S::PS was verified. Furthermore, the spatial and temporal changes of JA, SA, ABA, and IAA were analyzed.