Improved methodology for analysis of multiple phytohormones using sequential magnetic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry
Improved methodology for analysis of multiple phytohormones using sequential magnetic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry
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DOI：
10.1016/j.aca.2017.06.019
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发表时间：
2017-08-29
影响因子：
6.2
通讯作者：
Feng, Yu-Qi
中科院分区：
文献类型：
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作者：
Luo, Xiao-Tong;Cai, Bao-Dong;Feng, Yu-Qi
植物激素是一种特殊的小分子物质，在植物的生长发育过程中发挥着微量的重要作用。多种植物激素的定量研究将有助于研究植物激素通过协同或拮抗相互作用调节植物对生物和非生物胁迫反应的交叉对话。在目前的研究中，我们开发了一种在一个小样本中分析植物激素的方法，包括吲哚-3-乙酸、脱落酸、茉莉酸、赤霉素、细胞分裂素和油菜素内酯。采用序贯磁固相萃取法（MSPE）对上述植物激素进行纯化和分离，采用超高效液相色谱-电喷雾串联质谱法（UHPLC-MS/MS）对其进行分析。在优化的提取条件下，相关系数(r)在0.9961 ~ 0.9998范围内线性良好。检出限（lod, S/N = 3）为0.45 ~ 126.1 pg mL（-1）。加样回收率在85.0% ~ 116.2%之间。相对标准偏差（rsd）范围为2.7% ~ 16.1%。利用该方法，可以从一种植物提取物中纯化和分析23种植物激素。最后，在100 mg（鲜重）甘蓝型油菜花中检测到16种植物激素，包括IAA、ABA、JA、4种GAs、3种BRs和6种ck，浓度范围为0.09 ~ 305.23 ng g（-1）。(C) 2017 Elsevier B.V.版权所有
Phytohormones are special small molecules that play important role in plant growth and development at trace levels. Quantification of multiple phytohormones will be great helpful for researches about cross-talks that plant hormones regulate the responses of plants against both biotic and abiotic stresses by means of synergistic or antagonistic interactions. In the current study, we developed a method for profiling of phytohormones in one small sample, including indole-3-acetic acid, abscisic acid, jasmonic acid, gibberellins, cytokinins and brassinosteroids. These phytohormones mentioned above were firstly purified and separated by sequential magnetic solid-phase extraction (MSPE) and then analyzed by ultrahigh performance liquid chromatography-electrospray tandem mass spectrometry (UHPLC-MS/MS). Under the optimized extraction conditions, good linearity was obtained with correlation coefficients (r) ranging from 0.9961 to 0.9998. The limits of detection (LODs, S/N = 3) were ranged from 0.45 to 126.1 pg mL(-1). The recoveries were between 85.0% and 116.2%. The relative standard deviations (RSDs) were ranged from 2.7% to 16.1%. With the proposed strategy, 23 phytohormones could be purified and analyzed from a single plant extract. Finally, 16 phytohormones could be detected in 100 mg (fresh weight) flower of Brassica napus L., including IAA, ABA, JA, 4 GAs, 3 BRs and 6 CKs with the concentration ranged from 0.09 to 305.23 ng g(-1). (C) 2017 Elsevier B.V. All rights reserved.