Bosco Renato, Daeseleire Els, Van Pamel Els, Scariot Valentina, Leus Leen
Department of Agricultural, Forest and Food Sciences, University of Torino , Largo Paolo Braccini 2, 10095 Grugliasco, Torino, Italy.
J Agric Food Chem. 2014 Jul 9;62(27):6278-84. doi: 10.1021/jf5023884. Epub 2014 Jun 26.
This paper describes a method to detect and quantitate the endogenous plant hormones (±)-2-cis-4-trans-abscisic acid, (-)-jasmonic acid, and salicylic acid by means of ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) in hybrid rose leaf matrices. Deuterium-labeled [(2)H6] (+)-2-cis-4-trans-abscisic acid, [(2)H6] (±)-jasmonic acid, and [(2)H4]-salicylic acid were used as internal standards. Rose samples (10 mg) were extracted with methanol/water/acetic acid (10:89:1) and subsequently purified on an Oasis MCX 1 cm(3) Vac SPE cartridge. Performance characteristics were validated according to Commission Decision 2002/657/EC. Recovery, repeatability, and within-laboratory reproducibility were acceptable for all phytohormones tested at three different concentrations. The decision limit and detection capability for (±)-2-cis-4-trans-abscisic acid, (-)-jasmonic acid, and salicylic acid were 0.0075 and 0.015 μg/g, 0.00015 and 0.00030 μg/g, and 0.0089 and 0.018 μg/g, respectively. Matrix effects (signal suppression or enhancement) appeared to be high for all substances considered, implying the need for quantitation based on matrix-matched calibration curves.
本文描述了一种通过超高效液相色谱-串联质谱法(UHPLC-MS/MS)在杂交玫瑰叶片基质中检测和定量内源性植物激素(±)-2-顺式-4-反式脱落酸、(-)-茉莉酸和水杨酸的方法。氘代标记的[(2)H6](+)-2-顺式-4-反式脱落酸、[(2)H6](±)-茉莉酸和[(2)H4]-水杨酸用作内标。玫瑰样品(10毫克)用甲醇/水/乙酸(10:89:1)提取,随后在Oasis MCX 1 cm(3) Vac固相萃取柱上进行纯化。根据委员会第2002/657/EC号决定对性能特征进行了验证。在三种不同浓度下测试的所有植物激素的回收率、重复性和实验室内再现性均可接受。(±)-2-顺式-4-反式脱落酸、(-)-茉莉酸和水杨酸的决策限和检测能力分别为0.0075和0.015微克/克、0.00015和0.00030微克/克以及0.0089和0.018微克/克。对于所有考虑的物质,基质效应(信号抑制或增强)似乎都很高,这意味着需要基于基质匹配校准曲线进行定量。