Simultaneous determination of 13 phytohormones in oilseed rape tissues by liquid chromatography‐electrospray tandem mass spectrometry and the evaluation of the matrix effect

Abstract

In the experiment, a high-performance liquid chromatography and electrospray ionization-tandem mass spectrometry with selected reaction monitoring was used to simultaneously determine various classes of phytohormones, including indole-3-acetic acid, α-naphthaleneacetic acid, 2-chlorobenzoic acid, 4-chlorobenzoic acid, indole-3-butyric acid, gibberellic acid, 2,4-dichlorophenoxyacetic acid, 2-naphthoxyacetic acid, abscisic acid, 2,3,5-triiodobenzoic acid, uniconazole, paclobutrazol and 2,4-epibassinolide in rape tissues. The analyses were separated by an HPLC equipped with a reversed-phase column using a binary solvent system composed of methanol and water, both containing 0.1% of formic acid. The matrix effect was also considered and determined. The technology was applied to analyze rape tissues, including roots, stems, leaves, flowers, immature pods and rape seeds. The rape tissues were subjected to ultrasound-assisted extraction and purified by dispersive solid-phase extraction, and then transferred into the liquid chromatography system. The detection limit for each plant hormone was defined by the ratio of signal/background noise (S/N) of 3. The results showed perfect linearity (R(2) values of 0.9987-1.0000) and reproducibility of elution times (relative standard deviations, RSDs,<1%) and peak areas (RSDs,<7%) for all target compounds.

The role of matrix effects on the quantification of abscisic acid and its metabolites in the leaves of Bauhinia variegata L. using liquid chromatography combined with tandem mass spectrometry

“…In contrast to our results, Fan et al (2011) reported no matrix effects on the analysis of several phytohormones (including ABA) in canola. Their results are in accordance with our findings, because ABA showed a 6.6% matrix effect (Table 3), allowing its quantification by the solvent curve method (i.e., a matrix effect <10%).…”

“…When using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) for plant hormone analysis, co-extracted constituents in the matrix can interfere with the analysis and cause ionization enhancement or suppression (Fan et al, 2011). The type and magnitude of the matrix effect may vary for different concentration ranges (Zrostlíková et al, 2001); thus, the use of surrogates (typically deuterated phytohormones) cannot effectively correct for interference in all cases.…”

Aflatoxin Measurement and Analysis

“…LC-MS provides decisive advantages in performing identification as well as determination of analytes at trace levels. Matrix effects, however, limit the potential of LC-MS. Molecules originating from the sample matrix that coelute with the compounds of interest can interfere with the ionization process in the mass spectrometer, causing ionization suppression or enhancement, which is the so-called matrix effect (Fan et al, 2011). Ion suppression (or enhancement) might be encountered due to matrix components that co-elute with the analyte of interest.…”

Development of an Ultrahigh-Performance Liquid Chromatography–Electrospray Ionization–Tandem Mass Spectrometry Method for the Simultaneous Determination of Salicylic Acid, Jasmonic Acid, and Abscisic Acid in Rose Leaves

“…24 A rapid analysis of multiple hormones is possible through the relatively short dwell times typical for modern mass spectrometers. 25 In addition, the application of ultrahigh-performance liquid chromatography (UHPLC) 26 as well as solid-phase extraction (SPE)-based enrichment 27 may lead to the further improve-ment of sensitivity of phytohormone analysis.…”