Abstract
BackgroundBrassinosteriods (BRs) are a group of important phytohormones that have major effects on plant growth and development. To fully elucidate the function of BRs, a sensitive BR assay is required. However, most of the previously reported methods are tedious and time-consuming due to multiple pretreatment steps. Therefore, it is of great significance to develop a method to increase the throughput and detection sensitivity of BR analysis.ResultsWe established a novel analytical method of BRs based on magnetic solid phase extraction (MSPE) combined with in situ derivatization (ISD). TiO2-coated magnetic hollow mesoporous silica spere(TiO2/MHMSS) was served as a double identity- a microextraction sorbent and “microreactor” for the capture and derivatization of BRs in sequence. BRs were first extracted onto TiO2/MHMSS through hydrophilic interaction. The BR-adsorbed TiO2/MHMSS was then employed as a “microreactor” for the derivatization of BRs with 4-(N,N-dimethyamino)phenylboronic acid (DMAPBA). The MSPE-ISD method was simple and fast, which could be accomplished within 10 min. Furthermore, the derivatives of BRs showed better MS response because they were incorporated with tertiary amino groups. Uniquely, endogenous BRs were detected in only 100 mg fresh weight plant tissue.ConclusionOur proposed MSPE-ISD method for the determination of endogenous BRs is rapid and sensitive. It can be applied to the analysis of endogenous BRs in 100 mg fresh plant tissue (Brassica napus L. (B. napus L)). The proposed strategy for plant sample preparation may be extended to develop analytical methods for determination of a wide range of analytes with poor MS response in other complex sample matrices.
Highlights
Brassinosteriods (BRs) are a group of important phytohormones that have major effects on plant growth and development
For the analysis of plant samples, the compromised sensitivity is frequently caused by the signal suppression from complex sample matrix during mass spectrometry (MS) analysis
In hydrophilic interaction chromatography (HILIC), the high content of acetonitrile is normally used as the sampling solution
Summary
Brassinosteriods (BRs) are a group of important phytohormones that have major effects on plant growth and development. For the analysis of plant samples, the compromised sensitivity is frequently caused by the signal suppression from complex sample matrix during mass spectrometry (MS) analysis. The trace amounts of BRs in complex plant matrixes and their inherently low MS response makes reliable qualitative and quantitative analysis of BRs challenging. The current pretreatment methods of BRs to remove the sample matrix required the combination of two or more sample preparation processes, including SPE [7,8], LLE [9], MSPE [10] etc. To improve MS responses of BRs, a pre-column derivatization process was employed to incorporate ionized moieties into BRs before LC-MS analysis [7,11]. It is essential to develop a fast and sensitive BR assay
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