LC-ESI-MS/MS Method for the profiling of glycerophospholipids and its application to the analysis of tobacco hairy roots as early indicators of phenol pollution

Abstract

The growing impact of organic and inorganic pollutants on the environment has led to the development of numerous strategies to counteract this effect. Among all, green technologies such as phytoremediation, has won relevance for its low cost as well as being an eco-friendly and sustainable process. In the past years, tobacco hairy roots (HRs) have become a good example of the effectiveness of plants as a tool for removing different pollutants. In this sense, the understanding of the intracellular mechanisms involved in the phytoremediation process has become of great interest. Glycerophospholipids (GLPs) are a key factor on intracellular signaling, given that they can activate different pathways, despite there is not enough information available about their levels and fluctuation under certain stress conditions. The aim of this work was to develop of a new chromatographic method coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) with selected reaction monitoring (SRM) and data dependent scan detection for the evaluation of GLP profile after the exposure to phenol, to get a better understanding of lipids turnover and their effect on signaling pathways. This highly sensitive method allows the identification, separation and quantification in less than a 20 min-run of the eleven most relevant GPL species present in hairy tobacco root extracts after phenol exposure, and is suitable for evaluating small changes in GPL levels.