Ultra Performance Liquid Chromatography and High Resolution Mass Spectrometry for the Analysis of Plant Lipids

Jan Hummel,Yan Li,Shruthi Segu,Jessica Jueppner,Patrick Giavalisco,Susann Irgang

doi:10.3389/fpls.2011.00054

Abstract

Holistic analysis of lipids is becoming increasingly popular in the life sciences. Recently, several interesting, mass spectrometry-based studies have been conducted, especially in plant biology. However, while great advancements have been made we are still far from detecting all the lipids species in an organism. In this study we developed an ultra performance liquid chromatography-based method using a high resolution, accurate mass, mass spectrometer for the comprehensive profiling of more than 260 polar and non-polar Arabidopsis thaliana leaf lipids. The method is fully compatible to the commonly used lipid extraction protocols and provides a viable alternative to the commonly used direct infusion-based shotgun lipidomics approaches. The whole process is described in detail and compared to alternative lipidomic approaches. Next to the developed method we also introduce an in-house developed database search software (GoBioSpace), which allows one to perform targeted or un-targeted lipidomic and metabolomic analysis on mass spectrometric data of every kind.

Highlights

Holistic analysis of a cellular metabolome, the complement of all small molecules within a cell (Oliver et al, 1998), is still quite complicated due to the huge complexity and the large chemical heterogeneity of all the contained molecules
In this report we describe a versatile and reproducible ultra performance liquid chromatography (UPLC)-based separation system, coupled to a high resolution mass spectrometer operating in mass spectrometry (MS) as well as all-ion fragmentation mode
The extracted lipids were analyzed on a C8 reversed phase UPLC column, using 1.7 μm particles (Rainville et al, 2007), in a 22 min method

Summary

Introduction

Holistic analysis of a cellular metabolome, the complement of all small molecules within a cell (Oliver et al, 1998), is still quite complicated due to the huge complexity and the large chemical heterogeneity of all the contained molecules. In the automated approach the molecular masses, retention time, and associated peak intensities for the three replicates of each sample were extracted from the raw files, which contained the full scan MS and the all-ion fragmentation MS data.

Results

Conclusion