New procedures for rapid screening of leaf lipid components from Arabidopsis

Abstract

Four procedures have ben developed to simplify routine analysis of large numbers of plant lipid samples. First, a rapid extraction method has been devised in which the key step consists of washing propan-2-ol-chloroform extracts with an aqueous phase impregnating solid Sephadex® G-25-300. The extracts are then separated by high performance liquid chromatography (HPLC) with evaporative light-scattering detection, using a complex gradient that permits separation of most of the important simple lipid, glycolipid and phospholipid classes in sequence. An example of an application to the analysis of a newly generated mutant is illustrated. In addition, a novel solid-phase extraction procedure with a bonded propylamine phase allows separation of four broad classes, simple lipids, glycolipids, zwitterionic phospholipids and acidic phospholipids (plus sulphoquinovosyldiacylglycerol), for further analysis. Finally, base-catalysed transesterification followed by HPLC analysis, permits identification of those lipids that do not contain conventional ester bonds.