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-olchloroform extracts with an aqueous phase impregnating solid Sephadex TM 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. #

Microalgae Lipid Characterization

“…The flow rate was 0.5 mL/min. The solvent composition and gradient program were modified on the basis of previous studies. ,, The solvent system consisted of isopropanol/hexane (2:98, by vol, with 0.1% of stoichiometric triethylamine/formic acid, solvent A1), isopropanol/acetonitrile/chloroform/acetic acid (84:8:8:0.025, by vol, solvent B1), and chloroform/isopropanol/methanol (39:21:40, by vol, solvent A2). The gradient elution program was as follows: 0–2.5 min, 100% A1; 2.5–40 min, 0–100% B1.…”

A Compartmental Comparison of Major Lipid Species in a Coral-Symbiodinium Endosymbiosis: Evidence that the Coral Host Regulates Lipogenesis of Its Cytosolic Lipid Bodies

“…Total lipids from the three collected fractions and cultured Symbiodinium (n = 4 for each of the four treatments/compartments) were first extracted by the Bligh and Dyer procedure [ 20 ]. The extracted lipids were then analyzed by an HPLC instrument equipped with an evaporative light scattering detector (HPLC-ELSD) [ 21 ]. A Hitachi Model L7100 HPLC pump equipped with an auto-sampler (L7200, Hitachi, Japan) was used with a Sedex 80 ELSD (Sedere, France).…”

Fatty Acid and Phospholipid Syntheses Are Prerequisites for the Cell Cycle of Symbiodinium and Their Endosymbiosis within Sea Anemones

“…The extracted lipids were analyzed by a high-performance liquid chromatography equipped with evaporative light scattering detector (HPLC-ELSD) [26]. A Hitachi Model L7100 HPLC pump, equipped with an auto-sampler (L7200, Hitachi, Japan), was used with a Sedex 80 evaporative light-scattering detector (Sedere, France).…”