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New procedures for rapid screening of leaf lipid components from Arabidopsis
Author(s) -
Christie William W.,
Gill Samantha,
Nordbäck Johan,
Itabashi Yutaka,
Sanda Sherrie,
Slabas Antoni R.
Publication year - 1998
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/(sici)1099-1565(199803/04)9:2<53::aid-pca391>3.0.co;2-u
Subject(s) - chemistry , chromatography , high performance liquid chromatography , extraction (chemistry) , glycolipid , chloroform , phospholipid , solid phase extraction , transesterification , organic chemistry , biochemistry , membrane , methanol
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 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. © 1998 John Wiley & Sons, Ltd.