CULTIVAR IDENTIFICATION IN BRASSICA-NAPUS L BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF ETHANOL EXTRACTS

A method has been developed to identify cultivars of oilseed rape (Brassica napus L.), an outcrossing species, by reversed-phase high-performance liquid chromatography (HPLC) based on the separation of ethanol-extractable seed components. Defatted flour was extracted into 70% ethanol in water, and chromatography was carried out in a reversed-phase C18 column. Optimum results were achieved with water solvent and a gradient to 50% acetonitrile, and peak detection at 210 nm and 240 nm. Trifluoroacetic acid (0.1%) was added to dissociate proteins. Extraction and chromatographic conditions were reproducible, and significant differences were found in the chromatograms of the 29 cultivars studied. The chromatographic differences between these cultivars were stable over a range of growing environments. It was possible to allocate unknown cultivars to their correct classes with the HPLC method together with discriminate analysis. This test has potential as a method of characterization for cultivar registration.