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A new size-exclusion chromatography method for fast rapeseed albumin and globulin quantification

Defaix, Claire, Aymes, Arnaud, Slabi, Sara Albe, Basselin, Mélody, Mathe, Christelle, Galet, Olivier, Kapel, Romain
Food chemistry 2019
Helianthus annuus, absorbance, acetonitrile, albumins, densitometry, gel chromatography, globulins, high performance liquid chromatography, linseed, polyacrylamide gel electrophoresis, rapeseed, storage proteins
The method described in the article aims at the quantification of both main storage proteins, globulins and albumins, in aqueous extract from rapeseed, as an alternative to the current reference methods, Kjeldahl and SDS-PAGE electrophoresis. The new method lies on the analytical separation of extracted compounds by Size-Exclusion High Performance Liquid Chromatography (SE-HPLC) (Biosep-SEC-s2000, 5 µm). The elution of rapeseed extracts with water/acetonitrile/trifluoroacetic acid (45/55/0.1 % v/v) during 30 minutes yields two distinct peaks for the main proteins of rapeseed. Based on the protein extinction coefficients, a calibrationless methodology was developed for their quantification on the basis of the UV signal. The SE-HPLC method was successfully compared to references: Kjeldahl and SDS-PAGE densitometry for the determination of the proportion of each protein. Then, it was successfully applied on two other oleoproteagineous plants, linseed and sunflower.