High Performance Liquid Chromatography of Molecular Species from Free Sterols and Sterylglycosides Isolated from Oat Leaves and Seeds1

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Abstract
Free sterols and sterylglycosides (SG) from oat leaves and seeds were isolated by conventional thin layer chromatography (TLC) and subjected to high performance liquid chromatography (HPLC) for resolution of molecular species. Acylsterylglycosides, isolated by TLC, were converted to SG by mild alkaline hydrolysis and determined as SG. Sterols and SG were injected onto the column without any chemical treatment and the separated species were detected at 200 nm. The separation of SG-species follows exactly the separation of free sterols. Though gas liquid chromatography still is the method of choice, advantages of HPLC is to analyse directly the SG-species without hydrolysis and derivatization as compared to GLC. After TLC the sterol- and the SG-fraction are injected directly onto the column. This is extremely important for labile sterylglycosides or sterols, as demonstrated for the avenasterols.