Analytical and Bioanalytical Chemistry · 6 authors, 1 centre
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This paper describes the development and validation of a novel countercurrent chromatography (ccCCC) method for preparing free sterol fractions from vegetable oils. The method allows for the processing of large sample quantities (~800 mg per injection) with good reproducibility and can also recover free tocochromanols.
The method utilized a solvent system of n-hexane/ethanol/methanol/water (34:11:12:2, v/v/v/v) and introduced two UV-active internal standards (8-phenyl octanoic acid n-hexyl ester and 3-phenyl propionic acid n-propyl ester) to mark the fractionation window. The ccCCC method was validated by analyzing five vegetable oils in duplicate (chili seed oil, high oleic sunflower oil, palm oil, soybean oil, and corn oil). The method also enabled the detection of free tocochromanols (tocopherols) without antioxidant protection. A key limitation noted is the higher solvent requirement compared to conventional methods, though solvent reduction measures were implemented. The overall sample preparation time per oil was one hour.
