Rapid Separating and Enrichment of 4,4′-Dimethylsterols of Vegetable Oils by Solid-Phase Extraction

Phytosterols are separated into three classes: 4-desmethylsterols, 4-monomethylsterols and 4,4′-dimethylsterols. 4,4′-Dimethylsterols are used to detect vegetable oil adulteration and some compounds from this class can have anti-inflammatory and anticancer properties. There are methods such as thin layer chromatography (TLC) and solid phase extraction (SPE) used to separate phytosterol classes from each other. However, in some cases, separation of all three classes is not required. In addition, TLC has some drawbacks such as low recovery and it is time consuming. An SPE method has previously been used, but it was necessary to use high volume of solvents with this method to avoid coelution of phytosterol classes. In this study, an SPE (silica, 1 g) method was developed to separate and enrich only 4,4′-dimethylsterols from unsaponifiables of vegetable oil samples using 25 mL n-hexane and diethyl ether (95:5, v:v). This method was applied to hazelnut and olive oils and results were compared with those of TLC and the previously developed SPE method. Recovery of 4,4′-dimethylsterols was two times higher with the new SPE method compared with the TLC method. The newly developed SPE method generally gave a similar recovery compared with the previously developed SPE method. Moreover, the SPE method developed in this study has the advantage of using a 3.5 times lower volume of solvent than previously developed SPE methods. Because the newly developed SPE method has a single step requiring a low volume of solvents, it is rapid and simple, and can easily be used to detect olive oil adulteration with hazelnut oil and to analyze and quantify effective nutritional compounds in the 4,4′-dimethylsterols class.