Simultaneous Determination of Tocopherols, Phytosterols, and Squalene in Vegetable Oils by High Performance Liquid Chromatography-Tandem Mass Spectrometry

Vegetable oil is one of the principal sources of minor bioactive substances such as tocopherol, phytosterol, and squalene. In order to obtain the distribution of these substances in vegetable oil, this study developed and validated high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) method for the simultaneous analysis of tocopherols (α-, β-, γ-, and δ-tocopherol), phytosterols (β-sitosterol, stigmasterol, brassicasterol, and campesterol), and squalene in vegetable oils. An automatic gel permeation chromatography (GPC) system was used to remove triglycerides from oils, avoiding the heat treatment of traditional saponification. Method validation revealed the accuracy and reliability of this method. The current approach achieved the successful separation of β- and γ-tocopherol, which solved the problem of coelution and inaccurate quantification for β- and γ-tocopherol in previous study. Additionally, this method was applied to detect tocopherols, phytosterols, and squalene in 9 categories of vegetable oils.