Comparison of reversed-phase liquid chromatography–mass spectrometry with electrospray and atmospheric pressure chemical ionization for analysis of dietary tocopherols

ESI and APCI ionization techniques in both negative and positive ion modes were evaluated for simultaneous LC–MS analysis of the four tocopherol homologues (α, β, γ and δ). The ESI and APCI ionization of tocopherols in positive ion mode was not efficient and proceeded via two competitive mechanisms, with the formation of protonated pseudo-molecular ions and molecular ions, which adversely influenced the repeatability of MS signal. Ionization in negative ion mode in both ESI and APCI was more efficient as it only produced target deprotonated pseudo-molecular ions. The APCI in negative ion mode showed larger linearity range, lower detection limits and was less sensitive to the differences in chemical structure of analytes and nature of applied solvents than negative ion ESI. Negative ion APCI was, therefore, chosen for the development of LC–MS method for simultaneous determination of the four tocopherols in foods. A baseline separation of the tocopherols was achieved on novel pentafluorophenyl silica-based column Fluophase PFP. The use of methanol–water (95:5, v/v) as a mobile phase was preferable to the use of acetonitrile–water due to considerable gain in MS signal. The limits of quantifications were 9 ng/mL for α-tocopherol, 8 ng/mL for β- and γ- and 7.5 ng/mL for δ-tocopherol when 2 μL was injected. This method was successfully applied to determination of tocopherols in sunflower oil and milk.