A method for the simultaneous extraction and quantitation of lipophilic antioxidants in Triticum sp. by HPLC-DAD/FLD-MSn

A method for the simultaneous determination of carotenoids, tocopherols, tocotrienols, alk(en)ylresorcinols, and steryl ferulates from whole grain wheat flour was developed and validated using HPLC-DAD/FLD-MSn. After comparing various solvents and extraction techniques, samples were extracted with acetone without previous saponification. Extraction recoveries ranged from 93.2 to 101.6% for carotenoids, tocopherols, and steryl ferulates, while 83.4-99.4% were determined for alkylresorcinols. The inter-day CV for all analytes was below 5%. HPLC separation was compared on porous particle C18- and C30- as well as a core-shell particle C18-column, achieving optimum separation on the latter within 40 min after gradient optimization. The limit of detection (LOD) ranged from 0.03 to 20.41 ng on column for lutein and tricosylresorcinol, respectively. Further validation parameters are reported in detail. HPLC-MSn supported the identification of minor compounds, when standards were unavailable. This method is proved to be suitable for high-throughput screening of a large number of lipophilic compounds in wheat.