Tocopherols in espresso coffee: Analytical method development and validation

The present paper reports the development and validation of an analytical micro-method for tocopherols quantification in espresso coffee by normal-phase HPLC with fluorescence detection. The proposed method consists in a liquid–liquid extraction with n-hexane:ethyl acetate, followed by a clean-up with dimethylformamide to eliminate co-eluting interferences. The method showed good intra- and inter-day precisions (coefficient of variation < 6.5%), good accuracies (98 ± 6%), and high correlation coefficients (r > 0.999) for standards subjected to the entire procedure. Only α- and β-tocopherols were identified in the brews. The detection and quantification limits were 0.5 and 1.4 ng/ml, for α-tocopherol, and 0.4 and 1.1 ng/ml, for β-tocopherol, respectively. A mean total tocopherol content (α + β) of 3.5 ± 0.9 μg in commercial espresso coffee blends (30 ml) was detected. The proposed method requires low solvent consumption and proved to be sensitive, precise and accurate.