Tocopherol and tocotrienol analysis in raw and cooked vegetables: a validated method with emphasis on sample preparation.

Vegetables can be important dietary sources of vitamin E. However, data on vitamin E in raw and cooked vegetables are in part conflicting, indicating analytical pitfalls. The purpose of the study was to develop and validate an HPLC-FLD method for tocochromanol (tocopherols and tocotrienols) analysis equally suitable for raw and cooked vegetables. Significant instability of tocochromanols was observed in raw broccoli and carrot homogenates. Tocochromanols could be stabilized by freeze-drying or ascorbic acid addition prior to homogenization. The optimized protocol for tocochromanol analysis included knife and ball milling of freeze-dried vegetable pieces. Direct acetone extraction of vegetable powders allowed for satisfactory recoveries and precisions. A significant decrease of tocochromanols in baked compared to raw vegetables was shown, the extent of which varied largely between vegetables. For some raw vegetables, such as spinach or broccoli, underestimation of vitamin E in nutrient databases cannot be ruled out and should be examined.