Sequential determination of fat- and water-soluble vitamins in green leafy vegetables during storage

The simultaneous analysis of fat- and water-soluble vitamins from foods is a difficult task considering the wide range of chemical structures involved. In this work, a new procedure based on a sequential extraction and analysis of both types of vitamins is presented. The procedure couples several simple extraction steps to LC–MS/MS and LC–DAD in order to quantify the free vitamins contents in fresh-cut vegetables before and after a 10-days storage period. The developed method allows the correct quantification of vitamins C, B1, B2, B3, B5, B6, B9, E and provitamin A in ready-to-eat green leafy vegetable products including green lettuce, ruby red lettuce, watercress, swiss chard, lamb's lettuce, spearmint, spinach, wild rocket, pea leaves, mizuna, garden cress and red mustard. Using this optimized methodology, low LOQs were attained for the analyzed vitamins in less than 100 min, including extraction and vitamin analysis using 2 optimized procedures; good repeatability and linearity was achieved for all vitamins studied, while recoveries ranged from 83% to 105%. The most abundant free vitamins found in leafy vegetable products were vitamin C, provitamin A and vitamin E. The richest sample on vitamin C and provitamin A was pea leaves (154 mg/g fresh weight and 14.4 mg/100 g fresh weight, respectively), whereas lamb's lettuce was the vegetable with the highest content on vitamin E (3.1 mg/100 g fresh weight). Generally, some losses of vitamins were detected after storage, although the behavior of each vitamin varied strongly among samples.

► A new method to sequentially analyze fat and water-soluble vitamins is presented.
► Ultrasound extraction and LC–MS/MS and LC–DAD are coupled.
► It allows the quantification of several free vitamins in green leafy vegetables.
► Vitamins content in fresh-cut vegetables before and after storage is assessed.