Ultra-High-Pressure Liquid Chromatographic method for the analysis of tocopherols in human colostrum and milk

A rapid, highly sensitive and direct Ultra-High-Pressure Liquid Chromatographic method was developed and validated for quantifying δ-, β + γ-, and α-tocopherol in human colostrum and milk. Two reversed-phase chromatographic columns and two detectors (Fluorescence Detector or FD and Photodiode Detector Array or PDA) were used and both methods were independently validated. Two internal standards were selected according to the detector used. Recoveries ranged from 96.71% to 103.55% and the relative standard deviations for the within-day precision were below 6% (PDA) and 3% (FD). Both approaches enabled to achieve low detection limits, on the order of ng (PDA) or pg (FD). Only 300 μL of sample and a chromatographic run of less than 1.6 min were enough to efficiently quantify the isomers in the colostrum and milk of Spanish women.