Technical note: Development and validation of an HPLC method for the quantification of tocopherols in different types of commercial cow milk

Abstract

In the present study, a methanol-fluorescence-based HPLC method was validated for its use to quantify α-tocopherol and γ-tocopherol in raw milk, whole UHT milk, partially skimmed UHT milk, whole pasteurized milk, and partially skimmed pasteurized milk. Repeatability and reproducibility, calculated as relative standard deviation of 10 measurements within the same day and 30 measurements across 3 d, respectively, were always below 5% for both tocopherols concentrations and retention times. Recovery was assessed through 3 spiking levels and it ranged from 89 to 107%. The method was able to detect the expected declines in tocopherols in milk exposed to UHT or skimming treatments. Vitamin E, calculated as the sum of α-tocopherol and γ-tocopherol, was similar in whole pasteurized and raw milk, averaging 1.57 and 1.56 mg/L, respectively, followed by whole UHT (1.33 mg/L), partially skimmed pasteurized (0.77 mg/L), and partially skimmed UHT milk (0.61 mg/L).