Simultaneous HPLC–DAD quantification of vitamins A and E content in raw, pasteurized, and UHT cow’s milk and their changes during storage
2014
Chotyakul, Nattaporn | Pateiro-Moure, Miriam | Saraiva, Jorge Alexandre | Torres, J Antonio | Pérez-Lamela, Concepción
An improved extraction and HPLC method for the simultaneous extraction and quantitation of retinol, α-tocopherol, α-tocotrienol, and β-carotene was developed to analyze commercial whole/semi-skim/skim samples of raw/pasteurized/UHT milk in transparent plastic/glass bottles and Tetra Brik™ containers. The sample preparation method required prior saponification at 40 °C for 15 min followed by n-hexane extraction. An isocratic acetonitrile/methanol (65:35 v/v) mobile phase, C₁₈analytical column, and UV detector were chosen for HPLC quantification. The liposoluble vitamin content in raw, pasteurized conventional/organic, and UHT milk ranged 0.055–5.540 (retinol), 0.135–1.410 (α-tocopherol), and 0.040–0.850 mg/L (β-carotene). No significant differences (p > 0.05) were observed on losses of retinol, α-tocopherol, and β-carotene content in UHT whole milk after 5 days at 4 °C in the dark. After 14 days at 4 °C in the dark, the contents of retinol, α-tocopherol, and β-carotene remained higher in milk with higher fat content and were higher in unopened containers. In UHT whole milk, samples containing 0.02 % NaN₃, retinol (33 %), and α-tocopherol (11 %) but not β-carotene (2 %) decreased significantly (p < 0.05).
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