The aim of this study was the determination of chromatographic profiles of major whey proteins (WP): α-lactalbumin (α-La), β-lactoglobulin A and B, (β-Lg A and B), bovine serum albumin (BSA) in dairy beverages based on Zonar milk serum. The studied WP were separated by high-performance liquid chromatography (HPLC) on Aeris XB-C18 column using gradient elution with 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 80% acetonitrile, at 214 nm detection. The HPLC method was validated for system suitability, selectivity/specificity, linearity (R2 ≥ 0.996), precision (RSD% ≤ 2.01), trueness (recovery 96.29%-102.08%), sensitivity (limits of quantification (LOQ) 1.35-10.08 μg/mL), and robustness. The total of studied whey proteins (WPT) in dairy beverages based on Zonar milk serum varied between 1.42 g/L and 3.04 g/L. The results obtained were evaluated by principal component analysis (PCA) for correlation between the types of studied dairy beverages (natural and with additives added), the HPLC dataset of four whey proteins and WPT (active variables) to differentiate type of dairy beverages samples. The obtained results confirm that the studied dairy beverages constitute a valuable source of bioactive components with benefits in human healthy nutrition and medical purposes.
Keywords: Dairy beverages; HPLC; validation; whey proteins.
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