Development and validation of UPLC method for quantitative determination of major bovine milk oligosaccharides and their heat stability

Abstract

Bovine milk oligosaccharides (BMOs) and human milk oligosaccharides (HMOs) have similar functions. In this investigation, a simple and rapid quantitative analysis method of UPLC for BMOs concentration was proposed, and the heat stability of BMOs was also revealed. BMOs were separated on an Acquity UPLC® Glycan BEH amide column (2.1 mm × 150 mm, 1.7 μm) using 100 mm ammonium formate (pH 4.5) and 100% acetonitrile as mobile phase for gradient elution at a flow rate of 0.45 mL min−1. The potential in quantification of BMOs were satisfactorily validated in terms of linearity (R2 ≥ 0.9992), repeatability and intermediate precision (RSD ≤ 4.46%), and spike recovery (≥90.27%). For all investigated oligosaccharides, LOD ranged from 1.97 to 9.38 μg L−1 and LOQ values ranged between 6.84 and 31.25 μg L−1. In addition, BMOs did not degrade after heating at 45–100 °C for 10–120 min.