Quantification of 14 Human Milk Oligosaccharides by Ultra-High Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry

Abstract:

[Objective] To establish a method for quantifying 14 human milk oligosaccharides based on ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry. [Method] The supernatant obtained by centrifugation of defatted human milk was loaded on an Waters ACQUITY UPLC^? BEH Amide column (2.1 mm×100 mm×1.7 μm) for gradient elution, with 25 mmol/L ammonium acetate solution and acetonitrile as mobile phases. Data of human milk oligosaccharides were collected by parallel reaction monitoring in the positive ion mode. Quantitative analysis was performed with the external standard method. [Result] The calibration curves of 14 human milk oligosaccharides were linear within the mass concentration range of 0.10~100.00 μg/mL, with the correlation coefficient≥0.980 2. The limits of detection and limits of quantitation were 0.05~1.00 μg/mL and 0.10~5.00 μg/mL, respectively. The recovery of each oligosaccharide added to human milk ranged from 78.03% to 105.23%, with the relative standard deviations ranging from 0.14% to 5.86%. [Conclusion] The results showed that this method could rapidly and effectively detect human milk oligosaccharides in the samples from different sources, providing technical support for the research on human milk oligosaccharides.