Quantitative nuclear magnetic analysis of human milk oligosaccharides 2’-fucosyllactose and 3-fucosyllactose in complicated food matrices

Abstract

Human milk oligosaccharides (HMOs) are the third most abundant solid component in breast milk, which is essential for the healthy growth of infants. 2′-fucosyllactose (2’-FL) and 3-fucosyllactose (3-FL) are the main components of HMOs. In this study, we developed and validated a quantitative nuclear magnetic resonance (qNMR) method for determining HMOs in different complex food matrices. The spectra of HMOs often overlap with impurities in the samples, which hampers quantification using the conventional integral method. We overcame this obstacle by building the Global Spectrum Deconvolution-area coefficients algorithm.

The method proved to be precise, yielding satisfactory results in terms of precision (RSD% 0.5–1.9), trueness (bias% 1.9–8.2), and recovery (90.5–106.6 %). The limit of quantification for 2’-FL was 0.10 mg/mL, and for 3-FL, it was 0.15 mg/mL. A comparison using Deming regression between the existing high-performance liquid chromatography method and the developed qNMR method for determining HMOs demonstrated the high accuracy of the qNMR method. The proposed method allows for high-throughput measurements of large-volume samples due to its simple and fast sample preparation. Therefore, the method is an essential tool for determining HMOs in various complex foods.