A Validated Method for Detection and Quantitation of 2’Fucosyllactose in Infant Formula Matrices

Abstract

Analytical methods to assess the content of free carbohydrates in solution range from simple tests of reductive power to combinations of chromatography and mass spectrometry. Soluble carbohydrates such as lactose, maltose, fructooligosaccharides, and galactooligosaccharides are commonly found in infant formulas either as sources of energy or soluble fibers. On the other hand, a rich repertoire of lactose-based carbohydrates occurs naturally in human milk. The advent of novel biosynthesis technologies resulted in the availability of human milk oligosaccharide structures that are being used as ingredients in infant formulas. Notably, 2’Fucosyllactose has been tested in preclinical and clinical studies to determine its safety and to explore its potential health benefits in the context of pediatric nutrition. Several chromatographic methods for the analysis of human milk oligosaccharides have been published and, the main challenge associated with 2’Fucosyllactose quantitation has been to improve its resolution from lactose, which is present at concentrations around 70 g/l in both, infant formula and human milk. We developed a high-performance anion exchange chromatography method to detect and quantify 2’ Fucosyllactose in the presence of lactose by expanding the elution time between these saccharides. We validated the analytical procedure which behaved linearly (average R=0.99951) at concentrations as low as 1.75 µg/ml (limit of quantitation) with an average limit of detection of 0.43 µg/ml.