Rapid capillary gel electrophoresis analysis of human milk oligosaccharides for food additive manufacturing in-process control

Abstract

Industrial production of human milk oligosaccharides (HMOs) represents a recently growing interest since they serve as key ingredients in baby formulas and are also utilized as dietary supplements for all age groups. Despite their short oligosaccharide chain lengths, HMO analysis is challenging due to extensive positional and linkage variations. Capillary gel electrophoresis primarily separates analyte molecules based on their hydrodynamic volume to charge ratios, thus, offers excellent resolution for most of such otherwise difficult-to-separate isomers. In this work, two commercially available gel compositions were evaluated on the analysis of a mixture of ten synthetic HMOs. The relevant respective separation matrices were then applied to selected analytical in-process control examples. The conventionally used carbohydrate separation matrix was applied for the in-process analysis of bacteria-mediated production of 3-fucosyllactose, lacto-N-tetraose, and lacto-N-neotetraose. The other example showed the suitability of the method for the in vivo in-process control of a shake flask and fermentation approach of 2′-fucosyllactose production. In this latter instance, borate complexation was utilized to efficiently separate the 2′- and 3-fucosylated lactose positional isomers. In all instances, the analysis of the HMOs of interest required only a couple of minutes with high resolution and excellent migration time and peak area reproducibility (average RSD 0.26% and 3.56%, respectively), features representing high importance in food additive manufacturing in-process control. Graphical abstract

Capillary electromigration methods for food analysis and Foodomics: Advances and applications in the period March 2021 to March 2023

“…In the worst case, when both systems have to be used in parallel, a full analysis of 12 samples was possible within less than 20 min with appropriate resolution and quantification features, demonstrating how multi‐CGE can be a powerful analytical tool in challenging applications. This approach can be also used not only for the characterization of samples but also for process control during the synthesis of HMOs [53].…”

Applications of capillary electrophoresis in the fields of environmental, pharmaceutical, clinical, and food analysis (2019–2021)

“…Metal ions [89], acid-base compounds [90,91], amino acids [92], peptides [93], proteins [94], and saccharides [95,96] are very important principal components in food, whose types and contents could affect food safety.…”

“…Metal ions [89], acid‐base compounds [90, 91], amino acids [92], peptides [93], proteins [94], and saccharides [95, 96] are very important principal components in food, whose types and contents could affect food safety. Recently, CE has been gradually applied for the screening of principal components in food to identify the types and authenticity of food, control food quality, and ensure food safety.…”

Capillary electromigration methods for food analysis and Foodomics: Advances and applications in the period February 2019–February 2021

“…This approach has been shown to be useful to describe the products resulting from the HMOs synthesis based on the use of Leloir‐glycosyltransferases, even attaining a good separation between Lacto‐ N ‐biose‐type and Lacto‐ N ‐neo‐type linkage isomers [74]. Similar GCE methods have been also employed to monitor the fermentation products from a modified HMO producer Escherichia coli in a bioreactor, and proposed for in‐process analytical control thanks to their excellent performance [75].…”