4.7 Article

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

Journal

ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 413, Issue 6, Pages 1595-1603

Publisher

SPRINGER HEIDELBERG
DOI: 10.1007/s00216-020-03119-0

Keywords

Human milk oligosaccharides; In-process control; Capillary gel electrophoresis

Funding

  1. University of Debrecen
  2. National Research, Development and Innovation Office of the Hungarian Government [BIONANO_GINOP-2.3.2-15-2016-00017, 2018-2.1.17-TET-KR-2018-00010
  3. 2020-4.1.1-TKP2020]
  4. New National Excellence Program Hungarian Ministry of Human Capacities [UNKP-20-5]
  5. Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences

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Industrial production of human milk oligosaccharides (HMOs) is gaining increasing attention, with challenges in analysis overcome by capillary gel electrophoresis. Commercially available gel compositions were successful in analyzing a mixture of synthetic HMOs and demonstrated feasibility in real-time process control examples.
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.

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