期刊
FOOD CHEMISTRY
卷 341, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2020.128200
关键词
Human milk oligosaccharides; Capillary electrophoresis; Borate complexation
资金
- V4-Korea Joint Research Program, project National Research, Development and Innovation Office (NKFIH) grants of the Hungarian Government [NN 127062]
- New National Excellence Program of the Ministry of Human Capacities [UNKP-19-3-I-DE-356]
- [BIONANO_GINOP-2.3.2-15-2016-00017]
This study presents an ultrafast high-resolution method for separating human milk oligosaccharides (HMOs) using multicapillary gel electrophoresis, which improves the analysis speed and resolution of HMOs.
There is recently growing interest towards synthesized human milk oligosaccharides (HMOs) as baby formula additives, and interestingly also as dietary supplements for adults. Currently quite a few manufacturers synthesize HMOs, however, their analysis is challenging, both in resolution and speed. In this paper an ultrafast high-resolution method is introduced for the separation of HMOs by multicapillary gel electrophoresis. Two gel compositions were evaluated with complementary resolving power. One was a conventionally used industrial standard carbohydrate separation matrix, resolving oligosaccharides according to their charge to hydrodynamic volume ratios. The other one was a borate-buffered dextran gel, which utilized the secondary equilibrium of the borate-vicinal diol complexation to enhance resolution. Considering the rapid analysis time and multiplexing (12-channel system), a 96 well sample plate can be analyzed in less than 80 min with the conventional type carbohydrate separation matrix and in less than one hour with the borate-buffered dextran gel. Exploiting the one fluorophore per molecule labeling stoichiometry, the limit of detection (S/N > 3) and limit of quantitation (S/N > 10) were determined as 0.025 and 0.100 mg/mL, respectively, with good linearity. Based on the calibration plot, the quantities of several low concentration HMOs were determined from a human milk sample.
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