A quantitative and comprehensive method to analyze human milk oligosaccharide structures in the urine and feces of infants

Leoz, Maria Lorna A. De; Wu, Shuai; Strum, John S.; Niñonuevo, Milady R.; Gaerlan, Stephanie C.; Mirmiran, M.; German, J. Bruce; Mills, David A.; Lebrilla, Carlito B.; Underwood, Mark A.

doi:10.1007/s00216-013-6817-1

Cited by 89 publications

(104 citation statements)

References 30 publications

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“…To identify the oligosaccharide structures, their mass and retention times were compared to a library previously developed for milk in our group [13,14], where a 15 ppm mass error was allowed. Moreover, comparisons to previous analyses of urine samples [23], which should have more similar oligosaccharide compositions to plasma than milk, were performed. Oligosaccharide compositions, retention times and volume were exported to csv-format.…”

Section: Methodsmentioning

confidence: 99%

Detection of milk oligosaccharides in plasma of infants

et al. 2014

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Human Milk Oligosaccharides (HMO) are one of the major components of human milk. HMO are non-digestible by the human gut, where they are known to play important functions as prebiotics and decoys for binding pathogens. Moreover, it has been proposed that HMO may provide sialic acids to the infant that are important in brain development, however this would require absorption of HMO into the bloodstream. HMO have consistently been found in the urine of humans and other mammals, suggesting systemic absorption. Here we present a procedure for the profiling of milk oligosaccharides (MO) in plasma samples obtained from 13 term infants hospitalized for surgery for congenital heart disease. The method comprises protein denaturation, oligosaccharide reduction and porous graphitized carbon solid phase extraction for purification followed by analysis using nHPLC-PGC-chip-TOF-MS. Approximately 15 free MO were typically observed in the plasma of human infants, including LNT, LDFP, LNFT, 3’SL, 6’SL, 3’SLN and 6’SLN, of which the presence was confirmed using fragmentation studies. A novel third isomer of SLN, not found in human or bovine milk was also consistently detected. Differences in the free MO profiles were observed between infants that were totally formula-fed and infants that received at least some part breast milk. Our results indicate that free MO similar in structure to those found in human milk and urine are present in the blood of infants. The method and results presented here will facilitate further research toward the possible roles of free MO in the development of the infant.

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Section: Methodsmentioning

confidence: 99%

Detection of milk oligosaccharides in plasma of infants

et al. 2014

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“…A comprehensive label-free procedure to identify and quantify milk oligosaccharides and other glycans using nano-liquid chromatography-time of flight MS was recently developed [113]. During this study, MALDI-Fourier transform ion cyclotron resonance (FTICR) was used for accurate mass measurement compositional analysis.…”

Section: Mass Spectrometry Methodsmentioning

confidence: 99%

Advances in Fractionation and Analysis of Milk Carbohydrates

Nagaraj¹,

Upadhyay²,

Nath³

et al. 2018

Technological Approaches for Novel Applications in Dairy Processing

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Lactose is a principal component of the milk obtained from the mammals. Milk also contains several other sugars such as monosaccharides and oligosaccharides in low concentrations. Lactose has reactive functional groups and can be converted to valuable food-grade derivatives for commercial applications through chemical and/or enzymatic reactions. Physical and chemical properties of carbohydrates determine the methods that can be used for their fractionation and purification. In this chapter, the advanced extraction techniques for fractionation and analytical methodologies applied for the determination of different carbohydrates of milk (lactose, lactulose, and oligosaccharides) are summarized.The main aim of this contribution is to provide the reader with a broad view on the recent fractionation and analytical techniques employed for the analysis of carbohydrates in dairy foods and their applications in food and pharmaceutical industry.

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“…Oligosaccharide Extraction-Free oligosaccharides were extracted from breast milk and infant feces following previously reported methods, with slight modifications (8,26,33). Fecal samples were weighed, diluted with nanopure water to 1 mg/10 l, and homogenized in the shaker overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…Along with the degraded HMO, intact HMOs have also been discovered in the feces of breast-fed infants (15,16,25,26). Computational, genomic, and in vitro methods have been combined to study bacterial consumption of HMOs and other carbohydrates.…”

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Identification of Oligosaccharides in Feces of Breast-fed Infants and Their Correlation with the Gut Microbial Community

Davis

Totten

Huang

et al. 2016

Molecular & Cellular Proteomics

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Glycans in breast milk are abundant and found as either free oligosaccharides or conjugated to proteins and lipids. Free human milk oligosaccharides (HMOs) function as prebiotics by stimulating the growth of beneficial bacteria while preventing the binding of harmful bacteria to intestinal epithelial cells. Bacteria have adapted to the glycanrich environment of the gut by developing enzymes that catabolize glycans. The decrease in HMOs and the increase in glycan digestion products give indications of the active enzymes in the microbial population. In this study, we quantitated the disappearance of intact HMOs and characterized the glycan digestion products in the gut that are produced by the action of microbial enzymes on HMOs and glycoconjugates from breast milk. Oligosaccharides from fecal samples of exclusively breast-fed infants were extracted and profiled using nanoLC-MS. Intact HMOs were found in the fecal samples, additionally, other oligosaccharides were found corresponding to degraded HMOs and non-HMO based compounds. The latter compounds were fragments of N-glycans released through the cleavage of the linkage to the asparagine residue and through cleavage of the chitobiose core of the N-glycan. Marker gene sequencing of the fecal samples revealed bifidobacteria as the dominant inhabitants of the infant gastrointestinal tracts. A glycosidase from Bifidobacterium longum subsp. longum was then expressed to digest HMOs in vitro, which showed that the digested oligosaccharides in feces corresponded to the action of glycosidases on HMOs. Similar expression of endoglycosidases also showed that N-glycans were released by bacterial enzymes. Although bifidobacteria may dominate the gut, it is possible that specific minority species are also responsible for the major products observed in feces. Nonetheless, the enzymatic activity correlated well with the known glycosidases in the respective bacteria, suggesting a direct relationship between microbial abundances and catabolic activity. Molecular & Cellular

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A quantitative and comprehensive method to analyze human milk oligosaccharide structures in the urine and feces of infants

Cited by 89 publications

References 30 publications

Detection of milk oligosaccharides in plasma of infants

Detection of milk oligosaccharides in plasma of infants

Advances in Fractionation and Analysis of Milk Carbohydrates

Identification of Oligosaccharides in Feces of Breast-fed Infants and Their Correlation with the Gut Microbial Community

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