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. 2013 May;405(12):4089-105.
doi: 10.1007/s00216-013-6817-1. Epub 2013 Mar 7.

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

Maria Lorna A De Leoz  1 , Shuai Wu, John S Strum, Milady R Niñonuevo, Stephanie C Gaerlan, Majid Mirmiran, J Bruce German, David A Mills, Carlito B Lebrilla, Mark A Underwood
Affiliations

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

Maria Lorna A De Leoz et al. Anal Bioanal Chem. 2013 May.

Abstract

Human milk oligosaccharides (HMOs), though non-nutritive to the infant, shape the intestinal microbiota and protect against pathogens during early growth and development. Infant formulas with added galacto-oligosaccharides have been developed to mimic the beneficial effects of HMOs. Premature infants have an immature immune system and a leaky gut and are thus highly susceptible to opportunistic infections. A method employing nanoflow liquid chromatography time-of-flight mass spectrometry (MS) is presented to simultaneously identify and quantify HMOs in the feces and urine of infants, of which 75 HMOs have previously been fully structurally elucidated. Matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance MS was employed for high-resolution and rapid compositional profiling. To demonstrate this novel method, samples from mother-infant dyads as well as samples from infants receiving infant formula fortified with dietary galacto-oligosaccharides or probiotic bifidobacteria were analyzed. Ingested oligosaccharides are demonstrated in high abundance in the infant feces and urine. While the method was developed to examine specimens from preterm infants, it is of general utility and can be used to monitor oligosaccharide consumption and utilization in term infants, children, and adults. This method may therefore provide diagnostic and therapeutic opportunities. VSports手机版.

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Figures

Figure 1
Figure 1
Workflow of the analysis of human milk oligosaccharides from biological samples of mother-infant dyads. *HMO library from [23, 24]
Figure 2
Figure 2
MALDI FT-ICR MS profiles in positive ion mode of the HMOs in milk (A), feces (B), and urine (C) of a mother-preterm infant dyad. Milk, feces and urine samples are 5, 1 and 150 ug, respectively. HMOs are marked with blue dots. Distributions of fucosylated and sialylated glycans are based on HMO intensities normalized against the total HMO intensities. Pie charts represent nanoLC MS data.
Figure 3
Figure 3
MALDI FT-ICR MS profiles of HMOs in mother’s preterm milk (A) and pasteurized concentrated donor human milk fortifier (B).
Figure 4
Figure 4
Annotated base peak chromatograms of the HMOs in preterm milk (A) and the pasteurized concentrated donor human milk fortifier (B). A total of 70 and 100 possible HMOs were found in preterm milk and human milk fortifier, respectively.
Figure 5
Figure 5
MALDI FT-ICR MS profiles of temporal fecal HMOs from one preterm infant. Concentrated donor human milk fortifier was added beginning week 4. Increasing amounts of donor human milk were given in weeks 5-7
Figure 6
Figure 6
Temporal nano-LC Chip/TOF MS base peak chromatograms of the MS spectra shown in Figure 5. The pie charts show the distributions based on abundances normalized according to the total HMO abundance per sample using nano-LC Chip/TOF MS
Figure 7
Figure 7
Fecal oligosaccharide MALDI FT-ICR MS profiles taken in the positive ion mode of an infant fed formula supplemented with A) GOS, and B) B. lactis
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References

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