Characterization of Oligosaccharides in Milk and Feces of Breast-Fed Infants by High-Performance Anion-Exchange Chromatography

Coppa, Giovanni V.; Pierani, Paolo; Zampini, Lucia; Bruni, Stefano; Carloni, Ines; Gabrielli, Orazio

doi:10.1007/978-1-4615-1371-1_38

Cited by 107 publications

(89 citation statements)

References 12 publications

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“…The individual variation of HMOs in breast milk and their recovery in feces has been reported previously [7,16,23,30]. However, in most cases the identification of HMOs was limited to the purified HMOs commercially available or connected with tedious preparative fractionation.…”

Section: Unknown Hmosmentioning

confidence: 92%

CE‐LIF‐MSⁿ profiling of oligosaccharides in human milk and feces of breast‐fed babies

et al. 2010

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Mixtures of the complex human milk oligosaccharides (HMOs) are difficult to analyze and gastrointestinal bioconversion products of HMOs may complicate analysis even more. Their analysis, therefore, requires the combination of a sensitive and high-resolution separation technique with a mass identification tool. This study introduces for the first time the hyphenation of CE with an electrospray mass spectrometer, capable to perform multiple MS analysis (ESI-MS(n)) for the separation and characterization of HMOs in breast milk and feces of breast-fed babies. LIF was used for on- and off-line detections. From the overall 47 peaks detected in off-line CE-LIF electropherograms, 21 peaks could be unambiguously and 11 peaks could be tentatively assigned. The detailed structural characterization of a novel lacto-N-neo-tetraose isomer and a novel lacto-N-fucopentaose isomer was established in baby feces and pointed to gastrointestinal hydrolysis of higher-Mw HMOs. CE-LIF-ESI-MS(n) presents, therefore, a useful tool which contributes to an advanced understanding on the fate of individual HMOs during their gastrointestinal passage.

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Section: Unknown Hmosmentioning

confidence: 92%

CE‐LIF‐MSⁿ profiling of oligosaccharides in human milk and feces of breast‐fed babies

et al. 2010

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“…To a small extent some HMO are absorbed intact and excreted in the urine [7], yet estimates of between 40-50% [8] and 97% [5] are recovered in the feces of infants consuming them. Beneficial physiological activities have been ascribed to HMO.…”

Section: Introductionmentioning

confidence: 99%

In vitro fermentability of human milk oligosaccharides by several strains of bifidobacteria

Ward

Niñonuevo

Mills

et al. 2007

Molecular Nutrition Food Res

222

209

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This study was conducted to investigate the catabolism and fermentation of human milk oligosaccharides (HMO) by individual strains of bifidobacteria. Oligosaccharides were isolated from a pooled sample of human milk using solid-phase extraction, and then added to a growth medium as the sole source of fermentable carbohydrate. Of five strains of bifidobacteria tested (Bifidobacterium longum biovar infantis, Bifidobacterium bifidum, Bifidobacterium longum biovar longum, Bifidobacterium breve, and Bifidobacterium adolescentis), B. longum bv. infantis grew better, achieving triple the cell density then the other strains. B. bifidum did not reach a high cell density, yet generated free sialic acid, fucose and N-acetylglucosamine in the media, suggesting some capacity for HMO degradation. Thin layer chromatography profiles of spent fermentation broth suggests substantial degradation of oligosaccharides by B. longum bv. infantis, moderate degradation by B. bifidum and little degradation by other strains. While all strains were able to individually ferment two monosaccharide constituents of HMO, glucose and galactose, only B. longum bv. infantis and B. breve were able to ferment glucosamine, fucose and sialic acid. These results suggest that as a potential prebiotic, HMO may selectively promote the growth of certain bifidobacteria strains, and their catabolism may result in free monosaccharides in the colonic lumen.

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“…HMO are the third most abundant component of human milk (9), and at least 130 different masses have been identified (14). In vitro, HMO are resistant to catabolism by host hydrolases (3,4), and based on the mass balance between consumption and excretion, Chaturvedi et al (1) calculated that 97% of HMO pass through infants undigested, while Coppa et al (2) estimated that 40 to 50% of HMO pass through infants undigested.…”

mentioning

confidence: 99%

In Vitro Fermentation of Breast Milk Oligosaccharides by Bifidobacterium infantis and Lactobacillus gasseri

Ward¹,

Niñonuevo²,

Mills

et al. 2006

Appl Environ Microbiol

266

229

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It has been proposed that human milk oligosaccharides (HMO) function as a prebiotic for bifidobacteria, yet this activity has not been adequately investigated. In this study, Bifidobacterium infantis was shown to ferment purified HMO as a sole carbon source, while another gut commensal, Lactobacillus gasseri, did not ferment HMO. Our results support the hypothesis that HMO selectively amplify bacterial populations in the infant intestine.Human milk is unique because of the high concentration and diversity of human milk oligosaccharides (HMO). HMO are the third most abundant component of human milk (9), and at least 130 different masses have been identified (14). In vitro, HMO are resistant to catabolism by host hydrolases (3, 4), and based on the mass balance between consumption and excretion, Chaturvedi et al.(1) calculated that 97% of HMO pass through infants undigested, while Coppa et al. (2) estimated that 40 to 50% of HMO pass through infants undigested.A prebiotic function was attributed to HMO based on studies done in the 1950s with Bifidobacterium bifidus subsp. pennsylvannicus (6, 7), yet the measurements were based on growth enhancement, not fermentability, as the media contained lactose. Direct fermentation of HMO by Bifidobacterium spp. and/or Lactobacillus spp. has not been demonstrated yet, and there are many questions about the metabolic fate of these molecules. Do subpopulations mediate specific functions, such as pathogen binding or prebiotic activity? Are specific bacterial species required for catabolism in the gut? The goals of this investigation were to determine the fermentability of HMO by two representative species of breast-fed infant microbiota, Bifidobacterium infantis and Lactobacillus gasseri (8,10) and to characterize the changes in the HMO after bacterial growth. Determination of the biological basis underlying the HMO abundance in human milk should be of general interest in human nutrition (15).Pooled milk was provided by the Mother's Milk Bank of San Jose, CA, and the Mother's Milk Bank of Austin, TX. Oligosaccharides were extracted as described by Gnoth et al. (5), with modifications. One liter of milk was centrifuged at 5,000 ϫ g for 30 min at 4°C, and the fat was removed. Ethanol (2 liters) was added, and the solution was incubated overnight at 4°C. The precipitate was removed by centrifugation at 5,000 ϫ g for 30 min at 4°C, and the solvent was removed by rotary evaporation. The concentration of the solution was adjusted to 0.05 M with phosphate buffer (pH 6.8), 3,000 U ␤-galactosidase (Kluyveromyces fragilis) was added, and the solution was incubated for 1 h at 37°C. Then the solution was extracted with 4 volumes of chloroform-methanol (2:1, vol/vol), and the aqueous layer was collected.As described by Redmond and Packer (12), monosaccharides and disaccharides were removed by selective adsorption of HMO, using solid-phase extraction with graphitized nonporous graphitized carbon cartridges (Supelco Inc., Bellefonte, PA). The oligosaccharides that were retained were eluted wit...

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Characterization of Oligosaccharides in Milk and Feces of Breast-Fed Infants by High-Performance Anion-Exchange Chromatography

Cited by 107 publications

References 12 publications

CE‐LIF‐MSⁿ profiling of oligosaccharides in human milk and feces of breast‐fed babies

CE‐LIF‐MSⁿ profiling of oligosaccharides in human milk and feces of breast‐fed babies

In vitro fermentability of human milk oligosaccharides by several strains of bifidobacteria

In Vitro Fermentation of Breast Milk Oligosaccharides by Bifidobacterium infantis and Lactobacillus gasseri

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Characterization of Oligosaccharides in Milk and Feces of Breast-Fed Infants by High-Performance Anion-Exchange Chromatography

Cited by 107 publications

References 12 publications

CE‐LIF‐MSn profiling of oligosaccharides in human milk and feces of breast‐fed babies

CE‐LIF‐MSn profiling of oligosaccharides in human milk and feces of breast‐fed babies

In vitro fermentability of human milk oligosaccharides by several strains of bifidobacteria

In Vitro Fermentation of Breast Milk Oligosaccharides by Bifidobacterium infantis and Lactobacillus gasseri

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Product

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CE‐LIF‐MSⁿ profiling of oligosaccharides in human milk and feces of breast‐fed babies

CE‐LIF‐MSⁿ profiling of oligosaccharides in human milk and feces of breast‐fed babies