Neutral and acidic oligosaccharides in Holstein-Friesian colostrum during the first 3 days of lactation measured by high performance liquid chromatography on a microfluidic chip and time-of-flight mass spectrometry

Department of Food Science and Technology, University of California, Davis 95616, USA.
Journal of Dairy Science (Impact Factor: 2.57). 09/2010; 93(9):3940-9. DOI: 10.3168/jds.2010-3156
Source: PubMed


Oligosaccharides (OS) from bovine milk are a class of bioactive molecules that are receiving increasing commercial attention for their potential health benefits. In the present work we measured, comprehensively and systematically, free milk OS in the colostrum of 7 Holstein-Friesian cows during the first 3 d of lactation in 12-h intervals by HPLC-chip/time-of-flight mass spectrometry to determine the biological variation of free milk OS in early lactation. The high sensitivity and resolution of the analytical technique made it possible to monitor all OS species, thus providing a comprehensive and quantitative analysis of OS variations during colostrum production. This study confirmed that although sialyllactose is the major OS in bovine colostrum, several neutral OS species are present in significant abundance even at the third day of lactation. Furthermore, variation in terms of OS species and relative abundances of OS between cows suggest individual animal variation. These variations are likely due to genetic factors because environmental factors such as nutrition, lactation number, and accommodation were the same for all cows. This investigation revealed that colostrum milk from Holstein-Friesian cows is a rich source of neutral and acidic OS for the food and pharmaceutical industries.

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Available from: Daniela Barile, May 14, 2014
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    • "In recent years, the use of nano-liquid chip-based technologies mainly coupled to MS or tandem MS (MS/MS) techniques have demonstrated to be extremely helpful for OS identification and it has been applied to milk characterization due to its high sensitivity and capacity for compositional verification [4]. Nano-LC-Chip technology coupled to time of flight (TOF) MS has been successfully used for OS analysis of human milk [24], porcine milk [25] and bovine milk [26] [27]. An exhaustive characterization of OS in goat's milks with and without the genetic ability to synthesize s1 -casein by nanoflow liquid chromatography–quadrupole-TOF MS (Nano-LC-Chip–Q-TOF MS) with a porous graphitized carbon column has been recently reported [4]. "
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    ABSTRACT: A detailed qualitative and quantitative characterization of goat colostrum oligosaccharides (GCO) has been carried out for the first time. Defatted and deproteinized colostrum samples, previously treated by size exclusion chromatography (SEC) to remove lactose, were analyzed by nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry (Nano-LC-Chip-Q-TOF MS). Up to 78 oligosaccharides containing hexose, hexosamine, fucose, N-acetylneuraminic acid or N-glycolylneuraminic acid monomeric units were identified in the samples, some of them detected for the first time in goat colostra. As a second step, a hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS) methodology was developed for the separation and quantitation of the main GCO, both acidic and neutral carbohydrates. Among other experimental chromatographic conditions, mobile phase additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry of target carbohydrates. Narrow peaks (wh: 0.2-0.6min) and good symmetry (As: 0.8-1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40°C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315mgL(-1) for neutral oligosaccharides and from 83 to 251mgL(-1) for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO.
    Journal of Chromatography A 10/2015; DOI:10.1016/j.chroma.2015.09.060 · 4.17 Impact Factor
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    • "Until recently, the analysis of oligosaccharides from bovine milk was hindered by the lack of effective analytical tools. Recently, the lower abundant oligosaccharides from bovine milk were characterized and shown to contain complex structures similar to HMO (Barile et al., 2010; Hua et al., 2012; Wickramasinghe et al., 2011). There are 40 unique BMO in bovine colostrum, including 24 acidic structures containing sialic acid, and 16 neutral oligosaccharide structures containing glucose, galactose, and N-acetylglucosamine (Tao et al., 2009). "

    Encyclopedia of Agriculture and Food Systems, Edited by Van Alfen NK, 07/2014: chapter Determining Functional Properties and Sources of Recently Identified Bioactive Food Components: Oligosaccharides, Glycolipids, Glycoproteins, and Peptides: pages 441-461; Academic Press., ISBN: 978-0-08-093139-5
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    • "Conversely, neutral oligosaccharides are more difficult to detect in negative mode because their ionization efficiency is lower, in fact neutral oligosaccharides have a low tendency to form [M-H]-. Therefore, to improve neutral oligosaccharides detection, we formed a metal-carbohydrate adduct using Sodium, and then performed the analyses in positive ion-detection mode, forming the adduct [M+Na]+. The spectrum of the neutral oligosaccharides found in the mother liquor powder is shown in Figure 2. Bovine oligosaccharides previously reported in whey [22] and milk/colostrum [16], [17], [19] were observed (i.e. those between 500–1000 m/z in Figure 2). "
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    ABSTRACT: Oligosaccharides are the third largest component in human milk. This abundance is remarkable because oligosaccharides are not digestible by the newborn, and yet they have been conserved and amplified during evolution. In addition to encouraging the growth of a protective microbiota dominated by bifidobacteria, oligosaccharides have anti-infective activity, preventing pathogens from binding to intestinal cells. Although it would be advantageous adding these valuable molecules to infant milk formula, the technologies to reproduce the variety and complexity of human milk oligosaccharides by enzymatic/organic synthesis are not yet mature. Consequently, there is an enormous interest in alternative sources of these valuable oligosaccharides. Recent research has demonstrated that bovine milk and whey permeate also contain oligosaccharides. Thus, a thorough characterization of oligosaccharides in bovine dairy streams is an important step towards fully assessing their specific functionalities. In this study, bovine milk oligosaccharides (BMOs) were concentrated by membrane filtration from a readily available dairy stream called "mother liquor", and analyzed by high accuracy MALDI FT-ICR mass spectrometry. The combination of HPLC and accurate mass spectrometry allowed the identification of ideal processing conditions leading to the production of Kg amount of BMO enriched powders. Among the BMOs identified, 18 have high-molecular weight and corresponded in size to the most abundant oligosaccharides present in human milk. Notably 6 oligosaccharides contained fucose, a sugar monomer that is highly abundant in human milk, but is rarely observed in bovine milk. This work shows that dairy streams represent a potential source of complex milk oligosaccharides for commercial development of unique dairy ingredients in functional foods that reproduce the benefits of human milk.
    PLoS ONE 05/2014; 9(5):e96040. DOI:10.1371/journal.pone.0096040 · 3.23 Impact Factor
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