Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1428, Issue -, Pages 143-153Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2015.09.060
Keywords
Goat colostrum; Oligosaccharides; Mass spectrometry; Hydrophilic interaction liquid chromatography; Nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry
Funding
- Junta de Andalucia [AGR2011-7626]
- CSIC [i-link0827]
- Comunidad Autonoma de Madrid (Spain)
- FEDER program (AVANSECAL-CM) [S2013/ABI-3028]
- Fundacion Ramon Areces
- UC Davis RISE program
- National Institutes of Health [R21AT006180, R01AT007079, R01AT008759-02]
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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 (w(h): 0.2-0.6 min) and good symmetry (A(s): 0.8-1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40 degrees C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315 mg L-1 for neutral oligosaccharides and from 83 to 251 mg L-1 for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO. (C) 2015 Elsevier B.V. All rights reserved.
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