Glycobiology Advance Access originally published online on February 22, 2006
Glycobiology 2006 16(6):477-487; doi:10.1093/glycob/cwj092
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Incorporation of orally applied 13C-galactose into milk lactose and oligosaccharides
2 Department of Pediatrics, University of Giessen, Feulgenstr. 12, 35392 Giessen, Germany; 3 Research Institute of Child Nutrition Dortmund, 44225 Dortmund, Germany; 4 Institute of Nutritional Sciences, University of Giessen, 35392 Giessen, Germany; 5 Institute of Medical Physics and Biophysics, University of Münster, 48149 Münster, Germany; 6 Institute of Physiological Chemistry, University of Bonn, 53113 Bonn, Germany; 7 Virchow-Clinics, 10117 Berlin, Germany; and 8 Center of Pediatrics, University of Bonn, 53113 Bonn, Germany
1 To whom correspondence should be addressed; e-mail: silvia.rudloff{at}ernaehrung.uni-giessen.de
Received on August 5, 2005; revised on February 14, 2006; accepted on February 15, 2006
Biosynthesis and functions of human milk oligosaccharides (HMO) are not well known. A typical housekeeping enzyme, ß1,4-galactosyltransferase, links galactose to glucose to form lactose which is then used as backbone for the assembly of HMO. We investigated whether milk lactose and HMO may be labeled in vivo by an orally given 13C-galactose bolus. Eleven exclusively breastfeeding mothers were given a 13C-galactose bolus at the end of their breakfast. Milk and urine samples from each nursing up to 36 h were analyzed for carbohydrate composition by high-performance thin-layer chromatography, high-pH anion-exchange chromatography, and fast atom bombardment mass spectrometry. 13C enrichment of milk fractions, urinary carbohydrates, lactose, and oligosaccharides as well as of breath CO2 was determined by isotope ratio mass spectrometry. Up to 10% of the orally given galactose bolus was directly transported to the mammary gland and incorporated into milk components. Characteristic for most milk samples was the appearance of two 13C-peaks, the first immediately after the 13C-bolus was taken and the second on the next morning. The highest 13C enrichment was found in lactose followed by neutral and acidic oligosaccharides. In breath samples, the 13C-excretion followed the same pattern as in milk. 13C nuclear magnetic resonance of isolated lactose revealed 13C only at C1-atom of galactose and C1-atom of glucose. This label was without any exception at the same position as the 13C-label of the orally applied galactose. Neutral and acidic HMO can easily be 13C-labeled in vivo which facilitates investigations of their metabolic fate in infants.
Key words: 13C-lactose / 13C-oligosaccharides / fast atom bombardment mass spectrometry / human milk / isotope ratio mass spectrometry