High-sensitivity qualitative and quantitative analysis of human, bovine and goat milk glycosphingolipids using HILIC-MS/MS with internal standards.

Glycosphingolipids (GSLs) in human milk regulate the immune system, support intestinal maturation, and prevent gut pathogens. The structural complexity and low abundance of GSLs limits their systematic analysis. Here, we coupled the use of monosialoganglioside 1-2-amino-N-(2-aminoethyl) benzamide (GM1-AEAB) derivatives as internal standards with HILIC-MS/MS to qualitatively and quantitatively compare GSLs in human, bovine, and goat milk. One neutral glycosphingolipid (GB) and 33 gangliosides were found in human milk, of which 22 were newly detected and three were fucosylated. Five GB and 26 gangliosides were identified in bovine milk, of which 21 were newly discovered. Four GB and 33 gangliosides were detected in goat milk, 23 of them newly reported. GM1 was the main GSL in human milk; whereas disialoganglioside 3 (GD3) and monosialogangloside 3 (GM3) were dominant in bovine and goat milk, respectively; N-acetylneuraminic acid (Neu5Ac) was detected in >88 % of GSLs in bovine and goat milk. N-hydroxyacetylneuraminic acid (Neu5Gc)-modified GSLs were 3.5 times more abundant in goat than in bovine milk; whereas GSLs modified with both Neu5Ac and Neu5Gc were 3 times more abundant in bovine than in goat milk. Given the health benefits of different GSLs, these results will facilitate the development of custom-designed human milk-based infant formula.

Novel Method for Adulterated Identification of Saneen Goat Milk Based on Free Oligosaccharides α3'-Galactosyllactose and N-Acetylhexaminyllactose as Marker Molecules.

Methods for the detection of adulterated milk are essential for assessing the quality of goat milk products. We hypothesized that goat milk oligosaccharides could provide a basis for this purpose and compared the levels of α3'-galactosyllactose (α3'-GL) and N-acetylhexaminyllactose (NHL) between goat milk and bovine milk oligosaccharides using reverse-phase high-performance liquid chromatography. The α3'-GL was detected to be three times more abundant in goat milk than in bovine milk, whereas NHL showed the opposite trend. Linear relationships were established between the relative proportions of α3'-GL and NHL levels for different ratios of bovine and goat milk, with a minimum detection limit of 2% bovine milk. The new method was validated by analyses of adulterants in eight commercially available goat dairy products. Overall, the degree of adulteration in goat milk products can be determined based on the relative proportions of α3'-GL and NHL.