Investigation on Oligosaccharides Composition of Farm milk and Processed Milk by Nano-HPLC/Chip Q-TOf Mass Spectoscopy

Allergic diseases cause severe problems in the United States. Therefore, a great deal of research was devoted to finding alternative solutions to reduce the allergic diseases manifestation. In 2006, a large European clinical trial called PARSIFAL (Prevention of Allergy - Risk factors for Sensitization In children related to Farming and Anthroposophic Lifestyle) discovered a significant inverse association between allergic diseases and all farm-produced products. Among those varied exposures, direct ingestion of unprocessed raw milk (farm milk) is considered the fundamental factor to reduce the risk of allergic diseases in farm children. Also, a growing body of epidemiological evidence suggests that consumption of unprocessed raw milk does not increase but rather decreases the risk of allergic disease sensitization. Currently, the specific constituent in raw milk responsible for the immune system modulation has not been clearly identified. Since analytical tools such as mass spectrometry have already demonstrated their power for characterizing milk constituents, the next step will be recognizing which one is the bioactive constituent in raw milk that is able to prevent allergic diseases. In this work, 60 unique samples from the PARISFAL clinical trial were analyzed; some of those samples proved to have a beneficial effect on the development of the immune system in infants from several rural regions in different European Countries. The main purpose of this research was to observe underlying differences in oligosaccharide compositions and abundances among the 60 samples. Another aim of this work was to evaluate and improve the existing methods for oligosaccharide extraction from bovine milk.;Our results by Gas Chromatography and Mass Spectrometry revealed that some neutral oligosaccharides, like 3Hex, were relatively consistent among 60 samples while acidic oligosaccharides displayed different abundance. The attempt of increasing the efficiency of the oligosaccharide extraction procedure resulted in a modified method that used less solvent and also reduced the extraction time.