Cow milk oligosaccharides (CMOs) are complex carbohydrates found in cow milk that resemble the oligosaccharides in human milk and are essential for regulating the immune system and forming the gut flora of infants. As prebiotics, they promote the growth of specific beneficial gut bacteria, such as Lactobacilli and Bifidobacteria, thus promoting the creation of short-chain fatty acids for gut health. Furthermore, CMOs correlate with enhanced infant immune system development, offering safeguards against pathogens and anti-inflammatory benefits. The results of recent CMO research are revealed in this review, together with their biological importance and potential applications. Their relevance to infant nutrition is highlighted, as is their potential to be used as bioactive ingredients in novel functional foods and nutraceuticals. This study also describes upcoming obstacles and opportunities for CMO research, such as understanding their structures and functions, improving extraction methods, and expanding applications to different age groups.

Milk, a cornerstone of neonatal nutrition and human health, harbors various components with antioxidant capabilities, including oligosaccharides. In this study, we explored the antioxidant potential of acetylated milk oligosaccharides (A-MOs) and deacetylated oligosaccharides (D-MOs) sourced from Rathi cow milk through assays, with a focus on their comparative efficacy. Concurrently, we evaluated the antioxidant activity of Ascorbic Acid (AA) for comparison. Results revealed significant dynamics within the antioxidant profiles of A-MOs and D-MOs. A-MOs demonstrated concentration-dependent antioxidant activity, with higher concentrations correlating with increased efficacy, while D-MOs displayed a distinct pattern, with varying antioxidant potential across concentrations. Notably, both A-MOs and D-MOs exhibited promising antioxidant activity, suggesting their potential as natural antioxidants. In contrast, AA exhibited a decline in antioxidant activity with increasing concentration, indicating a differing pattern from oligosaccharides. These findings underscore the significance of acetylation in modulating the antioxidant properties of milk oligosaccharides. Further investigation is warranted to elucidate the mechanisms underlying these observations and to explore the practical implications of A-MOs and D-MOs as functional food components or potential therapeutic agents. Overall, our study contributes to the understanding of milk oligosaccharides’ antioxidant properties and highlights the potential of acetylated and deacetylated oligosaccharides from Rathi cow milk as valuable sources of natural antioxidants with diverse applications in nutrition and healthcare.

Purification and characterization of milk oligosaccharides is a challenging process due to the complexity of the constituent oligosaccharides, which behave differently under various chemical treatment procedures and often lose their structural properties in the process. Rathi cow’s milk is widely used in the Rajasthan region of India for its nutritional and medicinal benefits. Here, we aim to present an optimized method for the purification and analysis of oligosaccharides present in Rathi cow milk. Contrary to the freeze transport methods used earlier, we treated the collected milk with ethanol for preservation, followed by microfiltration, lyophilization, and fractionation on silica gel (60–120 mesh size) column chromatography (CC) coupled with chloroform/methanol-mediated gradient elution. Fractions 31–45 (1.78 g), 71–80 (470 mg), and 106–120 (498 mg) from CC-1 and fractions 26–49 (1.14 g) from CC-2 were analysed for sugar content via the phenol–sulfuric acid method. Fraction homogeneity was confirmed using high-performance liquid chromatography. Isolated analytes were treated with acetic anhydride/pyridine (1:1, v/v) to form less polar oligosaccharide derivatives, which could then be easily visualized and semi-quantitated using partition chromatography (thin later and paper) with chloroform/methanol. Structural identities of the purified oligosaccharides were determined using a combination of mass spectrometry and NMR (¹H, ¹³C, HSQC, TOCSY, COSY, HMBC) techniques. Our results clearly demonstrate that the ethanol-based preservation, transport, and purification of oligosaccharides is a simple and robust method for the analysis of Rathi cow’s milk oligosaccharides. Furthermore, using the acetylation, purified oligosaccharides allow for rapid analysis on thin-layer chromatography, which is quite cost effective compared with other analytical methods.

Cow milk oligosaccharides (CMOs) have garnered significant attention due to their association with enhanced infant immune system development, defence against pathogens, and anti-inflammatory properties