An improved method for the purification of milk oligosaccharides by graphitised carbon-solid phase extraction

Abstract

Milk oligosaccharides (OS) are bioactive molecules that impart a variety of health benefits to the consumer. Techniques commonly used to analyse and quantify OS require optimised extraction methods to separate the OS from more abundant milk components. Solid phase extraction (SPE) is frequently used to isolate milk OS from lactose; however, the literature contains no formal studies on its efficacy in this application. In this study, established SPE conditions were modified to improve the technique’s effectiveness in purifying OS from lactose. Low concentrations of acetonitrile (ACN) and trifluoroacetic acid (TFA) were tested for solid phase washing. Lactose removal and retention of many OS were significantly improved when using 4% ACN/0.1% TFA compared with the more common water washing technique. Different behaviours between acidic and neutral OS were evident. The new SPE technique improves extraction efficiency for bovine milk OS in applications that do not require prior lactose hydrolysis.

Electrospray ionization mass spectrometry characterization of ubiquitous minor lipids and oligosaccharides in milk of the camel (Camelus dromedarius) and their inhibition of oxidative stress in human plasma

“…Free oligosaccharides are the most abundant solid constituent of milk, apart from lactose and lipids. Several oligosaccharides have been identified in milk from various sources, containing different structural moieties of sugars: lactose, glucose, N-acetyl glucosamine galactose, N-acetylgalactosamine, mannose, and fucose (Singh et al, 2016;Srivastava et al, 2016;Urashima et al, 2016;Gangwar et al, 2017;Robinson et al, 2018;Urashima et al, 2018;Tonon et al, 2019). Recently, a new oligosaccharide has been isolated from CM and named medalose (Gangwar et al, 2018).…”

Biopurification of Oligosaccharides by Immobilized Kluyveromyces Lactis

“…Hence, pilot-scale filtration or chromatographic processes have been studied for their industrial application [10][11][12]. Small-scale separation methods were also studied for the identification of oligosaccharides [13,14]. However, these methods are expensive and require complicated procedures to operate.…”

“…However, about 50 g/L of lactose has to be eliminated from bovine milk for practical application of BMOs. Hence, pilot-scale filtration or chromatographic manners have been studied for lactose removal [10][11][12][13][14]. A biological method using β-galactosidase (EC 3.2.1.23) was also studied [15].…”

Butyrate producing Clostridiales utilize distinct human milk oligosaccharides correlating to early colonization and prevalence in the human gut

“…Human milk oligosaccharides (HMOs) were purified from pooled human milk samples as previously described 47,48 . Milk fat was separated by centrifugation (10.000x g, 30 min at 4°C) and proteins were removed by ethanol precipitation (as above).…”