In this study, BMOs were concentrated by membrane filtration from a readily available whey stream, and analyzed by high accuracy mass spectrometry. Whey is the liquid part of milk that separates from the curd during cheese production. Whey is rich in lactose and protein, but its high biochemical and chemical oxygen demand make it expensive to dispose of within environmental regulations. Whey is a valuable dairy stream for extraction of useful proteins in the form whey protein concentrates by ultrafiltration. Whey UF permeate is either disposed of at a cost to the whey processor or used to produce food grade lactose by crystallization. The liquid separated from lactose crystals, known as Evodiamine mother liquor, is usually disposed of to sewage plants or sold as animal feed. Because mother liquor is already virtually protein-free and most of the lactose has already been removed, oligosaccharides can be purified by filtration, which allows retention of oligosaccharides and removes lactose and salts. Such mother liquor enriched in oligosaccharides would facilitate commercial use of these biofunctional ingredients for the food, beverage, and baby food industries, add value to the waste streams of dairy processing, and also reduce waste disposal costs for the dairy industry. The present study employed large-scale centrifugation and membrane filtration technology to produce powders enriched in BMOs using mother liquor as a low-cost starting raw material. Mother liquor was processed employing Tubeimoside-I membranes that fractionated and concentrated BMOs from the other molecules. Mass spectrometry analysis of oligosaccharides derived from this process revealed numerous BMOs including eighteen high-molecular weight species never previously described. Importantly, some of these novel BMOs contained fucose, a key component of HMOs. A main difference between HMOs and BMOs is the fact that most HMOs are highly fucosylated, while the presence of fucose in bovine milk has been controversial. Over 20 years ago Saito et al. reported a single fucosylated oligosaccharide from bovine colostrum. Since that time numerous studies of BMOs have been undertaken and have revealed 40 oligosaccharide species and novel structural detail on BMOs; however no fucosylated species were described. Indeed, the failure of numerous studies to identify fucosylated oligosaccharides in bovine milk led some to question the original observations by Saito et al. However, recently other studies reported the presence of fucosylated oligosaccharides in goat milk as well as bovine milk and colostrum.. It is well known that bovine colostrum oligosaccharides composition is different from that of mature bovine milk and it is reasonable to expect that it would be even more different in processed cheese whey obtained from late lactation milk. Furthermore, we hypothesize that the smaller oligosaccharides may have passed through the 1000 Da filtration membrane or even have co-crystallized with lactose during lactose crystallization in the production of mother liquor. These possibilities would explain why these oligosaccharides were not detected by mass spectrometric analysis in this work, whereas larger fucosylated oligosaccharides were identified.