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a b s t r a c tMilk oligosaccharides are believed to have beneficial biological properties. Caprine milk has a relatively high concentration of oligosaccharides in comparison with other ruminant milks and has the oligosaccharide profile closest to that of human milk. The first stage in recovering oligosaccharides from caprine milk whey, a by-product of cheese making, was accomplished by ultrafiltration to remove proteins and fat globules, leaving more than 97% of the initial carbohydrates, mainly lactose, in the permeate. The ultrafiltered permeate was further processed using a 1 kDa 'tight' ultrafiltration membrane, which retained less than 7% of the remaining lactose. The final retentate was fractionated by preparative scale molecular size exclusion chromatography to yield 28 fractions; oligosaccharide-rich fractions that were suitable for functionality and gut health promotion testing were detected between fractions 9/10 to 16/ 17. All fractions were evaluated for their oligosaccharide and carbohydrate profiles using three complementary analytical methods.

Milk oligosaccharides (OSs) confer unique health benefits to the neonate. Although human digestive enzymes cannot degrade these sugars, they support specific commensal microbes and act as decoys to prevent the adhesion of pathogenic micro-organisms to gastrointestinal cells. The limited availability of human milk oligosaccharides (HMOs) impedes research into these molecules and their potential applications in functional food formulations. Recent studies show that complex OSs with fucose and N-acetyl neuraminic acid (key structural elements of HMO bioactivity) also exist in caprine milk, suggesting a potential source of bioactive milk OSs suitable as a functional food ingredient.

The prebiotic effect of oligosaccharides recovered and purified from caprine whey, was evaluated by in vitro fermentation under anaerobic conditions using batch cultures at 37°C with human faeces. Effects on key gut bacterial groups were monitored over 24 h by fluorescence in situ hybridization (FISH), which was used to determine a quantitative prebiotic index score. Production of short-chain fatty acids (SCFAs) as fermentation end products was analyzed by high-performance liquid chromatography (HPLC). Growth of Bifidobacterium spp was significantly higher (P ≥ 0.05) with the purified oligosaccharides compared to the negative control. Lactic and propionic acids were the main SCFAs produced. Antimicrobial activity of the oligosaccharides was also tested, revealing no inhibition though a decrease in Staphylococcus aureus and Escherichia coli growth. These findings indicate that naturally extracted oligosaccharides from caprine whey could be used as new and valuable source of prebiotics.

The use of fecal inoculums for in vitro fermentation models requires a viable gut microbiota, capable of fermenting the unabsorbed nutrients. Fresh samples from human donors are used; however, the availability of fresh fecal inoculum and its inherent variability is often a problem. This study aimed to optimize a method of preserving pooled human fecal samples for in vitro fermentation studies. Different conditions and times of storage at −20 °C were tested. In vitro fermentation experiments were carried out for both fresh and frozen inoculums, and the metabolic profile compared. In comparison with the fresh, the inoculum frozen in a PBS and 30% glycerol solution, had a significantly lower (p < 0.05) bacterial count (<1 log CFU/mL). However, no significant differences (p < 0.05) were found between the metabolic profiles after 48 h. Hence, a PBS and 30% glycerol solution can be used to maintain the gut microbiota viability during storage at −20 °C for at least 3 months, without interfering with the normal course of colonic fermentation.

The functional food market is growing rapidly and membrane processing offers several advantages over conventional methods for separation, fractionation and recovery of bioactive components. The aim of the present study was to select a process that could be implemented easily on an industrial scale for the isolation of natural lactose-derived oligosaccharides (OS) from caprine whey, enabling the development of functional foods for clinical and infant nutrition. The most efficient process was the combination of a pre-treatment to eliminate proteins and fat, using an ultrafiltration (UF) membrane of 25-kDa molecular weight cutoff (MWCO), followed by a tighter UF membrane with 1-kDa MWCO. Circa 90 % of the carbohydrates recovered in the final retentate were OS. Capillary electrophoresis was used to evaluate the OS profile in this retentate. The combined membrane-processing system is thus a promising technique for obtaining natural concentrated OS from whey.

Cocoa and cocoa products have broad and wellproven health benefits, most of which are associated with the high antioxidant capacity of cocoa flavonoids. Most extraction methods for flavonoids in cocoa products use several steps including a defatting step and large amount of organic solvents. Moreover, they are labor-intensive and time-consuming. In this work, a pressurized hot water extraction (PHWE) method has been compared to conventional sonication extraction (CSE) method. The contribution of individual compounds to the total antioxidant capacity of the extracts was evaluated by developing an analytical technique consisting of high-performance liquid chromatography (HPLC) with photodiode array coupled to electrochemical and to charged aerosol detectors and HPLCmass spectrometry for the confirmation of the identity of compounds present. Additionally, procyanidins were analyzed by fluorescence detection. PHWE turned out to be more efficient in extracting phenolics and methylxanthines, as compared to the conventional method, in addition to being more Bgreenî n terms of using less organic solvents.

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