Commercial Galacto-oligosaccharides Research Articles

High purity galacto-oligosaccharides: Optimal process design and prebiotic effect

Prebiotic unpurified commercial galacto-oligosaccharides (GOS) often contain significant amounts of glucose, galactose and lactose. Current purification processes remove these digestible components, especially lactose with a high risk of affecting functionality.In the current study a high purity GOS (HP-GOS) was prepared by treating GOS with β-galactosidase to hydrolyze the remaining lactose into glucose and galactose. Subsequently mono sugars were removed by sequential simulated moving bed (SSMB) chromatography. To this end, a design of experiments (DOE) approach was applied to determine the dry matter content and β-galactosidase concentration that optimally support lactose hydrolysis in such a way that the detrimental effects on GOS composition are minimized to preserve its original oligosaccharide composition. The HP-GOS product obtained using the identified optimal settings for lactose hydrolysis contained more than 75% of the original DP2 GOS fraction while the fraction with a DP above 2 was unaffected by the purification. Using fecal fermentations, it was demonstrated that the GOS parts of the HP-GOS and the parent GOS have comparable effects on gut microbiota composition. Both GOS products equally increased the relative abundances of Bifidobacterium. Furthermore, there were no differences in alpha diversity and in the distribution of the different bifidobacterial species. Based on these outcomes it was concluded that the lactose hydrolysis step of the purification process did not influence gut microbiota modulating effects of GOS.

Qualitative and quantitative mass spectrometry comparison of characteristic galactosyl lactose isomers from goat milk at different lactation stages.

Galactooligosaccharides are composed mainly of galactosyl lactose, which is important for infant growth and as a functional food additive. Although galactosyl lactose is abundant in goat milk, its complex structure has hindered the separation and analysis of its isomers. In this study, 5 isomers of goat milk galactosyl lactose were separated by HPLC: β6'-galactosyl lactose (β6'-GL), α6'-galactosyl lactose (α6'-GL), β4'-galactosyl lactose (β4'-GL), α3'-galactosyl lactose (α3'-GL), and β3'-galactosyl lactose (β3'-GL). This composition differs from that of commercial galactooligosaccharide products, which comprise mainly β-configuration oligosaccharides. The isomers were then qualitatively and quantitatively compared at different lactation stages using online HPLC-mass spectrometry. Relative quantitative analysis showed that the total content of the 5 galactosyl lactose isomers was highest in transitional goat milk. Specifically, β3'-GL was the main isomer in colostrum and α3'-GL was the main isomer in transitional and mature milk. β6'-Galactosyl lactose and β4'-GL tended to increase and then decrease during lactation. Moreover, α3'-GL content was 2 times higher than in colostrum and 10 times higher in transitional milk than in mature milk; in contrast, for β3'-GL, the values were 5 and 2 times higher, respectively. Absolute quantitative analysis revealed that β3'-GL was the most abundant isomers in colostrum (32.3 mg/L), and α3'-GL was the most abundant in transitional milk (88.1 mg/L) and mature milk (36.3 mg/L). These findings provide an important quantitative basis for understanding the relationship between structure and function of galactosyl lactose in goat milk, as well as its exploitation as a functional food.

Simulated-moving-bed technology for purification of the prebiotics galacto-oligosaccharides

A commercial galacto-oligosaccharide (GOS) syrup with prebiotic properties was purified with simulated moving bed (SMB) chromatography to separate undesired side products induced by synthesis reaction mechanism. A promising polystyrene-divinylbenzene resin stationary phase has been characterized with regard to its selective adsorption behavior. The chosen chromatographic system allows a pseudo binary separation of the multi-component GOS syrup mixture exploiting a multi-column continuous chromatographic process. Suitable SMB operation parameters were estimated by means of the equilibrium (triangle) theory, further optimized by modelling the separation in more detail and finally validated experimentally. Changes of temperature, concentration of the feed as well as in the instrumental configuration (open loop, closed loop and 3 zones) were examined with a classical SMB plant in lab scale size. Highly purified GOS products could be obtained with purities up to >99% in the extract and raffinate fractions. A quantitative description of the transfer from batch chromatography to the continuous separation process via SMB technology is provided, indicated significant potential for a scale-up.

Characterization of galacto-oligosaccharides using high-performance anion exchange chromatography-tandem mass spectrometry.

The analysis of complex oligosaccharide mixtures remains a challenge in the field of analytical chemistry. In this work, two commercial galacto-oligosaccharides samples were characterized using high-performance anion exchange chromatography coupled to mass spectrometry. The isomeric oligosaccharides were resolved with high resolution. The structures of the individual isomers with a degree of polymerization up to 6 were analyzed using targeted selected ion monitoring with data-dependent tandem mass spectrometry, with additional in-source collision-induced dissociation.

Enrichment and evaluation of galacto-oligosaccharides produced by whole cell treatment of sugar reaction mixture.

A process was developed for enrichment of galacto-oligosaccharides (GOS), synthesized from a whole cell driven system, from a sugar reaction mixture (SRM) containing non prebiotic sugars (monosaccharides and disaccharides) as impurities. SRM containing 38% (w/w of total carbohydrates) of GOS was enriched by 7 and 27%, attaining a purity of 45 and 65% respectively using Saccharomyces cerevisiae followed by Kluyveromyces lactis var. lactis treatment. The two cell types could be recycled for consecutive 12 and 10 cycles respectively. The microbial purified GOS (MPG) was characterized by mass spectrometry and quantitated by HPLC. MPG was further evaluated for its prebiotic potential on Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei Shirotaand Saccharomyces boulardii. The growth profile and colony forming units were determined and compared with the profiles obtained on glucose, used as a control. MPG was efficiently utilized by L. acidophilus and L. plantarum which showed antimicrobial activity with zone of lysis (12 and 10mm) against Escherichia coli and Citrobacter (14 and 9mm) respectively and performed better than Vivinal (commercial GOS), fructo-oligosaccharides and inulin. The synergistic effect of the MPG with L. acidophilus and L. plantarum was found to be most effective against pathogens as compared to other tested commercial oligosaccharides.

Selective bioconversion with yeast for the purification of raw lactulose and transgalactosylated oligosaccharides

Raw transgalactosylated oligosaccharides (raw TOS) obtained by enzymatic synthesis with Aspergillus oryzae β-galactosidase were purified by selective bioconversion with Saccharomyces cerevisiae ATCC4126 and Kluyveromyces marxianus NRLLY-1109. Monosaccharides were completely removed by S. cerevisiae and K. marxianus from raw TOS without any nutrient supplementation. The effect of the biomass to carbohydrate mass ratio on the purity and specific productivity of removal of sugars was assessed with S. cerevisiae, where both effects were significant. Purity obtained with a collection strain of S. cerevisiae and bakers’ yeast was compared, the latter being higher after 24 h of bioconversion. A product free of monosaccharides was obtained with a purity (0.48; 28% lactulose and 20% oligosaccharides), similar to that of commercial galacto-oligosaccharide preparations. The possibility of yeast recovery and reuse in sequential batch operation was assessed: two batches could be performed before a significant reduction in monosaccharide removal was seen.

Stability of Oligosaccharides Derived from Lactose and Lactulose regarding Rheological and Thermal Properties

Prebiotic carbohydrates derived from lactulose (OsLu) are appealing ingredients that provide beneficial effects following their fermentation by the gut microbiota; however, more investigation is needed to evaluate their technological properties and applicability as a functional food ingredient in the market. In this paper, a comparative study on the rheological and thermal behavior of OsLu and a commercial mixture of galactooligosaccharides (GOS) was carried out. In both cases, there was a strong influence of temperature and shear rate on viscosity. Viscosity of OsLu was higher than that of commercial GOS and variations and rheopexy cycles were also higher for OsLu. The exponential increase of viscosity associated with the structural changes took place later in OsLu than in commercial GOS, suggesting more stability to the shear over time in the former. More stability of OsLu was also observed in the study of the effect of temperature on viscosity. In addition, the thermal study indicated different behavior of both prebiotics under the assayed conditions, showing that OsLu have lower values of glass transition temperature (Tg) than commercial GOS and that commercial GOS are more sensible against humidity. The results here obtained provide important information on the treatment and storage conditions of these prebiotic ingredients during the elaboration of functional foods.

Effect of Galacto-Oligosaccharides: Maltodextrin Matrices on the Recovery of Lactobacillus plantarum after Spray-Drying.

In this work maltodextrins were added to commercial galacto-oligosaccharides (GOS) in a 1:1 ratio and their thermophysical characteristics were analyzed. GOS:MD solutions were then used as matrices during spray-drying of Lactobacillus plantarum CIDCA 83114. The obtained powders were equilibrated at different relative humidities (RH) and stored at 5 and 20°C for 12 weeks, or at 30°C for 6 weeks. The Tgs of GOS:MD matrices were about 20–30°C higher than those of GOS at RH within 11 and 52%. A linear relation between the spin-spin relaxation time (T2) and T-Tg parameter was observed for GOS:MD matrices equilibrated at 11, 22, 33, and 44% RH at 5, 20, and 30°C. Spray-drying of L. plantarum CIDCA 83114 in GOS:MD matrices allowed the recovery of 93% microorganisms. In contrast, only 64% microorganisms were recovered when no GOS were included in the dehydration medium. Survival of L. plantarum CIDCA 83114 during storage showed the best performance for bacteria stored at 5°C. In a further step, the slopes of the linear regressions provided information about the rate of microbial inactivation for each storage condition (k values). This information can be useful to calculate the shelf-life of spray-dried starters stored at different temperatures and RH. Using GOS:MD matrices as a dehydration medium enhanced the recovery of L. plantarum CIDCA 83114 after spray-drying. This strategy allowed for the first time the spray-drying stabilization of a potentially probiotic strain in the presence of GOS.

Comparative structural characterization of 7 commercial galacto-oligosaccharide (GOS) products

Many β-galactosidase enzymes convert lactose into a mixture of galacto-oligosaccharides (GOS) when incubated under the right conditions. Recently, the composition of commercial Vivinal GOS produced by Bacillus circulans β-galactosidase was studied in much detail in another study by van Leeuwen et al. As a spin-off of this study, we used the developed analytical strategy for the evaluation of 6 anonymous commercial GOS products, in comparison with Vivinal GOS. These GOS products were first subjected to HPLC-SEC, calibrated HPAEC-PAD profiling (glucose units in relation to a malto-oligosaccharide ladder), and 1D 1H NMR spectroscopy. For a more detailed analysis and support of the conclusions based on the initial analysis, the GOS products were separated into DP-pure subpools on Bio-Gel P-2 (MALDI-TOF-MS analysis), which were subjected to calibrated HPAEC-PAD profiling and 1H NMR analysis. Unidentified peaks from different GOS products, not present in Vivinal GOS, were isolated for detailed structural characterization. In this way, the differences between the various GOS products in terms of DP distribution and type of glycosidic linkages were established. A total of 13 new GOS structures were characterized, adding structural-reporter-group signals and HPAEC-PAD based glucose unit G.U. values to the analytical toolbox. The newly characterized products enhance the quality of the database with GOS structures up to DP4. The combined data provide a firm basis for the rapid profiling of the GOS products of microbial β-galactosidase enzymes.

Evaluation of Prebiotic Effects of High-Purity Galactooligosaccharides in vitro and in vivo.

Galactooligosaccharides (GOS) are an important class of dietary prebiotics that exert beneficial effects on intestinal microbiota and gut barrier function. In this study, high-purity GOS (HP-GOS) were investigated in vitro and in vivo and confirmed as prebiotic ingredients in rat diet. HP-GOS were successfully produced using a two-step process, enzymatic hydrolysis and fermentation by yeast. They were found to serve as a good substrate and carbon source for supporting the growth of probiotic bacteria more effectively than other commercial GOS. Following administration of 1% (by mass) of HP-GOS to rats, the growth of Bifidobacterium bifidum and B. longum in the gut increased most rapidly up to 12 h, and thereafter the increase was slow. Therefore, 1% HP-GOS was found to be acceptable for the growth of probiotic bacteria. Groups of animals that were orally administered HP-GOS and bifidobacteria during the study, and the group administered HP-GOS during the 2nd (days 13-15) and 4th (days 28-30) period of the study had significantly (p<0.05) higher numbers of bifidobacteria in faeces than groups receiving a single dose of bifidobacteria. HP-GOS affected the expression of genes encoding glucagon-like peptide-1 (GLP-1) and peptide YY (PYY). There was a significant upregulation of GLP-1 and PYY mRNA with HP-GOS and bifidobacteria intake. We propose that the prebiotic properties of HP-GOS are potentially valuable for the production of functional foods for human consumption.

Stability of oligosaccharides derived from lactulose during the processing of milk and apple juice

The scientific evidence on the bioactivity of oligosaccharides from lactulose has encouraged us to study their physicochemical modifications during the processing of milk and apple juice. The carbohydrate fraction with a degree of polymerization ⩾3 was stable in milk heated at temperatures up to 100°C for 30min and in apple juice heated up to 90°C for 15min. An assessment of the Maillard reaction in heated milk pointed out a higher formation of furosine in milk with oligosaccharides from lactulose as compared to its counterpart without this ingredient, due to a higher presence of galactose. The organoleptic properties of juice with oligosaccharides from lactulose were acceptable and similar to those of apple juice with commercial galactooligosaccharides. The results presented herein demonstrate that oligosaccharides from lactulose can be used as prebiotic ingredients in a wide range of functional foods, including those intended for diabetics and lactose intolerant individuals.

Inhibition of the adhesion of Escherichia coli to human epithelial cells by carbohydrates

Mannose-containing carbohydrates provide an alternative therapeutic approach for controlling bacterial infections. The aim of this work was to compare the effects of depolymerised konjac glucomannan and other carbohydrates (including simple sugars) on the inhibition of the adhesion of Escherichia coli to human cheek epithelial cells. Sucrose, glucose, commercial galacto-oligosaccharides (GOS), maltodextrin, inulin, glucomannan hydrolysates (GMH) and mannose were assessed for this purpose. The average cell counts of E. coli adhering to the epithelial cells were found to be significantly higher (P<0.01) in sucrose, glucose, commercial galacto-oligosaccharides (GOS), maltodextrin and inulin compared to mannose or GMH. The use of mannose-rich carbohydrates as an approach to reduce bacterial infections could prove a successful approach although large scale studies in humans are required to characterise further the specific role of such carbohydrates with different pathogens.

Detection of adulteration in commercial galactooligosaccharide by flow-injection electrospray ionization tandem mass spectrometry

In recent years, a number of adulterants in foods, such as melamine in infant formula and horse meat in beef, have been reported. For ensuring the food safety, proper regulation of these adulterated foods is necessary. In this work, we have studied an adulteration of maltitol in a commercial galactooligosaccharide (GOS) product using the flow-injection electrospray ionization tandem mass spectrometry (ESI-MS/MS) and the MassBank database. Firstly, qualitative analysis of the adulterated maltitol in a GOS product was achieved by the flow-injection ESI-MS/MS in negative ion mode. Secondly, mass spectral data obtained from the ESI-MS/MS were compared with the MassBank database. Lastly, final confirmation and quantitative analysis of the adulterated maltitol was carried out by HPLC-RI using a reference material. As a result, content of the adulterated maltitol in a GOS product was determined to be 60.2%, which was not indicated on the label. On the other hand, unlike the labeled statement, content of raffinose was only 18.4% instead of labeled 100%. Although maltitol is generally considered to be safe, it is also known that large quantities of maltitol can have a laxative effect. Furthermore, consumers have the rights to know what are in the foods they eat. Therefore, food adulterations, including this case of maltitol, should be properly controlled by the regulatory authorities.

Galacto-oligosaccharides Derived from Lactulose Exert a Selective Stimulation on the Growth of Bifidobacterium animalis in the Large Intestine of Growing Rats

There is increasing interest in identifying novel dietary nondigestible carbohydrates capable of modulating the composition and/or metabolic activities of the gut microbiota. This work assessed the differential modulatory influence of novel galacto-oligosaccharides derived from lactulose (GOS-Lu) in comparison with commercial galacto-oligosaccharides derived from lactose (GOS-La) in gut microbiota of growing rats (5 weeks old). Rats were fed either a control diet or diets containing 1% (w/w) of GOS-Lu or GOS-La, and cecal and colonic contents were collected after 14 days of treatment. Compared to controls, GOS-Lu had significantly more bifidobacteria within the large intestine, showing a significant and selective increase of Bifidobacterium animalis in the cecum and colon; however, no significant differences in the number of bifidobacteria among GOS-Lu and GOS-La groups were observed. Both types of GOS significantly increased the number of the Eubacterium rectale / Clostridium coccoides group. These findings support a prebiotic role of galactosyl-fructoses in functional foods.

Utilization of galactooligosaccharides by Bifidobacterium longum subsp. infantis isolates

Prebiotics are non-digestible substrates that stimulate the growth of beneficial microbial populations in the intestine, especially Bifidobacterium species. Among them, fructo- and galacto-oligosaccharides are commonly used in the food industry, especially as a supplement for infant formulas. Mechanistic details on the enrichment of bifidobacteria by these prebiotics are important to understand the effects of these dietary interventions. In this study the consumption of galactooligosaccharides was studied for 22 isolates of Bifidobacterium longum subsp. infantis, one of the most representative species in the infant gut microbiota. In general all isolates showed a vigorous growth on these oligosaccharides, but consumption of larger galactooligosaccharides was variable. Bifidobacterium infantis ATCC 15697 has five genes encoding β-galactosidases, and three of them were induced during bacterial growth on commercial galactooligosaccharides. Recombinant β-galactosidases from B. infantis ATCC 15697 displayed different preferences for β-galactosides such as 4′ and 6′-galactobiose, and four β-galactosidases in this strain released monosaccharides from galactooligosaccharides. Finally, we determined the amounts of short chain fatty acids produced by strain ATCC 15697 after growth on different prebiotics. We observed that biomass and product yields of substrate were higher for lactose and galactooligosaccharides, but the amount of acids produced per cell was larger after growth on human milk oligosaccharides. These results provide a molecular basis for galactooligosaccharide consumption in B. infantis, and also represent evidence for physiological differences in the metabolism of prebiotics that might have a differential impact on the host.

A selected probiotic strain of Lactobacillus fermentum CM33 isolated from breast-fed infants as a potential source of β-galactosidase for prebiotic oligosaccharide synthesis

Lactic acid bacteria from healthy breast-fed infants were isolated and screened for β-galactosidase production in MRS broth. Among 49 isolates that exhibited the yellow clear zone on MRS agar supplemented with bromocresol blue, the isolate CM33 was selected as being the highest β-galactosidase producer and was identified as Lactobacillus fermentum based on its morphological characteristics and 16S rDNA nucleotide sequence. L. fermentum CM33 exhibited a good survival rate under the simulated stomach passage model, comparable to known probiotic strains L. gallinarum JCM2011 and L. agilis JCM1187. L. fermentum CM33 was antagonistic to pathogenic bacteria Listeria monocytogenes, Escherichia coli 0157:H7, Salmonella typhi, and Salmonella enteriditis, using the well diffusion method. In addition, the selected lactobacilli exhibited a high growth rate when cultivated in modified MRS containing commercial galactooligosaccharide (GOS) as a sole carbon source, as well as in glucose. A preliminary study on the enzymatic synthesis of oligosaccharide using crude β-galactosidase revealed the capability for oligosaccharide synthesis by the transgalactosylation activity.

Effect of prebiotic carbohydrates on the growth and tolerance of Lactobacillus

Resistance to gastrointestinal conditions is a requirement for bacteria to be considered probiotics. In this work, we tested the resistance of six different Lactobacillus strains and the effect of carbon source to four different gastrointestinal conditions: presence of α-amylase, pancreatin, bile extract and low pH. Novel galactooligosaccharides synthesized from lactulose (GOS-Lu) as well as commercial galactooligosaccharides synthesized from lactose (GOS-La) and lactulose were used as carbon sources and compared with glucose. In general, all strains grew in all carbon sources, although after 24 h of fermentation the population of all Lactobacillus strains was higher for both types of GOS than for glucose and lactulose. No differences were found among GOS-Lu and GOS-La. α-amylase and pancreatin resistance was retained at all times for all strains. However, a dependence on carbon source and Lactobacillus strain was observed for bile extract and low pH resistance. High hydrophobicity was found for all strains with GOS-Lu when compared with other carbon sources. However, concentrations of lactic and acetic acids were higher in glucose and lactulose than GOS-Lu and GOS-La. These results show that the resistance to gastrointestinal conditions and hydrophobicity is directly related with the carbon source and Lactobacillus strains. In this sense, the use of prebiotics as GOS and lactulose could be an excellent alternative to monosaccharides to support growth of probiotic Lactobacillus strains and improve their survival through the gastrointestinal tract.

Introducing Capillary Electrophoresis with Laser-Induced Fluorescence (CE−LIF) as a Potential Analysis and Quantification Tool for Galactooligosaccharides Extracted from Complex Food Matrices

The analysis and quantification of (galacto)oligosaccharides from food matrices demands both a reproducible extraction method as well as a sensitive and accurate analytical method. Three typical matrices, namely, infant formula, fruit juice, and a maltodextrin-rich preparation, to which a commercial galactooligosaccharide mixture was added in a product concentration range from 1.25 to 30%, served as model substrates. Solid-phase extraction on graphitized carbon material upon enzymatic amyloglucosidase pretreatment enabled a good recovery and a selective purification of the different galactooligosaccharide structures from the exceeding amounts of particularly lactose and maltodextrins. With the implementation of capillary electrophoresis in combination with laser-induced fluorescence (CE-LIF) detection, a new possibility facilitating a sensitive qualitative and quantitative determination of the galactooligosaccharide contents in the different food matrices is outlined. Simultaneous monitoring and quantifying prebiotic oligosaccharides embedded in food matrices presents a promising and important step toward an efficient monitoring of individual oligosaccharides and is of interest for research areas dealing with small quantities of oligosaccharides embedded in complex matrices, e.g., body liquids.

Prebiotic evaluation of a novel galactooligosaccharide mixture produced by the enzymatic activity of Bifidobacterium bifidum NCIMB 41171, in healthy humans: a randomized, double-blind, crossover, placebo-controlled intervention study

Background: Galactooligosaccharides are selectively fermented by the beneficial member of the colonic microflora contributing to the health of the host.Objective: We assessed the prebiotic potential of a novel galactooligosaccharide produced through the action of β-galactosidases, originating from a probiotic Bifidobacterium bifidum strain, against a galactooligosaccharide produced through the action of an industrial β-galactosidase and a placebo.Design: Fifty-nine healthy human volunteers participated in this study. Initially, the effect of the matrix on the prebiotic properties of a commercially available galactooligosaccharide (7 g/d) was assessed during 7-d treatment periods with a 7-d washout period in between. During the second phase, 30 volunteers were assigned to a sequence of treatments (7 d) differing in the amount of the novel galactooligosaccharide (0, 3.6, or 7 g/d). Stools were recovered before and after each intervention, and bacteria numbers were determined by fluorescent in situ hybridization.Results: Addition of the novel galactooligosaccharide mixture significantly increased the bifidobacterial population ratio compared with the placebo (P < 0.05), whereas 7 g/d of the novel galactooligosaccharide significantly increased the bifidobacterial ratio compared with the commercial galactooligosaccharide (P < 0.05). Moreover, a significant relation (P < 0.001) between the bifidobacteria proportion and the novel galactooligosaccharide dose (0, 3.6, and 7 g/d) was observed. This relation was similar to the effect of the novel galactooligosaccharide on the prebiotic index of each dose.Conclusions: This study showed that galactooligosaccharide mixtures produced with different β-galactosidases show different prebiotic properties and that, by using enzymes originating from bifidobacterial species, an increase in the bifidogenic properties of the prebiotic product is achievable.

Purification of oligosaccharides by nanofiltration

Nanofiltration (NF) of a model sugar solution and a commercial galacto-oligosaccharide mixture has been studied with a cross-flow filtration unit in an attempt to investigate the significance of the operating parameters and evaluate a possible purification process. The pressure, feed concentration and filtration temperature effects were studied in experiments carried out in full recycle mode of operation. The rejection factors of the sugar components present in the solutions increased with increasing pressure due to increased solvent flux and also compaction of the membranes (e.g. rejection factor increased from 0.10 to 0.42 for fructose), with this effect being greater for the low molecular weight sugars in the solutions. Increasing the total sugar concentration of the feed caused a decrease to the rejections of the sugars (e.g. from 0.58 to 0.43 for glucose). Increasing the filtration temperature caused a decrease to the observed rejections of the low molecular weight sugars of the solutions (e.g. from 0.72 to 0.48 for fructose). Continuous diafiltration (CD) purification using NF-CA-50 membranes (at 25 °C) and DS-5-DL membranes (at 60 °C), gave yield values of 14–18% for the monosaccharide, 59–89% for the disaccharide and 81–98% for the trisaccharide (oligosaccharide), respectively. The study clearly demonstrates the potential of cross-flow nanofiltration in the purification of oligosaccharide from mixtures containing contaminant monosaccharides.