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Galacto-oligosaccharides (GOS) and fructo-oligosaccharides (FOS) are food ingredients that improve human health, but their degradation throughout the human small intestine is not well understood. We studied the breakdown kinetics of FOS and GOS in the intestines of seven healthy Dutch adults. Subjects were equipped with a catheter in the distal ileum or proximal colon and consumed 5 g of chicory-derived FOS (degree of polymerization (DP) DP2–10), and 5 g of GOS (DP2–6). Postprandially, intestinal content was frequently collected until 350 min and analyzed for mono-, di-, and oligosaccharides. FOS and GOS had recoveries of 96 ± 25% and 76 ± 28%, respectively. FOS DP ≥ 2 and GOS DP ≥ 3 abundances in the distal small intestine or proximal colon matched the consumed doses, while GOS dimers (DP2) had lower recoveries, namely 22.8 ± 11.1% for β-D-gal-(1↔1)-α-D-glc+β-D-gal-(1↔1)-β-D-glc, 19.3 ± 19.1% for β-D-gal-(1 → 2)-D-glc+β-D-gal-(1 → 3)-D-glc, 43.7 ± 24.6% for β-D-gal-(1 → 6)-D-gal, and 68.0 ± 38.5% for β-D-gal-(1 → 4)-D-gal. Lactose was still present in the distal small intestine of all of the participants. To conclude, FOS DP ≥ 2 and GOS DP ≥ 3 were not degraded in the small intestine of healthy adults, while most prebiotic GOS DP2 was hydrolyzed in a structure-dependent manner. We provide evidence on the resistances of GOS with specific β-linkages in the human intestine, supporting the development of GOS prebiotics that resist small intestine digestion.

The human intestinal mucus layer protects against pathogenic microorganisms and harmful substances, whereas it also provides an important colonization niche for mutualistic microbes. The main functional components of mucus are heavily glycosylated proteins, called mucins. Mucins can be cleaved and utilized by intestinal microbes. The mechanisms between intestinal microbes and the regulation of mucin glycosylation are still poorly understood. In this study, in vitromucus was produced by HT29-MTX-E12 cells under Semi-Wet interface with Mechanical Stimulation. Cells were exposed to pasteurized nonpathogenic bacteria Akkermansia muciniphila, Ruminococcus gnavus, and Bacteroides fragilisto evaluate influence on glycosylation patterns. Following an optimized protocol, O- and N-glycans were efficiently and reproducibly released, identified, and semiquantified using MALDI-TOF-MS and PGC-LC-MS/MS. Exposure of cells to bacteria demonstrated increased diversity of sialylated O-glycans and increased abundance of high mannose N-glycans in in vitroproduced mucus. Furthermore, changes in glycan ratios were observed. It is speculated that bacterial components interact with the enzymatic processes in glycan production and that pasteurized bacteria influence glycosyltransferases or genes involved. These results highlight the influence of pasteurized bacteria on glycosylation patterns, stress the intrinsic relationship between glycosylation and microbiota, and show the potential of using in vitroproduced mucus to study glycosylation behavior.

In this study, we systematically investigated the structural characterization and in vitrofermentation patterns of crude black mulberry fruit polysaccharides (BMPs), either extracted by water (BMP) or by enzymatic treatment. Different enzymatic treatments were pectinase-extracted (PE)-BMP, pectin lyase-extracted (PL)-BMP, cellulase-extracted (CE)-BMP, and compound enzymes-extracted (M)-BMP (pectinase:pectin lyase:cellulase = 1:1:1). Our results show that enzymatic treatment improved the polysaccharide yield and led to a different chemical composition and structure for the polysaccharides. Change dynamics during the in vitrofermentation indicated that BMPs could indeed be degraded and consumed by human fecal microbiota and that different BMPs showed different degrees of fermentability. In addition, BMPs stimulated the growth of Bacteroidetes and Firmicutes, inhibited the growth of Fusobacteria and Proteobacteria (except for CE-BMP), and induced the production of short-chain fatty acids (SCFAs). Furthermore, we found that BMP and PL-BMP exhibited better fermentability and prebiotic potential than the other polysaccharides.

To follow human milk oligosaccharide (HMO) biosynthesis and in vivometabolization, mother milk and infant feces from 68 mother–infant dyads at 2, 6, and 12 weeks postpartum were analyzed, with 18 major HMOs quantitated. Fucosylated and neutral core HMO levels in milk were dependent on mothers’ Lewis/Secretor status, whereas most sialylated HMO levels were independent. Infant fecal excretion of HMOs gradually declined with age, especially for neutral core structures. Although decreasing in absolute concentrations in milk during lactation, the relative abundance of total fucosylated HMOs increased in both milk and feces. Mono-fucosylated HMOs were more consumed than those decorated with two fucose moieties. More (α2-3)-sialylated HMOs were degraded than (α2-6)-sialylated HMOs. The transition speed of HMO metabolization from nonspecific or structure-specific consumption stage to the complete consumption stage was individual-dependent. Variation was associated with mode and place of delivery, where caesarean section or early exposure to hospital environment delayed the transition.

The safety of a rhamnogalacturonan-I-enriched pectin extract (G3P-01) from pumpkin (Cucurbita moschatavar. Dickinson) was evaluated for use as an ingredient in food and dietary supplements. G3P-01 was tested in a battery of genetic toxicity studies including reverse mutagenicity and in vitromicronucleus assay. In addition, Sprague-Dawley rats were randomized and orally dosed with G3P-01 incorporated in animal diet at concentrations of 0, 9000, 18,000, and 36,000 ppm daily for 13-weeks (n=10/sex/group) in line with OECD guidelines (TG 408). The results of the in vitrobacterial reverse mutation assay and micronucleus assay in TK6 cells demonstrated a lack of genotoxicity. The 13-week oral toxicity study in Sprague-Dawley rats demonstrated that the test article, G3P-01 was well tolerated; there were no mortalities and no adverse effects on clinical, gross pathology, hematology, blood chemistry, and histological evaluation of the essential organs of the animals. The present study demonstrates that G3P-01 is non-genotoxic and is safe when ingested in diet at concentrations up to 36, 000 ppm. The subchronic no-observed-adverse-effect level (NOAEL) for G3P-01 was concluded to be 36,000 ppm, equivalent to 1,899 and 2,361 mg/kg/day for male and female rats respectively.

The aim of the current study was to investigate whether degradation of rapeseed meal (RSM) by a swine gut microbiota consortium was improved by modifying RSM by treatment with cellulase (CELL), two pectinases (PECT), or alkaline (ALK) compared to untreated RSM and to assess whether microbiota composition and activity changed. The predicted relative abundances of carbohydrate digestion and absorption, glycolysis, pentose phosphate pathway, and pyruvate metabolism were significantly increased upon CELL and ALK feeding, and CELL and ALK also exhibited increased total short-chain fatty acid (SCFA) production compared to CON. Megasphaera, Prevotella, and Desulfovibriowere significantly positively correlated with SCFA production. Findings were validated in ileal cannulated pigs, which showed that CELL and ALK increased fiber degradation of RSM. In conclusion, CELL and ALK rather than PECT1 or PECT2 increased fiber degradation in RSM, and this information could guide feed additive strategies to improve efficiency and productivity in the swine industry.

Galacto-oligosaccharides (GOS) are used in infant formula to replace the health effects of human milk oligosaccharides, which appear to be dependent upon the structure of the individual oligosaccharides present. However, a comprehensive overview of the structure-specific effects is still limited as a result of the high structural complexity of GOS. In this study, porous graphitic carbon (PGC) was used as the stationary phase during ultra-high-performance liquid chromatography–mass spectrometry (UHPLC–MS). This approach resulted in the recognition of more than 100 different GOS structures in one single run, including reducing and non-reducing GOS isomers. Using nuclear magnetic resonance-validated structures of GOS trisaccharides, we discovered MS fragmentation rules to distinguish reducing isomers with a mono- and disubstituted terminal glucose by UHPLC–PGC–MS. UHPLC–PGC–MS enabled effective recognition of structural features of individual GOS components in complex GOS preparations and during, e.g., biological conversion reactions. Hence, this study lays the groundwork for future research into structure-specific health effects of GOS.

To study the Chinese human milk N-glycome over lactation, N-glycans were released and separated from serum proteins, purified by solid-phase extraction, and analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In total, 66 different putative N-glycans were found in the colostrum (week 1) and mature milk (week 4) of seven Chinese mothers. A clear difference was observed between milk of five secretor and two nonsecretor mothers, based on the type and relative amounts of the individual N-glycans. The relative levels of the total neutral nonfucosylated and the fucosylated N-glycans in milk of five secretor mothers increased and decreased over lactation, respectively. This pattern could not be observed for the milk from the two nonsecretor mothers. Overall, this was the first study that provided detailed information on individual N-glycans in milk among mothers and over time as well as that fucosylation of N-glycans in milk was associated with the mother’s secretor status.

Corn fiber, a byproduct from the corn industry, would be a good source for bioethanol production if the hemicellulose, consisting of polymeric glucoronoarabinoxylans, can be degraded into fermentable sugars. Structural knowledge of the hemicellulose is needed to improve the enzymatic hydrolyses of corn fiber. Oligosaccharides that resisted a mild acid pretreatment and subsequent enzymatic hydrolysis, representing 50% of the starting material, were fractionated on reversed phase and size exclusion material and characterized. The oligosaccharides within each fraction were highly substituted by various compounds. Oligosaccharides containing uronic acid were accumulated in two polar fractions unless also a feruloyl group was present. Feruloylated oligosaccharides, containing mono- and/or diferulic acid, were accumulated within four more apolar fractions. All fractions contained high amounts of acetyl substituents. The data show that complex xylan oligomers are present in which ferulic acid, diferulates, acetic acid, galactose, arabinose, and uronic acids were combined within an oligomer. Hypothetical structures are discussed, demonstrating which enzyme activities are lacking to fully degrade corn glucuronoarabinoxylans.

The intermolecular distributions of amide groups within two commercial LMA pectins was studied after removal of the methyl esters followed by fractionation of the different populations by anion exchange chromatography. The populations obtained had almost equal degrees of amidation while the values of the degree of blockiness were not the same, indicating also a different intramolecular distribution of the substituents considered as semirandom. Populations from the methyl-esterified amidated pectins showed a rather random distribution for almost all populations. A striking difference between these different populations was that, despite the same level of substitution, the ratio between amide groups and methyl esters varied significantly, indicating a heterogeneous amidation process.

Investigations on the mode of action of Aspergillus niger pectin methylesterase (PME) towards differently C6- and C1-substituted oligogalacturonides (oligoGalpA) are described. De-esterification of methyl-esterified (un)saturated oligoGalpA proceeds via a specific pattern, depending on the degree of polymerization. Initially, a first methyl ester of the oligomer is hydrolysed, resulting in one free carboxyl group. Subsequently, this first product is preferred as a substrate and is de-esterified for a second time. This product is then accumulated and hereafter de-esterified further to the final product, i.e. oligoGalpA containing one methyl ester located at the non-reducing end residue for both saturated and unsaturated oligoGalpA, as found by post-source decay matrix-assisted laser-desorption/ionization–time-of-flight MS. The saturated hexamer is an exception to this: three methyl esters are removed very rapidly, instead of two methyl esters. When unsaturated oligoGalpA were used, the formation of the end product differed slightly, suggesting that the unsaturated bond at the non-reducing end influences the de-esterification process. In vivo, PME prefers methyl esters, but the enzyme appeared to be tolerant for other C6- and C1-substituents. Changing the type of ester (ethyl esterification) or addition of a methyl glycoside (C1) only reduced the activity or had no effect respectively. The specific product pattern was identical for all methyl- and ethyl-esterified oligoGalpA and methyl-glycosidated oligoGalpA, which strongly indicates that one or perhaps two non-esterified oligoGalpA are preferred in the active-site cleft.

A thorough investigation of the mode of action ofAspergillus niger(4M-147) pectin lyase A (PLA) on differently C6-substituted oligogalacturonides is described. PLA appeared to be very specific for fully methyl-esterified oligogalacturonides: removal of the methyl-ester or changing the type of ester (ethyl esterification) or transamidation resulted in (almost) complete loss of conversion. The PLA activity increased with increasing length of the substrate up to a degree of polymerization (DP) of 8 indicating the presence of at least eight subsites on the enzyme. Product analysis demonstrated the formation of several Δ4,5 unsaturated products and their saturated counterparts. The Δ4,5 unsaturated trimer was the main product up to DP 8. For DP 9 and 10 Δ4,5 unsaturated tetramer was the major product. Based upon the bond cleavage frequencies, a provisional subsite map was calculated, which supports the presence of eight subsites. By limited alkaline de-esterification of fully methyl-esterified pentamer and hexamer two sets of partially methyl-esterified pentamers (x and y methyl groups) and hexamers (a and b methyl groups) were prepared. Matrix-assisted laser desorption/ionization time of flight mass spectroscopy (MALDI-TOF MS) analysis demonstrated that the methyl-ester distribution was fully random. Using these partially methyl-esterified oligogalacturonides as substrates for PLA a 10-fold decrease in reaction rate was recorded compared with the fully methyl-esterified counterparts. Analysis of the methyl-ester distribution of the products showed that PLA tolerates carboxyl groups in the substrate binding cleft. At either subsite +2, +4, or −1 to −4 a free carboxyl group could be tolerated, whereas methyl-esters were obligatory at subsite +1 and +3. So PLA is capable to cleave the bond between a methyl-esterified and a non-esterified galacturonic acid residue, where the newly formed Δ4,5 unsaturated non-reducing end residue always contains a methyl-ester.

A method was developed that enables the study of the methyl ester distribution in the polymers of pectin on a molecular level. Endo-polygalacturonase was used to extensively degrade three 70% methyl esterified pectins. The molecular weight distribution of the non- and enzymatically degraded pectins was determined with high-performance size-exclusion chromatography. Next, the molecular weight distribution was converted into a degree of polymerization distribution of galacturonan fragments. Monte Carlo methods were employed for the reconstruction of the parental polymers from their enzymatic degradation products. The results for the random methyl esterified pectin revealed that the enzyme-degradable sites were indeed randomly distributed, which confirmed the correctness of the procedure developed. The two other pectins studied differed greatly in the amount of non-, low-, and high-esterified regions present in the reconstructed pectic molecules of a given molecular mass. That the approach developed is able to reveal such detailed information makes it unique. The information on the fragmental composition of pectic polymers obtained is an important addition to the study of the methyl ester distribution and the functional properties of pectin. © 2001 John Wiley & Sons, Inc. Biopolymers 58: 195–203, 2001

The methyl ester distribution of pectins was studied with a recently developed enzymatic method. Endopolygalacturonase of Kluyveromyces fragilis was used to degrade pectin and the composition of the degradation products was determined with high-performance anion-exchange chromatography at pH 5. Three characteristics indicative for the distribution of nonesterified galacturonic acid residues were obtained: the percentage of nonesterified galacturonic acid residues liberated of the total number of nonesterified galacturonic acid in the undigested polymer, the proportion of nonesterified mono-, di-, and trigalacturonic acid released, and the ratio of the sum of the peak areas of methyl ester containing oligomers divided by the sum of the peak areas of the nonesterified oligomers detected. From these characteristics and the degree of methyl esterification, the mean sequence similarity of the methyl ester distributions was calculated. Computational techniques commonly employed in the determination of the sequence similarity of DNA and proteins were used to discriminate the various types of distributions found and to construct a distance tree. In general, three types of methyl ester distributions could be discerned in pectin: random, high, and blockwise esterified. This report is the first to describe a parametric approach for the comparison of the substituent distribution in polymers. The importance of this novel approach in the study of the methyl ester distribution and the functional properties of pectin is discussed. © 2000 John Wiley & Sons, Inc. Biopoly 58: 1–8, 2001

Full-fat soya beans were separated into hulls and cotyledons. After separation the cell wall fraction was extracted from the cotyledons. These purified cell walls were sequentially extracted with 0.05 M cyclohexane-trans-1,2-diamine-N,N,N ',N '-tetraacetate (CDTA) + 0.05 M NH4 oxalate (extract 1), 0.05 M NaOH (extract 2), 1 M KOH (extract 3) and 4 M KOH (extract 4) to fractionate the cell wall into its polysaccharide fractions. The extraction procedure was designed to first extract the pectic fraction (extracts 1 and 2), after which more hemicellulose was extracted in subsequent steps (extracts 3 and 4). In addition to the polysaccharides solubilised during extraction, the residues after each extraction step were collected. Extracts and residues were analysed for their fermentation characteristics using an in vitro gas production procedure. Fermentability of the sequential extracts increased for each subsequent extraction step, as witnessed by an increasing rate of fermentation (from 2.2 to 10.0% h⁻¹) and decreasing half-time of gas production (from 56.0 to 18.7 h). Fermentability of the residues increased after pectins had been removed in the first two extraction steps, as witnessed by a shorter half-time of gas production (from 47.0 to 29.7 h). Fermentability was similar for residues 2 and 3, but decreased again for residue 4, which was deemed to consist mainly of cellulose. The different cell wall sugars were degraded at different rates, with a rapid rate of degradation for galactose and arabinose, an intermediate rate for xylose and uronic acids and a slow rate for glucose. The sugar degradation rates for extract 1 had a similar ranking. These results are discussed in light of the concept that pectins determine the pore size of the cell wall matrix (Carpita NC and Gibeant DM, Planta J3: 1–30 (1993) and the model of the cell wall architecture of legumes of Hatfield (Hatfield RD, in Forage Cell Wall Structure and Digestibility, Ed by Jung HG, Buxton DR, Hatfield RD and Ralph J. American Society of Agronomy/Crop Science Society of America/Soil Science Society of America, Madison, WI, pp 285–313 (1993)). © 2000 Society of Chemical Industry

The use of post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the structural analysis of ((partly) methyl-esterified) oligogalacturonides (oligoGalA) is described. The fragmentation behavior of purified (un)saturated oligoGalA (degree of polymerization 3–6), methyl-esterified and methyl-glycosydated oligoGalA was studied. General fragmentation patterns are described and used for the elucidation of the positions of methyl esters on partly methyl-esterified oligoGalA. This technique now permits the determination of the position of methyl esters or other substituents on pectic fragments, helping in understanding the mode of action of pectinolytic enzymes. Copyright © 2000 John Wiley & Sons, Ltd.

Pectins are a class of heterogeneous polysaccharides used in the food industry as a result of their ability to form gels. They are primarily composed of a (1ŕ4)‐α‐D‐galactopyranosyluronic acid backbone where the carboxylic acid group is methyl esterified at a level which depends on the source of the pectin and processing conditions used to isolate the material. Of considerable interest is the distribution of the free acid and methyl ester groups along this polymer chain. NMR spectroscopy, coupled with statistical analysis, is a powerful technique for the study of sequence distribution of monomers. Experimental conditions (temperature, pD) are reported which are appropriate for the analysis of pectic polysaccharides. Results are presented from a study of sequence distribution in native, modified, and fractionated pectins using 1H and 13C solution NMR methods. The triad sequence information was examined using Bernoullian and Markovian probability models in conjunction with continuous Gaussian distribution and discrete models. Intermolecular heterogeneity in pectins results in triad frequencies which reflect the distribution of acid and ester monomers for native and base saponified pectins. Fractionation of pectin through physicochemical methods and subsequent spectroscopic analysis provide insight into this heterogeneity. Segregation into discrete polymer populations shows a structural diversity best approximated by a 3‐component statistical model.

Zusammenfassung Es wurde ein Hemicellulose-Extrakt aus Äpfeln durch alkalische Extraktion der entpectinisierten alkoholunlöslichen Substanz gewonnen. Dieser Extrakt wurde fraktioniert mittels Ionenaustausch-Chromatographie und Gelfiltration vor und nach Behandlung mit einer reinen 1,4-ß-d-Glucanglucanohydrolase. Die Zusammensetzung der verschiedenen Fraktionen an Zuckerbausteinen und den glykosidischen Bindungen wurde bestimmt. Die Resultate deuten darauf hin, daß der Hemicellulose-Extrakt von Äpfeln Fucogalaktoxyloglucane, pektinassoziierte Arabane und Galaktane (Gluko)-Mannane, (Arabino)-Xylane und 1,3-Glucane enthält. Die Fucogalaktoxyloglucane und die Arabane und Galaktane wurden als solche und als Komplexe miteinander und mit (Arabino)-Xylanen isoliert. Der größte Teil der Mannane war nicht an andere Hemicellulosen gebunden.

Zusammenfassung Saure and neutrale Zuckerbausteine der Pplysaccharide wurden im Ethanol unlöslichen Rückstand verschiedener Frucht- and Gemüsearten bestimmt, sowie in der Pectin-, Hemicellulose- and Cellulose-Fraktion dieser Rückstände. Für die Uronsäure-Bestimmung mittels molekularer Absorptionsspektrometrie wurden die Resultate aus enzymatischer, saurer und alkalischer Extraktion verglichen. Die Gehalte an Neutralzuckern wurden gaschromatographisch bestimmt sowohl nach Hydrolyse mit Enzymen and Trifluoressigsäure (Konzentration 2 mol/1) als auch nach Saeman-Hydrolyse (Schwefelsäure, H+-Konzentration 0,8 mol/1 oder 2 mol/1). Uronsäuregehalte, mit der m-Hydroxydiphenyl-Reaktion und mittels Kupferionenbindung bestimmt, liegen in der gleichen Größen-ordnung. Die Zuckerzusammensetzung der Muster und Fraktionen weisen große Unterschiede auf.

A method has been developed for the rapid molecular mass determination and structural elucidation of mixtures of oligosaccharides derived from plant cell walls. The oligosaccharides were fractionated using gel permeation chromatography and ‘analytical’ high-performance anion-exchange chromatography (HPAEC), neutralized, dried and the mixtures of eluent salt and oligosaccharides were per-O-acetylated directly. The derivatized oligosaccharides were isolated by dissolution in dichloromethane and the salts were removed by aqueous partitioning. The per-O-acetylated oligosaccharides were analysed using electrospray (ES) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MS). Exploiting the fact that acid-catalysed per-O-acetylation of oligosaccharides can be achieved even under the extremely salty conditions that are found in post-column neutralized HPAEC fractions, and combining this derivatization step with off-line ESMS, allow rapid screening for molecular mass and thus yield information on the composition of the various oligosaccharides in these complex mixtures. Subsequent per-O-methylation of the per-O-acetylated, salt-free fractions and collision-induced dissociation tandem mass spectrometric analysis was used for additional sequence and branching determination of the oligosaccharides. © 1998 John Wiley & Sons, Ltd.

Zusammenfassung Apfelsaft-Modelle, bestehend aus Galakturoniden (Reaktionsprodukte des enzymatischen Pectinabbaus), Zuckern, Aminosäuren und organischen Säuren wurden einer Hitzebehandlung von 2 h in einem kochenden Wasserbad unterzogen. Dabei wurde die Abnahme der Galakturonid-Konzentration und die Entstehung von Reaktionsprodukten und Färbung ermittelt.

Zusammenfassung In verschiedenen Apfelsäften, die durch direkte Pressung, enzymatische Pulpbehandlung oder Verflüssigung und auf konventionelle Weise mittels Bentonit, Gelatine und Silicasol oder durch Ultrafiltration geklärt wurden, wurden der Zucker- und Säuregehalt, die Farbe, der Polyphenolgehalt, die Ultrafiltrations-Durchflußgeschwindigkeit und der Gehalt und die Zusammensetzung der Polysaccharide bestimmt. Die Menge und Zusammensetzung der gefundenen hochmolekularen Polysaccharid-Fraktionen zeigen deutlich die Bedeutung der Herstellungsmethode und der verwendeten Enzympräparate. Galakturonsäure und Arabinose waren die Hauptbestandteile der aus den Zellwänden gelösten Polysaccharide. Die Ultrafiltration ermöglichte die Abtrennung von hochmolekularen Polymeren; jedoch wurden auch in ultrafiltrierten Säften noch neutrale Arabinane nachgewiesen.

Two methods were developed to detect partially methyl-esterified galacturonic acid oligomers, generated by endopolygalacturonase treatment of a 30% methyl-esterified pectin. The enzyme digest was shown, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, to contain sodiated galacturonic acid oligomers with a degree of polymerization of 2–12, containing 0–6 methyl esters. Galacturonic acid (monomer) could not be detected because of matrix ions interference in the low mass region. Using high-performance anion-exchange chromatography, with a sodium acetate gradient at pH 5.0 and postcolumn sodium hydroxide addition to allow pulsed amplified detection, a complex elution profile was obtained with the endopolygalacturonase-treated 30% methyl-esterified pectin. All the components eluted before nonesterified tetragalacturonic acid. The partially methyl-esterified oligogalacturonic acids eluted in a discernible series of oligomers with an identical number of nonesterified carboxylic acid groups; the large, more esterified oligomers eluted before small, less esterified oligomers. The methyl esters may hinder the interaction of the neighboring carboxylic acid groups with the anion-exchange resin, thereby giving the components an apparent lower overall negative charge.

The polar fraction of virgin olive oil was separated into two main parts (A and B) using solid phase extraction. Analysis of individual components by RP-HPLC indicated that the part (A) contained only simple phenols and phenolic acids. Part (B) had a complex nature. The two parts tested for their antioxidant activity showed relatively high protection factors in safflower oil stored at 80°C. Part B was found to contribute more than part A to the stability of the oil. The antioxidant activity of both fractions was related to their content of total polyphenols and o-diphenols. Acidic and alkaline hydrolysis showed significant quantitative changes in the HPLC profiles indicating the presence of ether and ester bonds while high-performance anion exchange chromatography of sugars after hydrolysis gave evidence for the presence of only traces of glycosides. A first attempt to identify a characteristic chromatographic peak of part B by HPLC fractionation and mass spectrometry showed the presence of an ester of tyrosol most probably with a dicarboxylic acid. © 1997 SCI.

A solid-phase extraction procedure was optimized to extract 3-fucosyllactose and other human milk oligosaccharides (HMOs) from human milk samples separately, followed by absolute quantitation using high-performance anion-exchange chromatography-pulsed amperometric detection and porous graphitized carbon-liquid chromatography–mass spectrometry, respectively. The approach developed was applied on a pilot sample set of 20 human milk samples and paired infant feces collected at around 1 month postpartum. One-dimensional 1H nuclear magnetic resonance spectroscopy was employed on the same samples to determine the relative levels of fucosylated epitopes and sialylated (Neu5Ac) structural elements. Based on different HMO consumption patterns in the gastrointestinal tract, the infants were assigned to three clusters as follows: complete consumption; specific consumption of non-fucosylated HMOs; and, considerable levels of HMOs still present with consumption showing no specific preference. The consumption of HMOs by infant microbiota also showed structure specificity, with HMO core structures and Neu5Ac(α2-3)-decorated HMOs being most prone to degradation. The degree and position of fucosylation impacted HMO metabolization differently.