Hydrophilic interaction liquid chromatography with methanol-water eluent on a zeolite.

BACKGROUND: Hydrophilic interaction chromatography (HILIC) works with organic solvent-water mixtures as eluent and is based on the formation of a water enriched liquid phase on the surface of a hydrophilic stationary phase. Hydrophilic solutes are retained on that stagnant water-rich film depending on the difference of solvation compared to the mobile phase composition. However, the enhancement of selectivity by increasing the fraction of organic cosolvent is coupled with a limitation the analyte solubility, and the improvement of the HILIC principle by new hydrophilic stationary phases is the remaining option. RESULTS: Y-zeolite (faujasite, FAU type) in the Na+-form with an average particle diameter of 5 µm was used as packing material in a 125 mm long HPLC column. The chromatographic response of the column was tested in methanol-water mixtures as eluent after injection of several aliphatic alcohols, polyols and monosaccharides with eluent conditions where no separation occurs on diol functionalized silica. On the zeolite the retention time increases according to ethylene glycol < glycerol < erythritol < sorbitol < inositol. The separation principle is explained to be superposed by two effects: firstly, a partition equilibrium between the water-rich phase in the zeolite micropores exists, and secondly, selective interactions with the inner crystalline pore surface and fixed-position Na+ ions, both serving to enhance the selectivity. Furthermore, arabinose and fructose monosaccharides could be separated into their tautomeric forms. Only upon increasing the temperature from 20 to 60 °C the tautomeric pattern merges into a single peak. SIGNIFICANCE AND NOVELTY: Instead of the stagnant water rich surface layer, zeolite micropores now take over that function. As a result, the selectivity among polyols and between α/ß-arabinopyranose and ß-fructopyranose/ß-fructofuranose tautomers is extraordinary superior towards conventional hydrophilic interaction liquid chromatography (HILIC).

General cluster sorption isotherm: surface area determination.

The recently published General Cluster Sorption Isotherm is modified by separating forces lateral and vertical directed to the surface. Calculating the ratio of the infinitesimal contributions of adsorption in both directions as a function of the total adsorption results in an inflection point which is assigned to the position of the monolayer coverage. The method is applied to eight experimental high resolution type II sorption isotherms. The parameters are compared with those of the previous GCSI version and the surface areas are discussed together with those obtained by BET and the excess surface work method (ESW). For N2 and Ar sorption, the BET method is found to overestimate the actual surface area. Since the GCSI reduces to BET when the sorbate-surface interaction is strong enough, the GCSI derived surface area includes the BET surface as a special case. The application of GCSI for surface area determination requires a sufficiently broad range of experimental data points.

Modeling of type IV and V sigmoidal adsorption isotherms.

Based on the biochemical theory of multiple ligand-receptor complexes (Klotz (1946)) a sigmoidal proceeding adsorption isotherm is derived. The special case of an arbitrarily large number of equal interaction sites and a separate one yields an equation which corresponds to the ζ-isotherm, disparately derived by Ward et al. (2007) for surface tension of solid-fluid interfaces. From the mathematical point of view it is analogous to the BET-isotherm for a limited number of adsorption layers (1938). It is shown that the isotherm maps type IV and V isotherms. The isotherm is compared with others including the type IV isotherm of Do and Do (2009). The present isotherm is unified in contrast to existing hybrid models. It is successfully applied to numerous literature data concerning the adsorption of water on microporous carbon and aluminophosphate molecular sieves.

Chromatography of mono- and disaccharides on granulated pellets of hydrophobic zeolites.

The chromatographic response of sugars at granulated zeolite pellets in preparative scale liquid chromatography is analyzed with respect to the distribution equilibrium and mass transfer. In contrast to hydrophilic FAU type zeolites their hydrophobic dealuminated counterpart, used here, can separate disaccharides the retention of which can strongly exceed those of the monosugars. The retention is correlated with data of batch adsorption studies from the literature. Whereas the retention decreases with increasing temperature, the peak sharpness shows the opposite trend. The effective mass resistance is calculated for a series of mono- and disaccharides. It increases with the capacity factor. The diffusion coefficient of the trehalulose disaccharide is restricted by a factor of about 2 in the macropores and by a factor of more than 104 in the micropores.

Exo-enzyme like degradation of a polysaccharide by an inorganic solid acid catalyst.

Inulin is a ß(1 → 2) linked linear fructose polymer, the reducing end of which is capped by a glucose residue. Upon dissolution in aqueous HCl the polymer is statistically split. During the reaction progress the ratio of glucose to fructose increases and the concentration of sucrose, as an intermediate product, goes through a maximum. However, when the splitting is performed on a dealuminated faujasite zeolite, the ratio of glucose to fructose is constant and equal to the average degree of polymerization; sucrose does not appear as intermediate product. This behavior is explained by a sequential splitting starting from the end of the polymer (exo-mechanism). If the glucose or fructose end is preferred is unknown. Adsorption studies of the polymer fragments show that the polymer enters the pore mouth of the zeolite with a penetration depth limited to the length of maximal 3 saccharide units. The protons in that surface region are 4-5 times more catalytically active than those in HCl.

Hydrophobicity of carbohydrates and related hydroxy compounds.

The hydrophobic interaction of carbohydrates and other hydroxy compounds with a C18-modified silica gel column was measured with pure water as eluent, thereby expanding the range of measurements already published. The interaction is augmented by structure strengthening salts and decreasing temperature. Although the interaction of the solute with the hydrophobic interface is expected to only imperfectly reflect its state in aqueous bulk solution, the retention can be correlated to hydration numbers calculated from molecular mechanics studies given in the literature. No correlation can be established towards published hydration numbers obtained by physical methods (isentropic compressibility, O-17 NMR relaxation, terahertz spectroscopy, and viscosity). The hydrophobicity is discussed with respect to the chemical structure. It increases with the fraction and size of hydrophobic molecular surface regions.

Di-D-fructose dianhydride-enriched caramels: effect on colon microbiota, inflammation, and tissue damage in trinitrobenzenesulfonic acid-induced colitic rats.

In the present study we describe the preparation and chemical characterization of a caramel with a high (70%) content of difructose dianhydrides (DFAs) and glycosylated derivatives (DFAs). This product was obtained by thermal activation (90 degrees C) of highly concentrated (90% w/v) aqueous D-fructose solutions using the sulfonic acid ion-exchange resin Lewatit S2328 as caramelization catalyst. DFAs represent a unique family of cyclic fructans with prebiotic properties already present in low proportions (<15%) in commercial caramel. We report the antiinflammatory activity of the new DFA-enriched caramel in the trinitrobenzenesulfonic acid (TNBS) model of rat colitis, an experimental model that resembles human inflammatory bowel disease (IBD), and compare its effects with those obtained with a commercial sucrose caramel and with linear fructooligosaccharides (FOS). For this purpose, the effects on colon tissue damage, gut microbiota, short-chain fatty acid (SCFAs) production, and different inflammatory markers were evaluated. The administration of DFA-enriched caramel to colitic rats showed intestinal antiinflammatory effect, as evidenced macroscopically by a significant reduction in the extent of the colonic damage induced by TNBS. This effect was similar to that obtained with FOS in the same experimental settings, whereas commercial caramel was devoid of any significant antiinflammatory effect. The beneficial effect was associated with the inhibition of the colonic levels of the proinflammatory cytokines, tumor necrosis factor alpha (TNF alpha) and interleukin 1beta (IL-1beta), and the reduction in colonic myeloperoxidase (MPO) activity and inducible nitric oxide synthase (iNOS) expression. The DFA-enriched caramel also promoted a more favorable intestinal microbiota, increasing lactobacilli and bifidobacteria counts as well as inducing higher concentrations of SCFAs in the luminal colonic contents. These results reinforce the concept of DFAs and glycosyl-DFAs as dietary beneficial compounds with prebiotic properties and suggest that the novel DFA-enriched caramel here reported may be an interesting candidate to be explored for the dietary treatment of human IBD.