Effective Separation of Human Milk Glycosides using Carbon Dioxide Supercritical Fluid Chromatography

Carbohydrate purification remains problematic due to the intrinsic diversity of structural isomers present in nature. Although liquid chromatography-based techniques are suitable for analyzing or preparing most glycan structures acquired either from natural sources or through chemical or enzymatic synthesis, the separation of regioisomers or linkage isomers with a clear resolution remains challenging. Herein, a carbon dioxide supercritical fluid chromatography (SFC) method was devised to resolve 18 human milk glycosides: oligomers (disaccharides to hexasaccharides), fucosylated regioisomers (lacto-N-fucopentaose I, III, and V; lacto-N-neofucopentaose V; lacto-N-difucohexaose III; blood group H₁ antigen; and TF-LNnT), and connectivity isomers (lacto-N-tetraose/lacto-N-neotetraose and para-lacto-N-hexaose/para-lacto-N-neohexaose/type-1 hexasaccharide). The analysis of these glycosides represents a major limitation associated with conventional carbohydrate analysis. The unprecedented resolution achieved by the SFC method indicates the suitability of this key technology for revealing complex human milk glycomes.     

Comparison of the elution curves of pure components and their mixture and prediction of elution curves by a theoretical plate model in preparative gas-liquid chromatography

Summary Two close-boiling materials, diethyl ether (DEE) and dichloromethane (DCM), were separated to investigate the effects of the pure components and the mixture on elution in preparative gas-liquid chromatography.Nitrogen was used as the carrier gas, and the chromatographic column (1 cm I.D. and 0.75m length) was packed with Chromosorb A with different quantities of dinonyl phthalate stationary phase and particle sizes. Below ca. 7% (by wt.) of feed concentration, the experimental elution curves of pure DEE and DCM were almost the same as those of the mixture, and the theoretical plate model can be used successfully to predict the elution curves.     

Isolation and structure determination of minor components of teicoplanin

Summary Teicoplanin, a lipoglycopeptide antibiotic active against gram-positive bacteria, is produced as a complex mixture consisting of six major components and four chemically related minor cmmponents. Preparative HPLC was used to isolate small amounts of the pure minor components, in order to determine their structures. In the present paper the isolation procedures are presented, as well as the analytical HPLC conditions. The retention times of the minor components with respect to those of the more abundant compounds, permittedus to make hypotheses on their structures, which were then confirmed by NMR and FAB-MS investigations.     

Effects of encounters on phase separation dynamics from diffusion-controlled growth to coarsening

We investigate the dynamics of phase separations in quenched binary mixtures, such as binary alloys and polymer blends with low mobilities, at higher volume fractions. Computer simulations are performed using simple model equations in the presence of encounters of droplets. Four characteristics stages with three crossovers, including dynamical scaling behavior and temporal power laws, are found beyond a volume fraction of approximately 0.14, where each crossover is attributed to a transition from one growth mechanism to the other. The encounter effect is thus shown to strongly influence a transition from the diffusion-controlled (t^1/2) stage to the coarsening (t^1/3) stage. A simple analytical argument for the simulation results is also presented.     

The effects of pH and alcoholic organic modifiers on the direct separation of some acidic,basic and neutral compounds on a commercially available ovomucoid column

Summary The effects of alcoholic modifiers and pH on the chromatographic properties of an immobilized ovomucoid chiral stationary phase have been investigated using acidic, basic and neutral solutes. A series of primary, secondary and tertiary alcohols and pH's ranging from 3.5 to 6.0 were used in this study. The results indicate that both the shape and the hydrophobicity of the alcoholic modifier affect retention (k') and enantioselectivity (). In general, an increase in the hydrophobicity of the modifier results in a decrease in k's and 's. However, this is not the case whent-butanol is the modifier, suggesting that the size of the alkyl moiety attached to the carbinol carbon also contributes to the chromatographic results. The pH studies indicated that Coulombic interactions play a role in the retention of the acidic and basic solutes. The results also suggest that in addition to ethanol and 1-propanol,t-butanol should be considered during optimization and that maximum efficiencies may be obtained at pH 5.0.     

Size‐Selective Exclusion Effects of Liquid Crystalline Tactoids on Nanoparticles: A Separation Method

Liquid crystalline tactoids are anisotropic microdroplets existing in isotropic phases. We studied the structure and evolution of tactoids in the presence of doping nanoparticles by electron microscopy at the resolution of individual mesogens and observed size‐selective exclusion effects of liquid crystalline tactoids on foreign nanoparticles. We applied this principle to the separation of polymer nanospheres, gold nanoparticles, and magnetic nanoparticles by size. These results indicate a new way to size‐selectively separate nanoparticles using lyotropic liquid crystals, in which nanoparticles smaller than a threshold size will be selectively transferred from the disordered phase into the ordered phase by tactoids during the phase separation process.     

The enantioselectivity of modified cyclodextrins: Studies on interaction mechanisms

The separation of enantiomers of rather simple molecules, such as alkanes and alkanoic acid esters substituted at the 2-position, has been investigated on alkylated β-cyclodextrin stationary phases. By observing the effect of varying the substrate and the cyclodextrin alkylating agent it has been possible to propose a mechanism for the separation. The most important factors seem to be hydrophobic interactions and the ability of the cyclodextrin to change its conformation to accept the guest molecule.     

Analysis of nucleotides,nucleosides, nucleobases in cells by ion-pair reversed-phase HPLC

Summary HPLC methods for the separation of nucleotides, nucleosides and nucleobases by ion-pair reversed-phase are reviewed. The advantages of these are discussed versus anion-exchange and reversed phase separations. Extraction procedures for nucleotide determinations from cells and tissues are pointed out in detail. Extracts from red blood cells, Ehrlich ascites tumour cells, hepatocytes, intestine are used for determination of nucleotide concentrations by the methods described.     

Cyano-modified pre-coated plates — a medium polar modification for high performance thin-layer chromatography

Summary The introduction of a cyano-modified, pre-coated layer substantially widens the selectivity of stationary phases in thin-layer chromatography. This is a moderately polar sorbent based on silica gel 60, which can be used both in adsorption chromatography and in reversed-phase chromatography. This new pre-coated layer is particularly suitable for separation of steroids, alkaloids and derivatized amino acids. The possibility of separating habitforming drugs and preservatives in the presence of ionpair reagents is also discussed.