Chromatography polarity

In normal-phase chromatography, polar components are more strongly retained than nonpolar components. Thus, hydrocarbon carotenes elute quickly while xanthophylls are retained and separated. This approach provides a more complete separation of polar carotenoids and their geometric isomers. This protocol is useful to the analyst that is specifically interested in the xanthophyll fraction of a sample. 

In thin layer chromatography polar compounds will ... 

In normal-phase chromatography, polar stationary phases are employed and solutes become less retained as the polarity of the mobile-phase system increases. Retention in normal-phase chromatography is predominately based upon an adsorption mechanism. Planar surface interactions determine successful use of NPC in separation of isomers. The nonaqueous mobile-phase system used in NPC has found numerous applications for extremely hydrophobic molecules, analytes prone to hydrolysis, carbohydrates, and sat-urated/unsaturated compounds. In the future, with the advent of new stationary phases being developed, one should expect to see increasingly more interesting applications in the pharmaceutical industry. 

In normal-phase chromatography, polar compounds are separated on a polar stationary phase. In reversed-phase chromatography, nonpolar compounds are separated on a nonpolar stationary phase. The latter is more common ... 

Adsorption chromatography Polarity Medium-high Medium Medium... 

Normal-phase chromatography polar stationary phase and less polar solvent... 

Two groups of stationary phases are used in adsorption chromatography Polar and nonpolar adsorbents. 

If maltenes are subjected to liquid chromatography (see 2.1.2.4) the components eluted by the more polar solvents are called resins. Their composition, once again, depends on the procedure used. 

Liquid chromatography is preceded by a precipitation of the asphaltenes, then the maltenes are subjected to chromatography. Although the separation between saturated hydrocarbons and aromatics presents very few problems, this is not the case with the separation between aromatics and resins. In fact, resins themselves are very aromatic and are distinguished more by their high heteroatom content (this justifies the terms, polar compounds or N, S, 0 compounds , also used to designate resins). 

Liquid chromatography using a polar stationary phase and a nonpolar mobile phase. 

Liquid chromatography using a nonpolar stationary phase and a polar mobile phase. 

In reverse-phase chromatography, which is the more commonly encountered form of HPLC, the stationary phase is nonpolar and the mobile phase is polar. The most common nonpolar stationary phases use an organochlorosilane for which the R group is an -octyl (Cg) or -octyldecyl (Cig) hydrocarbon chain. Most reverse-phase separations are carried out using a buffered aqueous solution as a polar mobile phase. Because the silica substrate is subject to hydrolysis in basic solutions, the pH of the mobile phase must be less than 7.5. 

In liquid-solid adsorption chromatography (LSC) the column packing also serves as the stationary phase. In Tswett s original work the stationary phase was finely divided CaCOa, but modern columns employ porous 3-10-)J,m particles of silica or alumina. Since the stationary phase is polar, the mobile phase is usually a nonpolar or moderately polar solvent. Typical mobile phases include hexane, isooctane, and methylene chloride. The usual order of elution, from shorter to longer retention times, is... 

The most common mobile phase for supercritical fluid chromatography is CO2. Its low critical temperature, 31 °C, and critical pressure, 72.9 atm, are relatively easy to achieve and maintain. Although supercritical CO2 is a good solvent for nonpolar organics, it is less useful for polar solutes. The addition of an organic modifier, such as methanol, improves the mobile phase s elution strength. Other common mobile phases and their critical temperatures and pressures are listed in Table 12.7. 

For mixture.s the picture is different. Unless the mixture is to be examined by MS/MS methods, usually it will be necessary to separate it into its individual components. This separation is most often done by gas or liquid chromatography. In the latter, small quantities of emerging mixture components dissolved in elution solvent would be laborious to deal with if each component had to be first isolated by evaporation of solvent before its introduction into the mass spectrometer. In such circumstances, the direct introduction, removal of solvent, and ionization provided by electrospray is a boon and puts LC/MS on a level with GC/MS for mixture analysis. Further, GC is normally concerned with volatile, relatively low-molecular-weight compounds and is of little or no use for the many polar, water soluble, high-molecular-mass substances such as the peptides, proteins, carbohydrates, nucleotides, and similar substances found in biological systems. LC/MS with an electrospray interface is frequently used in biochemical research and medical analysis. 

The separation of the polysaccharide components utilizes their different solubUities, polar groups, extents of branching, molecular weights, and molecular flexibUities and may be accompUshed batchwise or with easUy automated column techniques such as column or high performance Uquid chromatography. These procedures have been summarized in several reviews (3,141—143). 

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