Selectivity of Stationary Phase Materials

Preparation, Testing, and Selectivity of Stationary Phase Materials... 

The selectivity of stationary phase materials can be understood if the method of their synthesis is understood. Differences in the same type of stationary phase material from different manufacturers or even from the same manufacturer depend on the synthetic methods and the quality control that has been employed. Details of the individual synthetic processes from different manufacturers have not been published, but are basically the same.1,2... 

An improvement in the resolution can also be achieved by increasing the separation of the peak (A/R) or narrowing the peak width (increased N). An increase in AtR depends on the selection of stationary phase materials and eluent a reduction of the peak width depends on improving the column... 

Gas chromatography is often divided into categories based on the type of stationary phase used. Gas-liquid chromatography (GLC) implements a porous, inert solid support that is coated with a viscous, nonvolatile liquid phase. On the other hand, gas-solid chromatography (GSC) uses a solid adsorbent as the stationary phase. Klee offers these general rules-of-thumb for selection of stationary phase materials use solid adsorbents to separate room-temperature gases, liquid stationary phases to separate room-temperature liquid and solid mixtures, polar phases for polar solutes, and nonpolar phases for nonpolar solutes. Table 1 lists common liquid- and solid-stationary phase materials available for use in capillary columns. Barry cross-refers numerous column materials from nine different manufacturers. ... 

Selection of columns and mobile phases is determined after consideration of the chemistry of the analytes. In HPLC, the mobile phase is a liquid, while the stationary phase can be a solid or a liquid immobilised on a solid. A stationary phase may have chemical functional groups or compounds physically or chemically bonded to its surface. Resolution and efficiency of HPLC are closely associated with the active surface area of the materials used as stationary phase. Generally, the efficiency of a column increases with decreasing particle size, but back-pressure and mobile phase viscosity increase simultaneously. Selection of the stationary phase material is generally not difficult when the retention mechanism of the intended separation is understood. The fundamental behaviour of stationary phase materials is related to their solubility-interaction... 

The physical and chemical aspects of liquid chromatography, in addition to mechanical aspects, are briefly described in this chapter. Theoretical approaches are explained in detail in later chapters. The effect of stationary phase materials on the chemical selectivity is described in Chapter 3, and the influence of the eluent components is covered in Chapter 4. The plate number theory is discussed in Chapter 5. Quantitative optimization is explained in Chapter 6. 

Affinity liquid chromatography and chiral separations (enantiomer separations) require similar analyte properties. The solutes may have interactions through hydrogen-bonding, ligand formation, or Coulombic forces with the surface of stationary phase materials or the sites of additives however, the selectivity is controlled by the steric effects of the structures of the analyte molecules and the recognition molecules (chiral selectors). 

The physical and chemical properties of stationary phase materials are described in Chapter 3 (including methods for their synthesis) to clarify the differences in similar stationary phase materials supplied from different manufacturers. A detailed selection guide to solvents is given in Chapter 4. The unlimited selection of eluent components and their concentrations is a powerful force in developing separations in liquid chromatography. Although this area seems rather complicated, it is easy to understand the selection of a suitable eluent when you first identify the molecular properties of the analytes and solvents. 

The narrower the particle size distribution, the higher in theory is the potential theoretical plate number. A rough sieving is achieved by a water flow, air flow, or a vibration method. A common sieving method is Hamiltonian water flow (Figure 3.4). The particle distribution can be controlled within + 1 jum by this method. A slurry of stationary phase material is allowed to float in the cylinder, and a solvent flows from the bottom to the top. The smaller and lighter particles float to the top of the cylinder and the larger and heavier particles sink to the bottom. The required particles are collected at the top of the cylinder. The selection of suspension solvent and control of the temperature are important. 

Many manufacturers sell the same types of stationary phase materials,6 but the most popular stationary phase materials, e.g. octadecyl-bonded silica gels, from different sources, even from the same manufacturer, often demonstrate different retention capacities and selectivities. Such differences are due to the aggressive reactivity of the silylation method and the different bonding reactions that are used, as described earlier. 

Example 2 Chromatography of nitroaniline isomers. The elution order of the nitroaniline isomers was ortho, meta, and para in normal-phase liquid chromatography using H-butanol-w-hexane mixtures as the eluent, when the stationary phase material was either silica gel, alumina, an ion-exchanger, polystyrene gel, or octadecyl-bonded silica gel. The results indicate that the separation of these compounds can be performed on a range of different types of stationary phase materials if the correct eluent is selected. The best separation will be achieved by the right combination of stationary phase material and eluent.68... 

Chromatography is a very versatile technique offering a wide range of solid phase materials and detector types which can deal with very complex mixtures. In practice all materials and conditions used in the instrument are carefully chosen to match the type of sample mixture involved. This includes selection of stationary phase (chemical and physical properties) column type and length sample pretreatment, operational temperatures, pressures, and flow rates physical and chemical nature of mobile phase detector type and so forth. Detection to nanogram level is quite common and some systems can detect to picogram level using very small volumes of sample. 

Almost any polar solid can be used, the most common choices being silica gel or alumina. A selection of stationary phases is listed in table 4.7 in order of decreasing adsorptive power. Silica gel and alumina are highly polar materials that adsorb molecules strongly. They are said to be active adsorbents. Activity is determined by the overall polarity and by the number... 

In searching for new separation processes, the chromato-grapher develops new types of stationary phase materials in order to gain improvements in selectivity. The development of zirconia-silica composites is an example which—although, at present, applications on these surfaces are limited to their development—typifies the ongoing search. Whether the separations achieved on the zirconia-silica composites are better or worse than those achieved on either zirconia or silica is immaterial to the fundamental process of discovery. The important factor is that the separation is different and, therefore, an alternative separation strategy may be employed. In addition to the differences in selectivity that may be found when new surfaces are developed, beneficial surface properties may also be realized. These may be related to aspects such as the pore stmcture, the acidic or basic properties, or the solubility of the support. 

Selection of stationary phases is, therefore, limited for these preparative techniques. The only materials which can be used are those for which an additional amount of binder endows the necessary stability and does not impair the separation these can be silica, kieselguhr, alumina, plaster of Paris (76), and their combinations. In general, silica gel of TLC quality (15 pm average particle size) with gypsum binder is used with additional binder, 3.5% calcium sulfate hemihydrate. If other types of silica without binder are used, approximately 20% of the binder must be added. It is reconunended that several layers be prepared together and stored in a safe place. They can be activated before use. Precoated glass rotors (1,2, or 4 mm layer thickness) are available from Analtech (Newark, Delaware, USA). 

A particular column can be used for different types of LC by changing the eluent components. For example, a column packed with RP-18 bonded silica gel can be used for SEC with THF, NPLC with n-hexane, and RPLC with aqueous acetonitrile. When separation cannot be achieved by improving the theoretical plate number of a column, it may be achieved by selection of an appropriate stationary phase material and/or eluent. 

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