Retention time hydrophobicity

Retention and stereoselectivity on the BSA columns can be changed by the use of additives to the aqueous mobile phase (30). Hydrophobic compounds generally are highly retained on the BSA, and a mobile-phase modifier such as 1-propanol can be added to obtain reasonable retention times. The retention and optical resolution of charged solutes such as carboxyUc acids or amines can be controlled by pH and ionic strength of the mobile phase. 

In 1981, a novel flotation device known as the air-sparged hydrocyclone, shown in Figure 3, was developed (16). In this equipment, a thin film and swid flotation is accompHshed in a centrifugal field, where air sparges through a porous wall. Because of the enhanced hydrodynamic condition, separation of fine hydrophobic particles can be readily accompHshed. Also, retention times can be reduced to a matter of seconds. Thus, this device provides up to 200 times the throughput of conventional flotation cells at similar yields and product quaHties. 

The van der Waals volume can be related to the hydrophobicity of the solutes, and retention of molecular compounds can be predicted from their van der Waals volumes, 7r-energy, and hydrogen-bonding energy effects [72-74], It should be noted that the isomeric effect of substituents cannot be predicted with good precision because this is not simply related to Hammett s a or Taft s other hand, the hydrophobicity is related to enthalpy [75], Retention times of non-ionizable compounds were measured in 70 and 80% acetonitrile/water mixtures on an octadecyl-bonded silica gel at 25-60°C and the enthalpy values obtained from these measurements. 

The most important parameter for the control of liquid chromatography is the composition of the eluent. Liquid chromatography is a powerful separation method with unlimited possibilities of eluent selection. However, it is not easy to choose a suitable eluent within a short time without a number of trial experiments. The crucial factor is to control the solubility of the analytes in the eluent. Increasing the solubility of analytes in the eluent decreases their retention times. The selection of the components of an eluent is described below, based on the properties of the analytes to be separated. The important properties are hydrophobicity, dipole moment, hydrogen bonding, ionization, and steric effects. 

Normal-phase liquid chromatography is thus a steric-selective separation method. The molecular properties of steric isomers are not easily obtained and the molecular properties of optical isomers estimated by computational chemical calculation are the same. Therefore, the development of prediction methods for retention times in normal-phase liquid chromatography is difficult compared with reversed-phase liquid chromatography, where the hydrophobicity of the molecule is the predominant determinant of retention differences. When the molecular structure is known, the separation conditions in normal-phase LC can be estimated from Table 1.1, and from the solvent selectivity. A small-scale thin-layer liquid chromatographic separation is often a good tool to find a suitable eluent. When a silica gel column is used, the formation of a monolayer of water on the surface of the silica gel is an important technique. A water-saturated very non-polar solvent should be used as the base solvent, such as water-saturated w-hexane or isooctane. 

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