Stationary phase retention parameter

Figure 7.2 shows that in CCC, the observed chromatogram significantly depends on the Us/Um phase ratio that is expressed using the stationary phase retention parameter Sf as... 

Recent studies have made it possible to classify water-organic solvent systems in CCC for separation of organic substances on the basis of the liquid-phase density difference, the solvent polarity, and other parameters from the point of view of stationary-phase retention in a CCC column [1,3-9]. Ito [1] classified some liquid systems as hydrophobic (such as heptane-water or chloroform-water), intermediate (chloroform-acetic acid-water and n-butanol-water) and hydrophilic (such as n-butanol-acetic acid-water) according to the hydrophobicity of the nonaqueous phase. Thirteen two-phase solvent systems were evaluated for relative polarity by using Reichardt s dye to measure solvachromatic shifts and using the solubility of index compounds [6]. 

CPCs for separation of proteins which are not soluble in the PEG-phosphate system. This two-phase solvent system consists of the PEG-rich upper phase and dex-tran-rich lower phase. The cross-axis CPC may be operated in four different elution modes PiHO, PuTO, PiTI, and PnHI. The parameters Pi and Pn indicate the direction of the planetary motion where Pj indicates counterclockwise and Pn clockwise when observed from the top of the centrifuge. H and T indicate the head-tail elution mode, and O and I the inward-outward elution mode along the holder axis. In mode I (inward), the mobile phase is eluted against the laterally acting centrifugal force, and in mode O (outward), this flow direction is reversed. These three parameters yield a total of four combinations for the left-handed coils. Among these elution modes, the inward-outward elution mode plays the most important role in the stationary-phase retention for the polymer-phase sys-... 

Figure 4,14. Diagram of the thermodynamic cycle used to explain retention in reversed-phase chromatography by solvophobic theory. Na = Avogadro number, AA = reduction of hydrophobic surface area due to the adsorption of the analyte onto the bonded ligand, y = surface tension, = energy correction parameter for the curvature of the cavity, V = molar volume, R = gas constant, T = temperature (K), Pq = atmospheric pressure, AGydw.s.i a complex function of the ionization potential and the Clausius-Moscotti functions of the solute and mobile phase. Subscripts i = ith component (solute or solvent), S = solute, L = bonded phase ligand, SL = solute-ligand complex, R = transfer of analyte from the mobile to the stationary phase (retention), CAV = cavity formation, VDW = van der Waals interactions, ES = electrostatic interactions.

The primary factors that govern retention are the distribution coefficient (K) and the volume of stationary phase (Vs)). It is now necessary to identify those parameters that control the magnitude of the distribution coefficient itself and the volume of available stationary phase in a column. A study of these factors will be the subject of the next chapter. 

Typical normal-phase operations involved combinations of alcohols and hexane or heptane. In many cases, the addition of small amounts (< 0.1 %) of acid and/or base is necessary to improve peak efficiency and selectivity. Usually, the concentration of polar solvents such as alcohol determines the retention and selectivity (Fig. 2-18). Since flow rate has no impact on selectivity (see Fig. 2-11), the most productive flow rate was determined to be 2 mL miiT. Ethanol normally gives the best efficiency and resolution with reasonable back-pressures. It has been reported that halogenated solvents have also been used successfully on these stationary phases as well as acetonitrile, dioxane and methyl tert-butyl ether, or combinations of the these. The optimization parameters under three different mobile phase modes on glycopeptide CSPs are summarized in Table 2-7. 

In a series of papers published throughout the 1980s, Colin Poole and his co-workers investigated the solvation properties of a wide range of alkylammonium and, to a lesser extent, phosphonium salts. Parameters such as McReynolds phase constants were calculated by using the ionic liquids as stationary phases for gas chromatography and analysis of the retention of a variety of probe compounds. However, these analyses were found to be unsatisfactory and were abandoned in favour of an analysis that used Abraham s solvation parameter model [5]. 

Apart from enabling rapid prediction of solute retention, the Soczewinski equation allows a moleeular-level scrutiny of the solute — stationary phase interactions. The numeiieal value of the parameter n from Equation 2.14, which is at least approximately equal to unity (n 1), gives evidence of the one-point attachment of the solute moleeule to the stationary phase surface. The numeiieal values of n higher than unity prove that in a given chromatographic system, solute molecules interact with the stationary phase in more than one point (the so-ealled multipoint attachment). 

Another fairly important stationary phase in straight phase PLC is aluminum oxide. Comparable with silica gel also in the case of aluminum oxides, hydroxyl groups at the surface of this adsorbent are responsible for the selective retention of sample molecules. The relevant physical parameters for the characterization of aluminum oxides suitable for straight phase PLC are the following ... 

In fact, the solute retention depends on the solubihty parameters of the solute, 8 , of the mobile phase, 8 , of the stationary phase, 8, and of the phase ratio given by Equation 4.7 [24] ... 

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