Column selectivity factor

Three separate factors affect resolution (1) a column selectivity factor that varies with a, (2) a capacity factor that varies with k (taken usually as fej). and (3) an efficiency factor that depends on the theoretical plate number. 

The relative selectivity of a chromatographic column for a pair of solutes is given by the selectivity factor, a, which is defined as... 

Equations 12.21 and 12.22 contain terms corresponding to column efficiency, column selectivity, and capacity factor. These terms can be varied, more or less independently, to obtain the desired resolution and analysis time for a pair of solutes. The first term, which is a function of the number of theoretical plates or the height of a theoretical plate, accounts for the effect of column efficiency. The second term is a function of a and accounts for the influence of column selectivity. Finally, the third term in both equations is a function of b, and accounts for the effect of solute B s capacity factor. Manipulating these parameters to improve resolution is the subject of the remainder of this section. 

According to Equation 3, the resolution of two peaks in column separation is controlled by three major variables retention defined in terms of the retention factor k column efficiency expressed as the number of theoretical plates N and selectivity characterized by the selectivity factor a [48] ... 

Fig. 3-18. Selectivity factors a determined for (3,5-dinitroben-zoyl)leucine diallylamide on CSP 15-24. Conditions analyte (3,5-dinitrobenzoyl)leucine diallylamide column 150 x 4.6 mm i.d. mobile phase 20 % hexane in dichloromethane flowrate 1 mL min f UV detection at 254 nm.

The competitive adsorption isotherms were determined experimentally for the separation of chiral epoxide enantiomers at 25 °C by the adsorption-desorption method [37]. A mass balance allows the knowledge of the concentration of each component retained in the particle, q, in equilibrium with the feed concentration, < In fact includes both the adsorbed phase concentration and the concentration in the fluid inside pores. This overall retained concentration is used to be consistent with the models presented for the SMB simulations based on homogeneous particles. The bed porosity was taken as = 0.4 since the total porosity was measured as Ej = 0.67 and the particle porosity of microcrystalline cellulose triacetate is p = 0.45 [38]. This procedure provides one point of the adsorption isotherm for each component (Cp q. The determination of the complete isotherm will require a set of experiments using different feed concentrations. To support the measured isotherms, a dynamic method of frontal chromatography is implemented based on the analysis of the response curves to a step change in feed concentration (adsorption) followed by the desorption of the column with pure eluent. It is well known that often the selectivity factor decreases with the increase of the concentration of chiral species and therefore the linear -i- Langmuir competitive isotherm was used ... 

Column selection remains the most important factor in successful enantiomeric separations. The CSPs most likely to be effective in SFC are those that have been employed under normal phase conditions in LC. In fact, the tremendous body of knowledge that has been accumulated for LC can also guide column selection in SFC [66]. The likelihood of success with a particular CSP can generally be gauged after one or two injections [67]. If no evidence of separation is observed, another CSP should be investigated. 

Table 3.42 lists the main factors influencing optimisation of SPE. When considering a specific extraction problem, many different aspects influence column selection, including nature of the analytes and of the sample matrix degree of purity required nature of major contaminants in the sample and final analytical procedure. Reversed-phase sorbents have nonpolar functional groups and preferentially retain nonpolar compounds. Thus, for a nonpolar analyte, to remove polar interferences using a polar sorbent phase, the sample... 

Retention distance (or time) is normally used to aid the identification of a component of a mixture, provided that a known sample of the component has been subjected to separation under identical conditions. Because of the variations that can occur in the retention time due to technical factors, e.g. fluctuations in flow rate, condition of the column, the relative retention or selectivity factor (a) is sometimes used. This expresses the test retention time as a ratio of the retention time of another component or reference compound when both are injected as a mixture ... 

Differential hydration of proteins has been little exploited as a selectivity factor in ion exchange, but it is simple to evaluate and can produce useful results. This technique relies on the preferential exclusion of certain solutes from protein surfaces to produce an exclusionary effect and favor their interaction with the column. Protein hydration is generally proportional to protein size and solubility. Among proteins of similar size, this predicts that retention will increase with protein solubility. Among proteins of similar solubility, retention increases with protein size.16... 

SRM 869a Column Selectivity Test Mixture for Liquid Chromatography [44] is composed of three shape-constrained PAHs (phenanthro[3,4-c]phenanthrene, PhPh l,2 3,4 5,6 7,8-tetrabenzonaphthalene, TBN and benzo[a]pyrene, BaP) and is routinely employed to evaluate the shape selectivity of stationary phases. The retention differences between the nonplanar TBN and planar BaP solutes (expressed as a selectivity factor axEN/BaP = provide a numerical assessment of... 

Katz, E.D. and Ogan, K.L., Selectivity factors for several PAH pairs on Cl8 bonded phase columns, J. Liq. Chromatogn, 3, 1151, 1980. 

The determination of endogenous compounds and drugs in biological matrices has always presented a formidable challenge as one has to consider various factors before attempting to develop a suitable HPLC assay. These include the physicochemical properties of the compound such as the pKa value, solubility, volatility, particular functional groups (e.g., possessing chromophores, fluorophores, or electroactive characteristics), potential metabolites, and the required sensitivity and specificity. All these aspects will determine the type of extraction processes, analytical column selection, and suitable detector systems to be used as part of the HPLC apparatus. 

As mentioned earlier as an axiom, separation between two components is only possible if they have different migration rates through the column. Selectivity or separation factor (a) is a measure of differential retention of two analytes. It is defined as the ratio of the capacity factors k ) of two peaks as shown in Figure 4. Selectivity must be >1.0 for peak separation. 

Equation 6 Calculation of optimum ratio of particle size and column length, with selectivity factor, a capacity factor of second component of critical pair under analytical chromatography conditions, fe 02 diffusion coefficient, (cm /s) (typical value for MW 1000 10 cm /s) viscosity, p (cP) specific permeability (1.2 X 10 for spherical particles), feo third term of the Knox equation, C and maximum safe operating pressure, Ap, (bar). 

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