Resolution column efficiency

Column efficiency N or iVefr defined by the number of theoretical plates or equilibrium steps in a column and reflects the column s ability to separate a mixture of components with good resolution. Column efficiency depends on the number of equilibrium steps in a column, length L, or planar system, smallest equilibrium steps or plate heights (//) give highest efficiencies and... 

Two methods for improving chromatographic resolution (a) Original separation showing a pair of poorly resolved solutes (b) Improvement in resolution due to an increase in column efficiency ... 

From equation 12.1 it is clear that resolution may be improved either by increasing Afr or by decreasing wa or w-q (Figure 12.9). We can increase Afr by enhancing the interaction of the solutes with the column or by increasing the column s selectivity for one of the solutes. Peak width is a kinetic effect associated with the solute s movement within and between the mobile phase and stationary phase. The effect is governed by several factors that are collectively called column efficiency. Each of these factors is considered in more detail in the following sections. 

Now that we have defined capacity factor, selectivity, and column efficiency we consider their relationship to chromatographic resolution. Since we are only interested in the resolution between solutes eluting with similar retention times, it is safe to assume that the peak widths for the two solutes are approximately the same. Equation 12.1, therefore, is written 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. 

Equation (16-168) shows that the resolution is the result of independent effects of the separation selectivity (ot), column efficiency [Np), and capacity (k ). Generally, peaks are essentially completely resolved when R, = 1.5 (>99.5 percent separation). In practice, values of R, 1, corresponding to 98 percent separation, are often considered adequate. 

The smallest size difference that can be resolved is related to the pore volume, the solute shape, and the efficiency of the column (see Fig. 2.6). However, this is at very low loadings. At higher loadings the sample volume will contribute to zone broadening and may, in some cases, be the dominating factor for resolution. Thus, for fractionation, an optimum exists with respect to column efficiency (represented by the flow rate as operational parameter) and sample volume for processing a particular volume of feed per unit time. As a rule of thumb this optimum can be found at a relative sample volume of 2-5% of the column volume (Hagel et al., 1989). 

Elow rate determines the separation time and can significantly affect resolution and efficiency. The effect of flow rate on HETP for TSK-GEL SW and TSK-GEL SWxi analytical columns is shown in Fig. 4.6. Resolution is typically higher at slower flow rates, although results shown in Fig. 5B indicate that, with increasing sample load, the faster flow rates can give higher resolution. 

For SEC separations of polymers, column efficiency is better characterized by specific resolution, R,p, and efficiency, T, than by theoretical plate count. Peak resolution, R is calculated according to (7) ... 

Improvement of column efficiency in terms of the number of theoretical plates realized by increasing column length often yields marginal increases in resolution, with a corresponding increase of analysis time to unacceptable levels. This... 

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] ... 

The use of both sub- and supercritical fluids as eluents yields mobile phases with increased diffusivity and decreased viscosity relative to liquid eluents [23]. These properties enhance chromatographic efficiency and improve resolution. Higher efficiency in SFC shifts the optimum flowrate to higher values so that analysis time can be reduced without compromising resolution [12]. The low viscosity of the eluent also reduces the pressure-drop across the chromatographic column and facilitates the... 

The curve exhibits a minimum, which means that there is an optimum mobile phase velocity at which the column will give the minimum HETP and consequently a maximum efficiency. In practice this usually means that reducing the flow rate of a column will increase the efficiency and thus the resolution. In doing so, however, the analysis time will also be increased. As seen in figure 5, however, there is a limit to this procedure, as reducing the column flow rate so that the mobile phase velocity falls below the optimum will result in an increase in the HETP and thus a decrease in column efficiency. 

The column that is required for an analysis must have the efficiency necessary to resolve the critical pair. However, it is necessary to decide what constitutes resolution, before the column efficiency can be calculated. How narrow must the peaks be maintained relative to their separation to permit an accurate quantitative analysis In figure 7, five pairs of peaks are shown, the area of the smaller peak being half that of the larger peak. 

Figure 1.9 illustrates the relationship between resolution, the separation factor, the average capacity factor and the column efficiency for some real chromatographic peaks [lOS]. The central portion of the figure illustrates how resolution increases with the capacity factor for a fixed separation factor and column efficiency. At first the resolution increases quickly as the... 

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