Chromatography column efficiency

Efficiency is mainly a column-specific parameter. In a gas chromatography column, efficiency is highly dependent on the flow rate. In HPLC, because of much higher viscosity, the applicable flow rate region is not so broad within this region, variations of the flow rate do not affect column efficiency significantly. 

Theory of chromatography column efficiency expanded Chromatography simulation software for method development Capillary gas chromatography (GC) columns updated and expanded Headspace, thermal desorption, and purge and trap GC analysis Fast gas and hquid chromatography... 

In their original theoretical model of chromatography, Martin and Synge treated the chromatographic column as though it consists of discrete sections at which partitioning of the solute between the stationary and mobile phases occurs. They called each section a theoretical plate and defined column efficiency in terms of the number of theoretical plates, N, or the height of a theoretical plate, H where... 

These factors make it necessary to reduce the amount of solvent vapor entering the flame to as low a level as possible and to make any droplets or particulates entering the flame as small and of as uniform a droplet size as possible. Desolvation chambers are designed to optimize these factors so as to maintain a near-constant efficiency of ionization and to flatten out fluctuations in droplet size from the nebulizer. Droplets of less than 10 pm in diameter are preferred. For flow rates of less than about 10 pl/min issuing from micro- or nanobore liquid chromatography columns, a desolvation chamber is unlikely to be needed. 

Displacement Development A complete prediction of displacement chromatography accounting for rate factors requires a numerical solution since the adsorption equilibrium is nonlinear and intrinsically competitive. When the column efficiency is high, however, useful predictious can be obtained with the local equilibrium theoiy (see Fixed Bed Transitions ). 

It appears that the equation introduced by Van Deemter is still the simplest and the most reliable for use in general column design. Nevertheless, all the equations helped to further understand the processes that occur in the column. In particular, in addition to describing dispersion, the Kennedy and Knox equation can also be employed to assess the efficiency of the packing procedure used in the preparation of a chromatography column. 

Using equation (10), the efficiency of any solute peak can be calculated for any column from measurements taken directly from the chromatogram (or, if a computer system is used, from the respective retention times stored on disk). The computer will need to have special software available to identify the peak width and calculate the column efficiency and this software will be in addition to that used for quantitative measurements. Most contemporary computer data acquisition and processing systems contain such software in addition to other chromatography programs. The measurement of column efficiency is a common method for monitoring the quality of the column during use. 

Most size exclusion chromatography (SEC) practitioners select their columns primarily to cover the molar mass area of interest and to ensure compatibility with the mobile phase(s) applied. A further parameter to judge is the column efficiency expressed, e.g., by the theoretical plate count or related values, which are measured by appropriate low molar mass probes. It follows the apparent linearity of the calibration dependence and the attainable selectivity of separation the latter parameter is in turn connected with the width of the molar mass range covered by the column and depends on both the pore size distribution and the pore volume of the packing material. Other important column parameters are the column production repeatability, availability, and price. Unfortunately, the interactive properties of SEC columns are often overlooked. 

Traditionally, column efficiency or plate counts in column chromatography were used to quantify how well a column was performing. This does not tell the entire story for GPC, however, because the ability of a column set to separate peaks is dependent on the molecular weight of the molecules one is trying to separate. We, therefore, chose both column efficiency and a parameter that we simply refer to as D a, where Di is the slope of the relationship between the log of the molecular weight of the narrow molecular weight polystyrene standards and the elution volume, and tris simply the band-broadening parameter (4), i.e., the square root of the peak variance. 

COLUMN EFFICIENCY AND PLATE HEIGHTS IN UNIFIED CHROMATOGRAPHY... 

There have been a few reports of column efficiency and reduced plate height measurements in several unified chromatography techniques. These have been based on the apparent plate height observed at the column outlet. In the notation used by Giddings (32) the apparent plate height, H, is given by the following ... 

Column efficiency (number of theoretical plates) As in batch chromatography, one needs to determine the efficiency of the column in order to evaluate the dispersion of the fronts due to hydrodynamics dispersion or kinetics limitations. The relationship of N proportional to L can be expressed in terms of the equation for height equivalent to a theoretical plate (HETP) as ... 

The effect of temperature on column efficiency, however, is frequently exploited, particularly in size exclusion chromatography (SEC). As has already been discussed, high efficiencies are essential in SEC due to the limited peak capacity of the column and consequently, the very small separation ratios. However the effect of temperature on column efficiency is not well understood by many analysts and consequently, will be discussed in some detail. It was shown on page... 

The injection device is also an important component in the LC system and has been discussed elsewhere (2,18). One type of injector is analogous to sample delivery in gas chromatography, namely syringe injection through a self-sealing septum. While this injection procedure can lead to good column efficiency, it generally is pressure limited, and the septum material can be attacked by the mobile phase solvent. 

Figure 2 shows the effect of flow rate on column efficiency using the SW-2000 column with cytochrome C. The column efficiency expressed as the number of theoretical plates (N) was dependent on flow rate, a result typical of size exclusion chromatography. 

The separation nuaber is the only column efficiency par2uaeter that can be deterained under teaperature progr2uued conditions [45,46]. The critical parameters that aust be standardized to obtain reproducible SM values for coluans of different length are the carrier gas flow rate and the temperature program. The SN is widely used as part of a standardized test method to evaluate the quality of open tubular columns for gas chromatography (section 2.4.3). 

The highest column efficiency will be obtained at In practice, higher values are frequently used to minimize the separation time. For gas chromatography a velocity of about 2 u, called the practical operating gas velocity, is frequently recommended [67]. Provided that the ascending portion of the van Deemter curve is... 

In addition to water, virtually any organic polar modifier may be used to control solute retention in liquid-solid chromatography. Alcohols, alkyl2aiines, acetonitrile, tetrahydrofuran and ethyl acetate in volumes of less than one percent can be incorporated into nonpolar mobile phases to control adsorbent activity. In general, column efficiency declines for alcohol-moderated eluents cogqpared to water-moderated eluent systems. Many of the problems discussed above for water-moderated eluents are true for organic-moderated eluents as well. 

Golshan-Shirazi, S. and Guiochon, G., Theory of optimization of the experimental conditions of preparative liquid chromatography optimization of column efficiency, Anal. Chem., 61, 1368, 1989. 

Warren, Jr., F. V. and Bidlingmeyer, B. A., Influence of temperature on column efficiency in reversed phase liquid chromatography, Anal. Chem., 60, 2821, 1988. 

Neddermeyer, P. A. and Rogers, L. B., Column efficiency and electrolyte effects of inorganic salts in aqueous gel chromatography, Anal. Chem., 41, 94, 1969. 

Nimura, N., Itoh, H., Kinoshita, T., Nagae, N., Nomura, M. (1991). Fast protein separation by reversed-phase high-performance liquid-chromatography on octadecylsilyl-bonded non-porous silica-gel — effect of particle-size of column packing on column efficiency. J. Chromatogr. 585(2), 207-211. 

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