Stationary-phase particles, chromatography

FIGURE 1.4 Optical micrograph of macroporous chromatographic column materials, (a) Monosized particles of 20 tm. (b) Commercial column filling of 12-28 tm. [Reprinted from T. Ellingsen et al. (1990). Monosized stationary phases for chromatography.7. Chromawgr. 535,147-161 with kind permission from Elsevier Science-NL, Amsterdam, The Netherlands.]... 

In exclusion chromatography, the total volume of mobile phase in the column is the sum of the volume external to the stationary phase particles (the void volume, V0) and the volume within the pores of the particles (the interstitial volume, Vj). Large molecules that are excluded from the pores must have a retention volume VQ, small molecules that can completely permeate the porous network will have a retention volume of (Vo + Fj). Molecules of intermediate size that can enter some, but not all of the pore space will have a retention volume between VQ and (V0 + Fj). Provided that exclusion is the only separation mechanism (ie no adsorption, partition or ion-exchange), the entire sample must elute between these two volume limits. 

Size exclusion chromatography. The separation occurs because the stationary phase particles are porous and the small molecules enter the pores and are slowed from passing through the column, while the large molecules pass through more quickly since they do not enter the pores. 

By studying the properties of polymer layers on soHd surfaces it soon became obvious that not only is the chemical composition of the immobihzed polymer cmcial for the performance of the material, but so is its morphology. This has been recognized in various fields of applications e.g. stabihzation of small particles suspensions by attached polymer brush-type layers [159, 160], control of adhesion [161] or friction [162] and tailored stationary phases for chromatography [163-165]. 

The general rate model of chromatography is the most complex of all the models used in this field. In this model, it is assumed that the mobile phase percolates through the interstitial volume between stationary phase particles, diffusion takes place from this stream into the particles and inside the pores of the stationary phase particles, where the mobile phase is stagnant, and adsorption-desorption takes place between the stagnant mobile phase within the pores and the adsorbent surface. 

Chromatography column packed with stationary phase particles... 

The term A is related to the flow profile of the mobile phase as it traverses the stationary phase. The size of the stationary phase particles, their dimensional distribution, and the uniformity of the packing are responsible for a preferential path and add mainly to the improper exchange of solute between the two phases. This phenomenon is the result of Eddy diffusion or turbulent diffusion, considered to be non-important in liquid chromatography or absent by definition in capillary columns, and WCOT (wall coated open tubular) in gas phase chromatography (Golay s equation without term A, cf. 2.5). 

Figure 23-19 Band spreading from multiple flow paths. The smaller the stationary phase particles, the less serious this problem is. This process is absent in an open tubular column. [Adapted from H m. McNair and E. J. Bonelli, Basic Gas Chromatography (Palo Alto. CA Varlan Instrument Division. 1968).]...

In chromatography, why is the optimal flow rate greater if the stationary phase particle size is smaller ... 

The use of hydrothermally formed retaining frits in capillary columns packed with stationary phase particles is an accepted limitation in CEC. The introduction of the frit to hold the packed bed is vital, yet introduces problems such as EOF and flow non-uniformities, compromised frit permeability [87], capillary fragility, increased likelihood of bubble formation [88] and a thermally induced modified frit surface chemistry which can detrimentally alter the chromatography [23]. Practical aspects to be considered include the appreciable effort and skill of the analyst who is required to repeatably manufacture capillaries of a particular phase and redevelop the fritting and packing methodology for each different stationary phase type. 

A high electroosmotic flow through the stationary-phase particles may be created when the appropriate conditions are provided. This pore flow has important consequences for the chromatographic efficiency that may be obtained in CEC. From plate height theories on (pressure-driven) techniques such as perfusion and membrane chromatography, it is known that perfusive transport may strongly enhance the stationary-phase mass transfer kinetics [30-34], It is emphasised... 

Fig. 12 Scanning Electron Microscopy images of MIP used as stationary phase in chromatography (1) MIP particles obtained by grinding, grading and sedimentation (2) beads prepared by an emulsion process (from [114])...

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