Preparative chromatography particle size

The agglomeration of the primary particles creates a secondary pore system which adds to the pore system of the primary particles. These agglomerated particles could be applied in preparative chromatography. The size of the agglomerates can be adjusted between 2 and 20 nm by changing various parameters such as flow rate of the silica suspension and the diameter of the jet nozzle. 

A novel cross-linked polystyrene-divinylbenzene copolymer has been produced from suspension polymerization with toluene as a diluent, having an average particle size of 2 to 50 /rm, with an exclusive molecular weight for the polystyrene standard from about 500 to 20,000 in gel-permeation chromatography. A process for preparing the PS-DVB copolymer by suspension polymerization in the presence of at least one free-radical polymerization initiator, such as 2,2 -azo-bis (2,4-dimethylvaleronitrile) with a half-life of about 2 to 60 min at 70°C, has been disclosed (78). 

Some authors have suggested the use of fluorene polymers for this kind of chromatography. Fluorinated polymers have attracted attention due to their unique adsorption properties. Polytetrafluoroethylene (PTFE) is antiadhesive, thus adsorption of hydrophobic as well as hydrophilic molecules is low. Such adsorbents possess extremely low adsorption activity and nonspecific sorption towards many compounds [109 111]. Fluorene polymers as sorbents were first suggested by Hjerten [112] in 1978 and were tested by desalting and concentration of tRN A [113]. Recently Williams et al. [114] presented a new fluorocarbon sorbent (Poly F Column, Du Pont, USA) for reversed-phase HPLC of peptides and proteins. The sorbent has 20 pm in diameter particles (pore size 30 nm, specific surface area 5 m2/g) and withstands pressure of eluent up to 135 bar. There is no limitation of pH range, however, low specific area and capacity (1.1 mg tRNA/g) and relatively low limits of working pressure do not allow the use of this sorbent for preparative chromatography. 

FIGURE 12.3 Particle sizes and particle size distribution of silica stationary phases for planar chromatography. (From Nyiredy, S., Preparative layer chromatography, in Handbook of Thin Layer Chromatography, 3rd ed., Vol. 89, Sherma, J. and Fried, B., lids., Marcel Dekker, New York, 2003, pp. 99-133. With permission.)... 

Figure 3.6 Silica spheres with monodisperse particle size are expensive analytical gels of top performance. Silica spheres with determined particle size ranges are ideally suited for preparative chromatography. (Photo courtesy of SiliCycle.)...

Larger diameter columns were also available for preparative chromatography. In later years, GPC analysis times were reduced and resolution was Improved by using shorter columns that were packed with smaller particle size material. A typical family of GPC columns that is available today contains 7pm particles... 

A ternary liquid-liquid system for partition chromatography is prepared from a mixture of 2,2,4-trimethylpentane, ethanol and water (34 5 1). The less polar upper layer is used as the stationary phase. A diatomaceous material, Hyflow Super Cel (particle size, 7-11 jam), is used as the solid support. The columns (40 cm X 4 mm I.D.) are packed by the slurry technique, and the support material is coated in situ with the liquid stationary phase as described earlier [54]. A pre-column is inserted in order to maintain equilibrium between... 

Ha et al. [93,94] prepared monodisperse polymer microspheres from 1 to 40 pm in diameter for medical diagnostic tests, as chromatography column packing and as calibration standards. The work deals with the synthesis of large and uniform poly (butadiene-styrene) latex. The ceramic SPG membrane, with a pore diameter of 1.6 pm, was employed. The uniform particle sizes were in the diameter range of 4-6 pm. 

Analytical thin layer chromatography (TLC) was conducted on pre-coated TLC plates, silica gel 60 F254, layer thickness 0.25 mm, manufactured by E. Merck and Co., Darmstadt, Germany. Silica Gel for flash column chromatography was obtained from Silicycle Chemical Division Silica Gel, 60 (particle size 0.040 - 0.063 mm) 230-240 mesh ASTM. All columns were prepared, loaded, and fractions collected according to the specification of Still.37 Ethyl acetate used for chromatography was dried over 4 A molecular sieves for at least 24 hours prior to use. Hexanes are the mixed hydrocarbon fraction (bp 60-70 °C), principally n-hexanes, which was purified as follows the commercial solvent was stirred concentrated sulfuric acid for at least 24 hours, decanted, stirred over anhydrous sodium carbonate for 6 hours, decanted, then distilled. 

High-performance liquid chromatography does similar things with more sophisticated instrumentation. It can separate closely related chemical compounds on a research scale or on a preparative scale liquid solvents, or mixtures of several solvents under positive pressure, replace the "carrier gas" of Fig. 11.3. The solid support must have small particle sizes (3- to 10-pm diameter), so that relatively high pressures can be sustained throughout the column, and it is at the interface between the liquid eluant and the solid particles that the chromatographic separation is accomplished. 

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