A Practical Handbook of Preparative HPLC by Donald A Wellings

By Donald A Wellings

A distillation of over two decades of the author's useful event of this strategy, this publication intentionally steers away from advanced theoretical points and concentrates extra at the 'black paintings' components of the options concerned. for this reason is a e-book for widespread use within the laboratory, in particular of curiosity to these within the pharmaceutical, agrochemical and biotechnology industries and to an individual excited about the research and purification of gear, small molecules, proteins and DNA.

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1. There are two transport mechanisms in progress. Firstly, the convectional flow around the particles; and secondly, the diffusion in and out of the pores of the stationary phase. 1 the efficiency of a chromatographic separation. Traditionally the term ‘number of theoretical plates’ is used to define the efficiency of the packed column bed. 3] and published in 1941. Why plates? This term actually originated in the petrochemicals industry and is derived from the oil refinery process, where an increased number of plates in a distillation column results in a more efficient separation.

4 in Chapter 1 shows a scanning electron micrograph (SEM) of typical irregular and spherical silica particles for comparison. Normal phase chromatography is typically, though not exclusively, performed in organic solvents. Retaining solvents are generally hydrocarbons, such as hexane, heptane or cyclohexane; or halocarbons including dichloromethane and chloroform. The eluting mobile phase is generally a miscible hydrophilic solvent, such as alcohols and esters. Organic acids are often added to minimize any cation exchange effects arising from acidic silanol groups on the silica surface.

As discussed in Chapter 2, van Deemter’s theory suggests that the most critical attribute of a stationary phase is the ease of mass transfer through the pores of the particle. The control of pore size and structure during preparation is a complex process that has been mastered by few manufacturers, but it is true to say that pore size distribution and uniformity is more readily achieved in silica particles than it is in polymerics. This is not to say that polymerics are inferior, because in contrast these stationary phases have a more uniform particle size.

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