By Donald A Wellings
A distillation of over two decades of the author's sensible event of this technique, this ebook intentionally steers away from complicated theoretical facets and concentrates extra at the 'black artwork' parts of the options concerned. accordingly is a e-book for common use within the laboratory, particularly of curiosity to these within the pharmaceutical, agrochemical and biotechnology industries and to a person thinking about the research and purification of substances, small molecules, proteins and DNA.
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Extra resources for A Practical Handbook of Preparative HPLC
A brief survey suggests that there are over five hundred different reversed phase materials commercially available. However, the eventual scale of operation may limit the number of suitable stationary phases to less than ten. Reversed phase media are designed to operate in aqueous buffers where the hydrophobic adsorption between analyte and stationary phase is disturbed by an increasing concentration of a miscible organic solvent. The most common application of reversed phase HPLC is in the purification of peptides and proteins where the analyte is often desorbed under gradient elution.
Cooling by conduction in the vicinity of the column walls reduces the viscosity of the solvent close to the wall in comparison to that at the centre of the column. Consequently, the solvent at the centre of the column is now travelling at a Fluid Dynamics, Mass Transport and Friction 27 higher flow rate than that at the column walls, resulting in a parabolic flow profile, and subsequently, to band broadening. In practice this column wall effect is particularly dominant in column diameters of 5 to 20 cm.
2]. 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.