By Donald A Wellings
A distillation of over two decades of the author's functional event of this approach, this e-book intentionally steers away from complicated theoretical points and concentrates extra at the 'black paintings' parts of the thoughts concerned. for this reason is a ebook for common use within the laboratory, in particular of curiosity to these within the pharmaceutical, agrochemical and biotechnology industries and to someone serious about the research and purification of gear, small molecules, proteins and DNA.
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Additional info for A Practical Handbook of Preparative HPLC
54⎜⎜⎜ 1 ⎟⎟ ⎜⎝ W1/2 ⎟⎠ 22 Fluid Dynamics, Mass Transport and Friction The separation factor ␣, is effectively a measure of the degree of separation of two components. This term is derived from the capacity factor, kЈ, of the corresponding peaks and is simplified to produce the following equation. ␣ϭ V2 V1 The measurements a, b, c and d on the chromatogram can be used to calculate peak asymmetry, A and resolution, R. Aϭ Rϭ b a 2(V2 Ϫ V1 ) (c ϩ d) Another term commonly employed is the height equivalent to a theoretical plate, HETP, which is simply the length of the column divided by the number of theoretical plates, N.
In addition, it can be a laborious process to optimize the crystallization process. 4]. 3 these has led to domination of the preparative market by two or three companies over the last 20 years. The interaction between the analyte and the stationary phase in chiral chromatography can involve characteristics of molecular size, hydrophilic, hydrophobic and ion-exchange mechanisms, often simultaneously. In general, the surface of the stationary phase is covered with a chiral ligand that will bind preferentially with one of the enantiomers of the mixture.
Modes of Chromatographic Separation 33 Pore structure, size and surface area The pore structure and size is critical for efficient chromatography and this is borne out by the fact that many supports of identical particle size, and nominally of similar pore size, often display different characteristics. 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.
A Practical Handbook of Preparative HPLC by Donald A Wellings