By Edwin S. Crawley, Henry B. Evans
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Nelis 61. Preparative and Production Scale Chromatography, edited by G. Ganetsos and P. E. Barker 62. Diode Array Detection in HPLC, edited by Ludwig Huber and Stephan A. George 63. Handbook of Affinity Chromatography, edited by Toni Kline 64. Capillary Electrophoresis Technology, edited by Norberto A. Guzman 65. Lipid Chromatographic Analysis, edited by Takayuki Shibamoto 66. Thin-Layer Chromatography: Techniques and Applications, Third Edition, Revised and Expanded, Bernard Fried and Joseph Sherma 67.
Size-exclusion LC is applied for the separation of (bio-)macromolecules in the molecular-mass range of 103-107 and for the characterization of the molecular-mass distribution of oligomers. For a homologous series of oligomers the retention volume is directly proportional to the logarithm of the molecular mass. 5 Other Modes of Liquid Chromatography Biopolymer separations The separation of biopolymers such as proteins and polynucleotides is of growing importance. The most widely used LC modes in protein separation are size-exclusion, ion-exchange, reversed-phase and hydrophobic interaction chromatography.
The flow-rate F, which can easily be measured in LC, through a column with an internal diameter dc is given by: Page 18 F = ¼pd2ceuu [Eq. 10] The average linear velocity through the column is related to the column pressure drop DP (in Pa) and the particle size dp of the stationary phase material (typically in the 510 mm ID range): [Eq. 11] where h is the liquid viscosity (in Pa s). The value 1000 in the denominator is an empirical factor, called the specific permeability coefficient. In LC, high pressure drops are needed, typically between 10 and 40 MPa.