Mobile-phase-viscosity dependence on DNA separation in slalom chromatography |
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Authors: | Peyrin E Guillaume Y C Grosset C Ravel A Villet A Garrel C Alary J Favier A |
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Institution: | Laboratoire de Chimie Analytique, UFR de Pharmacie, Domaine de la Merci, La Tronche, France. |
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Abstract: | Slalom chromatography (SC) is an alternative chromatographic procedure for the separation of relatively large double-stranded DNA molecules and is based on a new principle. The retardation of the DNA fragments from the cleavage of the Lambda DNA by the KpnI restriction enzyme was studied using an acetonitrile-phosphate buffer as a mobile phase with various concentrations of viscosity modifier (i.e. glycerol) and a C1 column as a stationary phase. The DNA molecule retention was accurately described over the glycerol concentration range using a model previously established. It was shown that the eluent viscosity increase enhanced the slalom chromatographic capacity to separate the DNA fragments. A connection between SC and 'hydrodynamic chromatography' processes was predicted to link the two processes in a global separation mechanism based on a non-equilibrium principle. |
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