Mobile-phase-viscosity dependence on DNA separation in slalom chromatography.
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.
Peyrin
E
E
Laboratoire de Chimie Analytique, UFR de Pharmacie, Domaine de la Merci, La Tronche, France.
Guillaume
Y C
YC
Grosset
C
C
Ravel
A
A
Villet
A
A
Garrel
C
C
Alary
J
J
Favier
A
A
D002853:Chromatography, Liquid / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*
D004247:DNA