Column efficiency and separation of DNA fragments using slalom chromatography: hydrodynamic study and fractal considerations.
Novel equations (Guillaume Y. C.; et al. Anal. Chem. 2000, 72, 853) were developed to describe the large double-stranded DNA molecule retention in slalom chromatography (SC). These equations were applied for the first time to model both the "apparent selectivity" and the resolution between two eluted DNA fragments on a chromatogram. A study of the column efficiency corroborated the fact that slalom chromatography is not based on an adsorption or equilibrium phenomenon, but can be attributed to a hydrodynamic phenomenon. Using a combination of the dynamics of DNA fragment progression in the column and fractal considerations, it was shown that the apparent selectivity depends both on the DNA fragment sizes and mobile-phase flow rate and therefore a balance between two hydrodynamic regimes. A chromatographic response function was also used to obtain the most efficient separation conditions for a mixture of DNA fragments in a minimum analysis time. The chromatographic data confirmed that in SC the flow rate can increase or maintain the separation efficiency with an associated decrease in the analysis time. This constitutes an attractive outcome in relation to the classical chromatographic separation.
Guillaume
Y C
YC
Laboratoire de Chimie Organique, Faculté de Medecine Pharmacie, Besançon, France. yves.guillaume@univ-fcomte.fr
Peyrin
E
E
Thomassin
M
M
Ravel
A
A
Grosset
C
C
Villet
A
A
Robert
J F
JF
Guinchard
C
C
D002845:Chromatography / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D017709:Fractals
D004247:DNA