Peculiarities of the retention mechanism of circular and linear DNA fragments using nonequilibrium chromatography

Citation
Yc. Guillaume et al., Peculiarities of the retention mechanism of circular and linear DNA fragments using nonequilibrium chromatography, J LIQ CHR R, 24(8), 2001, pp. 1061-1072
Citations number
23
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry & Analysis","Spectroscopy /Instrumentation/Analytical Sciences
Journal title
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES
ISSN journal
1082-6076 → ACNP
Volume
24
Issue
8
Year of publication
2001
Pages
1061 - 1072
Database
ISI
SICI code
1082-6076(2001)24:8<1061:POTRMO>2.0.ZU;2-5
Abstract
Two different methods have been used to investigate the retention mechanism of a series of DNA fragments in non equilibrium chromatography (NEC) over a range of column temperatures (T) acid with different mobile phase flow-ra tes (F). The first approach was the separate study of each factor affecting the retention mechanism; the second method was the simultaneous variation of all these factors. The apparent relative retention time of each DNA frag ment was used as a retention marker. The data obtained showed that the retention mechanism of the DNA fragment w as dependent on the F and T values, and also on the DNA form, i.e., linear or circular form. For low F values, the retention mechanism was based on th e classical hydrodynamic regime (HDC) specific to the circular form. On the contrary, for the linear forms. their retention mechanism was based on a s lalom chromatographic process (SC) specifically for high F values at low co lumn temperature. This confirms that the SC and HDC modes are interconnecte d and that the HDC reversible arrow SC transition exists and was clearly vi sualized. In addition, using a simplex process connected to the chemometric methodolo gy, the optimal conditions for F and T were calculated to obtain the more e fficient separation of the linear DNA forms in a minimum analysis time.