Alternative Approaches for the Estimation of the Band Broadening Parameters in Single-Detection Size Exclusion Chromatography

• Autores: Vega, Jorge Ruben; Schnöll Bitai, Irene
• Año de publicación: 2005
• Idioma: inglés
• Tipo de recurso: artículo
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

New approaches for the determination of the extent of symmetric and asymmetric band broadening (BB) in size exclusion chromatography (SEC) are presented. For this purpose raw data was simulated by starting with either a theoretical Poisson number chain length distribution (NCLD), or a log-normal weight chain length distribution (WCLD). Each distribution was first converted to a BB-free mass chromatogram, as typically obtained from a standard differential refractive index detector. Then, the broadened (or “measured”) chromatograms were simulated by convoluting the BB-free chromatograms with a BB function, which was assumed to follow symmetrical (Gauss) as well as unsymmetrical (exponentially modified Gauss) function. A broad range of BB parameters (standard deviation, sBB, and exponential decay, tBB) was used for the simulations. The approaches are based on the determination of the points of inflection belonging to the peak of the broadened chromatogram, and closed as well as empirically derived equations connecting the peak width, its variance, and the parameters sBB and tBB. The developed methods are applicable for Poisson distributions well above a peak chain length of 100.
Fil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Schnöll Bitai, Irene. University of Vienna; Austria