Estimation of band broadening in size-exclusion chromatography. I. A method based on analyzing narrow standards with a molar mass-sensitive detector.
A method is proposed for estimating the (asymmetrical and non-uniform) band broadening function (BBF) in size-exclusion chromatography (SEC). The following data are required: the molar mass calibration and the concentration- and molar mass chromatograms of a set of narrow standards. In the narrow range of each standard, the BBF is uniform but skewed. Each uniform BBF is estimated through a nonlinear optimization procedure that compares one (of the two) measured chromatograms with its theoretical prediction based on the other chromatogram. The method is validated with numerical examples that simulate the analyses of narrow standards exhibiting log-normal and Poisson weight chain length distributions. The BBF can be assumed of arbitrary shape, or represented by an exponentially-modified Gaussian (EMG). From the uniform BBF estimate, the true polydispersity of the standard can be determined. The global non-uniform BBF is obtained by interpolation between a set of uniform BBFs covering a wide range of elution volumes.
Yossen
Mariana M
MM
INTEC (Universidad Nacional del Litoral and CONICET), Güemes 3450, 3000 Santa Fe, Argentina.
Vega
Jorge R
JR
Meira
Gregorio R
GR
D000465:Algorithms; D002850:Chromatography, Gel / Q000379:methods*; D008962:Models, Theoretical; D008970:Molecular Weight; D016012:Poisson Distribution; D012015:Reference Standards