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Fractionating power and outlet stream polydispersity in asymmetrical flow field-flow fractionation. Part I: isocratic operation

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Abstract

Asymmetrical flow field-flow fractionation (As-FlFFF) has become the most commonly used of the field-flow fractionation techniques. However, because of the interdependence of the channel flow and the cross flow through the accumulation wall, it is the most difficult of the techniques to optimize, particularly for programmed cross flow operation. For the analysis of polydisperse samples, the optimization should ideally be guided by the predicted fractionating power. Many experimentalists, however, neglect fractionating power and rely on light scattering detection simply to confirm apparent selectivity across the breadth of the eluted peak. The size information returned by the light scattering software is assumed to dispense with any reliance on theory to predict retention, and any departure of theoretical predictions from experimental observations is therefore considered of no importance. Separation depends on efficiency as well as selectivity, however, and efficiency can be a strong function of retention. The fractionation of a polydisperse sample by field-flow fractionation never provides a perfectly separated series of monodisperse fractions at the channel outlet. The outlet stream has some residual polydispersity, and it will be shown in this manuscript that the residual polydispersity is inversely related to the fractionating power. Due to the strong dependence of light scattering intensity and its angular distribution on the size of the scattering species, the outlet polydispersity must be minimized if reliable size data are to be obtained from the light scattering detector signal. It is shown that light scattering detection should be used with careful control of fractionating power to obtain optimized analysis of polydisperse samples. Part I is concerned with isocratic operation of As-FlFFF, and part II with programmed operation.

The dash-dotted blue curve describes an assumed log-normal sample molecular weight distribution (right axis). The full black curves show predicted outlet polydispersity (left axis) at discrete elution times of 4.5, 6.0, 7.5, 9.0, and 10.5 minutes during an isocratic As-FlFFF analysis

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    P. Stephen Williams

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Williams, P.S. Fractionating power and outlet stream polydispersity in asymmetrical flow field-flow fractionation. Part I: isocratic operation. Anal Bioanal Chem 408, 3247–3263 (2016). https://doi.org/10.1007/s00216-016-9388-0

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