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High-Resolution Asymmetrical Flow Field-Flow Fractionation Data Evaluation via Richardson–Lucy-Based Fractogram Correction

High-Resolution Asymmetrical Flow Field-Flow Fractionation Data Evaluation via Richardson–Lucy-Based Fractogram Correction

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SCHMID, Marius, Benedikt HÄUSELE, Michael JUNK, Emre BROOKES, Jürgen FRANK, Helmut CÖLFEN, 2018. High-Resolution Asymmetrical Flow Field-Flow Fractionation Data Evaluation via Richardson–Lucy-Based Fractogram Correction. In: Analytical Chemistry. 90(23), pp. 13978-13986. ISSN 0003-2700. eISSN 1520-6882. Available under: doi: 10.1021/acs.analchem.8b03483

@article{Schmid2018-11-13HighR-44270, title={High-Resolution Asymmetrical Flow Field-Flow Fractionation Data Evaluation via Richardson–Lucy-Based Fractogram Correction}, year={2018}, doi={10.1021/acs.analchem.8b03483}, number={23}, volume={90}, issn={0003-2700}, journal={Analytical Chemistry}, pages={13978--13986}, author={Schmid, Marius and Häusele, Benedikt and Junk, Michael and Brookes, Emre and Frank, Jürgen and Cölfen, Helmut} }

Junk, Michael 2018-12-11T13:16:04Z 2018-11-13 Cölfen, Helmut Cölfen, Helmut Schmid, Marius Junk, Michael Schmid, Marius High-Resolution Asymmetrical Flow Field-Flow Fractionation Data Evaluation via Richardson–Lucy-Based Fractogram Correction Asymmetrical flow field-flow fractionation (AF4) is a chromatographic separation technique that can be used for a broad range of particles or macromolecules. As an orthogonal method to size exclusion chromatography (SEC) with a much broader separation size range (1–800 nm) AF4 is gaining importance. However, the data evaluation capacities are far behind in comparison to other techniques like analytical ultracentrifugation (AUC). A program for evaluation of data from AF4 with a coupled multiangle laser light scattering (MALLS) detector was developed that allows the determination of the distributions of diffusion coefficients (D), hydrodynamic radii (R<sub>h</sub>), molecular weights (M<sub>w</sub>), and relative concentrations (RC) of the obtained species. In addition, two algorithms to remove broadening effects via deconvolution were implemented and tested for their validity. The first is an extension of the known diffusion broadening correction applying the entire diffusion coefficient distribution instead of a single diffusion coefficient. The second applies the Richardson–Lucy algorithm for the deconvolution of overlapping signals from stars in astronomy. This program allows a reproducible strong enhancement of the fractogram resolution allowing for entire baseline separations of proteins. The comparison of the values for M<sub>w</sub> determined by a partial Zimm plot from each data point of the original fractogram and the deconvolved results shows that especially the Richardson–Lucy algorithm maintains a high degree of data robustness. 2018-12-11T13:16:04Z Frank, Jürgen Frank, Jürgen Brookes, Emre eng Häusele, Benedikt Häusele, Benedikt Brookes, Emre

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