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Asymmetric flow field flow fractionation with light scattering detection - an orthogonal sensitivity analysis



Anne A. Galyean, James J. Filliben, Richard D. Holbrook, Wyatt N. Vreeland, Howard S. Weinberg


Asymmetric flow field flow fractionation (AF4) has several instrumental factors that may have a direct effect on separation performance. A sensitivity analysis was applied to ascertain the relative importance of AF4 primary instrument factor settings for the separation of a complex environmental sample. The analysis evaluated the impact of instrumental factors namely, cross flow, ramp time, focus flow, injection volume, and run buffer concentration on the multi-angle light scattering measurement of natural organic matter (NOM) molar mass (MM). A 2(5-1) orthogonal fractional factorial design was used to minimize analysis time while preserving the accuracy and robustness in the determination of the main effects and interactions between any two instrumental factors. By assuming that separations resulting in smaller MM measurements would be more accurate, the analysis produced a ranked list of effects estimates for factors and interactions of factors based on their relative importance in minimizing the MM. The most important and statistically significant AF4 instrumental factors were buffer concentration and cross flow. The least important was ramp time. A parallel 2(5- 2) orthogonal fractional factorial design was also employed on five environmental factors for synthetic natural water samples containing silver nanoparticles (NPs), namely: NP concentration, NP size, NOM concentration, specific conductance, and pH. None of the water quality characteristic effects or interactions were found to be significant in minimizing the measured MM; however, the interaction between NP concentration and NP size was an important effect when considering NOM recovery. This work presents a structured approach for the rigorous assessment of AF4 instrument factors and optimal settings for the separation of complex samples utilizing efficient orthogonal factional factorial design and appropriate graphical analysis.
Journal of Chromatography A


Asymmetric flow field flow fractionation, Engineering nanoparticles, Environmental matrix, Light scattering, Orthogonal fractional factorial design, Natural organic matter


Galyean, A. , Filliben, J. , Holbrook, R. , Vreeland, W. and Weinberg, H. (2016), Asymmetric flow field flow fractionation with light scattering detection - an orthogonal sensitivity analysis, Journal of Chromatography A, [online], (Accessed May 24, 2022)
Created November 17, 2016, Updated August 10, 2020