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Assessment of the binding performance of histamine-imprinted microspheres by frontal analysis capillary electrophoresis
journal contribution
posted on 2023-05-19, 14:42 authored by Romano, EF, Joselito Quirino, Holdsworth, JL, So, RC, Holdsworth, CIFrontal analysis capillary electrophoresis was used to evaluate the binding performance of molecularly imprinted microspheres (MIM) toward its template histamine and analogs at pH 7, and compared to the high performance liquid chromatographic method. In both methods, batch binding was employed and the binding parameters were calculated from the measured concentration of unbound amine analytes and afforded comparable histamine equilibrium dissociation constants (Kd ∼ 0.4 mM). FACE was easily carried out at shorter binding equilibration time (i.e. 30 min) and without the need to separate the microspheres, circumventing laborious and, in the case of the system under study, inefficient sample filtration. It also allowed for competitive binding studies by virtue of its ability to distinctly separate intact microspheres and all tested amines which could not be resolved in HPLC. Kd’s for nonimprinted (control) microspheres (NIM) from FACE and HPLC were also comparable (∼ 0.6 mM) but at higher histamine concentrations, HPLC gave lower histamine binding. This discrepancy was attributed to inefficient filtration of the batch binding samples prior to HPLC analysis resulting in an over-estimation of the concentration of free histamine brought about by the presence of unfiltered histamine-bound microspheres.
History
Publication title
ElectrophoresisVolume
38Issue
9-10Pagination
1251-1259ISSN
0173-0835Department/School
School of Natural SciencesPublisher
Wiley-V C H Verlag GmbhPlace of publication
Po Box 10 11 61, Weinheim, Germany, D-69451Rights statement
Copyright 2017 WILEY-VCH Verlag GmbH & Co. KGaA, WeinheimRepository Status
- Restricted