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Simultaneous separation of nitrofuran antibiotics and their metabolites by using micellar electrokinetic capillary chromatography

Version 2 2024-09-17, 02:08
Version 1 2023-05-17, 06:56
journal contribution
posted on 2024-09-17, 02:08 authored by PU Wickramanayake, TC Tran, JG Hughes, Miroslav MackaMiroslav Macka, N Simpson, PJ Marriott
Mixtures comprising nitrofuran antibiotics (NFA) and nitrofuran metabolites (NFM) were resolved for the first time by using MEKC. Sodium deoxycholate (SDC) was chosen as the micelle-forming surfactant. Optimization of separation conditions was achieved by using a central composite experimental design (CCD) approach. Experimental parameters such as concentration ratio of borate to phosphate in the buffer, pH of the running electrolyte and voltage were investigated. The effect of concentration of the surfactant on resolution was significant. Under optimal conditions of 80 mM SDC, pH 9.0, (20 mM borate + 20 mM phosphate) and 16 kV, the resolution between eight consecutive peak pairs ranged from 1.9 to 11.8. Due to the absence of a UV-active chromophore in the metabolites, they were derivatized with 2-nitrobenzaldehyde (2-NBA). In order to mimic a proposed extraction procedure for the analysis of both NFA and/or derivatized NFM in a sample, aqueous samples (prederivatized with 2-NBA) were extracted by using C-18 SPE cartridges. After washing with H2O, the cartridges were eluted with a small portion of organic solvent with weak elution characteristics to remove excess 2-NBA (hexane was chosen). Target analytes were then recovered with ACN. Excellent reproducibility of migration time (t(mig)) was achieved for all analytes using the developed MECC approach, with absolute t(mig) < 1 % RSD and t(mig) ratio < 0.2% RSD, and peak area ratio was 4% RSD. The LOD for each compound, calculated by extrapolating to an S/N of 3, were found to be 0.19-2.0 mu g/mL.

History

Publication title

Electrophoresis

Volume

27

Issue

20

Pagination

4069-4077

ISSN

0173-0835

Department/School

Chemistry

Publisher

Wiley-V C H Verlag Gmbh

Publication status

  • Published

Place of publication

Po Box 10 11 61, Weinheim, Germany, D-69451

Socio-economic Objectives

280105 Expanding knowledge in the chemical sciences

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