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

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 desig...

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Bibliographic Details
Published in:Electrophoresis 2006-10, Vol.27 (20), p.4069-4077
Main Authors: Wickramanayake, Priyanga U., Tran, Tin C., Hughes, Jeff G., Macka, Mirek, Simpson, Nigel, Marriott, Philip J.
Format: Article
Language:English
Subjects:
Animals
Chromatographic exponential function
Chromatography, Micellar Electrokinetic Capillary - methods
Deoxycholic Acid
Derivatization
Hydrogen-Ion Concentration
Micellar electrokinetic chromatography
Nitrofuran antibiotics
Nitrofuran metabolites
Nitrofurans - isolation & purification
Nitrofurans - metabolism
Penaeidae - chemistry
Reproducibility of Results
Surface-Active Agents
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Summary: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 C18 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 (tmig) was achieved for all analytes using the developed MECC approach, with absolute tmig 
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.200600105