Simultaneous Determination of Doxycycline and Chlortetracycline in Real Samples by Europium-Sensitized Luminescence

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A simple luminescent methodology for the simultaneous determination of doxycycline and chlortetracycline in pharmaceutical preparations and human urine is proposed. Since the native fluorescence of both analytes is negligible, this method takes advantage of the lanthanide-sensitized luminescence, which provides increased sensitivity. Due to the strong overlapping between the luminescence spectra of both europium complexes, the use of luminescence decay curves to resolve mixtures of the analytes is proposed, particularly as these curves are more selective. A factorial design, with three levels per factor, coupled to a central composite design was selected to obtain a calibration matrix of 13 standards plus one blank sample, which were processed with a partial least-squares analysis. In order to assess the effectiveness of the proposed method, a prediction set of 10 synthetic samples was analyzed, and recovery percentages between 95 and 104% were obtained. Limits of detection, calculated by means of a new criterion, were 3.27 and 1.06 μg L−1 for doxycycline and chlortetracycline, respectively. The method was tested in three different pharmaceutical preparations containing the analytes, with average recovery percentages of 99.4 ± 1.8 for doxycycline and 100.5 ± 2.1 for chlortetracycline. Moreover, a central composite design was also developed to obtain a calibration matrix that made feasible the simultaneous determination of both tetracyclines in human urine samples. In this case, average recovery percentages were 98.0 ± 4.4 and 97.8 ± 4.6 for doxycycline and chlortetracycline, respectively. No extraction method or prior separation of the analytes was needed.

Keywords: Chlortetracycline; Decay curves; Doxycycline; Europium-sensitized luminescence; Human urine; Pharmaceutical preparations

Document Type: Research Article


Affiliations: Department of Analytical Chemistry and Foods Technology, University of Castilla-La Mancha, 13071 Ciudad Real, Spain

Publication date: April 1, 2013

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