A Sensitive Liquid Chromatographic Method for the Analysis of Clarithromycin with Pre-Column Derivatization: Application to a Bioequivalence Study

Authors

  • Amir Farshchi School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran|Young Researchers Club, Islamic Azad University, Kermanshah branch, Iran
  • Gholamreza Bahrami School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran|Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
  • Golbarg Ghiasi School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran|Young Researchers Club, Islamic Azad University, Kermanshah branch, Iran
Abstract:

Objective(s) A sensitive liquid chromatographic method for the analysis of clarithromycin- a macrolide antibiotic- in human serum, using pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) is described. Materials and Methods The method involved liquid-liquid extraction of the drug and an internal standard (amantadine) followed by pre-column derivatization of the analytes with FMOC-Cl. A mixture of 0.05 M phosphate buffer containing triethylamine (2 ml/l; pH 3.8) and methanol (17:83, v/v) was used as mobile phase and chromatographic separation was achieved on a Shimpack CLC-ODS column. The eluate was monitored by a fluorescence detector with respective excitation and emission wavelengths of 265 and 315 nm. Results The analytical method was linear over the concentration range of 0.025-10 pg/ml of clarithromycin in human serum with a limit of quantification of 0.025 pg/ml. The assay is sensitive enough to measure drug levels obtained in human single dose studies. Conclusion In the present method, sensitivity and the running time of analysis have been improved and successfully applied in a bioequivalence study of three different clarithromycin preparations in 12 healthy volunteers.

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Journal title

volume 12  issue 1

pages  25- 32

publication date 2009-01-01

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