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

Document Type : Original Article

Authors

1 School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran

2 Young Researchers Club, Islamic Azad University, Kermanshah branch, Iran

3 Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, 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.

Keywords

References

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