RAPID AND SENSITIVE HPLC METHOD FOR THE DETERMINATION OF SIROLIMUS WITH KETOCONAZOLE AS INTERNAL STANDARD AND ITS FURTHER APPLICATIONS
Abstract
Sirolimus and Ketoconazole are used in organ transplantation regimen and potential metabolic interactions of these drugs were reported when administered concomitantly. An analytical method based on high-performance liquid chromatography (HPLC) with photo diode array (PDA) detection was developed for quantification of sirolimus using ketoconazole as internal standard. Extraction was performed using dichloromethane under nitrogen atmosphere and the separation of sirolimus and ketoconazole was accomplished by reverse phase chromatography. The mobile phase consists of a combination of methanol, water and glacial acetic acid at 90:10:0.1% ratios run isocratically through a C18 (250mm × 4.6mm, 5µm) reverse phase analytical column. The PDA detection was done at 278nm with analytical run time less than 6 min. The average mean recovery was found to be 98.3% for 1, 3, 5, 10µg/ml concentrations. The assay exhibited good linear relationship. LLOQ was 10ng/ml with 0.84% and 1.28% of accuracy and precision over the concentration range of 0.1-10µg/ml. The method can be successfully applied for estimation of sirolimus from in-vitro elution studies of sirolimus eluting stents, simultaneous estimation of ketoconazole and sirolimus in therapeutic drug monitoring and other pharmacokinetic studies.
Keywords:
HPLC, Ketoconazole, Sirolimus, Therapeutic drug monitoringDOI
https://doi.org/10.25004/IJPSDR.2012.040112References
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