Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

New Analytical Method for the Determination of Metronidazole in Human Plasma: Application to Bioequivalence Study

Essam Ezzeldin^1,2 , Toqa M El-Nahhas^1,3

¹Drug Bioavailability Center, National Organization for Drug Control and Research (NODCAR), Cairo, Egypt; ²Drug Bioavailability Laboratory, College of Pharmacy, King Saud University, Saudi Arabia; ³Clinical Pharmacology Department, Queen Marry and Westfield College of London, University College of London, EC1M 6BQ.

For correspondence:- Essam Ezzeldin Email: ezzeldin24@hotmail.com Tel:+996597449358

Received: 17 December 2011 Accepted: 6 July 2012 Published: 18 October 2012

Citation: Ezzeldin E, El-Nahhas TM. New Analytical Method for the Determination of Metronidazole in Human Plasma: Application to Bioequivalence Study. Trop J Pharm Res 2012; 11(5):799-805 doi: 10.4314/tjpr.v11i5.14

© 2012 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To develop new sensitive, accurate, rapid and reproducible high performance liquid chromatography (HPLC) method to determine metronidazole levels in human plasma and to apply the method in a bioequivalence study.

Methods: Metronidazole was extracted from human plasma through one step of protein precipitation by methanol using carbamazepine as internal standard (IS). After centrifugation of the plasma sample, the supernatant layer was separated and injected into HPLC system using Eclipse XDB-phenyl column. The mobile phase consisted of phosphate buffer (pH 4.5): acetonitrile (95:5, v/v). The UV detector was set at 320 nm. The bioavailability of the test metronidazole product (Brand A) was compared to a commercial metronidazole brand as reference product in 24 healthy volunteers who received a single dose equivalent to 500 mg of the test and reference products in a randomized balanced two-way cross-over design separated by two-week wash-out period.

Results: Mean standard calibration curves of metronidazole over the concentration range of 0.05 – 30 μg/ml were linear. No significant differences were found based on analysis of variance of the pharmacokinetics parameters required for the assessment of bioequivalence of test and reference formulations. The mean value and 90 %CI of test/reference ratios for the derived parameters were: C_max, 9.64 vs. 8.38 (0.93 – 1.10), AUC_0-24, 124.6 vs.122.3 μg.h/mL (0.973 – 1.051) and AUC_0-∞, 140.9 vs. 128.4 h/mL (1.15 – 1.23).

Conclusion: The test metronidazole product was bioequivalent to the reference. The method is suitable for bioequivalence and pharmacokinetic studies in humans with a low limit of quantification of 0.05 µg/ml.

Keywords: Metronidazole, Bioequivalence, HPLC, Pharmacokinetics, Human plasma