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E-Journal of Chemistry
Volume 7, S1, Pages S363-S371
http://dx.doi.org/10.1155/2010/235186

UPLC Method for Simultaneous Separation and Estimation of Secnidazole, Fluconazole and Azithromycin in Pharmaceutical Dosage Forms

Ramesh Yanamandra,1 Avinash Chaudhary,1 Srinivasa Rao Bandaru,1 Balaram Patro,1 Y. L. N. Murthy,2 Parimi Atchuta Ramaiah,2 and C. S. P. Sastry3

1Analytical Development LaboratoryGVK Biosciences Private Limited No.28A, Street No.15IDA, Nacharam, Hyderabad-500 076, India
2Department of Organic Chemistry Food, Drugs and Water, A.U College of Science and Technology Andhra University, Visakhapatnam-530 003, India
3Prof (Retd.), Department of Organic Chemistry Food, Drugs and Water, A.U College of Science and Technology Andhra University, Visakhapatnam-530 003, India

Received 18 April 2010; Accepted 10 June 2010

Copyright © 2010 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A novel approach was carried out to develop and validate a rapid, specific, accurate and precise reverse phase ultra performance liquid chromatographic (UPLC) method for the simultaneous separation and quantification of secnidazole, fluconazole and azithromycin in pharmaceutical dosage forms. The developed analytical method is superior in technology to conventional HPLC with respect to time, resolution, solvent consumption and cost of analysis. Elution time for the separation was 10 min and ultra violet detection was carried out at 210 nm. Efficient separation was achieved on BEH C18 sub-2-μm UPLC column using 0.002 M Na2HPO4 and acetonitrile as organic solvent in a gradient program. Benzophenone was used as internal standard. Resolutions between secnidazole, fluconazole and azithromycin were found to be more than 4.8. The calibration graphs were linear for secnidazole, fluconazole, benzophenone and azithromycin. The method showed excellent recoveries for all dosage forms. The test solution was found to be stable in diluent for 72 h when stored in the refrigerator between 2 to 8 °C. The proposed UPLC method was validated with respect to linearity, accuracy, precision, specificity and robustness and can be used for the simultaneous estimation of secnidazole, fluconazole and azithromycin in tablet dosage forms available as a combi kit.