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Journal of Analytical Methods in Chemistry
Volume 2015 (2015), Article ID 892470, 12 pages
http://dx.doi.org/10.1155/2015/892470
Research Article

Development and Validation of an HPLC Method for Simultaneous Quantification of Clopidogrel Bisulfate, Its Carboxylic Acid Metabolite, and Atorvastatin in Human Plasma: Application to a Pharmacokinetic Study

1Faculty of Pharmacy, Department I of Pharmacy, University of Medicine and Pharmacy, Petru Rares Street, 200349 Craiova, Romania
2Faculty of Pharmacy, Doctoral School, University of Medicine and Pharmacy, Petru Rares Street, 200349 Craiova, Romania
3Faculty of Pharmacy, Department II of Pharmacy, University of Medicine and Pharmacy, Petru Rares Street, 200349 Craiova, Romania

Received 19 September 2015; Revised 23 November 2015; Accepted 30 November 2015

Academic Editor: Bengi Uslu

Copyright © 2015 Octavian Croitoru et al. 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 simple, sensitive, and specific reversed phase liquid chromatographic method was developed and validated for simultaneous quantification of clopidogrel, its carboxylic acid metabolite, and atorvastatin in human serum. Plasma samples were deproteinized with acetonitrile and ibuprofen was chosen as internal standard. Chromatographic separation was performed on an BDS Hypersil column (250 × 4.6 mm; 5 μm) via gradient elution with mobile phase consisting of 10 mM phosphoric acid (sodium) buffer solution (pH = 2.6 adjusted with 85% orthophosphoric acid) : acetonitrile : methanol with flow rate of 1 mL·min−1. Detection was achieved with PDA detector at 220 nm. The method was validated in terms of linearity, sensitivity, precision, accuracy, limit of quantification, and stability tests. Calibration curves of the analytes were found to be linear in the range of 0.008–2 μg·mL−1 for clopidogrel, 0.01–4 μg·mL−1 for its carboxylic acid metabolite, and 0.005–2.5 μg·mL−1 for atorvastatin. The results of accuracy (as recovery) with ibuprofen as internal standard were in the range of 96–98% for clopidogrel, 94–98% for its carboxylic acid metabolite, and 90–99% for atorvastatin, respectively.