Journal of Analytical & Bioanalytical Techniques
Open Access

Abstract

The aim of this study was to optimize and validate a simple and sensitive method for the analysis of atorvastatin acid, ortho - and para -hydroxyatorvastatin and atorvastatin lactone in rat plasma using liquid chromatography-tandem mass spectrometry. Rosuvastatin was used as internal standard. All five analytes were extracted from 500 μl of plasma by protein precipitation with acetonitrile. The chromatographic separation of analytes was achieved using a Shim-pack XR-ODS (100 mm x 3.0 mm, 2.2 μm). The mobile phase consisted of a gradient mixture of 0.1% v/v glacial acetic acid (solvent A) and acetonitrile (solvent B). All analytes were baseline-separated within 5.5 min using a flow rate of 0.4 mL/min. Column temperature was 25°C and samples were kept in autosampler on 4ºC. Mass spectrometry detection was carried out in positive electrospray ionization mode. The calibration curves for all analytes were linear (R2 ≥ 0.9975) over the concentration range of 0.1-30 ng/mL and with lower limit of quantitation of 0.1 ng/mL. Mean extraction recoveries were higher than 75%. Intra- and inter-run mean percent accuracy were between 85-115% and perecent imprecision was ≤15%. Stability studies revealed that atorvastatin acid and its three metabolites were stable in plasma during bench top (8 h on ice), at the end of three successive freeze and thaw cycles and at -20°C for three months. The method was successfully applied on plasma samples obtained from rats that received atorvastatin for 6 weeks to determine concentrations of atorvastatin and its metabolites.

Biography

Crevar Sakac Milkica is a postgraduate student at the Faculty of Pharmacy in Belgrade, Serbia. She works at the Department of Medicinal Chemistry as a teaching assistant in carrying out practical training of three courses of Medicinal Chemistry. She has published four papers in international journals