1. LC-ESI-MS/MS method for the quantification of entecavir in human plasma and its application to bioequivalence study.
- Author
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Challa BR, Awen BZ, Chandu BR, and Rihanaparveen S
- Subjects
- Analysis of Variance, Antiviral Agents blood, Antiviral Agents pharmacokinetics, Area Under Curve, Drug Stability, Guanine blood, Guanine pharmacokinetics, Humans, Lamivudine analysis, Linear Models, Reproducibility of Results, Sensitivity and Specificity, Solid Phase Extraction, Therapeutic Equivalency, Chromatography, Liquid methods, Guanine analogs & derivatives, Spectrometry, Mass, Electrospray Ionization methods, Tandem Mass Spectrometry methods
- Abstract
Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) was used for a quantitative estimation of entecavir (EV) in human plasma using lamivudine (LM) as internal standard (IS). The method herein described is simple, sensitive, and specific. Chromatographic separation was performed on XBridge-C18, 4.6 mm × 50 mm, 5-μm column with an isocratic mobile phase composed of 10 mM ammonium hydrogen carbonate (pH 10.5):methanol (85:15 v/v), pumped at 0.3 ml/min. EV and LM were detected using proton adducts at m/z 278.1→152.1 and 230.2→112.0 in multiple reaction monitoring (MRM) positive mode. Solid phase extraction method was employed in the extraction of EV and LM from the biological matrix. This method was validated over a linear concentration range of 50.0-20000.0 pg/ml with a correlation coefficient (r) ≥0.9983. Intra and inter-day precision of EV was found within the range of 1.2-4.2 for EV and 4.4-4.5 for LM. EV was stable throughout three freeze/thaw cycles, bench top and postoperative studies. This method was successfully used in the analysis of plasma samples following oral administration of EV (0.5 mg) in 26 healthy human volunteers., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2011
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