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DOI: 10.1055/s-0031-1296671
Development and Validation of a Highly Sensitive and Robust LC-MS/MS with Electrospray Ionization Method for Quantification of Rosuvastatin in Small Volume Human Plasma Samples and its Application to a Clinical Study
Publication History
Publication Date:
21 December 2011 (online)
Abstract
A high-throughput, simple, highly sensitive and specific LC-MS/MS method (liquid chromatography coupled with tandem mass spectrometry) has been developed for the estimation of rosuvastatin (CAS 287714-41-4, RST) with 100 µl human plasma using atorvastatin (CAS 134523-00-5) as an internal standard (IS). The API-4000 LC-MS/MS was operated under the multiple reaction-monitoring mode (MRM) using the electro spray ion-ization technique. The assay procedure involved direct precipitation of RST and IS from plasma with acetonitrile. Sample preparation with this method yielded clean extracts and consistent recoveries: 91.39 % for RST and 99.28 % for IS. The total chromatographic run time was 3.5 min and the elution of RST and IS occurred at 2.5 and 3.1 min, respectively; this was achieved with a mobile phase consisting of 0.05 mol/L formic acid: acetonitrile (20:80, v/v) at a flow rate of 0.50 ml/min on an Inertsil ODS-3 column (4.6 × 100 mm, 3.0 µm). The developed method was validated in human plasma with a limit of quantitation of 0.05 ng/ml.
A linear response function was established for the range of concentrations of 0.05 to 50.0 ng/ml with a correlation coefficient (r) of 0.999. The inter- and intra-day precision in the measurement of RST quality control (QC) samples at 0.05, 0.15, 25 and 40 ng/ml were in the range of 6.55 to 11.40 % relative standard deviation (RSD) and 1.76 to 11.17 % RSD, respectively. Accuracy in the measurement of QC samples for RST was in the range of 95.02 to 101.37 % of the nominal values. RST was stable in the battery of stability studies viz., bench-top, auto-sampler and freeze-thaw cycles. The stability of RST was established for 1 month at –80 °C. The application of the assay to a clinical study confirmed the utility of the assay to derive human pharmacokinetic parameters.