Simultaneous determination of atorvastatin and niacin in human plasma by LC‐MS/MS and its application to a human pharmacokinetic study

Abstract

A rapid, simple, sensitive and selective LC-MS/MS method has been developed and validated for quantification of the atorvastatin (AT) and niacin (NA) in 250 μL human plasma. The analytical procedure involves a liquid-liquid extraction method using nevirapine as an internal standard (IS). The chromatographic separation was achieved on a Hypurity Advance (4.6 × 50 mm, 5 µm) column using a mobile phase consisting of 0.1% formic acid buffer-acetonitrile (20:80, v/v) at flow rate of 0.8 mL/min. The API-4000 LC-MS/MS was operated in the multiple-reaction monitoring mode using electrospray ionization. The total run time of analysis was 3 min and elution of AT, NA and IS occurred at 1.06, 1.84 and 0.92 min, respectively. A detailed validation of the method was performed as per the US Food and Drug Administration guidelines and the standard curves found to be linear in the range of 0.10-30.0 ng/mL for AT and 20.2-6026 ng/mL for NA, with a coefficient of correlation of ≥ 0.99 for both the compounds. AT and NA were found to be stable in a battery of stability studies, viz. bench-top, autosampler, re-injection, wet-extract and repeated freeze-thaw cycles. The developed assay method was successfully applied to a pharmacokinetic study in humans.