Journal of Pharmaceutical Analysis (Feb 2014)

Simultaneous quantitation of atorvastatin and its two active metabolites in human plasma by liquid chromatography/(–) electrospray tandem mass spectrometry

  • Pankaj Partani,
  • S. Manaswita Verma,
  • Sanjay Gurule,
  • Arshad Khuroo,
  • Tausif Monif

DOI
https://doi.org/10.1016/j.jpha.2013.09.007
Journal volume & issue
Vol. 4, no. 1
pp. 26 – 36

Abstract

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A sensitive, accurate and selective liquid chromatography–tandem mass spectrometry method (LC–MS/MS) was developed and validated for the simultaneous quantitation of atorvastatin (AT) and its equipotent hydroxyl metabolites, 2-hydroxy atorvastatin (2-AT) and 4-hydroxy atorvastatin (4-AT), in human plasma. Electrospray ionization (ESI) interface in negative ion mode was selected to improve the selectivity and the sensitivity required for this application. Additionally, a solid phase extraction (SPE) step was performed to reduce any ion-suppression and/or enhancement effects. The separation of all compounds was achieved in less than 6 min using a C18 reverse-phase fused-core® column and a mobile phase, composed of a mixture of 0.005% formic acid in water:acetonitrile:methanol (35:25:40, v/v/v), in isocratic mode at a flow rate of 0.6 mL/min. The method has lower limit of quantitation (LLOQ) of 0.050 ng/mL for all analytes. The method has shown tremendous reproducibility, with intra- and inter-day precision less than 6.6%, and intra- and inter-day accuracy within ±4.3% of nominal values, for all analytes, and has proved to be highly reliable for the analysis of clinical samples. Keywords: Atorvastatin, LC–MS/MS, Solid phase extraction, Pharmacokinetics, Method validation