Validated HPLC–MS/MS method for simultaneous determination of simvastatin and simvastatin hydroxy acid in human plasma

Cholesterol lowering statin drugs are the most frequently prescribed agents for reducing morbidity and mortality related to coronary heart disease. This publication presents a validated, highly sensitive and selective isocratic HPLC method for the quantitative determination of the major statin drug simvastatin (SIM) and its metabolite simvastatin hydroxy acid (SIMA). Detection was performed on an electrospray ionization triple quadrupole mass spectrometer equipped with an ESI interface operated in positive and negative ionization mode. The multiple reaction-monitoring mode (MRM) was used to provide MS/MS detection. The linearity for the calibration curve in the concentration range of 0.10–16.00 ng/mL for SIM and 0.10–16.00 ng/mL for SIMA is presented. Inter- and intra-day precision and accuracy of the proposed method were characterized by relative standard deviation (R.S.D.) and percentage deviation, respectively; with both lower than 7% for all analytes. The limit of quantitation was 0.03 ng/mL for SIM and 0.02 ng/mL for SIMA. The devised method was employed in the pharmacokinetic study of SIM and the pharmacokinetic parameters of all analytes are also presented.