Simultaneous Determination of Rosuvastatin, Rosuvastatin-5 S-lactone, and N-desmethyl Rosuvastatin in Human Plasma by UPLC-MS/MS and Its Application to Clinical Study

Abstract

Objective A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay was developed and validated for the simultaneous quantification of rosuvastatin (RST), rosuvastatin-5 S-lactone (RSTL), and N-desmethyl rosuvastatin (DM-RST) in human plasma.

Methods Sample was prepared by liquid-liquid extraction with ethyl acetate from 100 μL acidulated buffered plasma. Then analytes were chromatographically separated using an Acquity UPLC HSS T3 column (3.0 mm×100 mm, 1.8 µm) by 0.1% formic acid and gradient acetonitrile at a flow rate of 0.30 mL/min. Three analytes and internal standards (carbamazepine) were eluted in 3.5 min. Mass spectrometry detection was performed through positive ion electrospray ionization (ESI).

Results The calibration curves for three analytes were linear (R≥0.9987, n=3) within the concentration range of 0.1–50 ng/mL for RST and RSTL, and 0.2–100 ng/mL for DM-RST. Mean extraction recoveries were enhanced by means of acidulated plasma using ammonium acetate of pH 4.0, which ranged within 75.3–98.8% for three analytes. Intra- and inter precision and accuracy were 88.2–96.4%.

Conclusions This present method was lower LLOQ, less time consuming (3.5 min), less plasma consuming (100 µL) and simpler sample preparation. And it was successfully applied to determine steady state concentrations of RST, RSTL and DM-RST in a clinical study of RST for patients with coronary artery disease (CAD).

Key words

Rosuvastatin and its metabolites - quantitative analysis - plasma - assaying

• References

• 1 McTaggart F, Buckett L, Davidson R. et al. Preclinical and clinical pharmacology of Rosuvastatin, a new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor. Am J Cardiol 2001; 87: 28B-32B
• 2 Jones A, Wang-Smith L, Pham T. et al. Pharmacokinetics of 3 times a day dosing of oral treprostinil in healthy volunteers. J Cardiovasc Pharmacol 2014; 63: 227-232
• 3 Adams SP, Tsang M, Wright JM. Lipid-lowering efficacy of atorvastatin. Cochrane Database Syst Rev 2015; DOI: 10.1002/14651858.CD008226.pub3. [doi]: CD008226
• 4 Ioannidis JP. More than a billion people taking statins?: Potential implications of the new cardiovascular guidelines. JAMA 2014; 311: 463-464
• 5 White CM. A review of the pharmacologic and pharmacokinetic aspects of rosuvastatin. J Clin Pharmacol 2002; 42: 963-970
• 6 Birmingham BK, Bujac SR, Elsby R. et al. Rosuvastatin pharmacokinetics and pharmacogenetics in Caucasian and Asian subjects residing in the United States. Eur J Clin Pharmacol 2015; 71: 329-340
• 7 Lee E, Ryan S, Birmingham B. et al. Rosuvastatin pharmacokinetics and pharmacogenetics in white and Asian subjects residing in the same environment. Clin Pharmacol Ther 2005; 78: 330-341
• 8 Ganga HV, Slim HB, Thompson PD. A systematic review of statin-induced muscle problems in clinical trials. Am Heart J 2014; 168: 6-15
• 9 Hansen KE, Hildebrand JP, Ferguson EE. et al. Outcomes in 45 patients with statin-associated myopathy. Arch Intern Med 2005; 165: 2671-2676
• 10 Echaniz-Laguna A, Mohr M, Tranchant C. Neuromuscular symptoms and elevated creatine kinase after statin withdrawal. N Engl J Med 2010; 362: 564-565
• 11 Hermann M, Bogsrud MP, Molden E. et al. Exposure of atorvastatin is unchanged but lactone and acid metabolites are increased several-fold in patients with atorvastatin-induced myopathy. Clin Pharmacol Ther 2006; 79: 532-539
• 12 Skottheim IB, Gedde-Dahl A, Hejazifar S. et al. Statin induced myotoxicity: The lactone forms are more potent than the acid forms in human skeletal muscle cells in vitro. Eur J Pharm Sci 2008; 33: 317-325
• 13 Li XN, Xu HR, Chen WL. et al. Pharmacokinetics of rosuvastatin in healthy Chinese volunteers living in China: A randomized, open-label, ascending single- and multiple-dose study. Clin Ther 2010; 32: 575-587
• 14 Macwan JS, Ionita IA, Akhlaghi F. A simple assay for the simultaneous determination of rosuvastatin acid, rosuvastatin-5 S-lactone, and N-desmethyl rosuvastatin in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Anal Bioanal Chem 2012; 402: 1217-1227
• 15 Yang DJ, Hwang LS. Study on the conversion of three natural statins from lactone forms to their corresponding hydroxy acid forms and their determination in Pu-Erh tea. J Chromatogr A 2006; 1119: 277-284
• 16 Jemal M, Ouyang Z, Chen BC. et al. Quantitation of the acid and lactone forms of atorvastatin and its biotransformation products in human serum by high-performance liquid chromatography with electrospray tandem mass spectrometry. Rapid Commun Mass Spectrom 1999; 13: 1003-1015
• 17 Baek IH, Kwon KI, Kim MS. Effect of food on the pharmacokinetics of rosuvastatin after administration of a single oral dose in dogs. Drug Res (Stuttg) 2013; 63: 145-149
• 18 Hull CK, Penman AD, Smith CK. et al. Quantification of rosuvastatin in human plasma by automated solid-phase extraction using tandem mass spectrometric detection. J Chromatogr B Analyt Technol Biomed Life Sci 2002; 772: 219-228
• 19 Huguet J, Lu J, Gaudette F. et al. No effects of pantoprazole on the pharmacokinetics of rosuvastatin in healthy subjects. Eur J Clin Pharmacol 2016; 72: 925-931