Stability Indicating Liquid Chromatographic Method for the Simultaneous Determination of Rosuvastatin and Ezetimibe from Pharmaceuticals and Biological Samples

Abstract

Fixed dose combinations of ezetimibe and rosuvastatin are well known to provide significantly superior efficacy to rosuvastatin alone in lowering total cholesterol, and triglyceride levels. Therefore, in this recent work, we prepared fixed dose (10mg/10mg) ezetimibe and rosuvastatin tablet formulations, and a simple, selective and we established and validated rapid chromatographic method for the sensitive and simultaneous determination of rosuvastatin and ezetimibe. We performed the separation on Zorbax SB-C18 (100 mm x 4.6 mm, 3.5 µm particle size) column at 30 oC with a mobile phase that has a composition as water containing 0.1% H3PO4:methanol:acetonitrile (50:25:25 v/v/v) at 1.2 mL/min flow rate. We further performed validation studies according to ICH guidelines. Furthermore, we exposed rosuvastatin and ezetimibe to degradation conditions. We plotted peak area vs. concentration graphs for rosuvastatin and ezetimibe and used for defining the linearity of the detector response. In this suggested method, we observed linear relationships varying from 0.05 to 50 µg/mL for ezetimibe concentrations and from 0.05 to 25 µg/mL for rosuvastatin concentrations. We found the limit of detection values for rosuvastatin and ezetimibe as 0.006 and 0.008 µg/mL, respectively. Moreover, we further used the established method for to the analysis of ezetimibe and rosuvastatin from real spiked samples of rabbit serum.

Keywords

• Rosuvastatin
• Ezetimibe
• Liquid Chromatography
• Validation
• Stability Indicating Methods

References

1. 1. Schultheis A. Hypercholesterolemia: prevention, detection and management. Nurse Pract. 1990;15:40–6.
2. 2. Ross R. The Pathogenesis of Atherosclerosis — An Update. Vol. 314, New England Journal of Medicine. 1986. p. 488–500.
3. 3. Ornish D, Brown SE, Billings JH, Scherwitz LW, Armstrong WT, Ports TA, et al. Can lifestyle changes reverse coronary heart disease?. The Lifestyle Heart Trial. Lancet. 1990 Jul 21;336(8708):129–33.
4. 4. Ulbricht TLV, Southgate DAT. Coronary heart disease: seven dietary factors. Lancet. 1991 Oct 19;338(8773):985–92.
5. 5. Chapman MJ, McTaggart F. Optimizing the pharmacology of statins: Characteristics of rosuvastatin. In: Atherosclerosis Supplements. 2002. p. 33–7.
6. 6. Kosoglou T, Statkevich P, Yang B, Suresh R, Zhu Y, Boutros T, et al. Pharmacodynamic interaction between ezetimibe and rosuvastatin. Curr Med Res Opin. 2004;20(8):1185–95.
7. 7. Gagné C, Bays HE, Weiss SR, Mata P, Quinto K, Melino M, et al. Efficacy and safety of ezetimibe added to ongoing statin therapy for treatment of patients with primary hypercholesterolemia. Am J Cardiol. 2002;90(10):1084–91.
8. 8. Bays HE, Moore PB, Drehobl MA, Rosenblatt S, Toth PD, Dujovne CA, et al. Effectiveness and tolerability of ezetimibe in patients with primary hypercholesterolemia: pooled analysis of two phase II studies. Clin Ther. 2001;23(8):1209–30.

9. 9. Nutescu EA, Shapiro NL. Ezetimibe: A Selective Cholesterol Absorption Inhibitor. Pharmacotherapy. 2003;23(11):1463–74.
10. 10. McTaggart F. Comparative pharmacology of rosuvastatin. In: Atherosclerosis Supplements. Elsevier Ireland Ltd; 2003. p. 9–14.
11. 11. McTaggart F, Buckett L, Davidson R, Holdgate G, McCormick A, Schneck D, et al. Preclinical and clinical pharmacology of rosuvastatin, a new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor. In: American Journal of Cardiology. Elsevier Inc.; 2001. p. 28–32.
12. 12. Stein EA. Management of hypercholesterolemia. Approach to diet and drug therapy. Am J Med. 1989;87(SUPPL. 4).
13. 13. Davidson MH. Rosuvastatin: A highly efficacious statin for the treatment of dyslipidaemia. Expert Opin Investig Drugs. 2002;11(1):125–32.
14. 14. Shah Y, Iqbal Z, Ahmad L, Khan A, Khan MI, Nazir S, et al. Simultaneous determination of rosuvastatin and atorvastatin in human serum using RP-HPLC/UV detection: Method development, validation and optimization of various experimental parameters. J Chromatogr B Anal Technol Biomed Life Sci. 2011 Mar 15;879(9–10):557–63.
15. 15. Nasir F, Iqbal Z, Khan A, Ahmad L, Shah Y, Khan AZ, et al. Simultaneous determination of timolol maleate, rosuvastatin calcium and diclofenac sodium in pharmaceuticals and physiological fluids using HPLC-UV. J Chromatogr B Anal Technol Biomed Life Sci. 2011;879(30):3434–43.
16. 16. Gomes FP, Garcia PL, Porto Alves JM, Singh AK, Kedor-Hackmann ERM, Santoro MIRM. Development and validation of stability-indicating hplc methods for quantitative determination of pravastatin, fluvastatin, atorvastatin, and rosuvastatin in pharmaceuticals. Anal Lett. 2009;42(12):1784–804.
17. 17. Sultana N, Arayne MS, Shah SN, Shafi N, Naveed S. Simultaneous determination of prazosin, atorvastatin, rosuvastatin and simvastatin in API, dosage formulations and human serum by RP-HPLC. J Chinese Chem Soc. 2010;57(6):1286–92.
18. 18. Kawashiri MA, Nohara A, Noguchi T, Tada H, Nakanishi C, Mori M, et al. Efficacy and safety of coadministration of rosuvastatin, ezetimibe, and colestimide in heterozygous familial hypercholesterolemia. Am J Cardiol. 2012;109(3):364–9.
19. 19. Ballantyne CM, Weiss R, Moccetti T, Vogt A, Eber B, Sosef F, et al. Efficacy and Safety of Rosuvastatin 40 mg Alone or in Combination With Ezetimibe in Patients at High Risk of Cardiovascular Disease (Results from the EXPLORER Study). Am J Cardiol. 2007;99(5):673–80.
20. 20. Yang YJ, Lee SH, Kim BS, Cho YK, Cho HJ, Cho KI, et al. Combination Therapy of Rosuvastatin and Ezetimibe in Patients with High Cardiovascular Risk. Clin Ther. 2017 Jan 1;39(1):107–17.
21. 21. Swartz ME, Murphy B. New frontiers in chromatography. Am Lab. 2005;37(3):22–7.
22. 22. Borman P, Elder D. Q2(R1) Validation of Analytical Procedures. In: ICH Quality Guidelines. John Wiley & Sons, Inc.; 2017. p. 127–66.
23. 23. Swartz ME, Krull IS. Analytical Method Development and Validation. 1997;3(4):96.
24. 24. The USP 40. The United States Pharmacopeia, The Official Compendia of Standards. MD, Rockville, USA.; 2017.
25. 25. Gumustas M, Kurbanoglu S, Uslu B, Ozkan SA. UPLC versus HPLC on drug analysis: Advantageous, applications and their validation parameters. Chromatographia. 2013;76(21–22):1365–427.
26. 26. Blessy M, Patel RD, Prajapati PN, Agrawal YK. Development of forced degradation and stability indicating studies of drugs - A review. J Pharm Anal. 2014;4(3):159–65.
27. 27. Swathi Sri D, Hemant Kumar T, Vara Prasada Rao K, Srinivasa Rao Y. Validated RP-HPLC method for simultaneous determination of Rosuvastatin Calcium and Ezetimibe in pharmaceutical dosage form. Int J Pharm Pharm Sci. 2015;7(4):209–13.
28. 28. Gajjar A, Gajjar AK, Shah VD. Development and Validation of a Stability-Indicating Reversed-Phase HPLC Method for Simultaneous Estimation of Rosuvastatin and Ezetimibe from Their Combination Dosage Forms Rational Drug Designing on Glycogen Synthase Kinase-3 beta View project Developme. Eurasian J Anal Chem. 2010;5(3):280–98.
29. 29. Mukthinuthalapati MA, Bukkapatnam V, Bandaru SPK. Stability indicating liquid chromatographic method for the simultaneous determination of rosuvastatin and ezetimibe in pharmaceutical formulations. Adv Pharm Bull. 2014;4(4):405–11.
30. 30. Kalia V, Garg T, Rath G, Goyal AK. Development and evaluation of a sublingual film of the antiemetic granisetron hydrochloride. Artif Cells, Nanomedicine Biotechnol. 2016;44(3):842–6.
31. 31. Varghese SJ, Ravi TK. Determination of rosuvastatin and ezetimibe in a combined tablet dosage form using high-performance column liquid chromatography and high-performance thin-layer chromatography. J AOAC Int. 2010;93(4):1222–7.