SIMPLE BIOANALYTICAL QUANTIFICATION METHOD FOR SIMULTANEOUS ESTIMATION OF SIMVASTATIN AND EZETIMIBE IN HUMAN PLASMA BY REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY TECHNIQUE
Objective: A bioanalytical quantification method was developed for the simultaneous estimation of simvastatin (SIM) and ezetimibe (EZE) from the human plasma.
Methods: The technique was developed and equipped with reverse-phase (RP) high-performance liquid chromatography, using RP-C18 column with an ultraviolet detector. For the estimation of SIM and EZE, the mobile phase (acetonitrile:acetate buffer pH 4.0, pH adjusted with acetic acid) was pumped at a flow rate of 0.8 ml/min in the ratio of 85:15% v/v and the eluents were monitored at 234 nm. A calibration graph to study linearity of the SIM and EZE in biological matrix was carried out in the concentration range of 400–4000 ng/ml for both these drugs SIM and EZE.
Results: The developed method was validated according to the US FDA and European Medicines Agency guidelines for sensitivity, accuracy, precision, and stability. The obtained statistical data of validation were found to be within prescribed limit assures rigidity of the method. Both the drugs in combined form were estimated in human plasma by the proposed method.
Conclusions: The developed method is free from solid-phase extraction so it becomes simple and economical. The method is efficient for precise and accurate quantification of SIM and EZE in plasma and hence applied for bioequivalence, bioavailability study in real clinical samples.
2. Beale JM Jr., Block JH. Wilson and Gisvold’s Textbook of Organic Medicinal and Pharmaceutical Chemistry. 12th ed. New Delhi: Wolters Kluwer (India) Pvt. Ltd.; 2011. p. 652-4.
3. Brayfield A. Martindale: The Complete Drug Reference. 38th ed. London: Pharmaceutical Press; 2014. p. 1379, 1489.
4. Lemke TL, Williams DA, Roche VF, Zito SW. Foye’s Principles of Medicinal Chemistry. 7th ed. New Delhi: Wolters Kluwer (India) Pvt. Ltd.; 2013. p. 827.
5. Dixit RP, Barhate CR, Padhye SG, Viswanathan CL, Nagarsenker MS. Stability indicating RP-HPLC method for simultaneous determination of simvastatin and ezetimibe from tablet dosage form. Indian J Pharm Sci 2010;72:204-10.
6. Sahoo NK, Sahu M, Rao PS, Vineela RS, Devi JN, Rani NS, et al. Validation of assay indicating method development of simvastatin in bulk and its tablet dosage form by RP-HPLC. J Appl Pharm Sci 2014;4:117-22.
7. Ali H, Nazzal S. Development and validation of a reversed-phase HPLC method for the simultaneous analysis of simvastatin and tocotrienols in combined dosage forms. J Pharm Biomed Anal 2009;49:950-6.
8. Nalaiya J, Sagineedu SR, Rajasingam R, Kassim Z, Pichika MR. A stability-indicating RP-HPLC method for simultaneous determination of simvastatin and niacin in a combined dosage form. Int J Pharm Pharm Sci 2014;7:262-8.
9. Novakova L, Satinsky D, Solich P. HPLC methods for the determination of simvastatin and atorvastatin. TrAC Trends Analyt Chem 2008;27:352-67.
10. Alakhali KM. Validation method for measuring simvastatin in human plasma by HPLC-UV and its application in study simvastatin stability in plasma and working solution. Asian J Pharm Clin Res 2014;7:131-3.
11. Lawson G, Cocks E, Tanna S. Bisoprolol, ramipril and simvastatin determination in dried blood spot samples using LC-HRMS for assessing medication adherence. J Pharm Biomed Anal 2013;81:99 107.
12. Barrett B, Huclová J, Borek-Dohalský V, Nemec B, Jelínek I. Validated HPLC-MS/MS method for simultaneous determination of simvastatin and simvastatin hydroxy acid in human plasma. J Pharm Biomed Anal 2006;41:517-26.
13. Selvan PS, Pal TK. Chromatography-tandem mass spectrometry method for the simultaneous quantitation of metoprolol succinate and simvastatin in human plasma. J Pharm Biomed Anal 2009;49:780-5.
14. Mishra R. Spherical crystallization a novel approach for solubility and dissolution enhancement of simvastatin. Asian J Pharm Clin Res 2016;9:65-72.
15. Virani P, Sojitra R, Raj H, Jain V. Chromatographic method for irbesartan and its combination with other drug. J Crit Rev 2015;2:7-11.
16. Hefnawy M, Al-Omar M, Julkhuf S. Rapid and sensitive simultaneous determination of ezetimibe and simvastatin from their combination drug products by monolithic silica high-performance liquid chromatographic column. J Pharm Biomed Anal 2009;50:527-34.
17. Raul SK, Aravelli RB, Jhansi D. RP-HPLC method development and validation for the simultaneous estimation of atorvastatin and ezetimibe in pharmaceutical dosage form. Asian J Pharm Clin Res 2015;8:178-81.
18. Bolla N, Atla SR. A validated stability indicating RP-HPLC method for the simultaneous determintion of atorvastatin calcium and ezetimibe hydrochloride in bulk and tablet dosage form. Int J Pharm Pharm Sci 2016;8:370-7.
19. Yulianita R, Sopyan I, Muchtaridi M. Forced degradation study of statins: A review. Int J Appl Pharm 2018;10:38-42.
20. Verma N. Introduction to hyperlipidemia and its treatment: A review. Int J Curr Pharm Res 2016;9:6-14.
21. Karanam SR, Katakam P, Chandu BR, Hwisa NT, Adiki SK. Simultaneous determination of ezetimibe and simvastatin in rat plasma by stable-isotope dilution LC-ESI-MS/MS and its application to a pharmacokinetic study. J Pharm Anal 2014;4:286-94.
22. Abdelwahab NS, El-Zeiny BA, Tohamy SI. Two spectrophotometric methods for simultaneous determination of some antihyperlipidemic drugs. J Pharm Anal 2012;2:279-84.
23. Sharma M, Mhaske DV, Mahadik M, Kadam SS, Dhaneshwar SR. UV and three derivative spectrophotometric methods for determination of ezetimibe in tablet formulation. Indian J Pharm Sci 2008;70:258-60.
24. Belal TS, Daabees HG, Abdel-Khalek MM, Mahrous MS, Khamis MM. New simple spectrophotometric method for determination of the binary mixtures (atorvastatin calcium and ezetimibe; candesartan cilexetil and hydrochlorothiazide) in tablets. J Pharm Anal 2013;3:118-26.
25. Artici E, Karliga B. Identification, synthesis and characterization of process related desfluoro impurity of ezetimibe and HPLC method validations. J Pharm Anal 2015;5:356-70.
26. Santa Z, Koti J, Szoke K, Vukics K, Szantay C Jr. Structure of the major degradant of ezetimibe. J Pharm Biomed Anal 2012;58:125-9.
27. Medicines and Healthcare Products Regulatory Agency. British Pharmacopoeia. London: Medicines and Healthcare Products Regulatory Agency; 2015. p. 812.
28. Government of India, Ministry of Health and Family Welfare. Indian Pharmacopoeia. 7th ed. Ghaziabad: The Indian Pharmacopoeia Commission; 2014. p. 1727, 2684, 2730.
29. Center for Drug Evaluation and Research. Guidance for Industry. Bio Analytical Method Validation. United States Department of Health and Human Services. Rockville: Food and Drug Administration. Center for Drug Evaluation and Research. Center for Biologics Evaluation and Research; 2013.
30. European Medicines Agency. Guideline on Bio Analytical Method Validation. Committee for Medicinal Products for Human Use. London: Churchill Place, EMEA/CHMP/EWP/192217/2009; 2011.
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.