METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF LYCOPENE AND UBIDECARENONE BY RP-HPLC IN COMBINED PHARMACEUTICAL DOSAGE FORM
Abstract
Lycopene is used for the treatment of cancer and cardiovascular disease and Ubidecarenone is used as a dietary supplement and categorized as cardiovascular agent used for the treatment of heart failure and cardiac disorder. A Reverse Phase Liquid Chromatography (RP–HPLC) method was developed and validated for the simultaneous estimation of Lycopene and Ubidecarenone in combined pharmaceutical dosage form. The different analytical parameters such as linearity, range, precision, accuracy, and robustness were determined according to the International Conference on Harmonization ICH Q2 R (1) guidelines. Chromatography was carried out by Isocratic technique at a flow rate of 1.5 ml/min on Waters Reliant C18 (150 X 4.6mm I.D., 5µm particle size) column at 50 The mobile phase consists of mixture of methanol:tetrahydrofuran:acetonitrile:water in the ratio of 25:15:58:2 % v/v and optimized depending upon the polarity. The UV detection wavelength was 400 nm and 50 µL of sample was injected. The retention times of Lycopene and Ubidecarenone were found to be 11.126 min and 20.504 min respectively. The calibration curves were linear over the concentrations 13-38 mcg/mL and 188-563 mcg/mL with correlation coefficient of 0.9986 and 0.9989, for Lycopene and Ubidecarenone. Percentage recovery obtained 98.58-100.15 % for Lycopene and 99.30-100.64 % for Ubidecarenone. The % RSD for Precision and Accuracy of the method was found to be NMT 2%. The proposed method was highly sensitive, precise and accurate. Hence, the method was successfully applied for the reliable quantification of active pharmaceutical ingredient content in in-house prepared tablet formulation.
Key words: Lycopene, Ubidecarenone, RP - HPLC, Validation, Simultaneous estimation, % RSD.
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2. Giovannucci E, Ascherio A, Rimm E B, Stampfer M J, Colditz G A, Willett W C. Intake of Carotenoids and retinol in relation to risk of prostate cancer, NCBI, 1995, 87, 1767–1776.
3. Müller et al. Author manuscript, J AOAC Int, PMC Feb 2012, 15.
4. Lycopene: Monograph. Alt Med Rev 2003, 8, 336-42.
5. Goodman and Gillman's, The Pharmacological basis of therapeutics, 10th ed. McGrawâ€Hill: London, 2001, 569â€620.
6. Tzouganaki Z.D, Atta- Politou, Koupparis M.A, Determination and Validation of liquid chromatographic method for the determination of Lycopene in plasma, Analytica Chimica Acta, september 2002, 467(1), 115-123.
7. Shantaram Nangude, Manisha Vite. A Simple and Sensitive RP-HPLC Method for Estimation of Lycopene in Pharmaceutical Solid Dosage Forms. Journal of Pharmaceutical Science and Bioscientific Research (JPSBR), 2013, 3(1), 16-19.
8. André Műller, Bernd Pletsch, Nicole Faccin, Joseph Schierle, Edwrd H. Waysek. Method for the determination of Lycopene in Supplements and Raw material by Reversed –Phase Liquid Chromatography: Single-Laboratory Validation. NIH-Public Access Author Manuscript. JAOAC Int. 2008, 91(6), 1284-1297.
9. Ligor M, KaváÄová J, Gadzala-Kopciuch R.M, StudziÅ„ska S, Bocian Sz, Lehotay J, Buszewski B, Behaviours in Analysis of Carotenoids. Chromatographia, 2014, 77, 1047-1057.
10. Thadikamala Sathish, Doradla Udayakiran, Kushala Himabindu, Patsamatla Lakshmi Durga Sridevi, Devarapalli Kezia, and Pullasi Bhojaraju. HPLC Method for the determination of Lycopene in Crude Oleoresin Extracts. Asian Journal of Chemistry, 2009, 21(1), 139-148.
11. Olives Barba A.I, Cámara Hurtado M, Sánchez Mata M.C, Fernández Ruiz V, Lòpez Sáenz de Tejada M. Application of a UV-Vis detection–HPLC method for a rapid determination of Lycopene and β-carotene in vegatables. Food Chemistry, 2006, 95, 328-336.
12. Roman KanÄár, Pavla Novotná, and Peter Drábková. Determination of Retinol, α-Tocopherol, Lycopene, and β-carotene in Human Plasma Using HPLC with UV-Vis Detection: Application to a clinical Study. Journal of Chemistry, 2013, 1-7.
13. Li Z., Deng Y.J., Wang X.M. and Lei G.F., HPLC determination of entrapment of coenzyme Q10, Chinese Journal of Pharmaceutical Analysis. 2006, 26(2), 149â€151.
14. Zhou T.Y. Sun H.D. Zhang D.W., Zheng L. and Zhang Q., Determination of total co-enzymes in plasma, Chinese Pharmaceutical Journal, 2002, 37(3), 189â€192.
15. Mailvelan R, Mounnissamy V M, Selvamani P, Rajesh J, Raviraj T. Development and Validation of UV Spectrophotometric methods for the simultaneous estimation of Ubidecarenone (Coenzyme Q-10) and Clomifene Citrate in Bulk and Tablet Dosage Forms, AJRCPS, 2013, 1(1), 23-30.
16. Mailvelan R, Rajesh J , Selvamani P, Latha S. HPLC Method Development and Validation for simultaneous estimation of Ubidecarenone and Clomifene Citrate in Bulk and Tablet Dosage Forms. International Journal of Research in Pharmaceutical and Nano Sciences, 2012, 1(1), 61-69.
17. Saravanamuthukumar M, Palanivelu M, Anandarajagopal K, Sridharan D. Simultaneous estimation and validation of Atorvastatin calcium and Ubidecarenone (Coenzyme Q10) in combined Table Dosage form by RP-HPLC. International Journal of Pharmacy and Pharmaceutical Sciences, 2010, 2(2), 36-38.
18. Kannapan K, Sasidharan D.K, Ramkumar P, Prajapati M.K, and Panda P.P. Analytical Method Development and Validation of Atorvastatin Calcium and Ubidecarenone Tablet by RP-HPLC. International Journal of Pharmaceutical Sciences and Research, 2011, 2(7), 1679-1682.
19. Karupinska J., Mikoluc B. and Piotrowska Jastrzebska J., HPLC method for simultaneous determination of retinol, alpha†tocopherol and coenzyme Q10 in human plasma. Journal of pharmaceutical and biomedical analysis, 1998, 17(8), 1345â€1350.
20. Anonymous. ICH Harmonised Tripartite Guidelines. Text on Validation of Analytical Procedures: Text and Methodology. Q2R (1). Geneva, 2005.
21. Beckett, A.H., Stanlake, J.B. Practical Pharmaceutical Chemistry II, 4th Edition.CBS Publishers and Distributors, New Delhi; 2007.
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