METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF LYCOPENE AND UBIDECARENONE BY RP-HPLC IN COMBINED PHARMACEUTICAL DOSAGE FORM
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 (RPâ€“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., 5Âµm 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 ÂµL 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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