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A simple, accurate and precise HPLC method for simultaneous determination of Trifluridine and Tipiracil in pure and tablet dosage form has been developed. To develop and validate analytical method for simultaneous estimation of Trifluridine and Tipiracil in pharmaceutical formulation by RP-HPLC. HPLC of Waters (Model: Alliance 2695) with Phenomenex Luna C18 (4.6 mm I.D. Ã— 250 mm, 5 Âµm) column was used for chromatographic separation. It contains waters injector and PDA Detector (Deuterium). Mobile phase consists of Methanol:Water (65:35% v/v) and flow rate adjusted was 1ml/min. Wavelength selected for detection was 220nm and injection volume was 10 Âµl. By using the developed method, retention time of Trifluridine and Tipiracil was found to be 3.2min and 5.4min respectively. The method has been validated forÂ linearity,Â accuracyÂ andÂ precision.Â Linearity of Trifluridine and Tipiracil were in the range of 75â€“375Î¼g/ml and 15â€“75Î¼g/ml respectively. The percentage recoveries obtained for Trifluridine and Tipiracil were found to be in range of 99.3 â€“ 99.6%. LOD and LOQ were found to be 12.5Âµg/ml and 38.1Âµg/ml for Trifluridine 3.7and 11.4Âµg/ml for Tipiracil
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