Background: 5-fluorouracil and resveratrol are the two most effective anticancer drugs. Combination therapy with these two drugs has shown promising results in cancer. The formulation containing 5-fluorouracil and resveratrol has been prepared, but no analytical method is available in the literature for their simultaneous estimation. However, several analytical methods are there for estimation of either 5-fluorouracil or resveratrol alone. Therefore, the present article is designed for the simultaneous estimation of 5-fluorouracil and resveratrol by HPLC and its application in the quantification of the drugs present in the formulated nanostructured lipid carrier (NLC). Methods: The method was developed using a C18 column (Purospher® STAR RP-18 endcapped (5 µm) Hibar® RT 250- 4.6) with acetonitrile and water as the mobile phase (25:75 v/v) and estimated at 272 nm. The currently developed method was further validated by the ICH guideline Q2 (R1). Combinatorial NLC of 5-fluorouracil and resveratrol was also prepared and characterized. Results: The LOD and LOQ were 8.22 and 24.91 µg mL-1 and 6.58 and 19.93 µg mL-1 , respectively. The precisions was under the acceptable limits of < 2% RSD. The content of 5-fluorouracil and resveratrol in NLC were found to be 65.558±1.343% and 96.326±1.421%, respectively. Conclusion: The findings showed that the developed and validated method was simple, fast, cost-effective and reproducible for the simultaneous estimation of both the drugs in the same formulation.
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