Abstract
The aim of the present study was to develop and validate an UHPLC-DAD method for quantification of cholesterylsuccinyl-5-fluorouracil conjugate in a standard sample, a lipid nanoemulsion and dissolved samples. The separation of the conjugate was carried out on Hypersil GOLD 50 X 2.1 mm RP C-18 column packed with 1.9 mu m packing as a stationary phase. The mobile phase was methanol:water (80:20 % v/v) at a flow rate of 0.3 ml/min with DAD detection at 276 nm. The proposed method was found to be precise, accurate, robust, sensitive and specific for quantification of the conjugate. The utility of the proposed method was checked by the assay of conjugate in a lipid nanoemulsion and in dissolved samples. High assay value of conjugate in the lipid nanoemulsion was observed (99.25 %). In vitro dissolution of the cholesteryl-succinyl-5-fluorouracil conjugate in the lipid nanoemulsion was observed as 78.1% after 24 h. The conjugate was found to be sufficiently degraded under acid, base and thermal stress conditions. The developed method successfully resolved the drug conjugate peak in the presence of its degradation products. These results indicated that the developed UHPLC-DAD method can successfully be used for routine analysis of drug conjugates in pharmaceutical formulations.