Development and Validation of UV Spectrophotometric Method For Simultaneous Estimation of Biochanin A and 5-Fluorouracil

Abstract

A validated UV spectroscopic method was established for the simultaneous estimation of Biochanin A and 5-Fluorouracil within a hydrogel system. The method was linear in the range of 0.1 to 1 µg/ml for Biochanin A at 229 nm and 1 to 10 µg/ml for 5-Fluorouracil at 265 nm with regression coefficient near to 1. The UV spectra of Biochanin A and 5-Fluorouracil revealed an isobestic point at 249 nm in phosphate buffer pH 5.5. The accuracy and precision results obtained using simultaneous equation method were observed within acceptable limits. The RSD (%) was found to be less than 2% for intraday and interday precision, demonstrating the acceptable precision of the developed method.  Biochanin A showed LOD and LOQ values of 0.064 µg/ml and 0.193 µg/ml at 229 nm respectively while 5-Fluorouracil exhibited corresponding values of 0.314 µg/ml and 0.953 µg/ml at 265 nm. The assay of 5-Fluorouracil and Biochanin A indicated % drug content to be 100.40 % and 103.10 % respectively, indicating accuracy and applicability of developed method.