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Background: Bergapten (BER) and taxifolin (TXN), a biomarkers which has many therapeutic activities such as anti-oxidant, anti-inflammatory, anti-microbial, and other pharmacological actions. This shows a new, simple, precise, and accurate High Performance Thin Layer Chromatography (HPTLC) and UV methods for estimating Taxifolin (TXN) and Bergapten (BER) as biomarkers in marketed formulations. Aim: The study determines simultaneous estimation and Q-analysis of UV-Vis spectrophotometry as well as to develop and validate method parameter in High Performance Thin Layer Chromatography (HPTLC). Objective: Application of developed method to quantify these biomarkers in commercial formulation. Materials and methods: The chemicals and reagents were purchased from Merck in India and were of analytical grade. The biomarkers were used of Tokyo Chemical Industry Co., Ltd. bergapten (98.0% w/w) and (+)-taxifolin (95.0%). The biomarkers were estimated and analysed with UV Probe 2.0 software and winCATS software relatively. Methods performed in UV-Vis spectrophotometry (i) Simultaneous estimation and (ii) Q-analysis whereas for HPTLC the biomarkers were simultaneously estimated on parameters of method development. Results: The method for UV-Vis spectroscopy were analysed with a LOD and LOQ of 0.5 and 1 respectively where the %RSD does not exceed more than 2%. The biomarkers were separated by chromatography on precoated aluminium plates as the stationary phase and where optimized by using mobile phase such as toluene:ethyl acetate:formic acid (4:6:0.1 v/v/v) at 306 nm densitometry, and where validated at separate zones. Taxifolin and Bergapten were resolved satisfactorily, with Rf values of 0.64 and 0.82, respectively, in accordance with ICH guidelines. Conclusion: The newly designed UV-Vis Spectroscopy and HPTLC technique has improved specificity, precision and accuracy. The quality of biomarkers containing a flavonoid and furanocoumarins were being validated and quantify these biomarkers in commercial formulation.
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