A Rapid High-Performance Thin-Layer Chromatographic Method to Estimate Quercetin inBenincasa hispida(Thunb.) Cogn. Fruit Pulp

Abstract

Ayurvedic medicines show great promise due to their holistic approach in the treatment of diseases. However, proper standardization is necessary for their integration into mainstream medicine. One such well-known Ayurvedic trailing herb is Benincasa hispida (Thunb.) Cogn. Its fruit contains numerous secondary metabolites, including quercetin, and is used to treat urinary calculi, blood disease, insanity, epilepsy, jaundice, dyspepsia, fever, and menstrual disorders. The current investigation was undertaken to develop and validate a rapid, sensitive, and reproducible method for quantifying quercetin in the hydroalcoholic extract of B. hispida fruit pulp (HABH). The pre-coated thin-layer chromatography (TLC) aluminum plates with silica gel 60 F254 were used with solvent system comprising toluene–ethyl acetate–formic acid (5:4:0.2, V/V). Determination and quantification were performed by densitometric scanning using a deuterium lamp in the absorbance mode at 262 nm. The validation of precision, accuracy, and reproducibility of the developed high-performance thin-layer chromatographic (HPTLC) method were done as per the International Conference on Harmonization (ICH) guidelines. The mobile phase used for the development of HPTLC/TLC plate yields a distinct band for quercetin (RF = 0.392). The limit of detection and limit of quantification for the method were found to be 20 and 60 ng per band, respectively. The quantified quercetin content was found to be 193.77 ± 2.86 μg in 10 mg of HABH, i.e., 1.94% w/w of HABH. This HPTLC method can be successfully employed for the standardization and quantitative analysis of quercetin in formulation containing B. hispida fruit pulp and it will be helpful in the quality control/assurance of such formulations.