Stability-Indicating Densitometric High-Performance Thin-Layer Chromatographic Method for the Quantitative Analysis of Biomarker Naringin in the Leaves and Stems of Rumex vesicarius L.

A simple, sensitive, and stability-indicating high-performance thin-layer chromatography (HPTLC)-densitometric method was developed for the quantification of biomarker naringin in the methanol extracts of stems and leaves of Rumex vesicarius. Chromatography was performed on glass-backed silica gel 60 F 254 high-performance thin-layer chromatography (HPTLC) plates with ethyl acetateglacial acetic acid-MeOH-H2O (30:10:5:1, v/v) as mobile phase. Scanning and quantification were done at 275 nm. The system was found to give compact spot for naringin at R-F = 0.46 +/- 0.001. The linear regression analysis data for the calibration plots showed good linear relationship with r(2) = 0.998 with respect to area in the concentration range of 100-1000 ng. The regression equation of standard was found to be Y = 3.438X + 38.485. Naringin was subjected to acid and alkali hydrolysis, peroxide oxidation, photodegradation, dry heat, moist heat, and ultraviolet (UV) treatment. The drug undergoes complete degradation under acidic treatment and mild degradation under basic and hydrogen peroxide treatment. The degraded products were well-separated from the pure drug. The statistical analysis proves that the developed method for quantification of naringin is reproducible and selective. Due to the ability of the method in separating naringin from other constituents including its degradation products, it can be employed as stability-indicating method for in-process as well as finished products in the market. It is for the first time that authors are reporting a complete stability-indicating densitometric HPTLC method for the estimation of biomarker naringin in the leaves and stems of R. vesicarius L.