ABSTRACT
INTRODUCTION: Thespesia populnea L. (Family: Malvaceae) is a well-known medicinal plant distributed in tropical regions of the world and cultivated in South Gujarat and indicated to be useful in cutaneous affections, psoriasis, ringworm, and eczema. Bark and fruits are indicated in the diseases of skin, urethritis, and gonorrhea. The juice of fruits is employed in treating certain hepatic diseases. The plant is reported to contain flavonoids, quercetin, kaempferol, gossypetin, Kaempferol-3-monoglucoside, ß-sitosterol, kaempferol-7-glucoside, and gossypol. T. populnea is a common component of many herbal and Ayurvedic formulation such as Kamilari and Liv-52. OBJECTIVE: The present study aimed at developing validated and reliable high-performance thin layer chromatography (HPTLC) method for the analysis of quercetin and kaempferol simultaneously in T. populnea. METHOD: The method employed thin-layer chromatography aluminum sheets precoated with silica gel as the stationary phase and toluene: ethyl acetate: formic acid (6:4:0.3 v/v/v) as the mobile phase, which gave compact bands of quercetin and kaempferol. RESULT: Linear regression data for the calibration curves of standard quercetin and kaempferol showed a good linear relationship over a concentration range of 100-600 ng/spot and 500-3000 ng/spot with respect to the area and correlation coefficient (R2) was 0.9955 and 0.9967. The method was evaluated regarding accuracy, precision, selectivity, and robustness. Limits of detection and quantitation were recorded as 32.06 and 85.33 ng/spot and 74.055 and 243.72 ng/spot for quercetin and kaempferol, respectively. CONCLUSION: We concluded that this method employing HPTLC in the quantitative determination of quercetin and kaempferol is efficient, simple, accurate, and validated.
ABSTRACT
In this study, a simple and rapid LC with tandem MS method was developed and validated for the simultaneous determination of kaempferol and quercetin in Thespesia populnea extract. The compounds were eluted using a Gemini C18 column (50 × 2.0 mm, 3 µm), with the mobile phase consisting of acetonitrile-0.3% formic acid in water at the flow rate of 0.400 mL/min. The assay exhibited a linear dynamic range of 25-2500 ng/mL for both kaempferol and quercetin. The values for intra- and interday precision and accuracy were well within the generally accepted criteria for analytical methods (<15%). Selectivity, linearity, LOD, LOQ, accuracy, and precision were evaluated for both analytes. The proposed method is accurate and sensitive and can be used for the routine quantification of kaempferol and quercetin in the herbal extract and in polyherbal formulations.
