Résumé
HPTLC and HPLC are two most widely accepted methods for determination of natural products. Present research work envisages microwave assisted extraction of quercetin from hydroethanolic extract of Cuscuta reflexa Roxb. (Cuscutaceae), an unusual parasitic vine. Further, chromatographic characterization of hydroethanolic extract of C. reflexa was carried out in terms of quercetin content using two validated methods (HPTLC and RP-HPLC). Confirmation of the presence of quercetin in the samples was carried out using Mass Spectrometry. HPTLC separation of quercetin was achieved on an aluminum-backed layer of silica gel 60 F254 using Toluene: ethyl acetate and formic acid as mobile phase while RP-HPLC was performed on Cosmosil C18-column (150 mm x 4.6 mm, 5 μm) using mobile phase comprising of 0.025 M NaH2PO4 buffer – ACN (pH - 2.6) at a flow rate of 1.2 mL/min. ICH guidelines were followed for validation of both the the chromatographic methods. Samples of C. reflexa collected from different regions of India and growing on different hosts were also screened for their quercetin content. The developed chromatographic methods described here were found to be simple, rapid, accurate and sensitive.
Résumé
Urtica dioica L. (Urticaceae, leaves) is commonly used in traditional systems of medicine for the treatment of a wide range of disorders. The present work emphasizes on a validated HPTLC method for estimation of ursolic acid from U. dioica leaves and its available formulation. Chromatographic separation was achieved on silica gel 60 F254 TLC plate with toluene: ethyl acetate: formic acid (7:3:0.1, v/v/v) as a mobile phase. Detection of ursolic acid was carried out by derivatizing the plate with Liebermann Burchard reagent at 110°C for 10 min. Camag TLC scanner 4 equipped with winCATS software was used for densitometric scanning at 366 nm. The accuracy of the method was checked by conducting various validation parameters according to ICH guidelines. The method was found applicable to evaluate the impact of regional variation on ursolic acid content in U. dioica leaves. The research also highlights estimation of ursolic acid from a marketed herbal formulation of U. dioica leaves. The described HPTLC method was found useful for quantitation of bioactive marker ursolic acid and can be used as a routine quality control tool for the assessment of botanicals.
Résumé
Many plants have been claimed to possess liver protecting activity. But, due to scarcity of potent modern medicine to treat severe liver diseases, many folk remedies of plant origin have been scientifically evaluated for their potential hepatoprotective activity in experimental animal models. Parmelia perlata (Huds.) Ach. (lichen, Parmeliaceae), commonly known as Dagad phool or stone flower in India, is mentioned in Indian Materia Medica as useful in treating a number of ailments. In the present study, hepatoprotective activity of aqueous slurry of P. perlata was investigated in CCl4 intoxicated Albino Wistar rats and the results were compared with, Silymarin. Intraperitoneal injection of CCl4, produced a marked elevation in the level of biochemical markers such as Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate Transaminase (SGPT), bilirubin, cholesterol and triglycerides. Oral administration of P. perlata aqueous slurry at 0.7 g / kg and 1.0 g / kg in CCl4 intoxicated rats showed marked decrease in the level of biochemical markers and results were at par with the effect shown by Silymarin. The results of histopathological analysis were in compliance with the findings of blood biochemical parameter analysis. This study confirms that P. perlata aqueous slurry has phytocomponents with hepatoprotective potential.