Bhavesh Tiwari

Estimation of Rhein from Cassia fistula Linn. using validated HPTLC method

Background: Rhein is the major anthraquinone found in Cassia fistula Linn, which is traditionally used as an important component of plant based medicine. Aims and Objective: To develop and validate a High Performance Thin Layer Chromatographic method for the estimation of Rhein from C. fistula Linn. Materials and Methods: Chromatographic separation was achieved on silica gel 60 F254 plates with n‑hexane: Ethyl acetate: Formic acid (5:5:0.2, v/v/v) as a mobile phase. Detection of Rhein was carried out by derivatizing the plate with 10% ethanolic KOH reagent followed by heating at 110°C for 10 min. CAMAG TLC scanner 4 equipped with winCATS software was used for densitometric scanning at 550 nm. The accuracy of the method was checked by conducting various validation parameters according to ICH (International Conference on Harmonization) guidelines. Results: The method was found applicable to evaluate the content of Rhein in different plant parts of C. fistula and the impact of different geographical regions on the Rhein content in fruit pulp of C. fistula was also studied. Estimation of Rhein from a herbal formulation containing C. fistula fruit pulp was also carried out using the validated HPTLC method. The developed method was found sensitive and accurate for the estimation of Rhein from various plant matrices. Conclusion: The developed method was found useful for the quantitation of bioactive marker Rhein and can be used as a routine quality control tool for the assessment of botanicals. Key words: Cassia fistula, fruit pulp, geographical variation, HPTLC, Rhein

Standardization of Shadbindu Taila: An Ayurvedic oil based medicine

Shadbindu Taila (ST) is an Ayurvedic formulation used as a remedy for loosening of tooth, weakness of the eyesight, loss of hair, diseases of head, etc., Present study is an attempt to develop some newer approaches for the quality control and standardization of ST. Standardized operating procedure for the preparation of ST was developed in accordance with Ayurvedic Formulary of India. Preliminary phytochemical, physicochemical, and chromatographic evaluation of ST was carried out. Safety of ST was evaluated in terms of skin irritation test and presence of heavy metals. Chemical characterization of ST was done on the basis of kaempferol using validated -High Performance Thin Layer Chromatographic (HPTLC) method. ST did not show presence of any of the heavy metals analyzed and was found non-irritant on rabbit skin. The quality control parameters resulted after scientific evaluation of ST can be used as reference standard for quality control/assurance laboratory of a pharmaceutical firm in order to have a proper quality check over its preparation and processing.

A High-Performance Thin-Layer Chromatographic Method for the Evaluation of Marmelosin from Aegle marmelos Corr. Extract and Its Traditional Formulation from Rat Plasma: Application to Pharmacokinetics

Marmelosin is one of the major phytochemical constituents of Aegle marmelos (L.) Corr. (Rutaceae). Till date, there are no methods reported for the estimation of marmelosin in plasma matrix using high-performance thin-layer chromatography (HPTLC) which is a versatile analytical technique promoting shorter analysis time and less consumption of solvents with efficient data acquisition and processing. This study is an attempt to develop a rapid and sensitive HPTLC method for the estimation of marmelosin from rat plasma and its application to evaluate pharmacokinetics of A. marmelos fruit pulp extract and a traditional Unani medicine Sharabat-e-Belgiri. Plasma processing involved liquid-liquid extraction with diethyl ether followed by HPTLC. Separation and determination of marmelosin were performed on silica gel 60 F254 TLC plate using toluene-ethyl acetate-glacial acetic acid as mobile phase. United States Food and Drug Administration (USFDA) guidelines for bioanalytical validation were followed to validate the HPTLC method. RF of marmelosin was found to be 0.57. The calibration curve was linear over the range of 1.0–150.0 µg mL−1 (r2 = 0.995) in rat plasma. Limit of detection and limit of quantitation were found to be 0.5 µg mL−1 and 1.0 µg mL−1, respectively. The extraction recovery from plasma was in the range of 90.07–98.22%. The intra-day and inter-day precisions were found to be 90.89–104.12%, and 92.82–104.86%, respectively. The absorption of marmelosin from extract was found significantly higher than the formulation. The results indicated that HPTLC is a suitable technique for the estimation and pharmacokinetic evaluation of marmelosin from plant extract and Unani formulation in rat plasma.