Kirti S. Laddha

In Asian countries, Centella asiatica is exploited for abundant types of pharmacological activities due to the presence of opulent phytochemicals. Asiaticoside, madecassoside, and their sapogenin triterpene acids such as asiatic acid and madecassic acid are the most noticeable triterpenes present in C. asiatica. To date, numerous techniques/methods are used to extract and isolate the different kinds of phytoconstituents from C. asiatica. Still, most methods require some special requirements, and some procedures are monotonous and time-consuming. Meanwhile, previously reported methods used for the extraction and isolation were not validated for large-scale production, yield, and purity. The study's primary goal is to develop the methodology for extracting and isolating the Asiaticoside and asiatic acid from C. asiatica at the minimum time with the highest yield and purity. Asiaticoside and asiatic acid extraction and isolation involved the acid hydrolysis method and recovered in alcohol. The Reverse Phase-High Performace Liquid Chromatography (RP-HPLC) method was developed and validated as per ICH guidelines for quantifying both compounds. The obtained results indicate that the developed method produces asiaticoside and asiatic acid with good purity. As per the ICH guidelines, the RP-HPLC was developed and validated. The proposed method can be used to isolate the asiaticoside and asiatic acid from C. asiatica. Some modification in this method leads to the large-scale production of highly pure asiatic acid and asiaticoside for their versatile application in the area of cosmetics and phytopharmaceuticals.

The goal of the study was to determine how much lupeol was present in different Bauhinia acuminata samples. Lupeol was analyzed using a unique (verified) high‐performance thin‐layer chromatography (HPTLC) method. Separation of various compounds was achieved on silica gel (66 F 254) plates with toluene and methanol (99:1) as the mobile phase, followed by derivatization with Liebermann–Burchard reagent (r² = 0.996). The proposed approach demonstrated a satisfactory linear relationship in the concentration range of 4–8 μg/mL. The limit of detection and limit of quantification were 63.84 ng/spot and 208.57 ng/spot, respectively. The proposed method was a simple and precise procedure that provided good resolution with other constituents of extracts. The validated HPTLC method described was simple, dependable, and practical and could be useful for standardizing and evaluating the quality of herbal material and formulations containing various Bauhinia species. The leaf, stem, unsaponified matter leaf, and stem petroleum ether extract had good amounts of lupeol (25.75, 38.25, 1.65, and 1.75, respectively). This research will be helpful for the quantitative determination of lupeol from herbal formulations as well as useful for further studies in this area.

A novel high-performance thin layer chromatography method for quantification of long chain aliphatic hydrocarbons from Cissus quadrangularis

The surge of interest in naturally occurring phytochemicals with high therapeutic potential has led to the discovery of many molecules, out of which naturally occuring coumarins such as marmelosin, umbelliferone and scopoletin present in Aegle marmelos (Bael) fruit shows good therapeutic potential. The aim of the present work is to develop and validate Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) method for simultaneous determination of marmelosin, umbelliferone and scopoletin in A. marmelos fruit extracts. The chromatographic separation was performed with isocratic elution of 55:45 (%, v/v) methanol–water containing 0.1 % acetic acid as mobile phase. The method used to analyse the extract of A. marmelos showed good resolution with retention time within 12 min. The relative concentrations of above phytoconstituent were determined in A. marmelos fruits. The method was found to give compact peaks for scopoletin, umbelliferone and marmelosin (R_t of 4.6, 6.5 and 11.3 min respectively) and were linear over the range 5–30 μg ml⁻¹ (R² = 0.9655), 2–10 μg ml⁻¹ (R² = 0.9964) and 2–10 μg ml⁻¹ (R² = 0.9862) respectively. The mean recoveries for marmelosin, umbelliferone and scopoletin at three concentrations were in the range of 98.8–102.9, 98.8–101.1 and 94.2–98.3 % respectively. The relative standard deviation of accuracy, precision and repeatability were within 2 %, indicating the method produced highly reproducible results. Therefore this simple, precise and accurate method enables simultaneous separation of this phytoconstituent and hence can be successfully applied in analysis and routine quality control of herbal material and formulation containing A. marmelos.

Embelin, a naturally occurring benzoquinone, is obtained from fruits of Embelia ribes, which plays a vital role in the Ayurvedic system of medicine. The benzoquinone has diverse pharmacological and therapeutic properties and now constitutes a part of modern medicine. The aim of our study was to observe the effect of various stress conditions on this potential drug candidate. Embelin was subjected to stress conditions of acid and alkaline hydrolysis, oxidation, photolysis and thermal degradation. A significant degradation was found to occur by acid hydrolysis, oxidation and to a lesser extent under thermal stress and alkaline hydrolysis; the compound was found to be stable to photolytic stress. Further, the degradation product resulting from the acid hydrolysis was isolated and its structure was elucidated using spectroscopic techniques. Stress degradation studies on embelin provide an insight for its stability and storage considering the formulation aspects of modern medicine.