Pharmacognostic analysis and bioactivity studies in Kamettia ca1yop Jyllata Roxb Nicolson Suresh

Abstract

Accurate recognition, uniformity, and quality assurance of medicinal plants are essential for ensuring the safety and effectiveness of herbal remedies. This study aims to establish a standard for Kamettia caryophyllata (Roxb.) Nicolson & Suresh, a climber belonging to the Apocyanaceae family, recognised in Indian alternative medicine for its efficacy in managing ailments like arthritis, scabies, and epilepsy, as detailed in Hortus Malabaricus. Until now, there has been no detailed scientific study on the pharmacognostic or chemical or bioactive properties of this plant. The objective of the study is to evaluate the quality, purity, identity, and bioactive principles of K. caryophyllata through pharmacognostic, physicochemical, phytochemical, and bioactivity analyses of the plant components. The study also seeks to validate the scientific evidence supporting traditional medicinal uses by isolating, purifying, and characterising a specific bioactive compound. Authentication of K. caryophyllata was achieved through morphological features and molecular methods using DNA barcoding with rbcL and matK gene sequences. Pharmacognostic features, including macroscopic, powder microscopic and anatomical characteristics and fluorescence colour behavioural patterns were documented. The physicochemical analysis revealed variations in total ash, water-soluble ash, acid-insoluble ash contents and extractive yields among different plant components, with the leaves showing superior characteristics, indicating greater concentration of phytoconstituents, suggesting their significant therapeutic potential than other components. The physicochemical characteristics combined with pharmacognostic, morphological, and molecular characteristics have contributed to create valuable reference markers for monograph preparation of the plant, which can serve as a reference standard for identification and detecting adulteration in crude drugs and guide traditional medicine practitioners in selecting the appropriate plant part of K. caryophyllata in herbal medicine. The preliminary phytochemical screening using various reaction tests and GC-MS and HR LC-MS screening for specific bioactive compounds revealed wide range of phytochemical groups with more diverse bioactive compounds in the methanolic extract of leaf, stem and root components with highest predominance in methanolic leaf extract. This inference was further substantiated by the quantitative estimation of secondary metabolites in the methanolic extracts which revealed that majority of these compounds, including terpenoids, alkaloids, and phenolics, were most concentrated in the leaf component compared to the stem and root. The analysis of various bioactivities such as antioxidant, antimicrobial, cytotoxic and anti-inflammatory properties of different component extracts revealed that the leaf methanolic extract has more promising potent activities than the stem and root extracts. The in-vivo anti-inflammatory studies using Carrageenan-induced and Formalin- induced inflammation models revealed, methanolic extract is very much effective in animal model. The study revealed that the phytochemical group terpenoids was one of the most abundant with highest dominance in leaf component. The phytochemical screening revealed the consistent occurrence of a terpenoid compound, squalene, in all the component extracts, which has diverse pharmacological properties including anti-inflammatory, antioxidant and antimicrobial. Through the isolation of this potential bioactive compound, squalene, scientific evidence and confirmation on the effectiveness of using K. caryophyllata in traditional medicine has been achieved. The study concludes that K. caryophyllata holds great potential as a natural medicine for preventing and managing various diseases. It provides a solid foundation for future research, which could lead to pioneering treatments derived from natural sources. However, further clinical studies are needed to validate the efficacy and safety of K. caryophyllata extracts for human use, paving the way for evidence-based therapeutic interventions.