Authentication of Cassia angustifolia and Cassia acutifolia in Herbal Medicines Employing SCAR Markers

According to the World Health Organization (WHO) estimates, herbal drugs are still the mainstay of 75 – 80% of the world population, mainly in the developing countries for the treatment of diseases, especially chronic ones due to their better compatibility with the human body and lesser side effects. Twelve percent of the population worldwide are affected with constipation and treated with herbal medicine prepared from either the pods and leaves of Cassia angustifolia and Cassia acutifolia or with their pharmacologically active ingredients i.e. Sennosoides. The therapeutic efficacy of these medicines, however, totally depends on the presence of authentic plant components in the formulation. Further, these medicines are often found to be adulterated with other species of Cassia i.e. C. sophera and C. tora. Various conventional taxonomic methods including macroscopy, microscopy and chemoprofiling are being used to determine the presence of genuine herbal components of C. angustifolia and C. acutifolia in herbal medicines, but these methods have limitations to distinguish genuine components from their adulterants. Thus, the therapeutic potential of adulterated medicines is either reduced or these medicines become ineffective due to adulteration. The aim of present work was, therefore, to develop SCAR markers for the genuine herbs, C. angustifolia and C. acutifolia and their adulterants, C. sophera and C. tora that can be used for the authentication of genuine herbal components in crude and processed drugs samples. In the development of SCAR markers, RAPD analysis was done first to identify and select putative species-specific amplicons in the genus of C. angustifolia and C. acutifolia as well as C. sophera and C. tora, which were then sequenced to design sequence specific internal pair of primers. These primer sets were referred as SCAR primers and able to amplify 829bp, 398bp, 589bp and 700bp fragments from the genomes of C. angustifolia, C. acutifolia, C. sophera and C. tora, respectively. These SCAR markers were finally used to authenticate market samples of the herbal medicines containing Cassia sp. The results showed that more than 50% of the market samples were adulterated and these markers were also to detect even 10% of the adulterant in the market samples of the medicines. The SCAR markers developed in this study, hence, could be employed by pharmaceutical companies and public agencies to monitor the quality of these herbal medicines. Acknowledgements: The financial support from Department of AYUSH, Ministry of Health and Family Welfare, Government of India for this work is greatly acknowledged. The research facilities developed from UGC-SAP (DRS-1) grant sanctioned to the Department of Biotechnology and used in this study are also thankfully acknowledged.