~ 51 ~  Journal of Medicinal Plants Studies 2015; 3(6): 51-55 ISSN 2320-3862 JMPS 2015; 3(6): 51-55 © 2015 JMPS Received: 22-09-2015 Accepted: 26-10-2015 V Baskar Ananda Raj Department of Pharmacognosy, Padmavathi College of Pharmacy and Research Center, Krishnagiri Main Road, Periyanahalli, Dharmapuri district, 635205, Tamil Nadu, India. S Suresh Kumar Department of Pharmacognosy, JKK Nataraja College of Pharmacy, Komarapalayam, Namakkal district, 638183, Tamil Nadu, India. KL Senthil Kumar Department of Pharmaceutics, Padmavathi College of Pharmacy and Research Center, Krishnagiri Main Road, Periyanahalli, Dharmapuri district, 635205, Tamil Nadu, India. Correspondence V Baskar ananda Raj Research Scholar, Department of Pharmacognosy, Padmavathi College of Pharmacy, Krishnagiri main road, Periyanahalli – 635205, Dharmapuri district, Tamilnadu, India. HPTLC Standardization and Quantification of Rhinacanthus nasutus V Baskar ananda Raj, S Suresh Kumar, KL Senthil Kumar Abstract Background: Ethnobiological status of the plant R.nasutus (Rhinacanthus nasutus) has been analyzed by HPTLC standardizing and quantifying of flavonoids metabolites. Methods: HPTLC method using 10µl of ethanol extract of R.nasutus at a concentration of 10mg/ml along with 10µl of each 1mg/ml standard concentration of Rutin and Quercetin were applied to the pre- coated silica gel F254 HPTLC plates, and the plate was developed in a twin trough TLC chamber saturated with Ethyl acetate: Formic acid: Glacial acetic acid: water (11:1.1:1.1:2.6). Evaluation by using a scanning TLC scanner was carried out in the absorbance mode at 254 and 366 nm and the result was visualized using CAMAG Visualizer. Result: The extract R.nasutus were estimated for the presence of standards Quercetin and Rutin by HPTLC method done by comparing the RF values and peak matching with HPTLC chromatogram and quantified to be Quercetin - 11.6% and Rutin - 5.4% respectively. Conclusion: Standardized criteria for studying the secondary metabolites such as flavonoids with reliable standard methods are often lacking. Efforts are underway to establish a pharmacoepidemiological evidence base regarding safety and practice of Ayurvedic medicines. Keywords: Rhinacanthus nasutus, traditional, Ayurveda, HPTLC, chromatogram, Quercetin, Rutin, Flavonoids and phytomedicine. Introduction In traditional medicine research, clinical experiences, observations or available data are starting points, which are issues related to their quality assurance remain vitally important. In many cases, the exact chemical constituents and percentage of active constituents present in the plant needed to give a definite and specific biological effect have yet to be verified. These constituents need to be limited to a biological effect that in turn is relevant to a therapeutic target. This approach may be obtained by determining the natural compounds present in the selected botanical, followed by biological testing, to identify biological fingerprints of the plant extracts and their effective concentration [1] . Although several studies have been well performed, there have been conflicting clinical data, suggesting a lack of specific activity or inappropriate preparation. Owing to these numbers of new chemical/molecular entities that are approved are declining. This situation calls for critical assessment of the current reduction strategies, where only new chemical entities are valued as potential new drugs. Currently, many pharmaceutical companies and research organizations’ have demonstrated renewed interest in investigating higher plants as sources for novel lead structures and also for the development of standardized phytomedicines with reasonable efficacy, safety and quality [2] . Botanical extracts can only be declared pharmacologically active if a reliable correlation can be made between a measurable active principle and its declared medicinal effect. Therefore, there is a requirement to look for secondary metabolites from plant sources. Consequently, this has led to the discovery of potentially useful preliminary active ingredients that can serve as a source and template for the synthesis of new drugs from the plant R.nasutus, thus by proving through quantification and standardization by using HPTLC method. Plant Source Rhinacanthus nasutus commonly known in English as “Snake Jasmine” due to the shape of its flowers [3] and as “Nagamalli” in Tamil belonging to family Acanthaceae. The plant is a small, slender shrub widely distributed in some parts of the sub-continent, in the region of Southeast