Original Article PHARMACOGNOSTICAL GENETIC BARCODING AND PHYTOCHEMICAL ANALYSIS ON LEAVES OF DALBERGIA VOLUBILIS ROXB.–AN EXTRA PHARMACOPOEIAL DRUG OF AYURVEDA ACHARYA RABINARAYAN * , JANI SWITU, C R HARISHA, SHUKLA VINAY Head of Dravyaguna Department, Room No. 329, 3 rd Floor, Institute Building, IPGT and RA, Gujarat Ayurved University, Jamnagar 361008 Email: drrnacharya@gmail.com Received: 19 Mar 2018 Revised and Accepted: 08 May 2018 ABSTRACT Objective: The leaves of Biraskala have been reported for its ethnomedicinal claims for the management of aphthae, heart pain, menorrhagia and post-partum problem. Botanically it has been identified as Dalbergia volubilis Roxb. This research article overcomes the lacuna and explores the anatomical, Deoxyribonucleic acid (DNA) barcoding and preliminary phytochemical analysis including High Performance Thin Layer Chromatography (HPTLC). Methods: After proper authentication, leaves were exposed to macroscopy, microscopy and micrometric analysis and physicochemical, qualitative, quantitative and HPTLC study as per of Ayurvedic Pharmacopoeia of India (API). DNA barcoding of the fresh tender leaves samples were carried out following standard methods. Results: Macroscopically leaves are compound, unipinnate, imparipinnate, rachis base pulvinus, leaflets 11-13. Microscopy of T. S. of rachis resembles anatomical structure of stem. Schematic outline of T. S. of petiolule is circular in shape with single layer epidermis is followed by ground tissue and centrally located vascular bundle. Schematic outline of T. S. of leaflet is bulged in the center and the extending arms of lamina are linear. Surface study reveals stomatal index to be 33.33. The genetic barcoding confirms the species D. volubilis. Loss on drying was found to be 8.78±0.57. HPTLC results shows 6 peaks and 7 peaks at 254 and 366 nm. Conclusion: D. volubilis, is a woody climbing unarmed inland shrub with stipules modified into hook and imparipinnately unipinnate compound leaves, are the key field identification character of the plant. Typical microscopical characteristic are presence of solitary crystal mostly embedded in sheath surrounding the vascular bundle and paracyctic type of stomata. Keywords: Dalbergia volubilis. DNA barcoding, Pharmacognosy, Anukta dravya © 2018 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) DOI: http://dx.doi.org/10.22159/ijpps.2018v10i6.26074 INTRODUCTION Medicinal plants used by traditional healers but not recorded in classical texts of Ayurveda are designated as Namatah Anukta dravya or extra-pharmacopoeial plants [1]. One of such folk plant is Biraskala, used by Tribals of Odisha. The leaves of this plant are used in aphthae [2] and also in heart pain, menorrhagia and post-partum problem [3]. The plant Biraskala is identified as Dalbergia volubilis Roxb. belonging to Fabaceae family. D. volubilis is woody climbing unarmed inland shrub with pinnatified leaves, leaflets ranging from 7-13 and pale blue flowers [4, 5]. Review of literature reveals that few work has been reported on Dalbergia genus which includes review on chemical constituents, ethanobotanical claims, pharmacology, biological activities [6, 7], Genetic barcoding of various other species of genus Dalbergia [8], pharmacognostical, phytochemical parameters of leaves of Dalbergia sissoo L [9] and anti-diabetic activity [10] has been reported. Though various parts of plants are used by tribals, the plant is not yet evaluated in a scientific way for its anatomical characters and phytochemical constituent. Hence the leaf of D. volubilis has been explored in this article to bring insight on the leaf anatomy, genetic barcoding and preliminary phytochemical constituents including High Performance Thin Layer Chromatography (HPTLC) profile. MATERIALS AND METHODS Chemical and reagents Chemicals utilized for pharmacognostical study and phytochemical study including HPTLC were procured from Rankem Mumbai and Analar India. Gallic acid standard was obtained from Loba chemia, Mumbai and Folin–Ciocalteu reagent was procured from SRL, India. All chemical used for analysis were of laboratory grade and chemical used for HPTLC were of analytical grade. Collection and authentication The plant samples were collected by the corresponding and second author from one of its natural habitat, Gandhamardana hills, Odisha, in month of September 2016 with help of local taxonomist. Herbarium was submitted to Pharmacognosy laboratory authentified by the Pharmacognosist of the institute and provided with herbarium reference no. Phm/6199/2016-17 for further reference. (fig. 1, A-D). Pharmacognostical study Macroscopic observations were made with naked eyes and centimeter scale (cm) was used to measure the leaf length and width. The leaves were washed and transverse sections were taken cleared with chloral hydrate to observe the anatomy of main rachis, petiolule and leaflet with help of Quasmo binocular compound microscope, India. Micro-photographs were taken with help of Kodak easy share C140, 8.2 megapixels 3x optical/5x digital zoom HD camera. For histochemical tests, the thick transverse sections of the leaf were exposed to Iodine (2 g iodine and 3 g potassium iodide in 100 ml water), Phloroglucinol (20 mg/ml alcohol) and Hydrochloric acid (6N) for observation of starch grain and lignified tissue. For powder microscopy, to obtain powder, shaded dried leaves were grounded by mechanical grinder and sieved through 80#. For micrometry, triplicate reading were recorded and mean value was taken into consideration along with standard deviation based on sample using Microsoft excel. The micrometric values are mentioned in micrometer scale (µm) [11]. International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 10, Issue 6, 2018