COMPARATIVE PHARMACOGNOSTICAL AND PHYTOCHEMICAL STUDY OF VARIOUS SPECIES OF THE CONTROVERSIAL DRUG ‘PUNARNAVA' Original Article BHAVIKA R VALA, KHYATI R BHALARA, DEVANG J PANDYA* School of Pharmacy, RK University Email: devang.pandya@rku.ac.in Received: 21 Sep 2015 Revised and Accepted: 25 Nov 2015 ABSTRACT Objective: Punarnava is a controversial drug i.e. its true botanical identity has not been ascertained, and several different species are used as Punarnava. It is a part of many medicinally important traditional formulations. Ayurvedic Pharmacopoeia of India describes three different species– Roots of Boerhaavia diffusa, Trianthema portulacastrum and Boerhaavia verticillata. The present work deals with the microscopic, macroscopic and phytochemical comparison of the roots of these species of Punarnava, so as to differentiate them and establish their quality parameters. Methods: Roots of these three species were collected, and their comparative morphological, microscopical (transverse section, powder study) and the phytochemical (screening, tannin estimation) study was performed. Results: Roots of Boerhaavia diffusa showed morphological characteristics which clearly distinguished it from the other species. They showed the presence of well developed and stratified cork, abnormal vascular bundles which were discontinuous, and a central cavity, presumably formed due to the disintegration of tissues. Boerhaavia verticillata showed the presence of xylem bundles arranged in the form of a ‘X’-shaped cross in the central region. Thick-walled cork cells and pitted xylem vessels were present only in Boerhaavia diffusa root powder, whereas bundles of fibrovascular tissue were present only in the powder of Boerhaavia verticillata. Tannin content was found to be highest in Boerhaavia diffusa roots. Conclusion: The present investigation will help herbal industries and traditional medicinal practitioners to detect adulteration of the medicinally important immunomodulator Boerhaavia diffusa by Trianthema portulacastrum and Boerhaavia verticillata, which are considered to have no immunomodulatory activity. Keywords: Boerhaavia diffusa, Boerhaavia verticillata, Controversial drug, Immunomodulator, Microscopy, Nyctaginaceae, Punarnava, Quality Control, Standardization, Trianthema portulacastrum © 2016 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/ ) INTRODUCTION Punarnava is a ‘controversial drug’ because its true botanical identity has not been ascertained yet. Various species are known by the name of ‘Punarnava’. Ayurvedic Pharmacopoeia of India describes three species–Roots of Boerhaavia diffusa, Trianthema portulaca strum and Boerhaavia verticillata. Roots of Boerhaavia diffusa (B. diffusa) syn. Boerhaavia repens, Boerhaavia procumbens (Family–Nyctaginaceae, also known as Rakta Punarnava) are traditionally used as an immunomodulator, adaptogenic, diuretic, anti-arthritic, analgesic, anti-fibrinolytic, blood purifier, hypotensive, hepatoprotective, anti-anemic, antioxidant, anti lymphoproliferative and in edema and dropsy [1, 2]. No work has been done so far to distinguish the three species from each other, which is crucial for ascertaining the actual species of ‘Punarnava’ and for its quality control. The present investigation deals with pharmacognostic and phytochemicals evaluation of roots of the three species, thereby setting parameters which will help herbal industries to detect adulteration of B. diffusa by T. portulaca strum and B. verticillata, thereby ultimately benefiting the patients. MATERIALS AND METHODS Dried roots of B. diffusa were obtained from Yucca Enterprise, Mumbai, whereas roots of T. portulacastrum and B. verticillata were collected from Punarnava roots available in the market are normally the roots of B. diffusa. Roots of Trianthema portulaca strum (T. portulaca strum) syn. Trianthema monogyna (Family–Ficoidaceace, also known as Varshabhu, Shwet Punarnava) are used traditionally as antipyretic, analgesic, spasmolytic, deobstruent, cathartic, anti-inflammatory, in diseases of liver and spleen, anaemia and edema [3, 4]. Roots of B. verticillata (Family–Nyctaginaceae, also known as Vrischiva, Shwet Punarnava) are used traditionally as digestive, appetizer, antipyretic, the antidote for poisonous bites of insects, anti-leprotic, anti-cough and in disorders of the heart, spleen and gastrointestinal tract [5, 6]. Roots of T. portulacastrum and B. verticillata are rarely available in the market, and none of them being immunomodulators, are considered to be adulterants of B. diffusa. the herbal garden of School of Pharmacy, RK University (22 ° 14' 27.1284'' N, 70 ° 54' 3.4308'' E) in flowering season in July, 2014. The samples were authenticated were authenticated by Dr. Kunjal Soni, Botanist, School of Science, RK University, and deposited in the institute repository Roots of B. diffusa were cylindrical, deep brown, thick, 6-8 cm X 0.5- 1 cm, with a rough surface and no lateral roots. Roots of T. portulacastrum were also cylindrical, creamish-white, very thin, 4-5 cm X 0.1-0.2 cm, with a smooth surface and many hairy secondary rootlets. Roots of B. verticillata were cylindrical, creamish-white, very thin, 6-10 cm X 0.1-0.4 cm, with a smooth surface and many hairy secondary rootlets (fig. 1). . The roots were compared morphologically and used for transverse section study. The roots were dried, powdered, stored in airtight containers and used for phytochemical studies. For microscopical studies, safranin was used for staining [7]. Photomicrography of the transverse sections and the powdered drug was performed using the camera. Phytochemical screening of both the species was performed using the appropriate extracts (aqueous and alcoholic) (table 2) [8-12]. Total tannins were estimated in all species [13]. RESULTS Macroscopy International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 8, Issue 1, 2016