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International Journal of Universal Pharmacy and Bio 148 | P a g e International Standard Serial Number (ISSN): 2319-8141 International Journal of Universal Pharmacy and Bio Sciences 5(1): January-February 2016 INTERNATIONAL JOURNAL OF UNIVERSAL PHARMACY AND BIO SCIENCES IMPACT FACTOR 2.093*** ICV 5.13*** Pharmaceutical Sciences RESEARCH ARTICLE …………!!! MICROMORPHOLOGICAL, MICROMETRIC INCLUDING HISTOCHEMICAL EVALUATION OF LEAF MADHAVILATA (HIPTAGE BENGHALENSIS) LINN. (KURZ) Madhavi Patel1, Harisha C.R2. 1Lecturer, Dravyaguna Department, Indian institute of Ayurvedic pharmaceutical sciences, Gujarat Ayurved University, Jamnagar, Gujarat, India. 2Head, of Pharmacognosy, I.P.G.T. & R.A., Gujarat Ayurved University, Jamnagar, Gujarat, India. ABSTRACT KEYWORDS: Hiptage benghalensis Linn. (Kurz). belonging to family Hiptage benghalensis Malpighiaceae. Commonly known as Madhavilata is a plant used by Linn. (Kurz.), Leaf, the indigenous people for the treatment of wound and obesity in Madhavilata, Malpighiaceae. India. Till date there is no pharmacognostical scientific work has For Correspondence: been done properly on its leaves. In the present study detailed Madhavi Patel * pharmacognostical evaluation has been done. Results showed that Address: Lecturer, Dravyaguna presence of rosette, cluster crystals and starch grains in petiole, Department, Indian single layer of palisade parenchyma, annular, pitted, spiral vessels in institute of Ayurvedic leaf, stomatal index i.e. 39 in lower epidermis. Powder microscopy pharmaceutical sciences, Gujarat Ayurved of sun dried powder showed cluster and rosette crystals, tannin University, Jamnagar, content and oil globules. Gujarat, India. Full Text Available On www.ijupbs.com 149 | P a g e International Standard Serial Number (ISSN): 2319-8141 INTRODUCTION: The exploration of traditional knowledge for cures to common diseases is attractive, but also overwhelming. Hiptage benghalensis Linn. (Kurz). commonly known as ‘Madhavilata’ is an important medicinal plant distributed throughout India, predominantly in Western ghats, Konkan, Deccan, Kumaon, Assam, Nepal and Andaman island, chiefly in damp places up to an altitude of 2000m[1,2]. It is a stout, high-climbing liana or large shrub, with white or yellowish hairs on the stem. Its leaves are lanceolate to ovate-lanceolate and approximately 20 cm (8 in) long, and 9 cm (4 in) broad; petioles are up to 1 cm long. It has scandent branches up to 5 m (16 ft) high. Flowers produce fragrant borne in compact ten-to-thirty- flowered axillary racemes. The flowers are pink to white, with yellow marks. Fruits are samaras with three spreading, papery oblanceolate to elliptic wings, 2–5 cm long, and propagate via wind or by cuttings[3,4]. The leaves of H. benghalensis Linn. (Kurz) are used in treating skin diseases in Burma. In India, it is widely used to treat cough, asthma, leprosy and also to quench thirst[5,6]. Although there are some pharmacognostical studies carried out by previous scholars regarding different parts of the plant. But those studies were mainly focused on morphological and microscopical aspect of the plant in general terms. In the present study of micrometric evaluation was very much encouraging and rationale in its own term. MATERIAL AND METHODS: Collection & authentication of raw drug: The fresh leaf of. H. benghalensis Linn. (Kurz).collected from natural habitat at Indian Institute of Ayurvedic Pharmaceutical Sciences, Gujarat Ayurved University, Jamnagar, Gujarat. Plant identification The correct identity of the species and its family were confirmed by studying its morphological characters and authenticated by comparing them with the characters mentioned in various floras and text book of botany and by subject expert. Collected samples were made into herbarium wide no. PHM. 6144/15 preserved in Pharmacognosy department, I.P.G.T. & R.A., Gujarat Ayurved University. Processing and preservation Genuine and good quality materials, required for the present study, free from any filth and worm infection were cut and separated, washed, dried in sun light, shade and stored in air Full Text Available On www.ijupbs.com 150 | P a g e International Standard Serial Number (ISSN): 2319-8141 tight dried container. Coarse powder of sufficient quantity was prepared and stored in air tight container. The powder was prepared for powder microscopy. Pharmacognostical analysis: Morphological : Under the Morphological study, the basic structure; the types of formation of a particular organ & their arrangement in different parts of H. benghalensis Linn. (Kurz). leaf was studied in detail. These observations were noted properly and then matched with the data available in taxonomic book & with the flora[2,7 ]. Macroscopic: The collected sample was identified and authentified by studying their characters were studied systematically as per the methods described in the textbooks of Pharmacognosy. The specimen was observed as such with necked eyes. The samples were cleaned properly and macroscopic study of the different leaf parts was carried out. The individual character of the leaf like petiole and their macroscopic characters like size, shape, and colour were noted in detail [8-9]. (plate 1b) Organoleptic: Evaluation of the raw sample was done by their various characters like, colour, texture, odour, taste etc.[7] Microscopic: Free hand sections of the plant material observed under distilled water under the microscope for the presence of primary and secondary metabolites, like starch grains which were confirmed by staining them with iodine. The sections cleared with chloral hydrate to observe the various ergastic cell contents like, crystals of calcium oxalate, calcium carbonate, and silica if present any. The natures of these crystals were also confirmed by performing some tests like, solubility of them in acids (HCl). The sections then stained with Phloroglucinol and HCl for detecting lignified elements like fibres, sclereids, xylem vessels, tracheids etc Repeat the same method for the powder samples [9]. Histochemical evaluation: Sample thick sections subjected to Histochemical tests to find starch grains, tannin, calcium etc. by treating various reagents[8]. Leaf surface preparation To make thin leaf transparent, cut a pieces of leaf into two portions, turn one piece upside down, add a few drops of chloral hydrate, and heat to boil directly on a slide. As soon as Full Text Available On www.ijupbs.com 151 | P a g e International Standard Serial Number (ISSN): 2319-8141 bubbles start to escape, remove the slide from the flame. When bubbles stop to appear, heat until the fragments are transparent. Add 1 or 2 drops of chloral hydrate solution, apply cover glass and observe under microscope. For thicker leaves that do not become transparent easily when prepared by this method clear by boiling with strong chloral hydrate solution in test tube, transfer a fragment onto a slide, cut into two portions, scrape the surface of the fragment by scalpel. Wash the epidermis by chloral hydrate solution. add 1 or 2 drops of chloral hydrate solution, cover it with a cover glass and observe under microscope[9]. Powder characters study method: For powder microscopy, slides were prepared by using water, chloral hydrate as a clearing agent, stained with phloroglucinol and conc. HCl for lignified tissues, iodine for starch grains and glycerin as mountant[8]. Micrometric evaluation: Under the micrometric study, the measurement of a particular character or a particular band/zone which was made up of similar kind of tissues/cells was taken into consideration, which was otherwise not possible to make them accountable aided with our naked eyes. These characters are microscopic in nature & when they are labelled with their measurement, those characters can be very effective tools to standardize a particular drug due to their specificity & preciseness[9]. RESULTS Macroscopic study Under the macroscopic study shape, size & color of the different parts of the leaf was measured & noted appropriately. Leaf is simple and the shape of the leaf was lanceolate to ovate. Margin entire, slightly wavy, apex acuminate, upper surface of leaf is dark green and glossy while lower surface is light in colour, venation alternate, veinlet is net like structure. Then the observations were documented in table no.1. Macroscopic study has been demonstrated in the fig (plate 1a-c). Microscopic study Different parts of the leaf like petiol, leaf, midrib were separated then transverse section of each individual part was studied thoroughly. The results obtained from transverse sections of various parts of the leaf were described below. Full Text Available On www.ijupbs.com 152 | P a g e International Standard Serial Number (ISSN): 2319-8141 Transverse section of Petiol : Petiol had multicellular, warty, covering trichome measuring 4.5mm in length & 0.5mm in width. Barrel shaped unicellular epidermis with thin cuticle. Epidemis was interrupted by both warty trichome, rarely interrupted by stomata. The total area of petiol was of 7X8mm in 40X. Rossate and cluster crystals of calcium oxalate embedded in cortical region and in epidermal layer. Prismatic crystal of calcium oxalate rarely seen in epidermal layer of the petiol. Below the epidermal layer single endodermal layer rich in starch grains. Cortex is embedded beneath the endodermis 2X2 mm in 40X. Two meristele with one major vascular bundle. 6X4mm in 40X Vascular bundle consists of radially arranged pericyclic fibre, parenchyma of xylem, uniserriate medullary rays. (plate. 2a,b) Localization: Tannin- epidermis,
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