PHARMA SCIENCE MONITOR AN INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES

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1 PHARMA SCIENCE MONITOR AN INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES PHARMACOGNOSTIC AND PHYTOCHEMICAL EVALUATIONS OF THE LEAVES OF HOLOPTELIA INTEGRIFOLIA ROXB. Dharmeshkumar Prajapati* and Natvarlal M. Patel Shri B M Shah College of Pharmaceutical Education &Research, College campus, Dhansura road, Modasa , Sabarkantha, Gujarat,India ABSTRACT Holoptelea integrifolia belongs to the family of Urticaceae. The bark and leaves are bitter, astringent, anthelmintic, diabetes, helminthiasis, skin disease, intestinal disorder, leprosy, rheumatism. Evaluation of the fresh, powdered and anatomical sections of the Holoptelia integrifolia leaves were carried out to determine the macromorphological, micromorphological, numerical and phytochemical profiles. Macroscopic studies indicated presence of the leaf of having cms and cms in size, apex is acute, oval in shape, entire margin, cordate base, reticulate pinnate types of venation, petiole is long, surface is glabrous, upper epidermis dark green in color, lower epidermis lighter in color with characteristic odour and bitter taste. In the microscopic studies, the leaves showed the presence of trichomes, collenchyma, vascular bundles, spongy parenchyma, palisade cells and stomata. Phytochemical evaluation revealed the presence of alkaloids, flavonoids, tannins, saponins, terpenoids, glycosides and absence of steroids and anthraquinones. The investigations also included leaf surface data quantitative leaf microscopy and fluoroscence analysis. The results of the study could be useful in setting some diagnostic indices for the identification and preparation of a monograph of the plant. Key words: Holoptelea integrifolia, Monograph, Microscopy. INTRODUCTION A medium sized to large glabrous deciduous tree, m in height with whitish or yellowish grey bark exfoliating in irregular flakes and with an offensive smell when freshly cut [1]. The bark and leaves are bitter, astringent, anthelmintic, diabetes, helminthiasis, skin disease, intestinal disorder, leprosy, rheumatism. The seeds and stem bark are used externally to treat ringworms [2,3]. Some drugs of plant origin in conventional medical practice are not pure compounds but direct extracts or plant materials that have been suitably prepared and standardized. The World Health 34

2 Organisation (WHO) has recommended the use of arthemisinin derivatives from Artemisia annua (Composite), a Chinese herb with established pharmacognostic data, as a first line drug in the treatment of malaria [4,5,6].Establishment of the pharmacognostic profile of the leaves of Holoptelea integrifolia will assist in standardization, which can guarantee quality, purity and identification of samples. MATERIALS & METHODS Collection of Plant materials Fresh leaves of Holoptelea integrifolia were collected from M.L.Gandhi Higher Education Society of college campus in Modasa City. Identification and confirmation were done by Dr.Jahagir, Botanist, Sir P.T.Scince College, Modasa. Where voucher specimens were deposited with the number, 08/BMCPER/PH. Macroscopy The following macroscopic characters for the fresh leaves were noted: size and shape, colour, surfaces, venation, presence or absence of petiole, the apex, margin, base, lamina, texture, odour and taste [7,8] Microscopy The outer epidermal membranous layer (in fragments) were cleared in chloral hydrate, stained with phloroglucinol and concentrated HCl and mounted with glycerin and observed under a compound microscope. The presence / absence of the following were observed: epidermal cells, stomata and epidermal hairs. The transverse sections of the fresh leaves through the lamina and the midrib as well as a small quantity of the powdered leaves were also cleared, mounted and observed.powder microscopy done with standard procedure [9]. Quantitative investigation Quantitative leaf microscopy to determine palisade ratio, stomata number, stomata index, vein islet number and veinlet termination number were carried out on epidermal strips. Other parameters determined for the powdered leaves were moisture content, total ash, acid insoluble ash, water soluble ash, alcohol (90 % ethanol) and water soluble extractive values [10]. Fluoroscence analysis of powder done by standard procedure [11]. 35

3 Phytochemical investigation Chemical tests were employed in the preliminary phytochemical screening for alcoholic extract. various secondary metabolites such as tannins, cardiac glycosides, alkaloids, Saponin glycosides, anthracene derivatives and Cyanogenetic glycosides detected by chemical test [12,13,14]. RESULTS Morphological features shows that leaf of Holoptelea integrifolia having cms and cms in size, apex is acute, oval in shape, entire margin, cordate base, reticulate pinnate types of venation, petiole is long, surface is glabrous, upper epidermis dark green in color, lower epidermis lighter in color with characteristic odour and bitter taste. TABLE 1: MORPHOLOGICAL CHARACTERS OF LEAF OF H. INTEGRIFOLIA ROXB. Morphological Observation parameter Condition Fresh Type simple or compound Size: Length Width cms cms Shape Oval Apex Acute Margin Entire Venation Reticulate pinnate Base Cordate Petiole Long Surface Glabrous Phyllotaxy Opposite Color: Outer Dark green in color Inner light green in colour Odour Characteristic Taste Bitter 36

Microscopical features revealed that upper and lower epidermis of thin walled polygonal cells with wavy margin.uniseriate covering and multicelular trichomes are present on both surfaces.

4 Microscopical features revealed that upper and lower epidermis of thin walled polygonal cells with wavy margin.uniseriate covering and multicelular trichomes are present on both surfaces. After epidermis chlolenchyma present. It is differentiated in to upper Palisade and lower spongy tissue. Transverse section of the leaf across the midrib shows xylem towards lower epidermal cells, which are highly lignified. Phloem towards lower epidermis.powder microscopic examination of the leaves revealed the presence of covering trichome, stomata, xylem vessel and epidermal cell. A-Upper Epidermis B- Palisade cell C- Trichome D- Multicellular Trichome E- Lower Epidermis F- Collenchyma G Meta xylem H- Phloem I- Xylem J - Collenchyma The numerical and quantitative values are presented in tables 2 and 3. The flouroscence analysis of powder shown in table no

5 TABLE 2: LEAF SURFACE DATA OF LEAVES OF H. INTEGRIFOLIA ROXB. Sr.No. Parameter Physicochemical constant 01 Total ash % 02 Acid insoluble ash % 03 Water soluble ash % 04 Sulphated ash 14.0 % 05 Loss on drying % 06 Alcohol soluble extractive value 39.5 % w/w 07 Aqueous extractive value 19.3 % w/w TABLE 3: QUANTITATIVE LEAF MICROSCOPY OF H. INTEGRIFOLIA ROXB. Sr.No. Parameter Range 01 Palisade ratio Stomata number Upper surface Stomata number Lower surface Stomata index Upper surface Stomata index Lower surface Vein islet number Veinlet termination number TABLE 4: FLUORESCENCE ANALYSIS OF POWDER OF H. INTEGRIFOLIA ROXB. Treatment H.integrifolia leaves UV short (254 nm) UV long (365 nm) As such Green Dark green Methanol Greyish green Green Methanol:NaOH(1:1) Green yellow Nil Conc.H 2 SO 4 Dark green Green black 38

6 Phytochemical investigation revealed the presence of alkaloids, glycosides, carbohydrates, steroids and flavonoids. These secondary plant metabolites are known to possess various pharmacological effects and may be responsible for the various actions of Holoptelea integrifolia. DISCUSSION Ethnomedically, the leaves and stem bark of this plant were used by local people for in the treatment of various disease conditions without standardization. The standardization of a crude drug is an integral part of establishing its correct identity. Before any crude drug can be included in a herbal pharmacopoeia, pharmacognostic parameters and standards must be established. Holoptelea integrifolia is a plant that has been confused with other species due to their relative similarities. The results of these investigations could, therefore, serve as a basis for proper identification, collection and investigation of the plant. The macro and micro morphological features of the leaf described, distinguishes it from other members of the genera. Leaf surface data, fluorescence analysis and quantitative leaf microscopy are parameters that are unique to the plant and are required in its standardization. CONCLUSION These parameters, which are being reported for the first time, could be useful in the preparation of the herbal section of Indian Herbal Pharmacopoeia. REFERENCES 1. Anonymous: The useful plants of India, Council of Scientific and Industrial Research, New Delhi, Publication and Information Directorate, CSIR, 1992:271, Prajapati N,Purohit SS, Sharma AK: A hand book of medicinal plants,agrobios publication, Jodhpur,2007: Prajapati N, Kumar U:Agro dictionary of medicinal plants, Agrobios publication, Jodhpur, 2003:60, Donald EC:Medicinal plant research in Nigeria: Retrospect and Prospect. In: Sofowora (Ed). The state of medicinal plants research in Nigeria,Nig. Soc. of Pharmacognosy, Ibadan University press, Nigeria, 1986: World Health Organisation (WHO):Antimalarials Drug Combination Therapy. Report of a WHO Technical Consultation. April 2001:

7 6. World Health Organisation (WHO). WHO urges Countries to act on New AntiResistance Malaria Medicines. Press release, WHO/31, April 2002: Trease GE, Evans WC:Trease and Evans Pharmacognosy: Harcourt brace & Co. Asia, Pvt. Ltd. and W.B. Saunders Company Ltd., Edition 15, 2002:32, 33, 95-99,512, Wallis TE: Textbook of Pharmacognosy, Published by SK Jain,1985: African Pharmacopoeia.General methods for Analysis.OAU/STScientific Publications, Lagos, vol.2.no. 21 st Ed. 1986: 1 5, , British Pharmacopoeia 11: Ash values, Acid insoluble Ash, Water soluble, Extractive and Alcohol soluble extractive. Appendix XI. Her Majesty s Stationery Office, London, 1980:A108, A Edwin S,Joshi SB,Jain DC:Comparative pharmacognostic studies on root powder of plumbago zeylanica and plumbago rosea. Indian journal of natural products,vol.xxiv,june,2008:(2), Brain KR and Turner TD. Practical evaluation of phytopharmaceuticals Wright Scientechnica, Bristol. 1 st Ed.: 1975, Ciulei I. Methodology for analysis of vegetable drugs. UNIDO Romania, 1981: Harborne JB. Phytochemical methods. A guide to modern technique of plant analysis. Chapman and Hill, London, 1992: 279. For Correspondence: Dharmeshkumar Prajapati Shri B M Shah College of Pharmaceutical Education &Research, College campus, Dhansura road, Modasa , Sabarkantha, Gujarat, India dharmesh09mpc@yahoo.co.in Cell:

Asian Journal of Biochemical and Pharmaceutical Research

ISSN: 2231-2560 Research Article Asian Journal of Biochemical and Pharmaceutical Research Pharmacognostical and Physico-chemical Standardization of Leaves of Sida acuta *Carol P. Macwan, A. J. Patel, M.