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Pharmacognostical Evaluation of Root of Gumma (Leucas Cephalotes Spreng.)

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Pharmacognostical Evaluation of Root of Gumma (Leucas Cephalotes Spreng.) Indian Journal of Natural Products and Resources Vol. 4(1), March 2013, pp. 88-95 Pharmacognostical evaluation of root of Gumma (Leucas cephalotes Spreng.) Mohammad Yusuf Ansari1, Abdul Wadud1*, Ehteshamudddin1 and Hamida Bano2 1Department of Ilmul Advia, National Institute of Unani Medicine, Kottigepalya, Magadi Main Road, Bangalore-560 091, Karnataka, India 2HAH Unani Medical College, Idgah Road, Dewas, Madhya Pradesh, India Received 20 June 2011; Accepted 6 August 2012 Gumma (Leucas cephalotes Spreng.) belonging to family Lamiaceae, primarily a folk drug, is also used in Ayurveda and Unani medicine in India and in adjacent countries for its varied therapeutic properties such as stimulant, diaphoretic, antiseptic, laxative, anthelminthic, insecticidal, germicidal, fungicidal, emmenogogue, expectorant and antipyretic. Its root is particularly useful in regularization of menstrual cycle, tuberculosis and dysentery. In view of insufficient data necessary to set up proper identification of the whole plant and its roots and to provide referential information for checking adulteration and substitution, root of this plant has been evaluated on pharmacognostical parameters by means of anatomical, physico-chemical and preliminary phytochemical studies, HPLC and UV-Vis spectrophotometry. Keywords: Gumma, Leucas cephalotes, Anatomical studies, Physico-chemical studies, HPLC, Spectrophotometery. IPC code;Int. cl. (2011.01)—A61K 36/00 Introduction some physical and chemical operations on the drug In spite of the reality that herbal drugs can be sample prior to the actual analysis. A blend of safely used only if their standards i.e. safety, efficacy classical methods and latest analytical techniques are and quality are up to the mark, these important values advantageous in the study of herbal drugs. have not been looked up appropriately. In view of the Gumma (Leucas cephalotes Spreng.) belonging growing interest in herbal drugs that has called for to the family Lamiaceae is an annual herb and greater exactitude in appraisal, it becomes crucial to an upland rainy season weed commonly found ascertain standard samples of single herbal drugs for in roadsides, meadows, waste lands and cultivated future reference1. The rapid development in different grounds throughout the greater part of India4-12 aspects concerning to crude herbal drugs, in recent (Plate 1). The plant as a whole and its various years, has necessitated a systematic approach to parts alone are used medicinally. It possesses stimulant, evaluate these drugs in modern pharmacognosy with a laxative13, diaphoretic14, antiseptic15, antihelmenthic16, methodical approach to qualitative and quantitative insecticidal17,18, germicidal19, fungicidal20, emmenogogue21, evaluation by means of appropriate methods as per expectorant and antipyretic22 properties. Powdered WHO guidelines2,3. root is used as one of the ingredient of compound Analytical instruments play important role in the remedy to treat tuberculosis. To treat dysentery it is production and evaluation of new products in the given with edible oil once daily23. Root of Gumma safety of consumers and the environment. The use of with Chota Kulpha [Trichodesma indicum (L.) R. Br.] sophisticated instruments, in present days, is a fascinating part of chemical analysis. Though, it is necessary to use several instrumental techniques to obtain the information required to solve analytical problems still worth of classical methods can’t be underestimated. Therefore, it is necessary to perform —————— *Correspondent author : E-mail: [email protected] Phone: 080-23584260 Fax: 080-23580725 Plate 1—a. Whole plant, b. Root ‘ ANSARI et al: PHARMACOGNOSTICAL EVALUATION OF ROOT OF GUMMA 89 and Kali Musli (Curculigo orchioides Gaertn.) is used Microscopic studies for the treatment of menorrhagia. Use of powder of Transverse section (T.S.) of the root was prepared 29 root with rice water regularizes menses23. Some recent according to the method described by Johnson . ethno pharmacological studies have shown the plant Small piece of the material was kept in a vial and to be hypoglycaemic, hypolipidaemic, antioxidant24, fixed in 50 % ethanol (v/v). After two days, washed anti-inflammatory25, antiprotozoal26 and hepatoprotective27. with the same fluid and dehydrated with ascending The whole plant reported to contain new labdane-, series of ethanol (50, 70, 95 and 100%), dipped in norlabdane- and abietane-type diterpenes named each series for about 2 h. Thereafter, passed through leucasdins A (1), B (2) and C (3), respectively, and the descending series of absolute alcohol and xylol two protostane-type triterpenes named leucastrins (75:25, 50:50; 25:75) and finally washed with xylol. A (4) and B (5) were isolated, together with a In xylol the infiltration was carried out by gradual known triterpene, oleanolic acid, five sterols, addition of paraffin wax at room temperature till the 7-oxositosterol, 7-oxostigmasterol, 7α-hydroxysitosterol, xylol was saturated. The vials were kept in an oven at o o 7α-hydroxystigmasterol and stigmasterol, and eight 35 C + 2 C, the contents were occasionally stirred and flavones, 5-hydroxy-7,4'-dimethoxyflavone, pillion, wax was added till the solution saturated. After 24 h, o o gonzalitosin I, tricin, cosmosin, apigenin 7-O-β-D- the temperature of the oven was raised to 60 C + 2 C, (6-O-p-coumaroyl) glucopyranoside, anisofolin A and and the addition of wax was continued till the smell of luteolin 4'-O-β-D-glucuronopyranoside28. xylol vanished. The mixture of xylol and wax was Though this plant possesses a number of poured off after three hours and pure melted wax was therapeutic properties, very little description is added. The vials were left inside the oven for 24 h and available in classical Unani literature and that too the material was tested for complete infiltration by compiled in Indian subcontinent. Also, it has not been cutting with a safety razor. The contents were poured studied scientifically to a great extent to support the into a dish containing melted wax and materials identification of genuine sample. Keeping these points arranged in rows. The dish was dipped into beaker in view present work was undertaken to study containing cold water. The rectangular blocks were pharmacognosy of root of Gumma. made after trimming suitable size. Soft wax applied on the both the surfaces and then fixed in block Materials and Methods holder. For sections the block was fixed in the block Collection and authentication of the plant holder which was fixed in clamp of microtome. The The fresh plant was collected from the forest of microscale of microtome was set at 10 micron. The Satpura range of Burhanpur (M P), India in the month wheel was moved with steady even stokes. The first of July. The plant was identified and authenticated by few sections were discarded until a ribbon was made. botanists of National Ayurveda Dietetic Research The slide was placed on the table and flooded Institute, Bangalore vide authentication certificate no. immediately with 03% formalin in distilled water. Drug Authentication/SMPU/NADRI/ BNG/2009-10/896. The ribbon was then kept on the slide with the help of Fresh material was used for anatomical studies a wet scalpel, and the slide was put on a warm plate. whereas shade dried material was powdered in The temperature was not allowed to exceed 43°C. electric grinder for physico-chemical, phytochemical, After few minutes the slide was removed and kept to HPLC and spectrophotometric studies. cool up to room temperature. The water was drained Preparation of extracts off by tilting the slide. After drying the slide, it was The study was carried out in the pharmacognosy placed in xylol for 5 minutes. The slides were then laboratory and Central Instrumentation Facility withdrawn slowly and transferred to a coupling jar, laboratory (CIF) of National Institute of Unani containing equal volumes of absolute alcohol and Medicine (NIUM). Coarse powder of air dried root xylol for five minutes then into 75:25 ethanol and (100 g) was subjected to Soxhlet apparatus for 8 h for xylol solution and finally in absolute alcohol solution continuous hot extraction with distilled water, later transferred to 95, 70, and 50% ethanol methanol, acetone, diethyl ether, petroleum ether and successively in each for five minutes. The section chloroform, separately. The extracts were filtered and were then kept in saffranin (in 50% ethanol) for five the filtrate was evaporated to dryness. The percentage minutes and transferred to 50, 70, 95 and 100% yield of each solvent was calculated with reference to ethanol in ascending order keeping each for the air dried drug. 2-3 minutes. The slides were again transferred to 90 INDIAN J NAT PROD RESOUR, MARCH 2013 clove oil and ethanol mixture (75:25) then in fast 0.5 mL/minutes using methanol: water (70:30) as green prepared in clove oil and xylol mixture (75:25) mobile phase solvent, under a pressure of 100 f/sq cm, for 2 minutes in each. It was washed through run time of 10 minute and an injection volume of xylol and finally mounted in DPX mountant. 20 µL. at 240, 205, 254, and 238 nm. Analyst The section observed under microscope with 1.4 software was used to control all the parameters. different magnifications i.e. 50, 100 and 450. Photomicrographs were taken by using digital camera Spectrophotometry (Sony, Japan) attached to microscope. Various cells Spectrum scan curves of aqueous and methanol were measured (micrometry) with the help of extracts of root were obtained by using UV-Vis micrometer (stage micrometer and ocular micrometer) Spectrophotometer 3000 (Labindia) at room under 100 magnifications by the method of Trease temperature according to the method given in the and Evans. Isolated elements were also studied by instruction manual. After pre-heat time, UV-Vis reported method of Trease and Evans 30. spectrophotometer was assessed to spectrum scanning mode. The parameters were set, the photometric Physico-chemical studies mode was assessed to Abs, scanning speed was For physico-chemical studies, ash and extractive set as middle and the wave length range was set to values and pH were measured by the method 190-660 nm.
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