Pharmacognostical Study of a Medicinal Plant of India – Sarcostemma acidum

Available online on www.ijppr.com International Journal of Pharmacognosy and Phytochemical Research 2014-15; 6(4); 690-697 ISSN: 0975-4873 Research Article

Pharmacognostical Study of a Medicinal Plant of India – Sarcostemma acidum *Bhavesh Kumar Dave1, Ronak Dhirawat1, Mukesh Kumawat2 Bhupal Nobles’ College of Pharmacy, Udaipur-313001 Lachoo Memorial College of Science and Technology, Jodhpur 1

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Available Online: 17th November, 2014 ABSTRACT Sarcostemma acidum is widely used by the folk people in the treatment of verity of diseases and disorders. The present study was therefore carried out to provide requisite pharmacognostic details about the plant. Methods: macroscopy, microscopy, physicochemical analysis, preliminary phytochemical screening and other WHO recommended parameters for standardization were performed. Result: A perennial leafless, jointed shrub with green, cylindrical, fleshy glabrous, twining branches having milky white latex, taste bitter; Flower white or pale greenish white with fragrant odour. The microscopy reveals dicotyledonous plant having circular vascular bundles in stem, absence of pith in root and anomocytic type of stomata on stem bark. Other physicochemical tests like moisture content, ash value, extractive values were determined and Preliminary phytochemical screening showed presences of carbohydrates, triterpenoids, phytosterols, saponins and lignins. Extraction of chemical constituents and chromatographically analysis (TLC) were performed. Conclusions: The microscopic using histological identification, microscopic constants, and other physico chemical examinations of Sarcostemma acidum can be used as a rapid, inexpensive and botanical identification technique is useful in standardization, hence would be of immense value in authentication of plan INTRODUCTION Sarcostemma acidum is an Indian traditional medicinal plant. It has been categorized as candidate of Soma plants by various authors. It was said that Soma (Somlata) was used to prepare ‘Som ras’ (Rejuvenating drink) by Aryans. The original source of 'Soma' plant is a mystery that has been debated by the vedic and botanical scholars for more than two & a half centuries.1 It is found wildly in India, Pakistan and Europe etc. It is distributed in various parts of India. It is found in dry rocky places in Bihar, Bengal, Konkan, Deccan, Tamil Nadu, Maharashtra, Madhya Pradesh and Kerala. The plant is bitter, acrid, cooling, alternate, narcotic, emetic, antiviral and rejuvenating. Chemical constituents - Malic acid, Succinic acid, Reducing sugar – sucrose, Traces of tannin, Alkaloids, Phytosterols, Alpha & beta amyrins, Lupeol & lupeol acetate, Beta sitosterol2,3. Chemical constituents of Sarcostimma acidum growing in China – Lignans ¬Sacidumlignan-A, Sacidumlignan-B, Sacidumlignan-C, Sacidumlignan-D, Degraded lignans derivativeSacidumol-A, Sacidumol-B, (+)- Pinoresinol, 9 alpha hydroxyl pinoresinol ,Perforatic acid, Peucenin -7 O methyl ether4. Sarcidumitol (2,6 dideoxy- disacchariditol)5 Antifertility, anti-microbial and in vitro antiinflammatory6,7,8 activities have been reported on this plant. Traditional uses in different areas of India  Latex is applied on wounds and cuts in Bidhar district of Karnataka.9

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 Three drops with honey taken orally thrice a day for chronic ulcer in Andra Pradesh Sirumalai Hills10  A dry powder of the plant is being used in the form of decoction to treat earache in babies. Milky latex is prescribed to the lactating mother in Purulia district of West Bangal.11  In digestive disorder, scabies, skin disease, poisoning, cardiac disease, fever, cold, thirst Mental disorders and dog bite in Karnataka12  The plant stem is used to cure bone fracture in Madhya Pradesh.13  Stem juice mixed with water is given in rheumatism, arthritis, and joints pain. Dry powder with Mustard oil is applied externally for treating earache in Nimar region of Madhya Pradesh14  Stem juice is used as ear drops during earache, for treating mad dog bite - extraction of stem and grass of garika (cynodon dacrylon) with turmeric is applied and tied with bandage over the bitten area twice for two weeks. Milky latex used as lotion by the ethnic people of Krishna district, Andhra Pradesh.15  Roots in snake bite and taken as an infusion in dog bite cases, latex to kill white ants (termites); dry stem as emetic in Thar Desert.16  Used as eye drops in Nallamala forests of Andra Pradesh.17  Water solution of pulpy mesocarp given as nasal drops (externally remedy) in epilepsy by yanadis community in Cuddapah district, Andra Pradesh.18

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Fig. 2: Flower parts, Flower diagram and Flower formula of Sarcostemma acidum

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Fig. 1: Plant and Flowers of Sarcostemma acidum

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Fig. 3: Transverse section of Sarcostemma acidum stem (Magnification-100x)

Extraction of chemical constituents and chromatography: According to preliminary phytochemical analysis, chemical constituents were extracted and performed thin layer chromatography. Extraction process: The powered drug was homogenized with methanol : water (4 : 1) for 30 minutes and filtered, the filtrate was acidifed (dilute sulphuric acid) and extracted exhaustively with chloroform then got two layer (acidic & chloroform) after 10 minutes the chloroform layer was separated with separating funnel and examined under thin layer chromatography(TLC)21. Solvent system: Firstly, TLC was performed in pure solvents based on trial and error method. Then different combinations of solvent systems were tried for best separation of constituents. By optimization of the solvent system, chloroform: methanol (9:1) was found the best system for the resolution of various component.

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MATERIAL AND METHOD Collection, identification and authentication of specimen: The plant specimen for the proposed study was collected from forest of Kailashpuri near Udaipuir & authenticated by Dr. Satish Kumar Sharma, assistant conservator of forest & wildlife – Fatahpura, Udaipur. Macroscopy: The collected plant was washed, cleaned and dried for further use. The following macroscopic characters of the fresh plant (stem, root and flower) were noted; color, odor, taste, size and shape and surface. Microscopicy: The free hand thin transverse section of stem, root and longitudinal section of bark of stem were treated with different staining agent and observed for general and specific microscopic characteristics.

Physicochemical parameter: The various physicochemical parameters such as total ash, acid insoluble ash, water soluble ash, moisture content, extractive value (water, alcohol, ether & decoction) have been studied20. Preliminary phytochemical analysis: For the preliminary phytochemical analysis, 10 gm of powdered stems was extracted with ethanol, chloroform & ether successively using soxhlet apparatus and the aqueous extract was prepared by decoction technique. The extracts were concentrated under vaccum rotary evaporator, dried and weight. Each extract was tested for presence of different phytochemicals20.

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Table 1: Physicochemical Parameters of Sarcostemma acidum stem Physicochemical Parameters (% W/W) Moisture Content 80.23 Ash Value Total ash 12.125 Acid insoluble 4.5077 Water soluble 1.2595 Extraction Value Alcohol soluble 7.424 Water soluble 12.4 Ether soluble 3.784 Decoction 13.66  100 ml of stem bark juice is given to cattle once in a day for 5–6 days as Galactagogue in Sariska region of Rajasthan19.

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Fig. 4: Transverse section of Sarcostemma acidum stem on various magnification levels Observation and interpretation of chromatogram: The white latex, leaves reduced to scales, opposite, color spots produced were observed in daylight, under short and green, shape - cylindrical, length – 2 to 4 meter, diameter long wavelength of ultraviolet light. RF values of spots of stem 0.5 to1cm; Root- depth only 5 to 8 inch with 3 to were calculated. 5 sub root branches, color- brownish; Flowersactinomorphic, color- white or pale greenish white, odorfragnant, in umbels on branch extremities, diameter- 0.8 to RESULT Macroscopy: It is a perennial leafless, jointed trailing 1cm. sepals(5), petals(5) attached with androceium shrub, fleshy glabrous, twining branches having milk (stamens-5), gyanoceium (overy-2), androcium and

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Fig.5: Transverse section of root of Sarcostemma acidum (100x & 400x)

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Stem- Transverse section of Sarcostemma acidum showed

Fig. 6: Longitudinal section of bark of Sarcostemma acidum (100x, 400x &1000x). Table 2: Phytochemicals present in different extractions of Sarcostemma acidum stem Phytochemicals Extractions Decoction Ethanol Chloroform Carbohydrates + + Phytosterols + + + Terpines + + + Saponins + + Lignins Present in T S of plant stem

Ether + + -

Solute run(cm) 6.0 7.50 9.6

RF value(cm) 0.48 0.60 0.77

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Color of Spots Blackish green Light yellow Brownish pink

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Table 3: Chromatographic study of Sarcostemma acidum stem S.No. Extraction Solvent System Solvent run(cm) 1 Isolated Chloroform : 2 chloroformic Methanol 12.4 3 extract (9 : 1)

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In U.V. light In Day light Fig. 7: Thin Layer Chromatographic plate with color spots of solute run gynocium are attached with the help of stigma (5) Figure three parts 1. Outer layer 2. Cortex 3. Vascular bundles. - 1 to 2. Outer most layer of stem (Epidermis) was seen as a thick Microscopy : Plant stem, root and bark were studied single cell layer. Collenchymal cells in 2 to 3 layers were microscopically. found underneath the epidermis, it similar to parenchymal cells. (Figure 4a) Thin walled polyhedral parenchymal

DISCUSSION The Pharmacognostic studies provide useful information to identifying and authenticating of medicinal plants. According to the microscopic characteristics of Sarcostmma acidum was found that it is dicotyledonous plant. These pharmacognostic details of Sarcostmma acidum would be helpful for further scientific studies. Sarcostmma acidum (somlata) is needed to be conserved because it is categorized in threatened vulnerable plant species. REFERENCES 1. Shah N C. The Mystery and Botanical identity of Soma plant: An Appraisal. Herbal Tech Industry 2011; 19-28. 2. Indian Medicinal Plants A Compendium of 500 Species. Orient Longman Private Limited, Chennai 2002; vol-5: 72-76. 3. Prajapati N D, Purohit S S, Sharma A K, Kumar T. Handbook of Medicinal Plants A Complete Source Book, Agrobios 2006; 417. 4. Gan Lishe, Yang Sheng-Ping, Fan Cheng- Qi, Yue Jian-min. Lignans and their degraded derivatives from

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Sarcostemma acidum. Journal of Natural Products 2005; 68(2): 221-225. 5.Gan Lishe, Xia zhao, Zhou Chang-xin. Sarcidumitol, A new naturally occurring 2,6-dideoxydisacchariditol from Sarcostemma acidum. Chemistry of Natural Compounds 2010; 46(1): 5. Verma P K, Sharma A, Mathur A, Sharma P, Gupta R S, Joshi S C, Dixit V P. Effect of Sarcostemma acidum stem extract on spermatogenesis in male albino rats. Asian Journal of Andrology 2002; 4(1): 43-47. 7.Anyonyya M, Surendar A, Sruthi G, Gulnaz A K. Antimicrobial activity of Sarcostemma acidum Voigt (Apocynaceae) stem. International Journal of Biological & Pharmaceutical Research 2012; 3(6): 752-757. 8.Gupta S, Kohli S, Sumeet D. In-vitro antiinflammatory activity of Sarcostemma acidum Wight. & Arn. Indian herb by human red blood cell membrane stabilization method. International Journal of Pharmacy Teaching & Practices 2011; 2(4): 184188. Kalmath S V, Patil M S, Kritika S K, Mahantesh S P, Patil C S. Existancy and survey of medicinal plants of Bidar District, Karnataka (India). World Research Journal of Medicinal and Aromatic Plants 2012; 1(1): 14-21. Karuppusamy S. Medicinal plants used by Paliyan tribes of Sirumalai hills of southern India. Natural Product Radiance 2007; 6(5): 436-442. Dey A, Nath DE J. Traditional use of medicinal plants in pediatric and maternal care practiced by ethnic group of Purulia district, West Bengal, India. International Journal of Medicinal Aromatic Plants 2011; 1(3): 189-194. Shiddamallayya N, Azra Y, Gopakumar k. Hundred common forest medicinal plants of Karnataka in primary healthcare. Indian Journal of Traditional Knowledge 2010; 9(1): 90-95. Patil D A. Indian ethnomedicines: origins in the perspective of doctrine of signature. Life Science Leaflets 2012; 6-15. Ray S, Sheikh M, Mishra S. Ethnomedicinal plants used by tribals of east Nimar Madhya Pradesh. Indian Journal of Traditional Knowledge 2011; 10(2): 367371. Reddy K N, Trimurthulu G, Reddy C S. Plants used by the ethnic people of Krishna district, Andhra Pradesh. Indian Journal of Traditional Knowledge 2010; 9(2): 313-317. Mohammed S, Kasera P K, Shukla J k, Unexploited plants of potential medicinal value from the Indian Thar Desert. Natural Product Radiance 2004; 3(2): 6974. Venkata R K, Reddy G T, Raju R R. Herbal remedies for eye infections used by the tribals of Nallamala forests, Andhra Pradesh. Indian Journal of Traditional Knowledge 2010; 9(4): 765-767. Sharma J, Gairola S, Gaur R D, Painuli R M, Siddiqi T O. Ethnomedicinal plants used for treating epilepsy by indigenous communities of sub-Himalayan region

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cells were arranged in 5 to 6 layers without intracellular space. A single layer of endodermis cells were seen separating cortex and vascular bundles. (Figure 3) Vascular bundles were seen in ring shape (Fig.4b) and cambium cells layers (2 to 3) were arranged separating phloem and xylem tissues. Starch grains were also present near phloem tissues. Medullary rays and pith were found to make central part of stem.(Figure 4 c & d) Root- Transverse section of Sarcostemma acidum showed three parts 1. Outer most layer 2. Cortex 3. Vascular bundles. Outer most layer ‘cork’ was thick walled and rectangular shape parenchymal cells layers (5 to 6) were seen underneath the cork which made cortex of root. Phloic fibers and phloem were arranged under parenchyma. Cambium layers were seen separating phloem and xylem. Xylem was scattered in between conjunctive tissues and covered large part of vascular bundles. Absence of pith and presence of secondary growth in root were also seen. (Figure 5 d & e) Bark Anomocytic or Ranunculaceous type stomata were found in longitudinal section of bark. Two guard cells and five subsidery cell were clearly seen.(Figure 6) Physicochemical parameter- Various physicochemical parameters are presented in table No.- 1 Preliminery physicochemical analysis- The aqueous extract (decoction) of stem shows presence of carbohydrates, sucrose, phytosterol, terpines and saponins. Ethanolic extract shows presence of carbohydrates ( redusing & nonredusing), phytosterol, triterpinoids. Chloroform extract shows presence of steroids & triterpinoids. In table No. - 2 Chromatographic study: Results of TLC was tabulated in Table No.-3

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Department of Ayurveda Yoga and Naturopathy, Unani, Siddha and Homoeopathy. The controller of publications civil lines, Delhi 2001; Part -1(1): 199, Part -1(3): 230 – 239. 21. Kokate C K, Purohit A P, Gokhale S B. Pharmacognosy. Nirali Prakashan 2010; 45th edition: A.1-A6.

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of Uttarakhand, India. Journal of Ethnopharmacology 2013; 150: 353–370. 19. Upadhyay B, Singh K P, A Kumar A. Ethnoveterinary uses and informants consensus factor of medicinal plants of Sariska region, Rajasthan, India. Journal of Ethnopharmacology. 2011; 133: 14–25. 20. The Ayurvedic Pharmacopoeia of India Government of India. Ministry of Health and Family Welfare

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