|Sanjeet Kumar1*, Padan Kumar Jena1, Monika Kumari4, Navyanita Patnaik2, Ashok Kumar Nayak3 and Prakash Kumar Tripathy1
|Corresponding Author: Sanjeet Kumar Dept. of Botany, Ravenshaw University, Cuttack Email: email@example.com|
|Received:15 january 2011 Accepted: 28 january 2011|
|Citation: Sanjeet Kumar, Padan Kumar Jena, Monika Kumari, Navyanita Patnaik, Ashok Kumar Nayak and Prakash Kumar Tripathy “Validation of tribal claims through pharmacological studies of Helicteres isora L. leaf extracts: an Empirical Research” Int. J. Drug Dev. & Res., January-March 2013, 5(1): 279-285.|
|Copyright: © 2013 IJDDR, Sanjeet Kumar et al. This is an open access paper distributed under the copyright agreement with Serials Publication, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.|
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Helicteres isora L. commonly known as “Mudmudika” is an important familiar shrub among the populace of Odisha, having potential to cure diabetic, weakness in new-born baby, diarrhoea and scabies. The leaf extracts were analyzed for qualitative screening of bioactive compounds, micronutrients, antimicrobial agent and their anti-microbial effect on known bacterial and fungal pathogens in conformity with the claim made by tribal communities of Odisha. Present study highlights the pharmacological importance of the plant with establishment of a correlation between the tribal claims and bioactive compounds present in leaf extracts and to justify through experimental results and published relevant literature.
Helicteres isora, Ethnobotany, Secondary metabolites, Antimicrobial activities, Validation, Tribal claims
Helicteres isora L. (Mudmudika) is a common subdeciduous shrub. It belongs to the family Sterculiaceae . In Odisha, it has many vernacular name(s) in the state, such as Kurkurbicha, Sinkri and Pita Baranda . It is very popular among the different tribal populace of Odisha due to its potent ethnomedicinal value(s). It is also known as “Indian Screw Tree” in the world . due to its unique shape like metallic screw. It is a shrub or small tree with hairy branches. Leaves orbicular, obviate or broadly ovate or oblong with cordate at base, irregular serrate and apex acuminate; 5-7-palmi-nerved and pubescent. Flowers brick-red, irregular and zygomorphic. Calyx orange. Fruits are woody greenish, 5 spirally rolled carpels on a very elongated gynophore, tardily follicular with brownish black when ripe and dehiscent along their inner edge. Seeds wrinkled, truncate and angular . It is widely distributed in Odisha; frequently available in costal forest block of the state and the hill slopes of Eastern Ghat. It is also distributed throughout the province from Champaran southwards, very common and often gregarious both in the valleys and especially on Northern Hills . It is also widely distributed throughout India, Malaysia, Sri Lanka, Java and Australia .
Fruits are demulcent, mildly astringent [4, 5], griping bowels, diarrheal diseases , refrigerant, stomatic , vulnerary flatulence of children, antispasmodic [7, 8] and colic . Also fruit is boiled with mustard oil, filtered and used for massaging legs of patients suffering from gout, twice a day for five days . Extracted juice from the raw fruit is mixed with equal quantity of mustard oil or fruit paste along with Cyanodon dactylon and mixed with turmeric paste is used for massaging the body of children to relieve them from profound weakness [5,9]. Bark of the plant are used for treating various diseases like treatment of diabetes , colic , human immunodeficiency virus , avian myeloblastosis virus [11, 12], hypoglycaemic , scabies , diabetes , diarrhoea and dysentery . The decoction of the root is mixed with turmeric powder and applied externally, to treat cuts and wounds, by the ethnic people of Rayalseema of Andhra Pradesh . There are reports on plant parts showing antimicrobial activities. The barks of H.isora showed activity against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli  and fruits against Candida albicans . But to the best our knowledge no or very little informations are available on the bioactive compounds and antimicrobial activity of leaf of Helicteres isora L. Keeping this in view an attempt was made to study the pharmacological values of Helicteres isora leaf.
An ethnobotanical survey was carried out from 2010 to 2012 in different forest and tribal blocks of the state. The informations on plants used as traditional medicine against different pathogens and disorders were collected through questioners with different tribal communities and their medical practitioners. The pharmacological and medicinal properties of the plant were confirmed by cross check with informants. The identified plant species was cofirmed with the Flora’s Book [1, 2].
The plant materials were collected from urban area of Bhubaneswar (near Nalco square), Dhaulagiri hills area (Near Lord Shiva temple) and Botanical Garden (near Orchid House). Collected plant materials were washed thoroughly by tap water followed by distilled water twice, were oven dried. The dried materials were crushed to powder with mechanical device and were kept it in air tight container for biochemical and antimicrobial analysis of the leaf of the plant.
Solvent extract was prepared using percolation method from 5gm of leaf powder which was macerated in solvent for 12 hrs in refrigerator. After 12 hrs sample was filtered and residue was again macerated in same solvent and for each solvent process was repeated thrice. Solvent extraction was started from methanol followed by acetone and chloroform. Aqueous extract was prepared separately by taking the powder in distiled water followed by filtration. Filtrate were dried and concentrated to get semisolid mass.
Phytochemical analysis were done with crude extract followed by [19, 20].
The antifungal activity (10 mg / ml crude extract concentration) was done using agar diffusion method followed by Scorzoni et al. (2007)  and antibacterial activity (10 mg / ml crude extract concentration) was done by Rios et al. (1988)  with standard (500μg / ml) taken as Spectinomycin (antibacterial) and Clatrimazole (antifungal).
0.7ml of the extracts was dissolved in 50ml of distilled water and was heated for 10 minutes. After cooling few drops of 1% ferric chloride was added. Colour of sample was changed from yellow to green and dark green precipitate was observed.
5ml of extract was dried and to it was add 1ml of Ethyl acetate. Ethyl acetate was removed and add distilled water and mixture was shaken vigorously and observed for persistent foam which lasted for at least 15 minutes.
Few amount of leaf extract was taken in a flask and dissolved in 10% NaOH. Few drops of HCl was added .Yellow colour turned to colourless.
1ml of extract was mixed with 400 μl chloroform. Then the mixture was added by drop of sulphuric acid. A reddish brown interface indicates terpenoid present.
400 μl acetic anhydride was added in 100microlitre leaf extract; cooled in ice. Then sulphuric acid was added. The colour change from violet to blue to green.
Few quantity of the each portion was stirred with 5 ml of 1% aqueous HCl on water bath and then filtered. Of the filtrate, 1 ml was taken individually into 2 test tubes. To the first portion, few drops of Dragendorff’s reagent were added; occurrence of orange-red precipitate was taken as positive.
2 ml of test solution / crude extract, added alcohol and then few drops of neutral ferric chloride solution were added. The test result was observed.
Quantitative estimation of biochemicals ware done followed by Lavekar, 2010 and Sadasivam & Manickam, 1991. [23, 24]
The ethno-botanical survey revealed the medicinal uses of H.isora in Odisha among the tribal group of the state (Table-1). The phytochemical screening as found in literatures and experimental results revealed the potent bio-active compounds present (Table-2) in the methanol and aqueous extract of leaves of H.isora. Phenolic compounds, Glycosides and saponins were present in methanol extract and phenolic compounds, Glycosides, Flavonoids and tannin were present in aqueous extract (Table- 2).Oliver-Bever (1986) and Okwu (2004) [25,26] had earlier reported that Saponins have antibiotic properties and so help the body to fight infections and microbial invasion. The antimicrobial and antifungal activity of the leaf extract was studied against known bacterial and fungal culture. The methanolic extract of the leaf was found to be more effective against E.coli than the aqueous extract with zone of inhibition 15.28 mm and 12.30 mm respectively (Fig-1). The anti-fungal activity of aqueous extract against A.niger (zone of inhibition) was 08.41 mm and methanol extract was 11.30 mm (Fig-1) in comparison with Clatrimazole (19.05 mm) (Fig-1). Anson et el., 2011 reported the presence of flavonoids, tannin and alkaloids in the fruits of Helicterous isora. Shriram et al., 2010 reported the antibacterial and anti-plasmid activities of Helicteres isora L. , Venkatesh et al., 2011 reported antidiabetic activity and anti-microbial activity of Helicteres isora root . The antibacterial activities observed could be due to the presence of secondary metabolites (Table-2), which have been reported as active constituents of the plant in confirmation of the result of the present experiments for validation (Fig-1) [29, 30, 31, 32]. Flavonoids are known to inhibit bacterial growth [33, 34] and are known to have activity against Gram-positive and Gramnegative bacteria, fungi and viruses . The presence of the phytochemical compounds (Table-2) could have attributed to the antibacterial activities observed in the present study (Fig-1). Previous studies have shown that tannins bind to the cell wall of bacteria, preventing growth and protease activity and can also be toxic to filamentous fungi, yeasts and ruminal bacteria [36, 37].Saponins are effective in the treatment of syphilis and certain skin diseases [20, 36]. Saponins were detected in all the extracts of the leaves in the present study (Table-2). Many pharmacological activities have been reported about saponins such as antibiotic, antifungal, antiviral, hepatoprotective anti-inflammatory and anti-ulcer [38, 39, 40, 41, 42, 43, 44, 45, 46] Some of the biochemical compositions of the leaves collected from three different localities of Bhubaneswar was studied. The result indicated that the plant is rich in biochemical constituents like calcium, magnesium and tannin (Table-3). However the percentage composition varied from locality to locality (Table-3). Based on the ethnobotanical survey, phytochemical screening and anti-microbial activity results of the present investigation, it could be said that the leaf extracts possess potent medicinal values and contain chemical constituents of pharmacological significance. The presence of these chemical constituents in this plant is an indication that the plant, if properly screened can yield compounds of pharmaceutical importance. The biochemical analysis revealed that plant leaf has anti-microbial compound like tannin in good amount. Further research can be carried out to isolate, purify and characterize the detail chemical constituents in the plant with a view to utilise these bioactive compounds in drug development. This study is an attempt to validate the tribal claims of medicinal value of H. isora through experimental observation.
Authors are thankful to Director Biotechnotric. Pvt. Ltd., Bhubaneswar
|Table 1||Table 2||Table 3|
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