Ambika Sharma et al. / Journal of Pharmacy Research 2011,4(6),1673-1674 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Pharmacognostic and Phytochemical Evaluation of Roots of Eclipta prostrata. Ambika Sharma*, Mittal Arun, Aggarwal Sushma, Gupta Anil Kumar, Sardana Satish. Department of Pharmacognosy, Hindu College of Pharmacy, Sonepat (Haryana). India. Received on: 11-02-2011; Revised on: 16-03-2011; Accepted on:21-04-2011 ABSTRACT Herbs are staging a comeback and have stood the test of time for their safety, efficacy, cultural acceptability and fewer side effects. Eclipta prostrata Linn. [Asteraceae] popularly known as “Bhringaraj” is a herb with a worldwide distribution. Extensive studies have been carried out to substantiate its various claimed pharmacologic activities. Present communication records the Pharmacognostic evaluation like Macroscopic and Microscopic characters, Extractive values, Moisture content, Volatile oil content, Ash value, Crude Fibre content, Bitterness value, Foaming Index and Swelling Index under Phsico-chemical parameters. Phyto-chemical investigations on this plant reveal the presence of Carbohydrates, Amino acids, Steroids, Saponins, Alkaloids, Flavonoids, Phenolic compound and Tannins. Chemo-profiling of Ethanolic extract of roots has also been carried out.

Key words: Eclipta prostrata, Pharmacognostic studies, Physicochemical parameters, Chemoprofiling.

INTRODUCTION NATURE always stands as a golden mark to exemplify the outstanding phenomenon of Foreign organic matter, Moisture content, Ash values, Extractive values, Volatile oil symbiosis. All the biotic and abiotic elements of nature are interdependent. The plants are content, Swelling index, Foaming index, Crude fibre content and Bitterness values, were indispensable to humans; as along with necessities of life, these also provide materials determined. having therapeutic potential [1]. (C) Chemo-profiling of extract: (a) Qualitative chemical examination [10], [11] of the extract was carried out for the identi- Eclipta prostrata Linn. [Asteraceae] is popularly known as “Bhangra” or “Bhringaraj” or fication of various active chemical constituents viz. Alkaloids, Glycosides, Steroids, Saponins, Flavonoids, Tannins and Phenolic compounds, Proteins and Carbohydrates. “Bhringaraja” (Figure. 1) in India. It is an herbaceous annual, about 30-50 cm. in height. [12] This plant has a world-wide distribution and can be found in many pacific islands. It (b) Fluorescence Analysis - Powder of roots, after treatment with various chemical grows mainly in muddy borders of ponds and rivers, ditches, and poorly drained areas [2]. reagents was examined under UV light (long and short wave length) and the visible light. (c) Chromatographic analysis [13] - Thin layer chromatography was performed to get the TAXONOMICAL CLASSIFICATION: chromatographic profile of phyto-constituents present in the Ethanolic extract of roots. Kingdom Plantae Division Magnoliophyta RESULTS: Class Magnoliopsida Macroscopically the roots are cylindrical in shape, with rough external surface. The roots Order Asterales are 7 to 10 cm in length and 3 to 5 mm in diameter, having easy and even fracture. The Family Asteracece fractured surface is somewhat light cream coloured. Results are also being presented in Genus Eclipta tabular form (Table: 1). Species alba Table 1: Macroscopic examination of roots of E. prostrata Fig.1 Aerial part of E. prostrata S. No. Parameters Inference Roots of Eclipta alba have been reported to contain thiophene acetylenes such as 5-senecioyl oxymethylene-2-(4-isovaleryloxybut-3-ynyl)-dithiophene, 5 tigloyloxymethylene-2 1. Shape Cylindrical 2. Size 3-4mm in diameter, 7-10cm in length (isovaleryloxybut-3-ynyl)-dithiophen and 2-(3-acetoxy-4-chloro-but-1-ynyl)-5-(pent-1, 3-diynyl) 3. Colour Greyish thiophene, hentriacontanol, stigmasterol, ecliptal, 14-heptacosanol [2]. In Ayurveda the plant is 4. Odour Slight, characteristic used as alterative anthelmintic, alexipharmic. It also cures inflammation, hernias, eye diseases, 5. Taste Bitter [3] 6. External surface Rough “Kapha” and “Vata”, bronchitis, asthma and leucoderma etc. . 7. Fracture Easy, even 8. Fractured surface Light Cremish colour Extensive studies have been carried out to substantiate the hepato-protective activity [2], anti microbial activity against Staphylococcus epidermis and Salmonella [3], anti- The roots and its powder are light brown, with slight characteristic odour and bitter taste hyperglycaemic [4], antiviral [5], wound healing [6] and anti-oxidant activity [7]. Leaves are as presented in Table 2. [2] most popularly known for its potential to promote hair growth . Table 2: Macroscopic examination of Root powder MATERIALS AND METHODS: Materials S. No. Parameters Inference The healthy plants of Eclipta prostrata (Figure-1) were collected from PUSA institute (New Delhi) and authenticated under a voucher specimen number-NISCAIR/RHMD/ 1. Color Light Brown 2. Odor Slight Consult/-2010-11/1552/150 dated oct.12, 2010. These were dried under shade and then 3. Taste Bitter powdered (#60) to obtain a coarse powder. Microscopy: The outer most layer is of cork cells; cork layer has only 3-5 rows of thin Extraction: walled cells which are tangentially elongated. Few stone cells are scattered in cork cells. The powder (300g) was macerated for 7 days using 900 ml of 95% Ethanol as menstrum. It is followed by outer secondary cortex, 1-2 layered, cells are tangentially elongated and Maceration was repeated, twice (7 days each time) by adding fresh menstrum, equal to the volume of extract obtained. Total extract was combined and concentrated, under vacuum; Cork to a semi solid consistency. The yield obtained was 5.6 % w/w. [9] (A) Pharmacognostic studies: [10], [11], [13] Outer secondary cortex Macroscopic examination of root and as well as root powder was performed. Transverse sections were observed for Microscopic characters. Inner secondary cortex [9] (B) Physiochemical parameters : Trichome

Pericyclic fibres

*Corresponding author. Secondary phloem Ambika Sharma Department of Pharmacognosy, Hindu Xylem College of Pharmacy, Sonepat (Haryana).India

Fig. 2.1.T.S Root (x600) Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1673-1674 Ambika Sharma et al. / Journal of Pharmacy Research 2011,4(6),1673-1674 has air spaces. Inner secondary cortex consists of comparatively bigger, irregular shaped Table 6: T.L.C. for ethanolic extract of roots parenchymatous cells with prominent air spaces. Even cortex has stone cells scattered in it as singles or in groups, groups having different shapes and sizes. Pericyclic fibres are S. No. Solvent System Rf value present as tangentially arranged bands. Secondary phloem consists of phloem fibres and 1. Hexane :Ethyl acetate (8: 2) multiserate phloem rays. Xylem consists of numerous xylem vessels, tracheids, xylem Detecting Agent = U.V. Long 0.28 0.39 fibres and xylem parenchyma immersed by xylem rays. (fig. 2.1). 0.47 0.92 The powder microscopy of roots shows the presence of almost rectangular stone cells 2. Toulene:Chloroform:Acetone (40:25:35) (fig.3.1), 2, celled uniseriate trichome with pointed end (fig.3.2) and long lignified fibres Detecting agent = U.V. Long 0.95 0.89 (fig.3.3) 0.82 3. Benzene:Pyridine:Formic acid (36:9:5) Powder Microscopy Detecting agent = U.V. Long 0.83 0.94

Chromatographic Profile

Fig. 3.1 Stone cells Fig. 3.2. Uniserate trichome Fig. 3.3. Lignified Fibres The physico-chemical analysis-Results of physico- chemical analysis have been tabu- lated (Table. 3). Table 3: Physiochemical parameters

S. No. Parameters Inference

1. Moisture content 7.2%w/w 2. Total ash 11.5%w/w 3. Water soluble ash 4.95%w/w 4. Acid insoluble ash 1.47%w/w 5. Alcohol extractive value 6.4%w/w 6. Water extractive value 12.8%w/w 7. Volatile oil content Traces of yellow coloured volatile oil 8. Swelling Index 6.2 9. Foaming index Less than 100 10. Crude Fibre content 55% 11. Bitterness value 71.42 Qualitative chemical examination The qualitative chemical examination showed the presence of Steroids, Saponins, Flavonoids, Tannins & phenolics as tabulated (Table.4).

Table 4: Phytochemical screening of ethanolic extract of root Fig. 4.1 Fig. 4.2 Fig. 4.3 S. No. Chemical constituents Ethanolic extract of leaves of any natural drug, it is must that its identity be undoubted and its standardized form be used. Hence the establishment of Pharmacognostic parameters and standards is prerequi- 1. Carbohydrates -VE site for use of drug as therapeutic agents. The results of present study can form the basis 2. Proteins -VE 3. Amino acids -VE for identification and standardization of ‘Bhringraj’ and the manufacturers can utilize them 4. Steroids +VE for identification and selection of the raw material for drug production. 5. Cardiac Glycosides -VE 6. Anthraquinone glycosides -VE 7. Saponins +VE ACKNOWLEDGEMENT: 8. Flavonoids +VE The author is thankful to Dr. S Sardana, Principal, Hindu College of Pharmacy 9. Alkaloids -VE 10. Tannins & Phenolics +VE Sonepat for funding research project and providing the necessary laboratory facilities for carrying out the experimental work. The fluorescence analysis of the powder of leaves of E. prostrata in various solvents and chemical reagents under UV (long and short) and normal day light is given in Table.5. REFERENCES: 1. Kokate CK, Purohit AP, Gokhale SB, Pharmacognosy, 12th Edition, Nirali Prakashan, 1999, pp-1. Table 5: Fluorescence Analysis of root 2. Jadhav VM, Thorat RM, Kadam VJ, Sathe NS, Eclipta alba Linn – ‘‘Kesharaja’’: A Review, Journal of Pharmacy Research, 2(8), 2009, 1236-1241. 3. Basu Kiritikar, Indian medicinal plants with illustrations, 2nd Edition, vol.6, 2003, pp 1882. S. No. Reagent Long UV Short UV Visible 4. Dalal S, Rana S, Sastry K, Kataria S, Wedelolactone as an antibacterial agent extracted from Eclipta alba, The internet journal of Microbiology, 7(1), 2009, 1-5. 1. Powder Brown Brown Brown 5. Bhattacharya SK, Kalkunte SS, Ghosal S, Antioxidant activity of glywithanolides from Withania 2. 1N Hcl Dark green Brown Green somnifera, Indian J. Exp. Biol, 35, 1997, 236-239. 3. 1N NaOH Brown Yellowish brown Greenish brown 6. Kannabiran Krishnan, Khanna VG, Hameed S, Antiviral activity of Dasyscyphin C extracted from

4. 50% HNO3 Dark brown Greenish brown Greenish brown Eclipta prostrata against fish nodavirus, Journal of Antivirals and Antiretrovirals, 2(1), 2010, pp-29- 32. 5. 50% H2SO4 Dark brown Brown Dark green 6. Methanol Light green Yellowish brown Greenish yellow 7. Sharma S, Sikarwar MS, Wound healing activity of ethanolic extract of leaves of Eclipta alba, 7. Acetic acid Light green Yellowish brown Green Pharmacognosy Magazine, 4 (13), 2008, 108-111. 8. Picric acid Black Yellowish brown Green 8. Rao Bhaskar D, Ch.Ravi Kiran, Y Madhavi, P Koteswara Rao, T Raghava Rao, Evaluation of antioxidant potential of Clitoria ternate L. and Eclipta prostrata L., Indian journal of Biochemistry 9. 5% Iodine Black Yellowish black Greenish black and Biophysics, 46, 2009, 247-252. 10. Extract Light green Brown Black 9. W.H.O. “Quality control methods for medicinal plant material”, Geneva, A.P.T.B.S. publisher and distributor, New Delhi, 2002, 22-37, 46. The Thin layer chromatographic analysis of ethanolic extract showed the presence of 10. Khandelwal KR, Practical Pharmacognosy, 8th Edition, Nirali Prakashan New Delhi, 2001, pp 11- phyto-constituents in different solvent systems with Rf values as shown in Table.6. (Fig. 16 & 157-160. 4.1), (Fig. 4.2), (Fig.4.3) 11. Kokate CK, Practical Pharmacognosy, 4th Edition, Vallabh Prakashan, Delhi, 2001, pp 7-9 & 107- 108. 12. Chase CR, Pratt R.J, Fluorescence analysis of powdered drugs with particular reference to devel- DISCUSSION AND CONCLUSION opment of a system of identification, J.American Pharm.Assoc, 38, pp. 324-331 Eclipta prostrata “Bhringraj” has value in the treatment of various diseases like inflam- 13. Mukherjee K. Pulok, “Quality control of herbal drugs”, 2nd Edition, Published by business horizons, mation, hepatitis, epilepsy and used for promoting hair growth. For reproducible efficacy New Delhi, 2003, pp- 9, 15, 536, 559, 564. Source of support: Nil, Conflict of interest: None Declared Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1673-1674