Pharmacognostic Study Of Plant Euphorbia hirta L Research Article Giram Ganesh Bhagwat 1 *,Chaulang Ganesh Mahadev 1,Ghodke Dhananjay Sahebrao 1, Patil Kundan Suresh 1,Yeole Pramod Govindrao 1,Ganjiwale Rajendra Onkar 1 1 Institute of Pharmaceutical Education and Research, Borgaon (Meghe) Wardha-442007 For correspondence:giram Ganesh Bhagwat, Institute of Pharmaceutical Education and Research, Borgaon (Meghe) Wardha-442007 E mail: ganesh_chaulang@rediffmail.com Received on: 22-05-2008; Accepted on :24-07-2008 Abstract: Euphorbia hirta L (Family: - Euphorbiaceae) is a folklore medicine of India used as antiasthmatic, anthelmetic, wound healing, antibacterial, antioxidant and diuretics. Powdered leaves of Euphorbia hirta linn were subjected to successive soxhelate extraction with petroleum ether (60-800C), chloroform, acetone (70%) and finally macerated with water so as to get respective extracts. Also cold maceration was done using water & Ethanol (70%). The extracts were screened for its physicochemical characteristics, preliminary phytochemical study of extracts and quantitative estimation of total phenolic and flavonoid content. The pharmacognostic study reveals that the plant Euphorbia hirta L contain phenolic and flavonoid constituents. Key words: Euphorbia hirta L, Extraction, saponins and flavonoids INTRODUCTION In the last few decades there has been an exponential growth in the field of herbal medicine. It is getting popularize in developing and developed countries owing to its natural origin and lesser side effects. In olden times, vaidyas used to treat patients on individual basis, and prepared drugs according to the requirement of the patients. But the scene has been changed now; herbal medicines are being manufactured on a large scale in mechanical units, where manufacturers are facing many problems such as availability of good quality raw material, authentication of raw material, availability of standards, proper standardization methodology of drugs and formulations, quality control parameters and etc[1&2]. Euphorbia hirta L is a tree belonging to the family Euphorbiaceae, common names include Euphorbia, (Pillbearing Spurge E. pilulifera) Asthma Weed and Catshair. The tree grows, widely in most parts of India and in tropical countries, especially on roadsides and on wasteland. Euphorbia forms dense thickets, cutting off access to waterways and dams, and smothering out native vegetation. Flowers are yellow and fragrant. Though the plant can flower through out the year, october is the main flowering period for this plant. The leaves of Euphorbia hirta have been used in Asia since antiquity as antiasthmatic, anthelmetic, wound healing, antibacterial, antioxidant and diuretics[3&4]. MATERIALS AND METHODS Plant material collection and authentication: The mature green leaves of Euphorbia hirta linn were collected locally from Wardha District, Maharashtra, India, in the month of August 2006. The plant was identified and authenticated by the Botanist, Post graduate Teaching Department of Botany, R. T. M., Nagpur University. A voucher specimen (No.9082) was deposited in the, Post graduate Teaching Department of Botany, Nagpur University. Drying and size reduction of leaves: The leaves of Euphorbia hirta were subjected to shed drying and further crushed to powder, then the powder is passed through the mesh 22 and stored in air tight container for further use. Determination of physicochemical parameters The leaves of Euphorbia hirta were subjected to fol- 39
lowing physicochemical parameters. Foreign organic matter, total ash value, acid insoluble ash value, water soluble ash value, loss on drying, alcohol soluble extractive, water soluble extractive and ph[2 &5]. The result of physicochemical parameters is reported in table 1. Extraction of Euphorbia hirta linn leaves The shade dried and 22 mesh powdered leaves of Euphorbia hirta linn, were subjected to successive soxhelate extraction with petroleum ether (60-800C), chloroform, acetone (70%) and finally macerated with water so as to get respective extracts. Also cold maceration was done with water & ethanol (70%). All extracts were filtered individually, evaporated to dryness and stored in freeze condition for further use.the details are given in Table no 2. Preliminary phytochemical screening of extracts Test for terpenoids (steroids), fats and oils, phenols, tannins, saponin glycosides, Flavonoids, Anthraquinone glycosides, alkaloids, proteins and sugars were carried out with procedure described earlier.[6,7,8&9], Results are displayed in table no.3 Quantitative estimation of active constituents Estimation of Total Phenolic content of the Extracts. The content of total phenolic compound in plant extracts were determined by Folin-ciocalteu reagent [10 &11]. The results are shown in table no. 4 Estimation of total flavonoid content The content of total flavonoid compound in plant extracts were determined by Aluminium Chloride Colorimetric Method [12 &13]. The results are shown in table no. 5 Table 1. Results of physicochemical parameters Parameters Values obtained * (%w/w) ± S.D Foreign organic matter 0.38 ± 0.01 Total ash value 5.17 ± 0.07 Acid insoluble ash value 2.75 ± 0.07 Water soluble ash value 3.00 ± 0.03 Loss on drying 3.05 ± 0.11 Alcohol soluble extractive 21.46 ± 0.46 Water soluble extractive 24.20 ± 0.22 *An average of three determinations 40
Giram et. al: Pharmacognostic Study Of Plant Euphorbia hirta L Table 2.Solvents, extraction methods and respective yield from leaves of Euphorbia hirta Linn. S.N. Extract Solvents Colour Nature % Yield w/w 1 Successive Petroleum ether Greenish brown Semisolid 2.2 2 Chloroform Greenish black Semisolid 3.1 3 Acetone (70%) Dark brown Solid 18.12 4 Water Brown Solid 11.28 5 Individual Water Dark brown Solid 24.5 6 Ethanol (70%) Dark brown Semisolid 21.32 Table 3. Results of preliminary phytochemical screening of Euphorbia hirta leaves extracts. Plant Constituents Petroleum Chloroform Acetone Aqueous Ethanol Total ether Extract Extract Extract Extracts Extract Aqueous Extract Steroids & + + - - - - Terpenoids Anthraquinone - - - + + + glycosides Fats and + + - - + - Oils Tannins - - + + + + Saponin - - + + + + glycoside Flavonoids - - + + + + Alkaloids - + - - + - Proteins - + + + + + Sugars - - + + + + Phenolics - - + + + + + = Positive, - = Negative 41
Table 4.Result of Total phenolic compound (mg/gm) of plant extracts Ethanol Absorbance Total phenolic extract compound, Mean* mg/gm of plant extracts 1 0.622 19.003 19.01 ± 0.064 2 0.625 19.096 3 0.620 18.940 Acetone Extract 18.76 ± 0.064 1 0.614 18.753 2 0.617 18.846 3 0.612 18.690 Total Aqueous Extract 18.37 ± 0.201 1 0.605 18.471 2 0.608 18.565 3 0.607 18.10 Aqueous Extract 14.81± 0.051 1 0.488 14.815 2 0.486 14.753 3 0.490 14.878 * Values are expressed in Mean ± S.D., Table5. Result of Total Flavonoids compound (mg/gm) of plant extracts S.No. Extract Absorbance Total Mean+ SD Flavonoids (mg/gm) of plant extracts 0.941 47.05 1 Ethanol 0.940 47.00 47.066+0.062 0.943 47.15 0.288 14.40 2 Acetone 0.276 13.80 14.07 ± 0.306 0.280 14.00 0.382 19.01 3 Total Aqueous 0.380 19.00 19.02 ± 0.022 0.381 19.05 0.363 18.15 4 Aqueous 0.360 18.00 18.06 ± 0.062 0.361 18.05 42
RESULT AND DISCUSSION The powdered leaves found to contain 0.38 ± 0.01 % w/w of foreign organic matter. Total ash, acid insoluble ash and water soluble ash values were found to be 5.17 ± 0.07 % w/w, 2.75 ± 0.07 % w/w and 3.00 ± 0.03 % w/w respectively. Loss on drying of the powder was 3.05 ± 0.11% w/w. Alcohol soluble extractive and water soluble extractive were found to be 21.46 ± 0.46% w/w and 24.20 ± 0.22% w/w respectively.with the successive solvent extraction, the yield was found to be 2.2 % w/w, 3.1 % w/w, 18.12 % w/w and 11.28 % w/w for petroleum ether, chloroform, acetone (70%) and water respectively. The maceration of plant with water and ethanol gave a yield of 24.50 % w/w and 21.32 % w/w respectively.the extracts so obtained were subjected to preliminary phytochemical screening for detection of chemical constituents present in them. The results of phytochemical screening revealed the presence of fats, steroid, alkaloids, carbohydrates, proteins and phenolic compound like flavonoids, tannins, and saponin. Present study also includes the estimation of total phenolic content and total flavonoid content of the isolated extracts. The total phenolic content was estimated by most popular Folin-Ciocalteu Reagent method. For the calculation of Phenolic content (mg/gm) calibration curve was plotted which has a regression equation y = 0.0032x, with a correlation coefficient, R2 = 0.993. Acetone and ethanol extract were found to contain a higher amount of phenolic compounds. References: [1] Ali M S., Ahmed F., Pervez M K., Azhar I., Ibrahim SA., Parkintin: a new flavanone with epoxy-isopentyl moiety from Parkinsonia aculeate Linn, Nat Prod Res, Jan;19(1), 2005, 53-60. [2] Agarwal, A., Critical issues in Quality Control of Herbal Products, Pharm Times, 37(6), 2005 09-11. [3] Aquil, M., and Khan IZ, Euphorbianin, a new glycoside from Euphorbia hirta Linn. Global J. pure and applied science 5 (3) 1999, 38. [4] Blanc, P., and De Saqui-Sannes G., Flavonoids of Euphorbia hirta.plantes Medicinales et Phytotherapie 6 (2): 1972,106. [5] Chakravarthy, B. K.,, Standardization of herbal products. Indian. J. Nat. Product, 9, 1993, 23-26. [6] Chen, L., Polyphenols from leaves of Euphorbia hirta Linn Zhonuo Zhongyao Zazhl 16 (1) 1991, 38. [7] Harborne, J. B., Phytochemical methods. A guide to Modern Techniques of Plant analysis. Charpman and hall, London, 1973, 279. [8] Indian Herbal Pharmacopoeia, Published by Indian. Drug Manufacturar Asso. Mambai, 2002, 429-447. [9] Indian Pharmacopoeia II Government of India Ministry of Human & Family Welfare Published by the Controller of Publication, Delhi, A: 1996, 88-89. [10] Jain, U. K. and Dixit, V. K., Spectrophotometric estimation of Tannins from Chyavanprash. Indian Drugs, 41 (8), 2004, 469-472. [11] Marinova, D., Ribarova, F., Atanassova, M., Total phenolics and total flavonoids in Bulgarian fruits and vegetables. J. the university of chemical technology and metallurgy, 40, 3, 2005, 255-260. [12] Miliauskas, G., Venskutonis, P. R., and van Beek, T. A., Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry, 85, 2004, 231-237. [13] Sabhi, M., Ramezanian, M., Jaffari, G., Haravi, G., Bahaeddini, F. and Aynehi, Y., Survey of Indian Plants for Saponins, Alkaloids, Flavonoids and Tannins, the Plant of Karman province (Capparidaceae), Int. J. Crude drug Research, 23(4), 1985, 165-177. 43