World Journal of Pharmaceutical Research

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1 World Journal of Pharmaceutical ReseaRch Mohanapriya et al. Volume 3, Issue 3, Research Article ISSN PRELIMINARY PHYTOCHEMICAL SCREENING OF MIMOSA PUDICA ROOT EXTRACTS AND COMPARATIVE ANALYSIS OF ITS FTIR PROFILE * Chelladurai Mohanapriya 1, Ramesh Rajprasanth 1, Vijayan Karthikeyan 1, Rangaraju Magesh 1 and Shanmugam Karthikeyan 1 1 Phytochemistry division, Biochemistry lab, Department of Biotechnology, KarpagaVinayaga College of Engineering and Technology,ChinnaKolambakkam, MaduranthagamTk, Kancheepuram Article Received on 05 March 2014, Revised on 28 March 2014, Accepted on 21 April 2014 *Correspondence for Author Chelladurai Mohanapriya Department of Biotechnology, KarpagaVinayaga College of Engineering and Technology, ChinnaKolambakkam, Kancheepuram, India, ABSTRACT Mimosa pudica L. (Mimosaceae)is a creeping annual or perennial herb. It is also referred to as touch me not, live and die and humble plant is a prostrate or semi-erect subshrub of tropical America and Australia, also found in India. Root decoction is efficacious in gravel and other urinary complaints. It also treats dysentery, fever, syphilis, leprosy, stomach worms, veneral diseases, insect bite, insomnia, nervousness, and piles. It appears to be a promising herbal candidate to undergo further exploration as evident from its pharmacological profile for its antibacterial, antivenom, antifertility, anticonvulsant, antidepressant, aphrodisiac and various other activities. This work is an attempt to explore and compare Mimosapudica root s secondary metabolites obtained by Direct extraction method with three different solvents using preliminary screening and Fourier Transform Infra-Red Spectroscopic techniques.the results revealed maximum number of phytochemicals and functional groups which corresponds toalkaloids, steroids, phenols and glycosides which in turn is responsible for the possible pharmacological property exhibited by the herb. Keywords: Mimosapudica, Phytochemicals, FTIR, Pharmacology, Solvents. INTRODUCTION Vol 3, Issue 3,

2 A number of Indian medicinal plants have been used for thousands of years in the traditional system of medicine. Amongst these are plants used for the management of neurodegenerative diseases such as Parkinson's, Alzheimer's, loss of memory, degeneration of nerves and other neuronal disorders by the Ayurvedic practitioners. Though the etiology of neurodegenerative diseases remains enigmatic, there is evidence, which indicates that defective energy metabolism, excitotoxicity and oxidative damage may be crucial factors [1]. Plants have played a significant role in maintaining human health and improving the quality of human life for thousands of years and have served humans well as valuable components of medicines, seasonings, beverages, cosmetics and dye. Herbal medicine is based on the premise that plants contain natural substances that can promote health and alleviate illness. In recent times, focus on plant research has increased all over the world and a large body of evidence has collected to show immense potential of medicinal plants used in various traditional systems [2]. Taxonomy Mimosa pudicalinn. Kingdom: Plantae Genus: Mimosa Subfamily: Mimosoideae Family: Leguminosae Species: M.pudica Order: Fabales [3] Occurrence The species is frost-sensitive. Mimosapudica (from Latin: pudica "shy, bashful or shrinking"; also called sensitive plant and the touch-me-not). This is a creeping annual or perennial herb often grown for its curiosity value [4]. Range The plant is a native of tropical America and naturalized nearly all through the tropical and subtropical parts of India. Sensitive plant was first described from Brazil and is perhaps native to much or all of the New World Tropics.Today it is pantropical in its distribution [5]. Exploitation of Mimosa Pudica (L) Vol 3, Issue 3,

3 It is used in wound healing, blood coagulation and to treat sexual weakness. All parts of the tree are considered to possess medicinal properties. Its roots has been used in traditional healthcare system in the treatment of leprosy, dysentery, vaginal and uterine complaints, inflammations, burning sensation, asthma, leucoderma, fatigue and blood diseases. The Mimosa Pudica, invites attention of the researchers worldwide for its pharmacological activities such as anti-diabetic, antitoxin, antihepatotoxin, antioxidant and wound healing activity. Mimosa Pudica(Family: Leguminosae) is a small or middle sized tree, about 1.5 m (5 ft) in height cultivated throughout India [2]. Phytochemicals Phytochemicals can be defined, in the strictest sense, as chemicals produced by plants. However, the term is generally used to describe chemicals from plants that may affect health, but are not essential nutrients. While there is ample evidence to support the health benefits of diets rich in fruits, vegetables, legumes, whole grains and nuts, evidence that these effects are due to specific nutrients or phytochemicals is limited. Because plant-based foods are complex mixtures of bioactive compounds, information on the potential health effects of individual phytochemicals is linked to information on the health effects of foods that contain those phytochemicals. Free radicals can damage the cells of our bodies and must be removed [6]. Metabolites identification techniques Secondary metabolites are organic compounds that are not directly involved in the normal growth, development, or reproduction of an organism. Unlike primary metabolites, absence of secondary metabolites does not result in immediate death, but rather in long-term impairment of the organism's survivability, fecundity, or aesthetics, or perhaps in no significant change at all. Secondary metabolites are often restricted to a narrow set of species within a phylogenetic group. Secondary metabolites often play an important role in plant defense against herbivory and other interspecies defenses. Humans use secondary metabolites as medicines, flavorings, and recreational drugs. Engineering modern techniques are used in various fields like drug and medicine. Many plants synthesize substances that are useful to the maintenance of health in humans and other animals. The use of plants as medicines predates written human history. Medicinal plants are in all way having its own immense importance in current era. All plants produce chemical compounds as part of their normal metabolic activities. These metabolic activities can be measured using various techniques such as GCMS, FTIR, TLCetc [7]. Vol 3, Issue 3,

4 MATERIALS AND METHODS Plant material Root Collection Mimosa pudicaroots were collected fromkancheepuram district, Tamil Nadu. Preparation of Mimosa pudicaroot powder The roots were carefully washed with tap water, rinsed with distilled water, and shade dried for two weeks, since certain compounds get denatured in sunlight; it is dried under shade to avoid decomposition. Then the roots were ground into fine powder and stored in air tight containers at room temperature. Chemicals Analytical grade chemicals supplied by Loba, Hi-Media, S.D.Fine Chemicals, E-Merck and Qualigens were used. Preparation of plant extract Direct extraction with different solvents Direct extraction with Ethanol, Ethyl acetate and cyclohexane was used as an extraction method [8].In this method, finely ground seed powder (1 gm) was extracted with 10 ml of Ethanol, Ethyl Acetate and cyclohexane in separate conical flasks in shaking condition with the help of a shaker. The process was repeated 3 times with the same material but using fresh solvent and each time the extract was decanted in to pre-weighed glass vials. The solvent in the extract was removed by condensation. The extracted residues were weighed and redissolved in different solvents to yield 10mg/ml solutions for further analysis. Preliminary phytochemical screening Qualitative phytochemical analysis has to be done for preliminary screening [9, 10]. Alkaloids (Mayer s Test) 0.5 g of the extract was stirred with few ml of dilute hydrochloric acid and filtered. To a few ml of filtrate, one or two drops of Mayer s reagent were added to the sides of the test tube. A white creamy precipitate demonstrated the test as positive. Vol 3, Issue 3,

5 Steroids (Salkowski s Test) 0.5g of the extract was dissolved in 2ml of chloroform. Sulphuric acid was then carefully added to form a lower layer. A reddish-brown colour at the interface indicated the presence of steroids. Flavonoids (Ferric chloride Test) 0.5 g of the extract was boiled with distilled water and then filtered. To 2 ml of the filtrate, few drops of 10% ferric chloride solution were then added. A green-blue or violet colouration indicated the presence of flavonoids. Tannins (Neutral Ferric chloride Test) 0.5 g of the extract was stirred with 10ml of distilled water, filtered, and ferric chloride reagent was added to the filtrate. A blue-black, green or blue-green-precipitate was taken as evidence for the presence of tannins. Phenols (lead acetate Test) 0.5 g of the extract was treated with lead acetate solution. Formation of precipitate indicated the presence of phenols. Glycosides (Salkowski s Test) 0.5 g of the extract was dissolved in 2ml of chloroform. Sulphuric acid was then carefully added to form a lower layer. A reddish-brown colour at the 0interface demonstrated the presence of glycosides. Saponin (Frothing Test) To 1g of the extract about 3 ml of distilled water was added and shaken vigorously for about 5 min. frothing which persisted on warming was taken as an evidence for the presence of saponins. Phlobotannins (Aqueous Hcl Test) Deposition of a red precipitate when an aqueous extract was boiled with 1% aqueous hydrochloric acid indicated the presence of phlobotannins. Anthraquinones (Borntrager s Test) Vol 3, Issue 3,

6 About 0.2 g of each portion to be tested was shaken with 10 ml of benzene and then filtered.5 ml of the 10% ammonia solution was then added to the filtrate. Appearance of a pink, red or violet colour in the ammoniacal (lower) phase was taken as the evidence for the presence of free anthraquinones. Fourier Transform Infra Red (FTIR) spectroscopy 2 mg of the ethanol extract of M.pudica roots were mixed with 200 mg KBr (FT-IR grade) and pressed into a pellet. The pellet was immediately put into the sample holder and FT-IR spectra were recorded in the range cm 1 for the sample and the results were recorded. All investigations were carried with a Bruker 55 model FT-IR spectrometer. RESULTS AND DISCUSSION Results of the present study were good as the tested root extract revealed the presence of phytoconstituents with good antioxidants and antimicrobial potentials. Secondary metabolite profiling by FT-IR also revealed good number of phytoconsituents. Qualitative Analysis of Phytochemicals Table.1 Qualitative analysis of phytochemicals in various extracts of M.pudicaroot Phytochemicals/Extracts Ethanol extract Cyclohexane extract Alkaloids Steroids Flavanoids Tannins Phenols Glycosides Saponins Phlobotannins Anthraquinones (+) Indicates the presence of phytochemicals (-) Indicates the absence of phytochemicals Ethyl acetate extract Table 1 demonstrates the results for phytochemical screening of three different root extracts of M.puidca namely Ethanol, Ethyl acetate and Cyclohexane extracts. The high polar ethanol extract indicated the presence of Alkaloids, Flavanoids, Phlobotannins, Tannins, Phenols, steroids, glycosides and Saponin. The medium polar Ethyl acetate extract revealed the presence of Alkaloids, steroids, phenols and glycosides. The low polar cyclohexane extract Vol 3, Issue 3,

7 showed maximum number of phytochemicals such as flavonoids, alkaloids, steroids, glycosides. Presence of tested secondary metabolites in the acetone extract of M.puidcaroot are in line with earlier reports [11].The phytoconstituents detected in the roots could be responsible for their medicinal property. Taking this into consideration we choose only the ethanol extract for further studies since this indicates maximum number of phytoconstituents. Fourier Transforms Infra-Red spectroscopy (FT-IR) Interpretation of infrared spectra involves the correlation of absorption bands in the spectrum of an unknown compound with the known absorption frequencies for types of bonds. Significant for the identification of the source of an absorption band are intensity (weak, medium or strong), shape (broad or sharp), and position (cm -1 ) in the spectrum. Theethyl acetate, ethanol and cyclohexaneextracts were passed into the FTIR analysis for the separation of the functional groups in the bioactive components based on its peak ratio and electron transition of compounds respectively SAMPLE - A 50 %T cm-1 Figure: 1. FTIR profile for the ethanol extract of Mimosa pudicaroot The results of FT-IR analysis confirmed the presence of the carboxylic acids, amines, esters, aldehydes, aromatic ring, alkanes and alkenes in all the three extracts. FTIR spectroscopy was useful for thecompound identification and when run under IR region in the range of cm-1 variation inthe peaks were observed for the corresponding extracts. Vol 3, Issue 3,

8 Table 2. Interpretation of the FTIR profile for the ethanol extract of M.pudica root S.NO Peak Value Cm -1 Intensity Of The Peak Possible Functional Groups Possible Compounds Strong,Broad O-H Monomeric-Alcohols,Phenols Strong,Broad O-H Monomeric-Alcohols,Phenols Medium N-H Amines Strong C-H Alkanes Strong C-H Alkanes Variable C N Nitriles Weak C C Aromatic Rings Strong NO 2 Nitro compounds Variable C-H Alkanes Variable C-H Alkanes Strong C-O Alcohols, Ethers, Carboxylic Acids,Esters Strong C-O Alcohols,Ethers,CarboxylicAcids,Esters Broad C-H Alkynes Fourier transform infrared spectrometry (FTIR) can be used to identify the structure ofunknown composition or its chemical group, and the intensity of the absorption spectraassociated with molecular composition or content of the chemical group [12,13]. At present, particularly in plant physiology research, FTIR has been used to identify the concrete structure of certain plant secondary metabolites. FT-IRSpectrum exhibits the characteristic finger print band features A %T cm-1 Figure: 2. FTIR profile for the ethyl acetate extract of Mimosa pudicaroot The 3911 and 3784 cm -1 is due to the stretching vibration of monomeric alcohols and phenols. The 2924, 2852, 1446,1351 and 664cm -1 bands represents alkanes which Vol 3, Issue 3,

9 corresponds to C-H bending with medium and strong intensity of absorption. In the other locations the 1261 and 1070 cm -1 represents the C-O stretching and bending vibration which corresponds to alcohols, ethers and esters. Table 3. Interpretation of the FTIR profile for the ethyl acetate extract of M.pudica root S.NO Peak Intensity Possible Possible Compounds Value Of The Functional Cm -1 Peak Groups Strong O-H Monomeric-Alcohols,Phenols Strong C-H Alkanes Strong C-H Alkanes Strong C-H Alkanes Weak C C Aromatic rings Weak C C Aromatic rings Weak NO 2 Nitriles Strong C-H Alkanes Variable C-H Alkanes Variable C-H Alkanes Strong C-O Alcohols, Ethers, Carboxylic Acids,Esters Strong C-O Alcohols,Ethers,CarboxylicAcids,Esters Broad C-H Alkynes Medium C-H Alkynes Medium C C Aromatic rings Strong C-H Alkynes Strong C-O Alcohols,Ethers,CarboxylicAcids,Esters Medium C-O Alcohols,Ethers,CarboxylicAcids,Esters Variable C-H Alkynes It is noticed that the bands at 3368 cm -1 is due to the N-H stretching vibrations with medium intensity of absorption. The intensity of absorption was found to be medium at 2328 cm -1 representative of nitriles with C=N stretching vibrations. The very strong 1612 cm -1 bands in the samples predict the presence of phenyl ring which corresponds to C-H bending [14] C %T cm-1 Vol 3, Issue 3,

10 Figure: 3. FTIR profile for the cyclohexane extract of Mimosa pudicaroot The 3391 and 2923 cm -1 is due to the stretching vibration of monomeric alcohols and phenols. The 1711, 1618, 1502, 1376 and 1223cm -1 bands represents alkanes which corresponds to C-H bending with medium and strong intensity of absorption. In the other locations the 889, 838 and 601 cm -1 represents the C-O stretching and bending vibration which corresponds to alcohols, ethers and esters. Table 4. Interpretation of the FTIR profile for the cyclohexane extract of M.pudica root S.NO Peak Possible Intensity Possible Compounds Value Functional Of The Cm -1 Groups Peak Strong,Broad O-H Monomeric-Alcohols,Phenols Strong,Broad O-H Monomeric-Alcohols,Phenols Medium N-H Amines Strong C-H Alkanes Strong C-H Alkanes Variable C N Nitriles Weak C C Aromatic Rings Strong NO 2 Nitro compounds Variable C-H Alkanes Variable C-H Alkanes Strong C-O Alcohols, Ethers, Carboxylic Acids,Esters Strong C-O Alcohols,Ethers,CarboxylicAcids,Esters Broad C-H Alkynes Medium C-H Alkynes Medium C C Aromatic Rings Strong C-O Alcohols,Ethers,CarboxylicAcids,Esters Strong C-O Alcohols,Ethers,CarboxylicAcids,Esters Medium C-O Alcohols,Ethers,CarboxylicAcids,Esters Medium C-H Alkynes Medium C-H Alkynes Variable C-H Alkynes The 3390 and 2917 cm -1 is due to the stretching vibration of monomeric alcohols and phenols. The 2917, 2849, 1712,1667 and 1615cm -1 bands represents alkanes which corresponds to C-H bending with medium and strong intensity of absorption. In the other locations the 833,756 and 720 cm -1 represents the C-O stretching and bending vibration which corresponds to alcohols, ethers and esters. CONCLUSION Plants have been used as medicines throughout history. Indeed, studies of wild animals show that they also instinctively eat certain plants to treat themselves for certain illnesses.medicinal plants are widely and successfully used on every continent. M.pudicais Vol 3, Issue 3,

11 one such plant in which itsroot extracts exhibits good medicinal properties so that it can be used in Pharmaceutical industries. Preliminary phytochemical screening reveals the presence of various bioactive compounds such as flavonoids, tannins, phenols, glycosides and sterols.ftir revealed the presence of various functional groups such as monomeric alcohols, alkanes, nitriles, nitro compounds and aromatic rings. In conclusion,the results presented here implied Mimosa pudicaroot extracts could be used in pharmaceutical industries for various purposes since it has wide range of applications. ACKNOWLEDGEMENT We thank IIT,Madras in helping us for processing the samples for FT-IR analysis. REFERENCE 1. Milind P., Anupam P. Aphrodisaic Activity of Root of Mimosa pudicalinn.ethanolic extract in mice. International journal of Pharmaceutical science and nanotechnology, 2009; 2(1): Azmi L., Manish KS & Ali KA.Pharmacological and biological overview on Mimosa pudicalinn.international Journal Of Pharmacy & Life Sciences, 2011; 3(1): Thomas MW.,Sulfin N., Murugan V. An insight into the Hypnotizing Herbs (VashiyaMooligaigal) as per the Siddha text PatharthaGunaSinthamani, 2010; 4. Srivastava V, Sharma A, AslamImran.A review on Ethnomedical and Traditional uses of mimosa pudica.international Research journal of pharamacy, 2012; 3(2). 5. Howard R.A. Flora of the Lesser Antilles, Leeward and Windward Islands. Dicotyledoneae, Jamaica Plain, MA: Arnold Arboretum, Harvard University., 1988; 4(1)673 p. 6. Nyam.Caribbean Food and Nutrition Institute, 2005, UWI Campus Mona, Kingston Thenmozhi M., Bavya P., Rajeswari, S. Compound identification using hplc and ftir in ecliptaalba and Emilia sonchifolia, International journal of engineering science and technology.pp: Eloff, J.N. A sensitive and quick method to determine the minimal inhibitory concentration of plant extracts for bacteria, 1988.PlantaMedica, 64, pp: Harborne JB. Phytochemicalmethods,London. Chapman and Hall, Ltd.1973;pp: Evans WC.Treasae and Evans pharmacognosy. 14 th Edition. Harcourt Brace and company,1997; Asia Pvt. Ltd. Singapore Vol 3, Issue 3,

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