PHYTOCHEMICAL SCREENING OF THREE MEDICINAL PLANTS NEEM LEAF (Azadirachta indica), HIBISCUS LEAF (Hibiscus rosasinensis) AND SPEAR GRASS LEAF (Imperata cylindrical)

Continental J. Pharmaceutical Sciences 4: 47 - 50, 2010 © Wilolud Journals, 2010

ISSN: 2141 - 4149 http://www.wiloludjournal.com

PHYTOCHEMICAL SCREENING OF THREE MEDICINAL PLANTS NEEM LEAF ( Azadirachta indica), HIBISCUS LEAF ( Hibiscus  Hibiscus rosasinensis ) AND SPEAR GRASS LEAF ( Imperata cylindrical ) K. E. Ayeni and Yahaya S,A Science Technology Department Federal Polytechnic, P.M.B 4 20, Offa. ABSTRACT This research work determined the bioactive constituents of some medicinal plants and their effectiveness in the area of pharmacology or pharmaceutical. The phytochemical screening carried on the leaves extract of   A. indica, H. rosasinensis, I. cylindrical revealed the presence of some active ingredients such as Alkaloids, Tannins, Saponins, and Phenols also glycosides, steroids, terpenoids, flavonoids and phlobatanins are present in this extract. The result demonstrated that the presence presence of  phytochemical components in the leaves extract. It was observed that Alkanoids has the following values, 0.52g in A. Indica, 0.51g in H. Rosasinensis, 0.42g in  I. cylindrical, while Tannins also has the following value 9.00g in A. indica, 8.40g in H. Rosasinensis, Rosasinensis, 9.20g in I. Cylindrical. Saponins has the value of 1.99 in  A. indica, 1.80 in  H. rosasinensis, 1.30 in  I. cylindrica. Flavonoids have have the value of  0.62 in  A. indica, 0.38 in  H. rosasinensis, 0.32 in  I cylindrica. Phenol has the values of 0.024 in A. A. Indica, 0.600 in H. rosasinensis , .050 in I. cylindrica. KEYWORDS: Alkaloid, glycoside, flavonoid, Tannins INTRODUCTION From the inception of the existence of earth, plant has been of great importance to the animal kingdom. It contributes either as a source source of food or shelter. In the primitive ages, plants plants were also used for clothing therefore provides the basic needs of humans humans and animals as well. Out of these plants medicinal plants are common and are source of much attention in Africa. (Akenova, et al., 1996). Uniformity of the quality of drug is also of great importance as far as the therapeutically active constituent of  raw material is concern. This makes it possible for pharmacist to prescribe Numerical values through which commodities are assessed and are used to ensure uniformity of standards.(Amen, 1996) Medical plant (Akinmoladun, et al., 2007) is defined as one, which contains substance that can be used for therapeutic purposes and its precursor for the synthesis of useful drugs. They contain nutrients that can heal the body (Frease and Evans, 1985). Material like these plants that has cellular structure is referred to as organized drug in pharmacy and those with non-cellular structure as unorganized or a cellular drug (Akinpelu, et al., 2006). Medicinal plants are termed as crude drugs of natural or biological origin b y pharmacists to describe whole plant or plant parts having medicinal properties (Okwu et al., 2006). COLLECTION AND PREPARATION OF PLANT SAMPLES The plant materials were brought from the school compound in Federal Polytechnic Polytechnic Offa, Kwara State. The plants were processed and analyzed. PROCESSING OF PLANT SAMPLES The leaves of the plant are properly washed in tap water and the rinsed in distilled water. water. The rinsed leaves are 0 dried in an oven at a temperature of 35 – 40 C for 3 days. The dried leaves of each plant are pulverized using a mortar and pestle, to obtain a powdered form. The powdered form of these plants is stored in airtight glass containers, protected from sunlight until required for analysis. PREPARATION OF AQUEOUS EXTRACT OF PLANT SAMPLES The aqueous extract of each plant sample is prepared by soaking 10g of powdered sample in 200 ml of distilled water for 12 h. The extracts are then then filtered using filter paper. The extracts are then concentrated to ¼ of the original extracts i.e. 50 ml. 47

K. E. Ayeni and Yahaya S,A: Continental J. Pharmaceutical Sciences 4: 47 - 50, 2010

PREPARATION OF ETHANOL EXTRACT OF PLANT SAMPLES The ethanol extract of each plant was prepared by soaking 10g of powdered samples in 100ml of ethanol for the same 12 h. The extracts are then filtered using filter paper. The extracts are then concentrated concentrated to 50ml of the extracts sample and stored in airtight container. PHYTOCHEMICAL SCREENING METHODS A portion of the concentrated extract was used for the screening tests, both qualitative analysis and quantitative analysis using standard method (Edcoga et al., 2005). QUALITATIVE ANALYSIS ON PHYTOCHEMICAL CONSTITUENTS (a) 0.5g of powdered sample of each plant is boiled in 20 ml of distilled water in a test tube and filtered 0.1% FeCl3 is added to the filtered samples and observed for brownish green or a blue black  colouration which shows the presence of T ANNINS. (b) Phlobatannins: 10ml of aqueous extract of each plant sample sample is boiled with 1% HCl acid in a test test tube or conical flask. If the sample of plant carried phlobatannins, a deposition of a red precipitate will occur and indicates the pr esence of phlobatannins. (c) Saponins: 2g of powdered sample of each plant is boiled together together with 20ml of distilled water in a water bath and filtered. 10ml of the filtered sample is mixed with 5ml of distilled water water in a test tube and shaken vigorously vigorously to obtain a stable persistent persistent froth. The frothing is then mixed with 3 drops of olive oil and for the for mation of emulsion which indicates the presence of saponins. (d) Flavonoids: A few chop of 1% NH3 solution is added to the aqueous extract of each plant sample in a test tube. A yellow coloration is observed if flavonold compound compound are present. (e) Terpenoids: 5ml of aqueous extract of each plant sample is mixed with 2ml of CHCl CHCl3 in a test tube 3ml of concentrated H2SO4 is carefully added to the mixture to form a layer. An interface with a reddish brown coloration is formed if terpenoid constituent is present. (f) Glycosides: 1ml of concentrated H2SO4 is prepared in test tube 5 ml of aqueous extract from each plant sample is mixed with 2ml of glacial CH3CO2H containing 1 drop of FeCl3. The above mixture is carefully added to 1ml of concentrated H 2SO4 so that the concentrated H 2SO4 is underneath the mixture. If cardiac glycoside is present in the sample, a brown ring will appear indicating the presence of the cardiac glycoside constituent. (g) Alkaloids: 5g o f the plant sample sample is prepared in a beaker and 200ml of 10% CH3CO2H in C2H5OH is added to the plant sample. The mixture is covered and and allowed to stand for 4 hours. The mixture is then filtered and the extract is allowed to become concentrated in a water bath until it reaches ¼ of the original volume. Concentrated NH4OH is added until until the precipitation is complete. The whole solution is allowed to settle settle and the precipitate is collected and washed with dilute NH 4OH and the filtered. The residue is alkaloid which is then dried and weighed. QUANTITATIVE ANALYSIS OF PHYTOCHEMICAL CONSTITUENT a) Phenols: The quantity quantity of phenols is determined using the spectrophotometer method. The plant sample is boiled with 50ml of (CH3CH2)2O for 15 minutes. 5mml of the boiled samples is then pipette into 50ml flask and 10ml of distilled water is added. After the addition of distilled water 2ml of NH4OH solution and 5ml of concentrated CH 3 (CH2)3 – CH2OH is added to the mixture. The sample is made up to mark and left for 30 minutes to react for colour development and measured at 505nm wavelength using a spectrophotometer. b)

Tannins: Quantity of tannins is determined by using the spectrophotometer method. method. 0.5g of plant sample is weighed into 50ml plastic bottle 50ml of distilled is added and stirred for 1h. the sample is filtered into a 50ml volumentric flask and made up to mark 5ml of the filtered sample is then pipetted out into test-tube and mixed with 2ml of 0.1m FeCl3 in 0.1m HCl and 0.008m K4Fe (CN)6 3H2O. The absorbance is measured with a spectrophotometer at 395nm wavelength within 10min.

c)

Saponins: The sample were ground and and 20g of each plant sample sample is put into a conical flask and 100ml of 20% C2H5OH is added to the the plant sample. The sample is heated over a hot water bath for 4 0 hours with continuous stirring at about 55 C. The mixture is then filtered and the residue re-extracted

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K. E. Ayeni and Yahaya S,A: Continental J. Pharmaceutical Sciences 4: 47 - 50, 2010

d)

with another 200ml of 20% C2H50H. Then the combined extracts extracts are reduced reduced to 40ml over a water 0 bath at about 90 C.

The concentrated is then transferred into a 250mlseparation funnel and 20ml of (CH 3CH2)2O is added to the extract and shaken vigorously. The aqueous is recovered while while the (CH 3CH2)20 layer is discarded and the purification process is repeated 60ml of N-C 4H9OH is added and combined n-C4H9OH extracts is washed twice with 10ml of 5% NaCl. NaCl. The remaining solution is then then heated in a water water bath and after evaporation: the samples are dried in the oven to a constant weight. e)

Flavonoids: 10g of plant sample sample is repeatedly extracted with 100ml of 80% aqueous methanol at room temperature. The whole solution is then filtered through filter paper and and the filtrate is later on transferred into a water bath bath and solution is evaporated into into dryness. The sample is then weighed weighed until a constant weight.

RESULTS AND DISCUSSION RESULTS The Table 1 is the results on the quantitative analysis of phytochemical constituents of three medicinal plants. Plants Tannins Phlobatannins Saponins Flavonoids Terpenoids + + + +  A. indica + + + +  H. rosasinensis + + + +  I. cylindrica Plants  A. indica  H. rosasinensis  I. cylindrica

Cardiac glycosides + -

+

Alkaloids

+ +

Phenol + +

Steroid  -

+

+

-

+ = The presence of phytochemical constituents, - =The absence of phytochemical constituents:Table two: QUANTITATIVE ANALYSIS OF PHYTOCHEMICAL CONSTITUENT (%) Plants Alkaloids Tannins Saponins Flavonoids Phenols 0.52 9.00 1.99 0.62 0.04  A. indica 0.51 8.40 1.80 0.38 0.600  H. rosasinensis 0.42 9.20 1.30 0.32 0.050  I. cylindrica DISCUSSION For the qualitative analysis results, below below is the discussion. The research work that was carried out on the three medicinal plants shows that phytochemical constituents are present in which it was summarized in Table 1. It shows that tannins, saponins, flavonoids, terpenoid and alkaloid, phenol were present in all the three plants. But phlobatannins were found to be absent absent in the three plants. plants. Cardiac glycosides were were present in  A. indica and  I. cylindrica and found to be absent in  H. rosasinensis . QUANTITATIVE ANALYSIS OF PHYTOCHEMICAL CONSTITUENTS The above results show in the table table reveal the five (5) major groups phytochemicals constituents constituents the medicinal plantsre shown in the Table 2.   A. indica and  H. rosasinensis have the highest yield of alkaloid which is 0.5g in which   I. Cylindrica has the lower yield of alkaloid which which is 0.4g. But   A. Indica and  I. cylindrica showed from the result to have highest yield of Tannins which is 9.00g. Moreover  A. indica, H. rosasinensis, and T. and  T. cylindrica have been founded to have similar yielded of saponins which are 1.99, 1.80 and 1.30 percentages respectively.

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K. E. Ayeni and Yahaya S,A: Continental J. Pharmaceutical Sciences 4: 47 - 50, 2010

 A. indica has the highest yield of flavonoids which is 0.62% followed by  H. rosasinensis which is 0.38% and I. cylindrica is found to have the least amount of flavonoid which is 0.32%.

Finally   H. rosasnensis has the highest yield of phenols quantity and followed by   A. indica and   I. cylindrica which are in similar or the same range. CONCLUSION This research work has revealed further potentials of these three plants in the area of pharmacology as potential source of useful drugs. T his study therefore has provided some biochemical basis for ethno pharmacological uses of these plants in the treatment and prevention of various diseases and disorders. RECOMMENDATION Since the results of the phytochemical screening have shown that the extract of the three samples contain alkaloids, tannin, saponin, phlobatannin, terpenoid, flavonoid, cardiac glycoside and phenol. Therefore I recommend that these extracts should be used in the production of drugs in the area of  pharmaceutical or pharmacology for the treatment of intestinal track, diarrhea, headache, skin disease and it should be used as a source of pesticide and insecticide, also herbicide in the agricultural area so as to improve the yield of crops and reduce pests, weeds and other organism which are competing with man. REFERENCES Akinpelu, D. A. and Onakoya, T.M. (2006). Antimicrobial activities of medicinal medicinal plant used in folk lore remedies in south-western. Afri. J. Biotechnol, Biotechnol, 5:1078-1081. Akinmoladun A. C., Ibukun E.O., Afor E., Obuotor, E.M. and Farombi E.O. (2007). Phytochemical constituent and antioxidant activity of extract from the leaves of Ocimum gratissimum. Sciences Research Essay 2: 163 166. Okwu, D. E. and Josiah, C. (2006), Evaluation of the chemical composition of two Nigerian Medicinal plants  Afri. J. Biotechnol. 5(4): 357-361. Oluronke, Taiwo ( ) Department of Applied Oral Science, Faculty of Denstistry, Dalhousie, University, Canada B3H315. A research article on antibacterial activities of extracts from Nigeria medicinal plants. Akenova, M.E. and Attackrah, A.N. (1986) Control of spear grass (imperata cylindrical {L} Bearuv in an alley cropping fallow. Nitrogen –fixing Tree research reports 4; 24-28. Amon, (1996) Imperats management for small holder an extensionists guide to rational imperata management for small holders. Received for Publication: 07/09/2010 Accepted for Publication: 19/10/2010 Corresponding Author K. E. Ayeni Science Technology Department Federal Polytechnic, P.M.B 420, Offa

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