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L.O. Ajala et al.

, IJSID, 2013, 3 (1), 109-116

ISSN:2249-5347

IJSID

International Journal of Science Innovations and Discoveries


Research Article
1Department 2Department *L.

An International peer Review Journal for Science

Available online through www.ijsidonline.info


O. Ajala1, M. C. Okafor2 and O. P. Ogunlowo3

EVALUATION OF PHYTO-CONSTITUENTS AND BACTERICIDAL POTENCY OF PHYLLANTUS NIRURI of Science Laboratory Technology, Akanu Ibiam Federal Polytechnic, Unwana, P.M.B. 1007, Afikpo, Nigeria; of Science Laboratory Technology, Akanu Ibiam Federal Polytechnic, Unwana, P.M.B. 1007, Afikpo, Nigeria; 3Department of Pharmacetical Microbiology, Igbinedion University, Okada, Nigeria. ABSTRACT phytochemical constituents and anti-bactericidal potentials. The mineral content of the pulverized plant was found to be 180.80, 195.30, 30.30, 10.40, 1.90, 10.60, 0.30, 0.70, and 0.30mg/100g on dry weight basis for sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc and manganese respectively. The proximate analysis of the powdered plant revealed 10.10% moisture, 7.70% ash, 8.70% protein, 25.20% crude Address: Name: L.O. Ajala Place: Nigeria E-mail: loajala@akanuibiampoly.edu.ng shown that alkaloids, glycosides and steroids were present in trace; saponins and tannins confirms the use of the plant in folk medicine in treatment of diverse ailments. constituents, Antimicrobial efficacy, Pathogenic bacteria. were detected moderately; phenols was absent while flavonoids was detected fully. The inhibition in all. The positive result for antimicrobial activities of the ethanolic extract Phyllantus niruri was assayed for its proximate compositions, mineral elements,

Received: 14-01-2013 Accepted: 17-02-2013


*Corresponding Author

fibre, 3.30% fat and 45.00% carbohydrate. Phytochemical constituents of the extract antimicrobial examinations of the plants extract on some selected bacteria showed KEYWORDS: Phyllantus niruri, Proximate compositions, Mineral elements, Phytochemical

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L.O. Ajala et al., IJSID, 2013, 3 (1), 109-116 diseases/ailments that are resistant to drugs. Nature has presented to humanity gift of vast therapeutic workshop with a wide The health benefits of medicinal plants are attributed in parts, to their unique phytochemical compositions maintaining human health [2,3]. documented diseases [4,5].
[3] [1].

varieties of medicinal plants. There are multitudes of potential and useful bioactive substance to be derived from these plants. phytochemicals are known to confer therapeutic properties on plants and they have made significant contribution in . The plant is popular in folk medicine. The whole parts of the plant are useful in treatment of multifarious

Medicinal plants have been gaining tremendous interest in the recent times in treatment of different kinds of INTRODUCTION

throughout the tropical and sub-tropical of both hemispheres. Other morphological appearances of the plant had been Many of the active metabolites are attributed to biologically active lignans, saponins, glycosides, flavonoids, alkaloids,

Among these valuable plants of therapeutic important is Phyllantus niruri. It is a small herb that is distributed

These

ellagitannins and phenylpropanoids found in the leaf, stem and root of the P. niruri. Common lipids, steroids and flavonols also occur in the plant [1,3,6,7]. Phyllantus niruri primarily contains lignans, alkaloids and flavonoids [8], glycosides and tannins [9]. P. niruri is of medicinal important and had proved to have a wide range of therapeutic effects as reported by many researchers elsewhere, among which are: hepatoprotective and antioxidant [10,11,12], hypolipemic [13], antilithic [14,15], anti-malarial [16,17,18]. There is at present little information documented in Nigeria despite wide-spread of this plant in the southern part of the niruri. The importance of such information cannot be over-emphasized, since herbal medicine has been shown to have genuine
[19].

country, concerning the chemical compositions, secondary metabolite constituents and antibacterial activities of Phyllantus utility and over 80% of the rural populace depends on it for primary health care delivery without proper evaluation before use to resistant nature of pathogenic microorganisms to the synthetic antibiotics. There is a need, therefore to examine and evaluate the phytochemicals, proximate compositions, mineral contents, bactericidal properties of this plant of hidden health serve as base-line for further studies on the plant therapeutic efficacy. institution. MATERIALS AND METHODS Efforts have been intensified by researchers towards a search for more alternative sources of antimicrobial agents owing

benefits with a view to ascertaining its potential as pharmaceutical raw material for drug formulation. The result obtained will Unwana, Nigeria. They were identified and authenticated at the department of Science Laboratory Technology of the same Preparation of sample: The uprooted whole plants were washed thoroughly and rinsed with distilled water. They were chopped to smaller size and sun-dried for two weeks and later pulverized with Thomas Wiley Milling Machine. The powder sample was preserved in clean polyethylene bottles at room temperature until needed for analysis. concentrate was preserved in an air-tight bottle for further analysis of phytochemical constituents and antimicrobial study. fibre contents were determined as described by
[20].

Plant collection and identification: Fresh whole plants of P. niruri were collected within Akanu Ibiam Federal Polytechnic,

Preparation of extract: One part of the powdered sample was soaked in methanol at room temperature for 48 hours, filtered Proximate studies: The other part of the powdery plant sample was used for proximate analysis. Ash, moisture and crude 60OC) for 6 hours. Total nitrogen was determined using the microjeldahl method and converted to crude protein content by International Journal of Science Innovations and Discoveries, Volume 3, Issue 1, January-February 2013

using sterile Whatman No1 filter paper and concentrated using a rotary evaporator with water bath set at 40 OC. The Crude fat was extracted by soxhlet method with petroleum ether (40-

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multiplying with a factor of 6.25. Carbohydrate content was determined by percentage difference of the other proximate parameters summed together. The results were expressed as averages of percentage values on dry weight basis. Mineral element determination: The mineral contents were analyzed after incineration in a muffle furnace and the ash determination of sodium and potassium by Flame Emission Photometry method. Calcium, magnesium, iron, copper, zinc, and Vanadomolybdate colorimetric method [21]. saponins, tannins, glycosides, flavonoids, phenols, and steroids using standard methods [22]. manganese by using Atomic Absorption Spectrophotometer (model 400 Perkin Elmer Analyst) and phosphorus by the obtained was dissolved in 2.0mol/dm3 HCl and diluted to 100cm3 with deionized water. The resulting extract was used for the

L.O. Ajala et al., IJSID, 2013, 3 (1), 109-116

Phytochemical analysis: Qualitative phytochemicals of the methanolic extract was tested for the presence of alkaloids, Collection of test organisms: The test organisms used for this study; Staphylococcus aureus, Escherichia coli, Pseudomonas College Hospital, Ibadan, Nigeria. Pure cultures were obtained by sub-culturing the isolated organisms. wire loop and inoculated into 5ml nutrient broth present in the test tubes and were incubated at 37 oC for 24 hrs. positive and negative control) were made on each plate with the aid of cork borer {6mm}. areoginosa, Neisseria meningitides and Bacillus subtilis were obtained from the Department of Medical Microbiology, University Preparation of Bacterial seeded plate: Dilution of 0.2ml of 10-6 each of these cultures was introduced into 20ml molten Muller Hilton agar in the bottles, it was mixed well and poured into plates and allowed to set. Three ditches (crude extract, antimicrobial activity using the seeded bacterial plates. The method employed was agar diffusion technique in which the

Bacterial cultures: The overnight bacterial culture used was prepared by sub-culturing, a loopful was streaked with a sterile

Antimicrobial sensitivity test: The crude extract was reconstituted with 30mg/ml, each extract was screened for extract was placed in plates containing a single microorganism. 40l of the extract was placed in the well with the use of pasture pipette. The plates were left on the bench for an hour at room temperature to allow diffusion of the extract into the inhibition was measured in millimeters. agar, incubated at 37oC for 24hrs. The negative control was Dimethylsulphoxide (DMSO) while gentamycin was used as positive control. After specified time of incubation, the plates were brought out of incubator and the diameter of zone of The results of the analysis for proximate compositions of the plants powder, some selected nutritional elements of RESULTS AND DISCUSSION Table 1: Result of proximate compositions of powdered sample of P. niruri on dry weight basis. Proximate composition Ash Fat Moisture 10.100.12 % weight 7.700.08 3.300.02 8.700.01 25.200.80 45.001.37

the pulverized plant, organoleptic properties of the plants extract, phytochemical constituents of the extract and antibacterial screening of the plants extract of Phyllantus niruri are presented in the tables 1 to 5 below.

Note: Data are means of replicate determinations Standard Deviation. International Journal of Science Innovations and Discoveries, Volume 3, Issue 1, January-February 2013

Crude protein Carbohydrate Crude fibre

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Table 2: Result of some nutritional elements of the pulverized sample of P. niruri Element Concentration(mg/100g) Potassium (K) 195.300.36 Sodium (Na) 180.801.10 Calcium (Ca) 30.501.00 Magnesium (Mg) 10.400.25 Phosphorus (P) 0.300.02 Iron (Fe) 1.900.02 Zinc (Zn) 0.300.00 Copper (Cu) 10..600.05 Manganese (Mn) 0.700.01 Note: Data are means of replicate determinations Standard Deviation. Parameter Observation Colour Dirty green Texture Smooth Odour Mild Table 4: Result of qualitative phytochemicals of the extract of P. niruri. Phytochemical Tannins Phenols Steroids Flavonoids Alkaloids Glycosides Saponins Inference ++ -+ +++ + + ++ Table 3: Result of organoleptic properties of the extract of P. niruri.

L.O. Ajala et al., IJSID, 2013, 3 (1), 109-116

Table5: Results showing zones of inhibitions of P. niruri extract, Gentamycin and Dimethylsulphoxide (DMSO) used as Bacteria S. aureus E. coli P. aeruginosa N. meningitides B. subtilis Extract (mm) 12.00 7.30 10.60 12.30 Gentamycin(25g/ml)(mm) 22 14 15 16 control. Dimethylsulphoxide Resistant Resistant Resistant Resistant Resistant

Legend:

++ means present moderately.

-- means absent completely.

+++ means present fully.

+ means present in trace.

(25.200.80%). The high portion which is mainly cellulose helps body to accelerate digestion and enhance easy stooling organisms, including the full components of enzymes involved in the metabolism during growth
[24].

Protein is also present in the plant. Proteins are constituents of plants and are of primary importance in both nutrition and health care delivery. They serve as components of nuclear and cytoplasmic structures that take part in maintaining cellular Fat is also present

The proximate compositions of P. niruri are presented in table 1. The plant contained high percentage level of fibre

12.90 15 Note: Data are means of replicate determinations

[23].

moderately with a value of 3.300.02%. Fat is an essential nutrient that provides energy. It contributes to the palatability of International Journal of Science Innovations and Discoveries, Volume 3, Issue 1, January-February 2013

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foods, acting as a carrier to fat soluble vitamins and supplies essential fatty acids. The low value of fat in this plant is acceptable, considering the adverse effects of high fat content in the humans body which is linked with obesity and mushroom by Anderson and Fellers (1942)
[26].

L.O. Ajala et al., IJSID, 2013, 3 (1), 109-116

arteriosclerosis [25]. The ash content of the plant was found to be 7.700.08%. Ash content of a food material is an indication of infection and reduces microbial attack [27]. 10.100.12% was obtained in this study. The moisture is related to the dry matter excellent (45.001.37%). The value obtained confirms the use of the plant as tonic [3]. indispensable for growth and well being of the body
[28].

the mineral elements present. The value obtained for this plant compared favourably with 7.0% reported for Lentinus edode

content and may be affected by season, location of the plant and stage of maturity [24]. The carbohydrate content of the plant is such minerals observed in the plant are interesting. The low concentrations of other elements apart from sodium and presence of magnesium serves as an activator of many enzymes and maintains the electric potential in nerves enough to assist in haemoglobin production in the body obtained for this plant is 0.300.02mg/100g.
[31]. [29]

The moisture content is an index of stability and susceptibility of fungal

potassium is justifiable, since minerals are often called micro nutrients because the body does not need a great deal of them, presence of calcium in ample quantity could help to excrete excess oxalate from the body. It is also important in solving the formation of high energy compounds (ATP) and involves in the synthesis of phospholipids and phosphoproteins
[31].

Table 2 revealed the concentrations of some selected mineral elements in the plant. The appreciable concentrations of

value is encouraging since potassium helps in dilation of blood vessels thereby reduces the risk of high blood pressure [25]. The problem of growth retardation and rickets[30]. Average iron concentration of 1.900.02mg/100g in the plant is appreciable Phosphorus functions as a constituent of bones, teeth, adenosine

The concentration of potassium is high (195.301.36mg/100g), this

and the

triphosphate (ATP), phosphorylated metabolic intermediate and nucleic acids. It serves as buffering action, functions in the are known to exhibit varied biochemical and pharmacological actions in animals when ingested. The table showed that the complex chemical compounds widely distributed in vascular plants. They have a large influence on the nutritive value of foods sense that they help in lowering cholesterol in the body and are analgesics
[33].

plant contained tannins in moderate proportion; therefore the fear of negative effects of high dosage is removed. Tannins are and rumen bacteria [32] . Saponins are terpene derivatives occurring widely in plant kingdom. They are beneficial to man in the stimulants and act to prolong the actions of several hormones like adrenaline and thyroid stimulating hormones and spices. They are important in medicine because of their action in the heart developing certain types of cancer [33].
[34]. [34].

Table 4 presents the phytochemical constituents of P. niruri. The plant is rich in secondary plant metabolites which

The value

eaten by man as they decrease organic matter and help in fibre digestion because of their interaction with cellulose enzymes

phytochemical compounds. They are complex nitrogen containing substances which are alkaline in nature. When

administered in small dose, they may provoke strong effects in the body and so are widely used in medicine. They are mild

Alkaloids comprise the largest class of

study, alkaloids were detected in trace. Cyanogenic glycosides on the other hand, are used in the preparation of many drugs

Flavonoids have antioxidant effects and protect body cells against damage from free radicals and tend to reduce the risk of with 12.3mm and 12.0mm respectively, it was only 7.3mm in P.aeruginosa, increasing the concentration of the extract would give higher zones of inhibition against the selected bacteria. In gentamycin, the inhibitory zone was highest with 22mm and International Journal of Science Innovations and Discoveries, Volume 3, Issue 1, January-February 2013 Shown in table 5 are the inhibitory actions of P. niruri extract against some selected bacteria. The result shows highest

Glycosides were also present in trace.

In this

zone of inhibition of 12.9mm against B. subtilis, and 7.3mm in E.coli, the activity was very close in S.aureus and N.meningitides

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least with 14mm against S.aureus and E coli respectively. P.aeruginosa and B. subtilis showed the same result of 15mm while it was 16mm against N.meningitides. bacteria and fungi nodiflora
[32,41,42,43]. [38],

L.O. Ajala et al., IJSID, 2013, 3 (1), 109-116

papaya against some selected microorganisms Pseudomonas pyocyania and Proteus

The inhibitory action of the extract compared favourably with the result obtained for Ocimum gratissimum and Carica Baphia nitida leaf against selected bacteria and fungi vulgaris[37].
[35,36]. [39].

complexes of -naphthol derivatives against B. subtilis and E. coli [40]. The plant extract is more potent than those of Synderella papaya but similar to the action of aqueous extract of the same pawpaw seeds against some bacteria and fungi treatment of diseases. CONCLUSION the high potency of P. niruri against these bacteria gives credence to it scientific claims for its uses in folk medicine for the The presence of rich nutrients and medicinally active compounds in the plant as revealed in this study has therefore
[34,44].

It was also discovered that the extract of P. niruri was less potent to ethanolic extract of seeds of Carica Finally,

It also had inhibitory action similar to that of Ipomea batatas leaf against It also compared with some metal ion

It is also in consonance with that of honey against Salmonella spp.,

strengthened various claims made by other researchers as to its medicinal and nutritional values. From all indications, P. niruri contained multifarious bioactive secondary metabolites which are the basis for plants medicinal starting materials. faked and adulterated. P. niruri had proved (from this investigation) to be useful to inhibit the growth of both gram positive ingredients that are responsible for the antimicrobial potency of the plants extract in the formulation of new drugs. 1. 2. 3. 4. 5. 6. 7. 8. REFERENCES Clerodendoron spendens. E-journal of Chemistry. 6(2):553-560. typhi. African J. Biomed. Res. 11:215-219. Press. Nowadays, pathogenic bacteria are more resistant to synthetic medicines that are often unavailable, expensive and frequently the potency against the pathogenic bacteria studied. In other word, further work should be done to elucidate the bioactive and gram negative bacteria. This plant extract can therefore be used in developing new therapeutic antibiotic drugs. This study has provided essential information in the selection of plant extract for further isolation of active agents responsible for

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