World Journal of Pharmaceutical Sciences ISSN (Print): 2321-3310; ISSN (Online): 2321-3086 Published by Atom and Cell Publishers © All Rights Reserved Available online at: http://www.wjpsonline.org/ Original Article
Phytochemical and Antimicrobial studies of stembark extracts from Uapaca pilosa (Hutch.)
George I. Ndukwe, Muhammed S. Sallau and Ayodele J. Atibioke
Department of Chemistry, Ahmadu Bello University, Zaria, Nigeria
Received: 06-02-2016 / Revised: 21-04-2016 / Accepted: 28-04-2016 / Published: 28-04-2016
ABSTRACT
Objectives: To determine the antimicrobial activity and to also detect the phytoconstituents that are present in the various fractions of the crude extract of Uapaca pilosa. Methods: The antimicrobial and medicinal properties of Uapaca pilosa stembark extracts was examined through phytochemical screening and bioassay on some selected microorganisms: E. coli, S. typhill, S. aureus, B. sbstilis, S. dysenteriae, C. albicans and C. krusei. Result: The preliminary phytochemical screening of the plant extracts were assessed using standard methods. Test for the following active components: alkaloid, anthraquinones, carbohydrate, flavonoids, saponins, glycosides and tanins. All were present. The n-hexane extract show more activity against most of the selected microorganisms than other fractions of the plant extracts measuring clear zones of inhibition of between 10mm and 18mm. but none of the extracts show activity against C. albican and C. krusei. This shows that the bioactive components present in Uapaca proves potency in the treatment of some bacterial infections. Conclusion: the results could justify the use of the plant Uapaca pilosa in traditional herbal medicine for the treatment of bacterial infections.
Keywords: Antimicrobial activity, hexane extract, phytochemical studies, Uapaca pilosa.
INTRODUCTION. important compounds like alkaloid, anthraquinones, flavonoids, phenolic compounds, From the time being world traditional medicine saponins, steroids, tannins, terpenoids etc, [4]. have being known to take it source from the higher plants and their extracts in the treatment of diseases Uapaca pilosa is a small to medium-sized tree and infections [1]. Traditional medicinal practice under phyllanthaceae family and belongs to the has been establish for centuries in many parts of the order malpighiales. The plant is distributed in world. Diverse plants and herbs are used all over tropical Africa and Madagascar. Other member of Nigeria by traditional medicine practitioners. These the family include Uapaca togoensis, Uapaca practice is known to be vary from one country to guinessis, Uapaca bail, Uapaca paludosa, Uapaca another [2]. Extracts from the various part (leaves, le-testuana, Uapaca lebrunii, Uapaca littoralis, stem bark and roots) of various higher plants are Uapaca paludosa, etc. Uapaca species are part of used in herbal medicine production [3]. Plant ethno medicinal plants used by Africans in treating extracts are giving singly or as concoctions for various diseases. The family phyllanthaceae various ailments. In actual sense more than 75% of comprises of about 60 species worldwide, of which the Nigeria population depend on these various 49 are confined to continental Africa [5] include forms of concoctions and herbal decoction for the the genus among those genera whose species treatment of some infections. Phytochemical composition require re-consideration. Some of the constituent are the basic raw material source for the species in the genus are represented in West Africa establishment of several pharmaceutical industries. [6, 7, and 8]. The constituents present in the plant play a vital role in the crude drugs identity. Phytochemical Two of the species are commonly used in folkloric screening is very important in identifying new medicine in the West African sub region [9]. sources of therapeutically and pharmacologically Uapaca heudelotii is medicinally useful for treating
*Corresponding Author Address: Dr. Ayodele J. Atibioke, Department of Chemistry, Ahmadu Bello University, Zaria, Nigeria; E-mail: [email protected] Atibioke et al., World J Pharm Sci 2016; 4(5): 200-204 skin infections, female sterility and pile, Antimicrobial Assay rheumatism, as an emetic, mouthwash and gargle Test organisms: 24hour cultures of six human for tooth-troubles and as a source of highly-priced pathogenic bacteria made up of gram positive hard timber [10, 11]. Uapaca togoensis is used for (Staphylococcus aureus and Bacillus substilis), treating female infertility and as a restorative wash gram negative (Shigella dysentriae, Escherichia against fatigue and for making charcoal [12]. The coli and Salmonella typhi) bacteria and (Candida wood of Uapaca staudtii is termite-proof and albicans) fungi were used for the in-vitro difficult to work upon because of its chemical antibacterial assay. All microorganisms were composition and strength; it is used for making obtained from the laboratory stock of Microbiology furniture, railway sleepers and barrel staves [10]. Department, Faculty of Science Ahmadu Bello Uapaca paludosa and Uapaca vanhouttei have also University Zaria. The culture media used for the been reported for making charcoal and used as study include Mueller Hinton agar (MHA), Mueller firewood [10]. Hinton broth (MHB), Potato dextrose agar (PDA) and Nutrient agar (NA). The above mentioned To the best of our knowledge there was no media were used for sensitivity test, determination phytochemical study on any part of this plant. of minimum inhibitory concentration (MIC) and Hence this work has been aimed at studying the minimum bactericidal concentration (MBC). All phytochemical and antimicrobial pattern of the media were prepared according to manufacturer plant by extracting and carryout both instructions and sterilized by autoclaving at 1210c phytochemical and antimicrobial studies of the for 15 minutes. various extracted fractions from this important medicinal plant. Antimicrobial agents: Amoxillin, 30µg/ml (Mecure industries limited Lagos Nigeria.) was EXPERIMENTAL SECTION used as standard antimicrobial reference drug.
Plant material: the fresh whole plant of Uapaca Determination of Inhibitory Activity using agar pilosa vernacular name Obloblo (Idoma) was diffusion method: The disc diffusion assay as collected from Benue state North-central part of describe by [15, 16] was used to screen for Nigeria. The botanical identification of the plant antibacterial activity. An overnight broth culture was carried out in the biological science herbarium used to obtain Standard inoculum (0.5 McFarland Ahmadu Bello University Zaria. The sample was standards) was introduced onto the surface of the air dried and the dried stembark was crushed to sterile Mueller Hinton Agar (MHA) plates and a powder so as to enhance effective contact of sterile glass spreader was used for even distribution solvent with sites on the plant material. over the media. Four holes 6mm respectively were bored in each of the plates 9cm diameter with an Extraction and Isolation: Dry powdered stembark aseptic cork borer, when seeded plates had material was extracted using maceration method solidified; 100mg/ml, 50mg/ml, 25mg/ml with methanol the crude methanol extract was and12.5mg/ml of extract were prepared in partitioned using n-hexane, chloroform, dimethylsulphoxide (DMSO) by preparing a stock ethylacetate, respectively. The crude methanol, n- solution and carrying out double fold dilutions on hexane, ethlacetate and chloroform extracts were it. And with the aid of a Syringe, the wells were filtered and concentrated under a reduced pressure filled with 0.25 ml (5drops) of different dilutions of using Rota vapor and air dried. the extract while the centre well was filled with 30μg/ml of amoxillin (also dissolved in DMSO). Preliminary phytochemical screening: In getting Diameters of zones of inhibition were determined information about the class of chemical compound after incubating plates at 37°C for 24h for existing in plant material the importance of bacteria. This test was conducted first on the crude phytochemical screening cannot be over look in extracts and the solvent dimethylsulphoxide was that it help the researcher to decide which extract to used as negative control while amoxillin was used be further isolated. The phytochemical screening of as positive control [17]. the plant stem bark were carried out using the standard procedures [13, 14]
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RESULTS AND DISCUSSION
Phytochemical Screening
Table 1: Result of phytochemical screening of stembark extracts from Uapaca pilosa Secondary Methanol Ethylacetate Chloroform n-hexane Metabolites Alkaloids + + - + Anthraquinines + + - + Carbohydrate + + + + Flavonoids + + - - Glycosides + + + + Saponins + - - + Tannins + - + + Key: +=present, -=absent.
Antimicrobial studies
Table 2: result of Antimicrobial screening of stembark extracts from Uapaca pilosa Table 2.1: diameter of zone of inhibition (ZI) mm Test organism Methanol Ethylacetate N-hexane Chloroform Amoxillin S. aureus 14 17 14 13 28 S. typhil 10 10 11 - 20 S.dysentriae 13 15 12 12 19 B. subtilis 11 14 10 - 22 E. coli 15 15 18 17 15 C. albicans - - - - -
Table 2.2: minimum inhibitory concentration (MIC) mg/ml Test organism Methanol Ethylacetate Chloroform N-hexane S. aureus 6.25 12.50 12.50 12.50 S. typhil 0.00 0.00 0.00 0.00 S.dysentriae 25.00 25.00 0.00 0.00 B. subtilis 0.00 6.25 0.00 0.00 E. coli 12.50 12.50 12.50 12.50
Table 2.3: minimum bactericidal concentration (MBC) mg/ml Test organism Methanol Ethylacetate Chloroform N-hexane S. aureus 12.50 25.00 25.00 25.00 S. typhil 0.00 0.00 0.00 0.00 S.dysentriae 50.00 50.00 0.00 0.00 B. substilis 0.00 12.50 0.00 0.00 E. coli 25.00 25.00 25.00 25.00
DISCUSSION methanol extracts but absent in the chloroform and hexane extracts, the presence of saponins was The phytochemical screening of all the extracts was observed in hexane and methanol extracts but analysed and their results are shown in table-1. In absent in ethylacetate and chloroform extracts. It is the present study carbohydrate and glycosides were expected that the identification of these noted to present in all four (hexane, chloroform, phytochemicals will help to draw a guidelines to ethylacetate and methanol) extracts of tested plant, ascertain the quality of the tested plant. while tannins was showed to be present in three Phytochemicals detected in this plant, possess (hexane, chloroform and methanol) extracts but various biological activities which may help in absent in ethylacetate extract, alkaloids and fighting against chronic diseases for example anthraquinones were present in three (hexane, alkaloids have anti-microbial, anti-hypertensive, ethylacetate and methanol) extracts but absent in anti-arrhythmic, anti-malarial, and anti-cancer chloroform extract, whereas flavonoids were effects [18]. Tannins in traditional medicine used showed to be present in the ethylacetate and against diarrhoea, duodenal ulcer, anti- 202
Atibioke et al., World J Pharm Sci 2016; 4(5): 200-204 inflammatory, anti-septic, haemostatic, astringent with the standard drug as can be seen in the tables and as diuretic. Flavonoids (hydroxyl phenolic (2.1, 2.2 and 2.3) above which make them good for compounds) showing activities, such as anti- consideration in the manufacture of anti-infections microbial, antioxidant, anti-tumour, anti- drugs. The inhibitory effect of the extract of U. inflammatory, estrogenic, anti-allergic, enzyme pilosa against several bacterial species is an inhibition and vascular activities. Saponins have indication of broad spectrum antimicrobial been known to possess anti-oxidant, anti-microbial, potential, thus introducing the plant as a potential and hypoglycaemic in activities. Cardiac substance for drug development for the treatment glycosides used as antiarrhythmic agents and the of infectious diseases caused by these pathogens. treatment of proliferative diseases as well as for the inhibition of several cancers, such as lung, prostate, CONCLUSION breast and leukaemia. Carbohydrates or reducing sugar known to provide useable energy to living The study shows that the stembark of Uapaca cells [19]. Therefore, use of this plant in the pilosa is a potential anti infections agent, based on production of drugs is well justified to achieve the the detection of various phytoconsistuents like desired effect in therapeutic processes. [20, 21 and alkaloids, flavonoids, anthraquinones, cardiac 22]. glycosides, carbohydrate, saponins and tannins. The presence of these secondary metabolites have The antimicrobial screening of all the extracts been established for various pharmacological (hexane, chloroform, ethylacetate and methanol) activities such as antimicrobial, anti-malaria, shows relevant antimicrobial activities, comparable anticancer and analgesic etc. The antimicrobial with that of the standard drug used. The zone of studies of the extracts validate the plant potential to inhibitions showed by the various extract has a serve as an alternative to some known antibacterial significant effect which can be compare to the drugs used in the treatment of bacterial infections. standard drug. The ethylacetate extract show a These observed activities justify reasons why the very good inhibiting strength against most of the plant should be consider in the manufacture of selected microorganisms with an inhibition zones antibacterial drugs. between 10-17mm. Salmonella aureus had the highest zone of inhibition whereas Staphiloccocus Recommendation typhil had the lowest as shown in table 2.1. From table 2.2 the MIC of the ethylacetate extract range There is need for further study, isolation and between 0-12.5mg/ml with no effect on characterization of compounds from the plant in Staphiloccocus typhil with 0 value while Shigella other to establish the active constituents of drug for dysentriae showing MIC value of 25mg/ml. Also easy syntheses. the MBC ranged between 0.00-50.00mg/ml with Eschrichia coli and Staphiloccocus aureus having ACKNOWLEDGEMENT MBC value of 25mg/ml, Bacillus subtilis showed the lowest MBC value of 12.5mg/ml by this it Thanks to God Almighty for his grace, favour, shows that the microorganism Bacillus subtilis can knowledge and strength to complete this work. My be kill completely at such lower concentration profound gratitude goes to my supervisors; Prof. G. compared to others with higher concentration. Also I. Ndukwe and Dr M. S. Sallau for their patience, like ethylacetate extract other extracts (methanol, encouragement and support during the period of hexane and chloroform) showed some antibacterial this work. potentials which to certain extent can be compare
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