Phytochemical and Antimicrobial Analyses of Extracts of Peperomia Pellucida (L) Edewor-Kuponiyi Theresa

Edewor-Kuponiyi / Journal of Pharmacy Research 2012,5(5),2934-2937 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Phytochemical and antimicrobial analyses of extracts of Peperomia pellucida (L) Edewor-Kuponiyi Theresa. Ibibia Department of Pure and Applied Chemistry, Ladoke Akntola University of Technology, Ogbomoso, Nigeria Received on:11-01-2012; Revised on: 17-02-2012; Accepted on:19-04-2012 ABSTRACT The leaves of Peperomia pellucida were studied with the aim of investigating the antimicrobial and phytochemical properties of the species that thrives in Ogbomoso, Nigeria. The phytochemical screening of the extracts revealed presence of flavonoids and saponins while alkaloids, steroids and tannins were absent in the methanolic and ethyl acetate extracts. All the phytochemicals screened were absent in the n-hexane extract. The antimicrobial activity of the extracts was tested against seven bacteria and two fungal strains. It showed inhibitory activity against Pseudomonas aeruginosa, Salmonella typhii, and Shigella. dysenteriae was inactive against Staphylococcus aureus, Klebsiella pneumoniae, Proteus mirabilis and Proteus vulgaris with minimum inhibitory concentration ranging from 1.25 – 2.50 mg/mL. It did not exhibit any inhibitory activity against the screened fungi, candidida albicans and Aspergillus niger. The results from this present study indicate that the observed phytochemicals are responsible for its medicinal properties; which is an affirmation of the use of this plant in the management of abdominal ailments only. The additive or synergistic action of these phytochemicals at target sites associated with physiological process may be responsible for the beneficial effects exerted by Peperomia pellucida. KEY WORDS: Peperomia pellucida (L); Phytochemical; Antimicrobial activity; TLC INTRODUCTION There is a worldwide increase in life threatening infections caused by patho- a medicinal in tropical Africa. In Nigeria (Ogbomoso). It is used mainly for genic microorganisms (Khan et al., 2010). These microorganisms are becom- the treatment of abdominal pains. Literature of many researchers prove that ing resistant to known antimicrobial agents (Ghani et al., 1998). Therefore the plant contain alkaloids, saponins, tannins and cardenolides (Khan et al., there is an increased interest in the search for antimicrobial compounds. This 2010) flavonoids, essential oils and carotol (Khan et al., 2002). This paper has led to the pharmacological and chemical investigations of medicinal plants. describes the phytochemical and antimicrobial study of the leaves of These investigations have provided important advances in the therapeutic Peperomia pellucida L that thrives in Ogbomoso, Nigeria. approach to several pathogens. MATERIALS AND METHODS Peperomia pellucida is an annual, shallow-rooted herb that belongs to the family Piperaceae (Ghani, 1998). It is commonly known as shiny bush. It is General procedure found in Nigeria, Sierra Leone and Ghana. Other places where it is found Thin layer chromatography was carried out on precoated plates with silica include China, Brazil and DR Congo. Other places where found are southern gel 60 (Merck 0059381). All solvents were routinely distilled prior to use. America and Asian countries (Ghani et al., 1998; Bayma et al., 2000; Santos Other chemicals were of commercial grade and used without further purifica- et al., 2001; Mde et al., 2002; Arrigoni-Blank et al., 2004). It grows in clumps, tion. thrives in loose, humid soils, tropical and subtropical climate. It usually grows to a height of about 15 to 45 cm and is characterized by succulent PREPARATION AND EXTRACTION OF PLANT MATERIAL stems, shiny, heart-shaped, fleshy leaves and tiny, dot-like seeds attached to Fresh plants were collected from its natural habitat in Ogbomoso, Nigeria and several fruiting spikes (Dos-Santos et al., 2001). The ethnomedical proper- taken to the Department of Pure and Applied Biology, Ladoke Akintola ties of P. pellucida are well known. It is used for treating abdominal pain, University of Technology, Ogbomoso, Nigeria for identification. The plant abscesses, acne, boils, colic, fatigue, gout, headache, renal disorders, and was pulverized using a sterilized food grinder. The pulverized plant was rheumatic joint pain (Ragasa et al., 1998; Khan and Omoloso, 2002). In extracted using solvents of different polarities (n-hexane, chloroform, ethyl Bolivia, the whole plant is used to stop hemorrhages by Alteños Indians. The acetate and methanol). 500 g of the plant was soaked in each of the solvents roots are used to treat fevers and the aerial parts are used as dressing for for 24 hours (3X), filtered through Whatman No.1 filter paper into screwed wounds (Munoz et al., 2000). In northeastern Brazil, the plants have been vials and concentrated using a rotary evaporator bath under reduced pres- used as a hypocholesteremic agent (Bayma et al., 2000). It is a popular cough sure. Some of the extracts were diluted with dimethylsulphoxide (DMSO) suppressant, emollient, and diuretic in Guyana and the Amazon as well as and used for antimicrobial screening. effective in the treatment of proteinuria (Arrigoni-Blank et al., 2002; De Fatima Arrigoni-Blank et al., 2004). A decoction of the plant is used in the PHYTOCHEMICAL ANALYSIS Philippines to decrease uric acid levels and to treat renal problems. It is also used topically for skin disorders such as acne and boils. It is widely used as The method used is that described by Harborne (1993). Test for flavonoids: 1 mL of NaOH was added to 3 mL of each extract. A *Corresponding author. yellow colouration indicates that flavonoids are present. Theresa. Ibibia Department of Pure and Applied Chemistry, Test for steroids: 1 mL of concentrated sulphuric acid was added to 2 mL of Ladoke Akntola University of Technology, each of the extracts. Observation of a red colouration indicates presence of Ogbomoso, Nigeria steroids. Journal of Pharmacy Research Vol.5 Issue 5.May 2012 2934-2937 Edewor-Kuponiyi / Journal of Pharmacy Research 2012,5(5),2934-2937 Test for saponins: 2 mL of distilled water was added to 2 mL of each extract Disc diffusion test and shaken vigorously. A persistent frothing indicates presence of saponins. Disc diffusion method by Bauer et al., 1963 was employed. This involved the use of filter paper disc as carrier for the antibacterial agents. Sterilized Test for tannins: 2 mL of 5% ferric chloride was added to 1 mL of each discs cut from Whatman no. 1 filter paper were impregnated with solutions extract. Observation of a greenish precipitate indicates the presence of tannins. of the antibacterial agents. The solvent was evaporated and the disc dried Test for alkaloids: 1 mL of 1% HCl was added to 3 mL of each extract in a test properly. The nutrient agar medium was inoculated with the test organism tube. The mixture was heated for 20 mins, cooled and filtered. The filtrate and the impregnated disc placed on the surface of the nutrient agar. The was used for the following test: 2 mL of Meyer’s reagent was added to 1 mL antimicrobial agent upon contact with the agar diffused into all directions. of the filtrate. Observation of a creamy precipitate indicates the presence of The ability of the test organism to grow or not in the presence of the test alkaloids. sample was then determined within 24 hours by measuring the zones of inhibition. The plates were incubated upside down at 37oC. All tests were CHROMATOGRAPHY done in quadruplicate and the antimicrobial activity was expressed as a mean of inhibition diameters (mm) produced by the plant extracts and Streptomy- Spotting of samples cin, standard antibiotic. Precoated Merck silica gel plates were used for the qualitative analysis. The plate was marked lightly with two pencil lines 0.5 cm from each end. A micro Minimum inhibitory concentration (MIC) drop of a solution of the extract in redistilled solvent (methanol, chloroform The minimum inhibitory concentration (MIC) of the extracts against the and ethyl acetate) was placed precisely on one of the two lines with the aid sensitive microorganisms was determined using the disc diffusion method. of a micro-capillary tube. Serial dilutions of the isolated compounds were prepared (10.0, 5.0, 2.5, 1.25, 0.625, 0.313 and 0.156 mg/ml). Each of the innocula was poured into Development of plate each Petri-dish and the agar was later poured and allowed to set. Wells were After spotting the sample on the plates, it was left for 5mins for the solvent made using the sterile 3 mm cork borer. Serial dilutions of the isolated com- to evaporate off and was subjected to linear ascending development to a pounds were added into the marked wells. The plates were incubated at 37oC distance of about 90 mm in different solvent systems of ethyl acetate and for 24 h. The growth was observed to determine the sensitivity of the micro- chloroform in the ratio of 9:1 (v/v) and 1:9 (v/v ) using a chromatographic organism using clear zones of no microbial growth. The least concentration of developing tank. The tank was left undisturbed until the rising solvent front the extracts that had inhibitory effect was taken as the minimum inhibitory has reached the upper line. The plate was then removed and the solvent concentration (MIC) of that extract against such microorganism. allowed to evaporate from it. RESULTS AND DISCUSSION Visualization The plate is placed in an iodine tank to visualize the colorless components. Phytochemical analysis The iodine vapor is absorbed on the plate wherever there is concentration of The phytochemical analysis of the leaf extracts showed presence of alkaloids organic material and produces a brown spot. After the colour has developed and flavonoids in the methanol and ethyl acetate extracts only. Tannins, the plate is removed and a circle penciled around each spot. On exposure to saponins and glycosides were observed to be absent in these. All the screened air the brown iodine spots evaporate gradually.

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