Antibacterial Screening of Crude Fractions of Calotropis Procera (Linn) and Gc-Ms Profile of Fractions Obtained from the Extracts of Calotropis Procera

Traditional medicine also known as Indigenous or folk medicine comprises of knowledge systems that developed over generations within various societies before the era of modern medicine. The World Health Organization (WHO) defines traditional medicine as the sum of total of the knowledge, skills, and practices based on the theories, beliefs and experiences indigenous to different cultures, whether explicable or not, used in the maintenance of health as well as in the prevention, diagnosis, improvement or treatment of physical and mental illness. The column chromatography was used to collect the crude fractions; solvents like petroleum ether, chloroform and methanol were used. The antibacterial activities of the crude fractions of Calotropis procera (leaf and stem) were evaluated in this study using some selected microorganisms like Escherichia coli, Shigella dysenteriae ATCC 24162, Salmonella typhi and Klebsiella pneumoniae ATCC 34089, whereby the plant was extracted using (acetone, methanol and aqueous). The paper disc method was used after which the zone of inhibition around the discs was estimated. The results showed that in the leaf of the plant; E. coli with Aqueous-methanol fraction and K. pneumoniae ATCC 34089 with acetone-petroleum ether fraction had the highest yield of 30mm respectively while in stem; K. pneumoniae ATCC 34089 with methanol-methanol fraction had the highest yield of 25mm. The structural elucidation of the bioactive compounds in the extracts were evaluated using Gc-Ms (Gas Chromatography-Mass Spectophotometry) which reveals the chemical compounds like phenol, methyl palmitate, Phthalic acid, 9-octadecenoic acid and other compounds known for valuable antimicrobial, biological activities and antioxidant properties. The leaves and the stem are having good chemical compounds that can be responsible for the antimicrobial and antioxidant properties.


Introduction
The use of herbal medicine as alternative therapy has prevalent throughout the world due to the growing resistance of pathogens to conventional antibiotics De Smet [1]. The need for more potent, safe and affordable drugs has led to intensified research into herbal drugs, the result of which is the introduction of new herbal preparation for therapeutic uses Griggs et al. [2]. Medicinal plants are frequently used as remedies for many infectious diseases Ahmed et al. [3]. The treatment and control of diseases by the use of the available medicinal plants in a locality have been helpful and of a priority to majority of urban and rural dwellers in healing various diseases because of the reliability and stability in plant products for healing Akharaiyi et al. [4] Calotropis procera (Apple of Sodom) is a shrub or small tree, which has become a serious weed in pastures and overgrazed rangelands. It is a native to West Africa as far as south as Angola, North and East African, Madagascar, the Arabian Peninsula, Southern Asia, India and China to Malaysia. Calotropis was formerly placed in the family of Asclepiadaceous (the milkweed family), which is now considered a subfamily of the Apocynaceae

Collection of Plant Samples
Apparently healthy plant namely C. procera were collected from Ado-Ekiti, Ekiti State Nigeria.

Preparations of Plant Extracts
The plants parts leaves and stem were air-dried for 5 weeks at room temperature (25+2 o C) and then ground to powder with a mechanical grinder (Thomas Wiley machine, model 5 USA).
Powders (200gs) of each plant were extracted with 1litre of sterile aqueous water, ethanol, methanol and acetone separately at room temperature (25+2 o C). They were labeled as crude extracts.

Antibacterial Screening of the Crude Fractions
The evaluation of antimicrobial activity was performed for all fractions by the paper disc agar diffusion method following the

Column Chromatography and Fraction Extracts
Glass wool was placed at the outlet of a column after which, one gram of plant extract powder was weighed into it and subjected to column chromatography (30 x 8cm column) using 60g of silica gel 60 F254 (Merck, 0.020mm thickness). The column was successively eluted first with petroleum ether (150ml) and then with chloroform (80ml): methanol (2ml) (40: 1) and finally with 100% methanol (150ml). Each 100 ml eluent was collected into a round bottom flask (250ml) capacity and distilled to obtain fractions. The fractions collected were numbered. Gc-Ms analysis was carried out in GCMS-QP2010 PLUS shamadzu.

Antibacterial Screening of Crude Fractions
The Antibacterial screening of the crude fractions of

Discussion
Antibacterial activities of the crude fractions showed that this plant can be used for curing many diseases such as pneumonia, dysentery etc. The results demonstrated that the crude extracts were more efficient than the fractions, this findings might suggest that Phytochemical constituents in combination may be having synergy in their efficacy, which is in agreement with many other report that have shown higher antibacterial potency of crude extracts as compared to the fraction Martins et al. [6]. The difference of various plant extracts in the antimicrobial activity is expected, as the activity is based not only on the different structures of microorganisms but also on their susceptibilities Zhang et al. [7].
This inhibitory action could be attributed to the phytochemical constituents, since these constituents are well established as antimicrobial agents Nenaah [8]. In the present study, in the agar diffusion assay the methanol and chloroform extracts showed the larger inhibition halo enabling to observe the extraction potential

Conclusion
The present study has investigated the efficacy of Calotropis procera which can be considered in folklore or traditional medicine, edible vegetable and animal forage. The overall results showed that the crude fractions of this plant have an appreciable antibacterial activity on the selected microorganisms and also the chemical compounds present in the plant simplifies that C. procera extracts could maximally serve as alternative to highly rated synthesized drugs whose costs are unaffordable by the common man.