# I. Introduction

ungi are rich sources of bioactive compounds. The medicinal chemists have always tried to design drug substance possessing maximum therapeutic application and minimum toxicity. Soil is traditionally the main source of fungal genetic resources for bioprospection programs. To expand the search for pharmacologically active agent, soil sample from virgin forest of 80 years old in Ikota, Nigeria was examined.  


# II. Methodology


# b) Isolation of the fungi

One gram of soil sample was transferred to a sterile Erlenmeyer (EM) flask containing 50ml sterile water. The flask was shaken on rotary shaker for 30 minutes for the detachment of the spore chains. The flask was kept aside for 30 minutes to settle down the particulate matter. The clear supernatant was diluted with sterile water (dilutions 10 -1 -10 -3 ) was used on innocular. One ml of each of these dilutions was pipette out into the medium, plated into Petri dishes 6mm diameter and incubated at 28 0 C for 2-4 weeks and potato dextrose agar was used.


# c) Identification

Cultural observation: using the natural eyes and microscope at low power magnification (x40), parameters such as, colony color, color change in the medium, characteristic of the submerged hyphae whether rhizoid, spiral or regular and characteristic shape of mature fruiting bodies are strictly observed.


# d) Microscopic observation

A small piece of mycelium free of medium was transferred using inoculating needle on to a glass slide containing a drop of cotton blue in loctophenol and the mycelium was spread properly with another needle. The preparation was covered with a cover slip and observed under medium power (x100) and later at high power (x400) magnifications. Details of spore colouration, shape, septation and surface marking were studied and P. Notatumwas identified and confirmed by professor of microbiology in Microbiology laboratory.


# e) Culturing the fungi

The fungi with strong antagonistic efficacy were culture in the laboratory for maximum yield of bioactive compounds using the fabricated fermenter, five ml of already cultured fungi was put into sterilized potato broth of 500ml and poured into the fermenter and allowed to excrete maximum yield for two weeks, the air pump supplied continuously sterilized air and the culture being mixed together using powered mixer.


# f) Extraction and Purification

The fungi cultures were centrifuged and extraction of compounds from multiplied fungi was carried out in a separating funnel using ethyl acetate. The extract was concentrated using rotary evaporator. The fraction was eluted using mixture of 50% ethyl acetate and 50% hexane through column chromatography and the fractions was concentrated using rotary evaporator. g) Gas Chromatography-Mass Spectrophotometer (GC-MS) Analysis was conducted using an HP (Hewlett Packard, 5890 series ? GC hyphenated with 5989 Mass Spectrometer). MS conditions were as follows: Detector mass spectrometer voltage 70eV and its source temperature was 300oC. The injector temperature was 240 0 C and the split less mode 0.5?L injection. The HP 55% dimethyl-95% diphenylpolysiloxane non-polar column was performed with length 30 cm x 0.25 mm, coating thickness film 0.25 ?m. The oven was adjusted at 100 0 C for 1 min and initial time 1.5 min with 40 0 C which ended by a final temperature of 300 0 C and 4 min hold time where the total run time was 45 min. The components were identified by comparing their retention times with those of authentic samples, as well as by comparing their mass spectra with those of (NIST).   Several naphthalene containing drugs are available, such as nafacillin, naftifine, tolnaftate, Also identified in the fraction was P-cymene having retention time of 5.170 minutes, percentage of total 16.439 % was revealed, this compound has just been patented and useful as nutraceutical composition for cognition -cognitive functions and psycho-social status, such as learning, memory and alertness, psychotic stability and maintenance (Ann, 2010 ). Benzene 1, 2, 4, 5tetramethyl having retention time 5.117 minutes and percentage of total 6.505 % was identified. Many compounds that have benzene ring have been synthesized and possess strong pharmaceutical activities such as Aspirin, sulfanilamide, amphetamine, acetaminophen.


# III. Results

2-isopropylidene-5-methylhex-4-enal having retention time 5.508 minutes and percentage of total 5.446 %. From the figure 3 showing the chromatogram of the fraction, it is found that p-cymene having % of total 16.439% was the highest followed by Naphthalene of % total 11.369 %, n-Nonadecanol-1 of % total 10.302 %. Understanding the biochemistry of alcohol metabolism has helped to develop treatments for alcohol abuse. For example, the drug Antabuse inhibits aldehyde dehydrogenase allowing a toxic accumulation of acetaldehyde to occur when alcohol is consumed (Brick, 2003;Brick and Erickson, 1999;Crews,2003;Pohorecky, L. and Brick).Trans-Decalin, 2methyl of % total 9.468 %. These compounds have been found to have various pharmaceutical applications, this include as anagelsic, antimicrobial activities and as composition in the formulation of drugs and other industrial compounds.


# V. Conclusion

From the above analysis, the fungus, P. Notatum has been found to be a reservoir of many bioactive compounds. if these compounds could be isolated and characterized using various spectroscopic techniques, novel and lead candidates compounds may be discovered.
1![Figure 1: Showing the map of study area Author ?: Department of Chemical Sciences, Afe Babalola University, Ado-Ekiti, Nigeria. Author ? ?: Department of Biological Sciences, Afe Babalola University, Ado-Ekiti, Nigeria. e-mail: adewolen50@yahoo.com](image-2.png "Figure 1 :")
2![Figure 2 : Showing the flowsheet for the process a) Soil sample collection Soil sample collection was done in July 2013 in Ikota, Ondo State, Nigeria. Random sampling method was used in collecting soil sample directly and the collection was done using soil auger at a depth of 10cm. The collected soil was put into polythene bag and stored inside refrigerator.](image-3.png "Figure 2 :")
3![Figure 3 : Showing the chromatogram of the extract](image-4.png "Figure 3 :B")
6![Figure 6 : Showing spectral of compound identified](image-5.png "Figure 6 :BB")
12![Figure 12 : Showing spectral of compound identified](image-6.png "Figure 12 :B")
![](image-7.png "")
			Volume XV Issue IV Version I Year 2015 © 2015 Global Journals Inc. (US)
		
		
			
			
			

				


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		Publication date: 2010-09-23) Patent application number
		
			AnnFowler
		
		
			20100240768
			Rheinfelden, CH) Regina Goralczyk
		
	




* 
	
		The Characteristics of Alcohol: Chemistry, Use and Abuse
		
			JBrick
		
	
	
		Medical Consequences of Alcohol and Drug Abuse
				
			JBrick
		
		
			Haworth Medical Press
			2003
			
		
	




* 
	
		Drugs, The Brain and Behavior: The Neuropharmacology of Abuse and Dependence
		
			JBrick
		
		
			CErickson
		
		
			1999
			Haworth Medical Press
			New York
		
	




* 
	
		The Effects of Alcohol Abuse on the Brain
		
			FCrews
		
	
	
		Medical Consequences of Alcohol and Drug Abuse
				
			JBrick
		
		
			Haworth Medical Press
			2003
			
		
	




* 
	
		Pharmacology of Alcohol
		
			LPohorecky
		
		
			JBrick
		
	
	
		International Encyclopedia of Pharmacological Therapeutics: Psychotropic Drugs ofAbuse
				
			Pergamon Press
			1990
			
		
	




* 
	
		
			YRokade
		
		
			RZSayyed
		
		Naphthalene derivatives; rasayan journal of chemistry
				
			2009
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