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             \author[1]{Adewole  E}

             \affil[1]{  Afe Babalola University, Ado-Ekiti (Abuad), Nigeria}

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\date{\small \em Received: 12 April 2015 Accepted: 2 May 2015 Published: 15 May 2015}

\maketitle


\begin{abstract}
        


Penicillium notatum has been found years ago to be one of the leading novel candidates of fungi for the discovery of novel antibiotics. The fungus was isolated from eighty years old virgin forest Ikota, Ondo state. The fraction was obtained through column chromatography using mixture of solvents, after culturing and purification process of the extract. The fraction was characterized using gas chromatography mass spectrophotometer, the Gc-Ms results revealed Benzene, 1,2,4,5-trimethyl- having retention time 5.117,% of total 6.505%, pcymene having retention time 5.170 minutes, % of total 16.439%, Trans-Decalin, 2 methyl with retention time 5.235 minutes and % of total 9.468 %, also n-Nonadecanol-1 having retention time 5.301 minutes and 13.302 %, Naphthalene with retention time 6.161 minutes, % of total 11.369 %, 5, 8,11,14-Eicosatetraenoic acid, methyl ester, having retention time of 6.381 minutes and % of total 2.544 %. Literature reports had indicated the physiological activities of these compounds and of interest p-cymene and Naphthalene derivative to possess strong pharmaceutical uses.Penicillium notatum has been found years ago to be one of the leading novel candidates of fungi for the discovery of novel antibiotics. The fungus was isolated from eighty years old virgin forest Ikota, Ondo state. The fraction was obtained through column chromatography using mixture of solvents, after culturing and purification process of the extract. The fraction was characterized using gas chromatography mass spectrophotometer, the Gc-Ms results revealed Benzene, 1,2,4,5-trimethyl- having retention time 5.117,% of total 6.505%, pcymene having retention time 5.170 minutes, % of total 16.439%, Trans-Decalin, 2 methyl with retention time 5.235 minutes and % of total 9.468 %, also n-Nonadecanol-1 having retention time 5.301 minutes and 13.302 %, Naphthalene with retention time 6.161 minutes, % of total 11.369 %, 5, 8,11,14-Eicosatetraenoic acid, methyl ester, having retention time of

\end{abstract}


\keywords{penicillium notatum, gas chromatography mass spectrophotometer, retention time, % of total.}

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\let\tabcellsep& 	 	 		 
\section[{I. Introduction}]{I. Introduction}\par
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.   
\section[{II. Methodology}]{II. Methodology} 
\section[{b) Isolation of the fungi}]{b) Isolation of the fungi}\par
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. 
\section[{c) Identification}]{c) Identification}\par
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. 
\section[{d) Microscopic observation}]{d) Microscopic observation}\par
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. 
\section[{e) Culturing the fungi}]{e) Culturing the fungi}\par
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. 
\section[{f) Extraction and Purification}]{f) Extraction and Purification}\par
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. 
\section[{III. Results}]{III. Results}\par
2-isopropylidene-5-methylhex-4-enal having retention time 5.508 minutes and percentage of total 5.446 \%. From the figure \hyperref[fig_2]{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 \hyperref[b1]{(Brick, 2003;}\hyperref[b2]{Brick and Erickson, 1999;} {\ref Crews,2003;}\hyperref[b4]{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. 
\section[{V. Conclusion}]{V. Conclusion}\par
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.\begin{figure}[htbp]
\noindent\textbf{1}\includegraphics[]{image-2.png}
\caption{\label{fig_0}Figure 1 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{2}\includegraphics[]{image-3.png}
\caption{\label{fig_1}Figure 2 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{3}\includegraphics[]{image-4.png}
\caption{\label{fig_2}Figure 3 :B}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{6}\includegraphics[]{image-5.png}
\caption{\label{fig_3}Figure 6 :BB}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{12}\includegraphics[]{image-6.png}
\caption{\label{fig_4}Figure 12 :B}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{}\includegraphics[]{image-7.png}
\caption{\label{figure7}}\end{figure}
 			\footnote{Volume XV Issue IV Version I Year 2015 © 2015 Global Journals Inc. (US)} 		 		\backmatter  			 			 			  				\begin{bibitemlist}{1}
\bibitem[Brick and Erickson ()]{b2}\label{b2} 	 		\textit{Drugs, The Brain and Behavior: The Neuropharmacology of Abuse and Dependence},  		 			J Brick 		,  		 			C Erickson 		.  		1999. New York: Haworth Medical Press.  	 
\bibitem[Pohorecky and Brick ()]{b4}\label{b4} 	 		‘Pharmacology of Alcohol’.  		 			L Pohorecky 		,  		 			J Brick 		.  	 	 		\textit{International Encyclopedia of Pharmacological Therapeutics: Psychotropic Drugs ofAbuse},  				1990. Pergamon Press. p. .  	 
\bibitem[Fowler (20100240768)]{b0}\label{b0} 	 		\textit{Publication date: 2010-09-23) Patent application number},  		 			Ann Fowler 		.  		20100240768. Rheinfelden, CH) Regina Goralczyk.  	 
\bibitem[Rokade and Sayyed ()]{b5}\label{b5} 	 		 			Y Rokade 		,  		 			R Z Sayyed 		.  		\textit{Naphthalene derivatives; rasayan journal of chemistry},  				2009. 2 p. .  	 
\bibitem[Brick ()]{b1}\label{b1} 	 		‘The Characteristics of Alcohol: Chemistry, Use and Abuse’.  		 			J Brick 		.  	 	 		\textit{Medical Consequences of Alcohol and Drug Abuse},  				 			J Brick 		 (ed.)  		2003. Haworth Medical Press. p. .  	 
\bibitem[Crews ()]{b3}\label{b3} 	 		‘The Effects of Alcohol Abuse on the Brain’.  		 			F Crews 		.  	 	 		\textit{Medical Consequences of Alcohol and Drug Abuse},  				 			J Brick 		 (ed.)  		2003. Haworth Medical Press. p. .  	 
\end{bibitemlist}
 			 		 	 
\end{document}
