Phytochem & BioSub Journal Vol 7(3) 2013 .pdf



Nom original: Phytochem & BioSub Journal Vol 7(3)-2013-.pdf
Titre: Microsoft Word - PCBS-J-Vol 7-Pgarde.doc
Auteur: CHERITI

Ce document au format PDF 1.6 a été généré par PScript5.dll Version 5.2.2 / Acrobat Distiller 11.0 (Windows), et a été envoyé sur fichier-pdf.fr le 25/11/2014 à 11:22, depuis l'adresse IP 197.203.x.x. La présente page de téléchargement du fichier a été vue 1206 fois.
Taille du document: 1.3 Mo (38 pages).
Confidentialité: fichier public


Aperçu du document


PhytoChem & BioSub Journal
Peer-reviewed research journal on Phytochemistry & Bioactives Substances
ISSN 2170 - 1768

PCBS Journal
Volume 7 N° 1, 2 & 3

2013

PhytoChem & BioSub Journal
ISSN 2170-1768

ISSN 2170 – 1768

PhytoChem & BioSub Journal (PCBS Journal) is a peer-reviewed research journal
published by Phytochemistry & Organic Synthesis Laboratory. The PCBS Journal publishes
innovative research papers, reviews, mini-reviews, short communications and technical notes
that contribute significantly to further the scientific knowledge related to the field of
Phytochemistry & Bioactives Substances (Medicinal Plants, Ethnopharmacology,
Pharmacognosy, Phytochemistry, Natural products, Analytical Chemistry, Organic Synthesis,
Medicinal Chemistry, Pharmaceutical Chemistry, Biochemistry, Computational Chemistry,
Molecular Drug Design, Pharmaceutical Analysis, Pharmacy Practice, Quality Assurance,
Microbiology, Bioactivity and Biotechnology of Pharmaceutical Interest )
It is essential that manuscripts submitted to PCBS Journal are subject to rapid peer review and
are not previously published or under consideration for publication in another journal.
Contributions in all areas at the interface of Chemistry, Pharmacy, Medicine and Biology are
welcomed.
Editor in Chief
Pr Abdelkrim CHERITI
Phytochemistry & Organic Synthesis Laboratory

Co-Editor
Dr Nasser BELBOUKHARI
Bioactive Molecules & Chiral Separation Laboratory
University of Bechar, 08000, Bechar, Algeria
Editorial Board
Afaxantidis J. (France), Akkal S. (Algeria), Al Hamel M. (Morocco), Al Hatab M. (Algeria), Aouf N. (Algeria),
Asakawa Y. (Japan), Atmani A. (Morocco) , Awad Allah A.( Palestine), Azarkovitch M. ( Russia), Baalioumer A.
(Algeria), Badjah A.Y. ( KSA), Balansard G. (France), Barkani M. (Algeria), Belkhiri A. (Algeria), Benachour D.
(Algeria), Ben Ali Cherif N. (Algeria), Benayache F. (Algeria), Benayache S. (Algeria), Benharathe N. (Algeria),
Benharref A. (Morocco), Bennaceur M. ( Algeria), Bensaid O. (Algeria), Berada M. ( Algeria), Bhalla A. ( India),
Bnouham M. (Morocco), Bombarda E. (France), Bouchekara M. (Algeria), Boukebouz A. (Morocco), Boukir A.
(Morocco), Bressy C. (France), Chehma A. (Algeria), Chemat F. (France), Chul Kang S. (Korea), Dadamoussa B.
(Algeria), Daiche A. (France), Daoud K. ( Algeria), De la Guardia M. ( Brazilia), Dendoughi H. (Algeria), Derdour
A. (Algeria), Djafri A. (Algeria), Djebar S. (Algeria), Djebli N.(Algeria), Dupuy N. (France), El Abed D. (Algeria),
EL Achouri M. (Morocco), Ermel G. ( France), Esnault M. A. ( France), Govender P. (South Africa), Jouba M.
(Turkey), Hacini S. (Algeria), Hadj Mahamed M. (Algeria), Halilat M. T. (Algeria), Hamed El Yahia A. ( KSA),
Hamrouni A. ( Tunisia), Hania M. ( Palestine), Iqbal A. (Pakistan), Gaydou E. (France), Ghanmi M. (Morocco),
Gharabli S. (Jordan), Gherraf N. ( Algeria), Ghezali S. (Algeria), Gouasmia A. (Algeria), Greche H. (Morocco),
Kabouche Z. (Algeria), Kacimi S. (Algeria), Kajima J.M. (Algeria), Kaid-Harche M. (Algeria), Kessat A.
(Morocco), Khelil-Oueld Hadj A. (Algeria), Lahreche M.B. (Algeria), Lanez T. (Algeria), Leghseir B. (Algeria),
Mahiuo V. (France), Marongu B. ( Italia), Marouf A. (Algeria), Meddah B.( Morocco), Meklati F. (Algeria),
Melhaoui A. ( Morocco), Merati N. (Algeria), Mesli A. ( Algeria), Mushfik M. ( India), Nefati M. (Tunisia),
Ouahrani M. R. (Algeria), Oueld Hadj M.D. (Algeria), Pons J.M. ( France), Radi A. (Morocco), Rahmouni A.
(Algeria), Raza Naqvi S. A. (Iran), Reddy K.H. ( South Africa), Reza Moein M. (Iran), Rhouati S. (Algeria),
Roussel C. (France), Saidi M. (Algeria), Salgueiro L.D (Portugal), Salvador J. A. (Spain), Seghni L. (Algeria),
Sharma S. ( India), Sidiqi S. K. ( India), Souri E. ( Turkey), Tabti B. (Algeria), Taleb S. (Algeria), Tazerouti F.
(Algeria), Vantyune N. (France), Villemin D. (France), Yayli N. (Turkey), Youcefi M. (Algeria), Ziyyat A.
(Morocco), Zouieche L. (Algeria), Zyoud H. (Palestine).

Guidelines for the publication of manuscripts in
PhytoChem & BioSub Journal (ISSN 2170 – 1768)
PhytoChem & BioSub Journal (PCBS Journal) is a periodical dedicated to the publication of
original scientific work, reviews, and communications in the field of of Phytochemistry & Bioactives
Substances. Contributions in all areas at the interface of Chemistry, Pharmacy, Medicine and Biology
are welcomed.
Submission of an article to the PCBS Journal implies that the work described has not been published
previously (except in the form of an abstract or as part of a published lecture or academic thesis), that
it is not under consideration for publication elsewhere, that its publication is approved by all authors.
The PCBS Journal reserves the right to submit all received manuscripts to ad hoc referees, whose
names will be kept confidential, and will have the authority to decide on the pertinence for acceptance.
Referees may send back manuscripts to Editor-in-Chief, for transmission to the author(s) with
suggestions for necessary alterations, which are to be made in order to conform to the standards and
editorial rules of the Journal. All manuscripts should be prepared in MS-Word format, and submitted
online to Phytochem07@yahoo.fr. Upon receipt of paper submission, the Editor sends an E-mail of
confirmation to the corresponding author within 1-4 working days. The Editors reserve the right to edit
or otherwise alter all contributions, but authors will receive proofs for approval before publication.
If you have any questions, please contact with the editor of the journal at the same E mail
The manuscript should be on A4 size paper, double spaced using Times New Roman size 12 font,
fully justified, with margins of 2 cm and should be arranged in the following order:
Title: Concise and informative, in accordance with the contents of the article. ( Times New Roman;
Size: 14, Blod.)
Author’s names and affiliations: Please indicate the given name and family name clearly (Times
New Roman; Size-12; Italic). Present the authors' affiliation addresses below the names (Times
New Roman; Size-11; Italic). Provide the full postal address of each affiliation, including the
country name, and, if available, the e-mail address, and telephone number of each author.
Abstract: A concise and factual abstract is required with 200 or less words highlighting the most
important information, including the methodology, results, and conclusions that allows readers
to evaluate their interest in the article and thus avoiding the reading of the full work (Times New
Roman; Size-12; Italic).
Keywords: Immediately after the abstract, provide a maximum of 7 keywords, avoiding general and
plural terms and multiple concepts (Times New Roman; Size-12; Italic).
Introduction: Should clearly establish the objectives of the work and its relationship with other works
in the same field
Material and Methods: Description of the Material and the Methods used should be brief, and clear enough
to make possible the comprehension and the reproducibility of the work.

Results and Discussion: Should be presented with a personal discussion or interpretation, and whenever
possible, be accompanied by adequate tables and figures and the discussion must be restricted to
the significance of the data presented. Figures, Tables and Structural Formulas are included in
the text.
Acknowledgements: (optional item)
References: Should be standardized to conform to the requirements of the journal. Preferentially use
references that can be accessed by the readers worldwide.

PhytoChem & BioSub Journal
Peer-reviewed research journal on Phytochemistry & Bioactives Substances

ISSN 2170 - 1768

PCBS Journal

Volume 7 N° 3
POSL

2013

Edition LPSO
Phytochemistry & Organic Synthesis Laboratory
http://www.pcbsj.webs.com , Email: phytochem07@yahoo.fr

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

PhytoChem & BioSub Journal

2013
Vol. 7 No. 3

ISSN 2170-1768

Contents
PhytoChem & BioSub Journal Vol. 7 N° 3
A. CHERITI

A stupid plagiarism in the article « Ethnopharmacology and phytochemical screening of
bioactive extracts of Limoniastrum feei ( plombagenaceae) » Asian Journal of Natural &
Applied Sciences, Vol. 2(1), 5-9, 2013.

88

M. A. GACEM, A. OULD EL HADJ-KHELIL, S. HADEF, A. BOUDARHEM & A. N. DJERBAOUI

Phytochemical screening and antibacterial activity of aqueous extracts of Citrullus
colocynthis seeds

90

F. Z. OUASTI, M. HAMADOUCHE & D. EL ABED

Synthesis of 2 (1,2,3)-Triazolines via 1,3-dipolar cycloaddition between organic azides and
1-morpholinocyclopentene derivatives

95

A. LARBI, N. BELBOUKHARI, A.CHERITI & A. ZANOUN

Comparative study on the vibrational IR spectra of N-aryl imidazoline-2-(thio) one
derivatives by various semi-empirical methods

103

D. AICI, A. CHERITI, Y. BOURMITA & N. BELBOUKHARI

Antimicrobial activity of essential oils of Bubonium Graveolens (Forssk.) and Anvillea
Radiata (Coss.).

87

116

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

PhytoChem & BioSub Journal

2013
Vol. 7 No. 3

ISSN 2170-1768

A stupid plagiarism in the article « Ethnopharmacology and phytochemical
screening of bioactive extracts of Limoniastrum feei ( plombagenaceae) »
Asian Journal of Natural & Applied Sciences, Vol. 2(1), 5-9, 2013
EDITORAL

Abdelkrim CHERITI
Phytochemistry & Organic Synthesis Laboratory
University of Bechar, 08000, Algeria
Received: May 19, 2013; Accepted: June 21, 2013
Corresponding author Email karimcheriti@yahoo.com
Copyright © 2013-POSL

It’s well known that plagiarism is considered misconduct research, unethical, perturb
and damage the integrity and knowledge of the scientific community. In addition that is a
form of violation copyright law and is therefore illegal. The scientific community is no
exception to this new disease of plagiarism (copy – past) became prevalent in the era of new
technologies and the Internet. Plagiarism affects all disciplines, sciences, humanities and
medicine, written as figures. The situation becomes even more serious when, supposedly
anonymous editors of journals, closing their eyes on plagiarism to "fill" the pages of journals
and make a few dollars. Thus, none reviewed online publication would become common
merchandise. In these disastrous practices, add the shameful "looting" of others works,
practices that engage without remorse. We must fight against plagiarism today that the
development of digital technologies not only tends to make it easier, but to trivialize it and
thus make it almost normal. We have plenty of examples showing that plagiarism is
increasingly commonplace in the word. Whereas matching an integral article (Copy and
paste) from another scientific journal is a potential plagiarism qualified unjustified attitude
and a stupid practice.
The most striking case, is the copy and paste of our article published since 2012 in this journal
(PhytoChem & BioSub Journal Vol. 6 N° 2, 83-87, 2012, http://pcbsj.webs.com/pcbs-j-vol6-2012 ) by another “ scientists” - teaching in Tlemcen and Bechar universities, L. Ziane, H.
A. Lazouni, A. Moussaoui, N. Hamidi - and published in Asian Journal of Natural & Applied
Sciences, Vol. 2(1), 5-9, 2013 (www. leena-luna.co.jp). A journal with editorial board but the
editor chief is anonymous.
A simple search of fragments of sentences of the article on Google quickly allowed us to see
that this article is a gross plagiarism. The article (text, figure, and picture) is the integral
copying of our works with slightly modification of title and introduction. As an anecdote,
even the non-specialist reader can easily notice the stupidity of the authors of flight that
indicated below:
Page 5: Introduction:
“ The aim of this studies is to evaluate the bioactives extracts of Limoniastrum feei plant of
the Southeast of Algeria ( saoura region of Bechar ) northern Africa [1-2] by phytochemical
screening. In earlier work we have reported the antimicrobial activity of aerial part crude
88

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

extracts from Limoniastrum feei (Belboukhari et al 2005). In continuation of our studies on
medicinal plants, we isolate tow flavonoids from methanolic extract of Limoniastrum feei. [34] “
Page 7: Ethno pharmacology Screening
“According to the results of ethno pharmacology study to several medical plants that are used
in Bechar (Southeast of Algeria) in traditional medicine, it carries out by laboratory LPSO
(1998). We are interested to deepen the investigation of this specie.”
Page 8: Phytochemical Screening
“According to the results of biological screening of several extracts of three parts of
Limoniastrum feei (leaves, stem and twig) [3- 4].The methanolic and heptan extracts of these
parts of Limoniastrum feei were selected ( table1 ) and were carried out a phytochelical
screening on the active extracts for this plants.”
Page 9: Refrences
On the 10 references cited, 6 are our publications!!!!!
[1] Cheriti, A. (2000). Rapport CRSTRA, plantes médicinales de la région de Bechar, sud ouest Algérie
(Ethnopharmacological studies) Bechar, Algeria.
[3] Belboukhari, N. & Cheriti, A. (2005). Antimicrobial activity of aerial part of limoniastrum feei. Asian
Journal of plant sciences, 4(5), 496-498.
[4] Belboukhari, N. & Cheriti, A, (2007). flavonoid of limoniastrum feei, Research journal of phytochemistry,
74-78.
[8] Belboukhari, N. (2002). Communication, 6 ieme Congrés de la S.A.Chem, University Ferhat Abbass Setif,
14-16.
[9] Belboukhari, N., Cheriti, A. & Hacini, S. (2002).Communication, 5 ieme SNCOIP, S.A.Chem, 22- 23.
[10] Cheriti, A., Belboukhari, N. & Hacini, S (2005). CO, Séminaire international sur la valorisation des plantes
médecinales dans les zones arides, Ouargla 01-03 fevrier, Algeria; p-12. Nd.

I think this practice can not be ignored. It is a case of plagiarism rather disconcerting. But this
example illustrates the potential dangers of the online diffusion of the article and the need for
vigilance that editors of journal should keep. The academic and scientific communities must
work together against all forms of plagiarism. They must do so not only in the prevention but
also by adequate sanctions against proved plagiarism
Thus, any one, authors or publisher not condemn and promote these practices should be
automatically disqualified from scientific research. The responsibility of editor against
plagiarism must be engaged, at least by the retracted of article in question.
Reference:
Armstrong J. D., 1993, “Plagiarism: What Is It, Whom Does It Offend, and How Does One Deal with
It?”, AJR; 161: 479.
Baždarić K., 2012, “Plagiarism detection – quality management tool for all scientific journals”, Croat
Med J. ; 53: 1.
Derbala A. , 2013, ‘’Le développement de la recherche scientifique en Algérie’’, Le Quotidien d'Oran
du 18 juin 2013
Fanelli D., 2009, “How many scientists fabricate and falsify research? A systematic review and metaanalysis of survey data”. PLoS ONE, 4(5): e5738.
Fanga F. C., Steenc R. G., Casadevalld A., 2012, “Misconduct accounts for the majority of retracted
scientific publications,” Proc Natl Acad Sci USA, 109(42): 17028.
Horrom T. A. , 2012, “The perils of copy and paste: Plagiarism in scientific publishing”, J Rehabil
Res Dev., 49( 8): vii.
Macilwain C., 2012, “the time is right to confront misconduct.”, Nature;488 :7.
Marušić A., Petrovečki M., 2012, “In: Science publishing: How to stop plagiarism.” Nature; 481:21.
Rouajia A., 2012, ‘’La banalisation du plagiat et le triomphe de la médiocrité au sein de l’université
algérienne ‘’, Le Quotidien d'Oran du 28 février.

89

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
 

PhytoChem & BioSub Journal

2013 
Vol. 7  No. 3  

ISSN 2170‐1768 

Phytochemical screening and antibacterial activity of aqueous extracts of
citrullus colocynthis seeds
GACEM Mohamed Amine1*, OULD EL HADJ-Khelil-Aminata1, HADEF Sawsen2,
BOUDARHEM Amel1 and DJERBAOUI Amina Nesrine1
1

Laboratoire de protection des écosystèmes en zone aride et semi-aride.
Université de Kasdi Merbah-Ouargla, Ouargla 30000, Alegria.
2
Laboratoire de Biotechnologie, Environnement et Santé,
Université de Jijel, Jijel 18000, Alegria.

 
Received: April 14, 2013; Accepted: July 09, 2013
Corresponding author Email biologieamine@yahoo.fr
Copyright © 2013-POSL
 

Abstract- Skin, gynecological and lung infections are caused by microorganisms exist in the world.
The treatment of these infections is mainly based on the use of synthetic drugs that become, in recent
years, ineffective, due to the resistance of bacterial strains and the proliferation of opportunistic fungi.
The aim of this study is to test in vitro the antibacterial activity of aqueous extract from seeds of
Citrullus colocynthis which was detected phytochemical families existing. These antimicrobial powers
are measured using the micro-dilution method against the Gram-negative bacteria (Escherichia coli
ATCC 25922 and Pseudomonas aeruginosa ATCC 27853) and Gram-positive (Enterococcus faecalis ATCC
29212 and Staphylococcus aureus ATCC 25923). The results showed that seeds of C. colocynthis are rich
by bioactive substances. All tested extracts of C. colocynthis showed antimicrobial activity against all
strains tested.. The folk medicinal use as a broad-spectrum antimicrobial agent is validated.
Key words: Citrullus colocynthis Schrad ; Phytochemical family ; Antibacterial activity.

 

INTRODUCTION  
In some underdeveloped country, plants are the main source of drugs to treat infectious
diseases due to the availability and the economic climate. However, only about 20% of plants
were subjected to pharmacological studies or laboratory tests, despite the large number of new
derivatives of natural resources or semi-synthetic is introduced on the market [1] antibiotics.
The natural substances from plants have multiple interests put to use in industry, for food,
cosmetics and pharmacology. Among these compounds are found largely secondary
metabolites which are mainly used in therapy.
The pharmacy still uses a high proportion of plant and research is directed towards the
discovery of new bioactive molecules drugs or raw materials for the semi synthesis [2].
Citrullus colocynthis Schrad. (Cucurbitaceae) grows increasingly in drylands is an endemic
plant in southern Algeria[3].
This herb is widely used in traditional Algerian medicine for the treatment of many diseases
such as arthritis, hypertension, and various infectious diseases, including dermatological and
90 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
 

gynecological problems, urinary and pulmonary infections [4, 5, 6]. It has other therapeutic
properties purgative, anti-tumor and immunostimulatory [7, 8], anti-inflammatory and
antioxidant [9, 10], antirheumatic [11], laxative and against leukemia, jaundice, fever, ascites,
biliary disorders and hemorrhoids. [12] It is also used against liver disease [13]. The ethanol
extract of the fruit of bitter gourd has an anti-microbial effect on Pseudomonas aeruginosa and
Staphylococcus epidermidis and antifungal effect of several types of fungi [14]. The present
study aims to investigate in vitro antibacterial activity of the seeds of this plant using a series
of micro-broth dilution after extraction and phytochemical study
MATERIEL AND METHODS
Fruit of Citrullus colocynthis
Matured fruits of Citrullus colocynthis was taken from the region of Oued N'sa located about
70 km from the town of Ouargla during the month of December 2010. The identification of
plant species is performed at the Laboratory of Plant Physiology at the University of Saida by
Dr. Hasnaoui on the support of his experience and some documentation regarding the
taxonomy of this species in the plant kingdom [3].
Aqueous extraction
The aqueous extract was prepared by maceration in cold distilled water at 20% for three
hours. The mixture is then centrifuged at 3600 g for 30 min. The supernatant was recovered
and filtered through paper filter Whatman No. 01. The fraction obtained is collected in a vial
and stored at 4 °C in the dark until their use. [15] In order to calculate the yield of extraction,
the aqueous extract is recovered by evaporation of water in an oven at 50 °C.
Phytochemical screening
The extract was subjected to phytochemical screening to identify existing phytochemical
families. The tannins are detected by the method of Karumi et al. [16] Coumarins, steroids
and anthocyanins were found using the technique of Bruneton [17]. Flavonoids have been
detected by the method of Malec and Pamelio [18], anthraquinones by the method of Oloyede
[19], the saponins by the method of Koffi et al [20] and finally the alkaloids are revealed by
the method of Mojab et al [21].
Antibacterial activity and determination of MIC and MBC
Four reference strains were selected for antibacterial investigation: Gram-positive cocci
(Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 25923) and Gramnegative (Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853).
The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration
(MBC) were measured by the method of micro-broth dilution using 96-well microplates,
following the procedure of Berche et al. [22]. The extract solutions were prepared by
dissolving in 10% dimethyl sulfoxide (DMSO). The extract concentrations tested ranged from
0.003 to 1.600 mg / ml. The (MIC) of each aqueous extract was defined as the lowest
concentration that inhibits bacterial growth after incubation at 37 ° C between 18 and 24 h.
The minimum bactericidal concentration (MBC) was determined by subculture on blood agar
at 37 ° C between 18 and 24 h. Levofloxacin was used as antibacterial positive control.
RESULTS
Extraction yields and phytochemical study
91 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
 

The calculation of the yield relative to the total weight of the dry powder of C. colocynthis
seeds used in aqueous extraction shows that the plant gave masses greater than 1 g/100 g seed
powder.
Table 1. Extraction yield and phytochemical study of
the aqueous extract of C. colocynthis seeds.
Extracts
types

Extraction
yields

Aqueous
extract

1.83 %

Polyphenols
Tannins

Anthraquinons

Flavonoids

Anthocyans

Coumarins

+

-

+

+

-

Saponosids

Terpens
Steroids

+

+

Alka
loids
+

Chemical characterization of the aqueous extract showed the presence of polyphenols in the
form of hydrolysable tannins, flavonoids, saponins and anthocyns while anthraquinoes and
coumarins are entirely absent. The same extract reacted positively vis-à-vis the chemical test
seeking steroids (belonging to the group of terpenes) and alkaloids (Table 1).
Antibacterial activity
The aqueous extracts of C. colocynthis seeds showed a good antibacterial activity against four
strains selected (Figure 1).

*CMI en jaune et CMB en vert

Figure 1. Determination of MIC and MBC of the aqueous extract
of C. colocynthis seeds on different bacterial strains selected.
The concentration range selected for the determined MIC and MBC varies between 0.003 to
1.6 mg/ml. The highest inhibitory activity was obtained against Echirichia coli, the MIC was
0.2 mg/ml while the CMB is 0.4 mg/ml. For the three other bacterial strains, the MIC was 0.4
mg/ml, while the CMB is 0.8 mg/ml (Figure 1).
DISCUSSION
Medicinal plants have long been used for the treatment of various diseases. Currently,
they are used for the treatment of difficult infections to treat by drugs. The antimicrobial
92 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
 

activity of the organs of C. colocynthis is elucidated in several studies [6, 10, 14]. For cons,
the phytochemical and antimicrobial screening of the aqueous extract of C. colocynthis seeds
from the region of Ouargla is not clarified.
This study yielded several results such as aqueous extraction yield, the phytochemical study
and antimicrobial activity. The extraction yield depends on the solvent and the same method
by which the extraction is performed, and the effectiveness of the extract depends on the type
of extraction solvent, active substances and other non-active therefore incapable to made
desired biological activity. The effectiveness also depends on the selected bacterial strains and
plant organ tested. For bacterial strains, antimicrobial activity may be different from one
bacterium Gram- to another Gram+ [23].
The good MIC of the aqueous extract of C. colocynthis seeds recorded against Pseudomonas
aeruginosa is a very good result, because this bacteria is among the most common causes of
nosocomial infections [24]. This reveals a new confirmation of antibacterial properties of C.
colocynthis seeds. The recorded as antibacterial activity against Escherichia coli class C.
colocynthis seeds among the most important medicinal plants used against digestive problems
caused by this bacteria [23].
The results give an indication that the plant is producing certain chemicals and compounds
toxic to microorganisms. Furthermore, the literature has shown that plant extracts have great
potential inhibitor against resistant bacterial strains [25, 26].
CONCLUSION
The results presented in this study indicate that the analyzed natural products are a great
choice for the development of new treatment strategy for gastrointestinal and pulmonary
dermatological, gynecological infections. The use of this plant in traditional medicine for the
treatment of infections is confirmed by the results obtained in this study. This plant can also
be used as an alternative to other chemical formula in order to increase their activity and
effectiveness. In another study will be made to purify and identify the chemical compounds of
this antimicrobial extract
REFERENCES
[1] Mothana R. A., Lindequist U., 2005. Antimicrobial activity of some medicinal plants of the island
Soqotra. J. Ethnopharmacol. 96, 177–181.
[2] Bahorun T., 1997. Substances naturelles actives: la flore mauricienne, une source
d’approvisionnement potentielle. Food. Agri. Resh council. 2, 83-93.
[3] Pottier-Alapetite G., 1981. Flore De La Tunisie, Angiospermes-Dicotylédones: Gamopétales. Ed.
Imprimerie officielle de la république tunisienne, Tunis, Tunisia, pp. 930.
[4] Boukef M. K., 1986. Médecine traditionnelle et pharmacopée. Les plantes dans la médecine
traditionnelle tunisienne. Ed. Agence de coopération culturelle et technique, Paris, pp.165.
[5] Le Flock E., 1983. Contribution à une étude ethnobotanique de la flore tunisienne. Ed. Imprimerie
officielle de la république tunisienne, Tunis, Tunisia, pp. 241–244.
[6] Marzouk B., 2008. Etude biologique et activités pharmacologiques de Citrullus colocynthis
Schrad. Master Thesis Biology & Health Care. University of Monastir, Monastir, Tunisia.

[7] Abdel Hassan I., abdel-barry J. A. and Mohammeda S. T., 2000. The hypoglycaemic and
antihyperglycaemic effect of Citrullus colocynthis fruit aqueous extract in normal and
alloxan diabetic rabbits. J. Ethnopharmacol. 71, 325-330.
[8] Bendjeddou D., Lalaoui K. and Satta D., 2003. Immunostimulating activity of the hot water
soluble polysaccharide extracts of Anacyclus pyrethrum, Alpinia galangal and Citrullus
colocynthis. J. Ethnopharmacol. 88, 155-160.
[9] Al-Ghaithi F., El-Ridi M. R., Adeghate E. and Amiri M. H., 2004. Biochemical effects of Citrullus
colocynthis in normal and diabetic rats. Mol. Cell. Biochem. 30, 1-7.
93 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
 

[10] Marzouk B., Marzouk Z., Haloui E., Fenina N., Bouraoui A. and Aouni M., 2010. Screening of
analgesic and anti-inflammatory activities of Citrullus colocynthis from southern Tunisia. J.
Ethnopharmacol. 128, 15-19.
[11] Boukef K. and Souissi H. R., 1982. Contribution à l’étude des plantes médicinales en médecine
populaire en Tunisie. Rev. Soc. Pharm. 3, 34- 35.
[12] Ziyyat A., Legssyer A., Mekhfi H., Serhrouchni M. and Benjelloun W., 1997. Phytotherapy of
hypertension and diabetes in oriental Morocco. J. Ethnopharmacol. 58, 45-54.
[13] Gebhardt R., 2003. Antioxidative, antiproliferative and biochemical effects in Hep G2 cells of a
homeopathic remedy and its constituent plant tinctures tested separately or in combination.
Arzneimittelforschung. 53, 823-830.
[14] Gurudeeban S., Rajamanickam E., Ramanathan T. and Satyavani K., 2010. Antimicrobial activity
of citrullus colocynthis in gulf of mannar. Int. J. Curr. Res. 2, 078-081.
[15] Senhaji O., Faid M., Elyachioui M. and Dehhaoui M., 2005. Antifungal activity of different
cinnamon extracts. J. Med. Mycol. 15: 220–229.
[16] Karumi Y., Onyeyili P. A. and Ogugbuaja V. O., 2004. Identification of active principals of M.
balsamina (Balsam apple) leaf extract. J. Med. Sci. 4, 179-182.
[17] Bruneton J., 1999. Pharmacognosie, Phytochimie, plantes médicinales. 3eme Ed, Tec &Doc
Lavoisier, Paris, pp.1120.
[18] Malec L. S. and Pamilio A. B., 2003. Herbivory effects on the chemical constituents of Bromus
pictus. Mol. Med. Chem. 1, 30-38.
[19] Oloyede O. I., 2005. Chemical profile of Unripe Pulp of Carica papaya, Pak. J. Nutr. 4, 379-381.
[20] Koffi N., Beugré K., Guédé N., Zirihi D. and Laurent A., 2009. Screening phytochimique de
quelques plantes médicinales ivoiriennes utilisées en pays Krobou (Agboville, Côte-d’Ivoire). Sci.
Nat. 6 (1), 1-15.
[21] Mojab F., Kamalinejab M., Ghaderi N. and Vahidipour H. R., 2003. Phytochemical screening of
some species of Iranian plants. Iranian. J. Pharm. Res. 77-82.
[22] Berche P, Gaillard JL, Simonet M., 1991. Les bactéries des infections humaines. Editeur
Flammarion, Médecine et Sciences, pp. 660.
[23] Marzouk B., Marzouk Z., Mastouri M., Fenina N. and Aouni M., 2011. Comparative evaluation
of the antimicrobial activity of Citrullus colocynthis immature fruit and seed organic extracts. Afr.
J. Biotech. 10(10) 2130-2134
[24] Carmeli Y., Troillet N., Eliopoulos G. M., Samore M. H., 1999. Emergence of Antibioticresistant Pseudomonas aeruginosa: Comparison of risks associated with different antipseudomonal
agents. Antimicrob. Agents Chemother. 43 1379-1382.
[25] Mshvildadze V., Favel A., Delmas F., Elias R., Faure R., Decanosidze Q., Kemertelidze E. and
Balansard G., 2000. Antifungal and antiprotozoal activities of saponins from Hedera colchica.
Pharmazie, 55 325-326.
[26] Abdel Ghani S. B., Weaver L., Zidan Z. H., Hussein M. A., Keevil C. W. and Brown R. C. D.,
2008. Microware-assisted synthesis and antimicrobial activities of flavonoid derivatives. Bioorg.
Med. Chem. Lett. 18 518- 522.

94 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

PhytoChem & BioSub Journal

2013 
Vol. 7  No. 3  

ISSN 2170‐1768 

Synthesis of 2 (1,2,3)-Triazolines via 1,3-dipolar cycloaddition
between organic azides and 1-morpholinocyclopentene derivatives
Fatima-Zahra OUASTI, Mohammed HAMADOUCHE & Douniazad EL ABED
Laboratory of Fine Chemistry
Department of Chemistry, Faculty of exact and applied Sciences
University of Oran, BP 1524 El Menouar, Oran 31005, Algeria.
Received: April 10, 2013; Accepted: July 20, 2013
Corresponding author Email hamadouchemed@yahoo.fr
Copyright © 2013-POSL
 

Abstract- The synthesis of some bicyclic 2-1,2,3-triazolines performed by 1,3-dipolar cycloaddition
reaction between cyclopentenic enamines and arylazides, at room temperature in ether, led to the
expected triazolines with yields which vary according to the structure of the arylazide and enamine.
The structure of the obtained triazolines was determined by the usual spectroscopic methods: (IR, 1H
NMR, 13C NMR).
Key words: 1,3-dipolar cycloaddition, 1-morpholinocyclopentene, Organic azides, Triazolines
Secondary amine, Heterocycles, Biological activities

 

1-INTRODUCTION
Heterocycles are a class of compounds commonly found in various natural and
pharmaceutical products. They play an important role in the field of plastics, agricultural
products, dyes industries, cosmetics ... and pharmaceuticals. They have various biological and
therapeutic properties [1]. The nuclei triazolines and triazoles, five membered heterocycles
containing tree nitrogens, are known for their multiple and diverse biological activities:
antibacterial, fungic, anti-inflammatory, antiallergic and ... inhibitor of HIV [2]. Associated
with other structures, they find their use in the pharmaceutical industry. As examples, we cite
fluconazole as an antifungal, the Tazobactam and cefatrizine as antibiotics [3] and Ribavirin
as antiviral [4]. The reaction of 1,3-dipolar cycloaddition in organic synthesis is one of the
most widely methods used for the construction of five-membered heterocyclic. Thus, the
addition of 1,3-dipole such as azides, nitrile oxides, diazo compounds or nitrones on multiple
bonds: alkenes, alkynes, …. leads to the formation of triazolines, isoxazole, pyrazoline, and ...
isoxazolines [5].
The synthesis method we adopted consists in reacting enamines of cyclopentanone or 2carboxylate cyclopentanone 1 with arylazides 2 lead to variously substituted bicyclic
triazolines. The reaction is carried out in ether at room temperature.

95 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

O
O
N

+

X

N3

N

r.t

R

R

2

1

X

N

Ether

N

3

N

ou 4

R = H, p-NO2, p-OMe, 2-Cl 4-NO2, m-CF3, p-F

X = H, CO2CH3

 

The arylazides used were prepared according to the method of Noelting and Michel [6] and
improved by Ranu [7]. This path consists in preparing beforehand the diazonium salt from the
substituted aniline on which the sodium azid reacts.

NH2

NaNO2, HCl, NaN3
O-5°C

R

N3
R

R = H, p-NO2, p-OMe, 2-Cl 4-NO2, m-CF3, p-F

2

The physical characteristics of arylazides 2 are summarized in Table 1.
Table 1. Yields and melting points of arylazides 2
N3
R

2

Entry

R

Yield %

m.p(°C)

2a

H

85

liq

2b

p-NO2

83

69-71

2c

2-Cl, 4-NO2

82

70-72

2d

p-OMe

93

<25

2e

m-CF3

88

liq

2f

p-F

81

liq

2-RESULTS AND DISCUSSION
Addition of enamines of cyclopentanone or 2-methyl carboxylate cyclopentanone 1 and
arylazides 2, in ether at room temperature, in a single step leading to triazolines 3, 4 with
yields which vary according to the structure of arylazides and cyclopentenic enamines [8].
The reaction times and yields of bicyclic triazolines obtained by 1,3-dipolar cycloaddition are
summarized in table 2 for the heterocycles 3 and table 3 for heterocycles 4.
96 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

Table 2. Reaction time and yields bicyclic triazolines 3
Entry

R

Time

Yield %

m.p(°C)

3a

H

5d

98-100

82

3b

p-NO2

24h

192-194

86

3c

2-Cl, 4NO2

24h

114-116

73

3d

p-OMe

5d

Liq.

88

3e

m-CF3

7d

86-88

88

3f

p-F

5d

80-82

90

O

H

N
N

N

N

3

R

Table 3. Reaction time and yields triazolines bicyclic 4
Entry

R

Time

Yield %

m.p(°C)

4a

H

6d

108-110

19

4b(i)

p-NO2

24h

190-192

45

4c(i)

2-Cl, 4NO2

24h

116-118

36

4d

p-OMe

3d

Liq.

14

4e(ii)

m-CF3

5d

84-86

51

4f(ii)

p-F

7d

94-96

53

O

CO2CH3

N
N

N

N

4

R

(i) The reactions carried out with p-chloro-and nitrophenylazide, provided unexpected triazolines 3b
and 3c. (ii) The spectra of the reaction products with fluorinated azides are not usable.

It was noted that the reaction of 1,3-dipolar cycloaddition carried out in ether led to bicyclic
triazolines 3 expected with yields varying between 73 and 90% at a time ranging from 24
hours to 7 days. Monofluorinated azide leads to better yield.
The structures of various bicyclic triazolines 3, 4 were determined by NMR spectroscopy. The
chemical shift of the proton in position 4 of pentagonal heterocycle is shown in Table 4.
Table 4. Chemical shift of H4 and coupling constants of the triazolines 3

Entry

R

δ of H4 (ppm)
(dd)

J (Hz)

3a
3b
3c
3d
3e
3f

H
p-NO2
2-Cl,4-NO2
p-OMe
m-CF3
p-F

4,77
4,95
4,84
4,74
4,82
4,78

3,66; 5,10
3,78; 5,01
3,77; 5,47
3,10; 5,20
3,51; 5,52
3,40;5,47

97 
 

         
O

N

5 4
N1 2 3N

N

R

H

chemical shift
H4 (ppm) in CDCl3

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

3 - CONCLUSION
The application of 1,3-dipolar cycloaddition reaction involving enamines of
cyclopentanone or 2-carboxylate cyclopentanone and arylazides in ether at room temperature,
allowed us to prepare five-membered heterocycles in mild conditions.
4- MATERIALS AND METHODS
Cyclopentanone was distilled before use. Melting points were determined using a Banc
Kofler and were not corrected.
The infrared spectra were recorded on a FTIR spectrometer Alpha Diamond ATR (Bruker
Optics). Sample liquids are examined in KBr film, while solids are recorded on a
spectrometer Infrared Fourier Transform Infrared (FTIR) Thermo-Nicolet IR200 controlled
by the EZ OMNIC 7.2a software. The absorption frequencies are expressed in cm-1 to their
maximum intensity and the intensities are denoted as follows: FF very strong, strong F, mean
m and f low.
The thin-layer chromatography (TLC) was performed on silica plates Merck 230-400 mesh
silica. The elution solvents are mixtures of ethyl acetate and petroleum ether, and revealed in
most cases by a Ultra-violet lamp.
The nuclear magnetic resonance (1H, 13C, DEPT) were recorded with a Bruker AC-300 (300
MHz) or AC-400 aircraft. The internal standard is chloroform (7.26 ppm) and the proton
resonance (77.0 ppm) for the resonance of carbon.
Chemical shifts are given in δ scale and expressed in parts per million (ppm) and refer to the
residual solvent peak. All spectra were carried out in deuterated chloroform.
The functions of signals for carbon spectra were recorded by DEPT (Distortion Less
Enhancement by Polarization Transfer), which differentiates the CH and CH3 CH2, allows
the assignment of signals.
Abbreviations s, d, dd, t, td, m and adopted respectively mean singlet, doublet, doublet of
doublet, triplet, triplet dedoubled and multiplet.
Coupling constants (J) are expressed in Hertz (Hz). * Indicates an international agreement
possible.
General procedure for the synthesis of triazolines
Cyclic enamines 1 and organic azides RN3 2 are mixed into equimolar quantity in ether
with stirring. The reaction mixture was kept at room temperature and monitored by TLC.
The chemical shift values of different triazolines are in good agreement with the proposed
structure.


4-(3-phényl-3,3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol-3a-yl)morpholine 3a
17

8

7

O
9

18

16

N
12

6

5

N1

14

2

N

10
11

15

4

13

3a

3

H
N

Appearance : brown solid
yield : 82% in ether
m.p : 98-100°C
1
H NMR (300 MHZ, CDCl3), δ: 1,22-1,23 (m, 1H16b), 1,55-1,65

(m, 1H16a), 1,88-1,98 (m, 1H18b), 2,01-2,14 (m, 2H17), 2,172,29 (m,1H18a), 2,41 (t, J= 4,39 4H6,7), 3,64 (t, J= 4,39 4H8,9),
4,77 (dd, J= 3,66;J=5,10 1H4), 7,04 (t, J= 7,03 1H15), 7,30 (t, J=
7,60 2H13,14), 7,61 (d, J= 8,4 2H11,12).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 23,26; C16: 32,29
; C18: 33,44; C6,7: 46,41; C8,9: 66,87; C4: 78,00; C5: 91,07 ;
C11,12: 116,66; C15: 123,17 ; C13,14: 129,00 ; C10: 139,38.
98 

 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768



4-(3-(4-nitrophényl)-3,
(3b)

3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol-3a-yl)morpholine

 
17

8

7

O

18

16

N

9

12

5

6

N1

14

2

3

H

(m, 1H16a), 1,91-2,00 (m, 1H18b), 2,11-2,17 (m, 2H17), 2,302,32(m,1H18a),2,34-2,39(m,2H6),2,44-2,49(m,2H7),
3,67-3,69
(m,4H8,9), 4,95 (dd, J= 3,78;J=5,01 1H4), 7,78 (d, J= 9,25
2H11,12), 8,23 (d, J= 9,25 2H13,14).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 23,75; C16: 32,74 ;
C18:33,55; C6,7: 46,78; C8,9: 67,14; C4: 79,73; C5: 91,03 ; C11,12:
115,49; C15: 199,32 ; C13,14: 125,79 ; C10: 144,80. 

N

N

10

O2N

4

11

15

13

3b



4-(3-(2-chloro,4-nitrophényl)-3,3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol-3ayl)morpholine (3c)
 
 
17

8

7

O

18

16
5

N

9

12

6

N1

14

15

13

11

4

2

3

H

Cl

3c 
 



4-(3-(4methoxyphényl)-3,3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol-3ayl)morpholine(3d
 
17

8

7

O
9

18

12

6

15

11
13

3d 



5

N
10

MeO

16

N

14
19

appearance : Light yellow solid
yield : 73% in ether
m.p : 114-116°C
1
H NMR (300 MHZ, CDCl3), δ: 1,46-1,56 (m, 1H16b), 1,65-1,72

(m, 1H16a), 1,75-1,82 (m, 1H18b), 1,98-2,10 (m, 2H17), 2,20-2,28
(m,1H18a), 2,48 (t, J= 4,72 4H6,7), 3,74 (t, J= 4,72 4H8,9), 4,84 (dd,
J= 3,77;J=5,47 1H4), 8,06 (d large, J= 9,06 1H12), 8,14 (dd, J=
2,64;J=6,42 1H14), 8,34 (d, J=2,45 1H13).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 24,24; C16: 32,62 ;
C18: 33,28; C6,7: 46,82; C8,9: 67,13; C4: 78,41; C5: 93,38 ; C12:
122,78; C14: 123,09 ; C13: 127,57 ; C11: 129,22; C15: 142,44; C10:
144,85.

N

N

10

O2N

appearance: yellow solid
yield: 86% in ether
m.p : 192-194°C
1
H NMR (300 MHZ, CDCl3), δ: 1,26-1,41(m, 1H16b), 1,66-1,72

1

4

2

N

3

H
N

appearance: brown oil
yield : 88% in ether
1
H NMR (300 MHZ, CDCl3), δ: 1,19-1,39 (m, 1H16b), 1,58-1,69

(m, 1H16a), 1,94-1,98 (m, 1H18b), 2,03-2,10 (m, 2H17), 2,13-2,18
(m,1H18a),2,45-2,47(m,4H6,7),3,67(t,J=4,44H8,9), 3,79 (s,3H19),
4,77 (dd, J= 3,10;J=5,20 1H4), 6,88 (d, J= 9,15 2H11,12), 7,52 (d,
J= 9,15 2H13,14).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 23,73; C16: 32,45 ;
C18: 33,29; C6,7: 46,32; C19: 54,85; C8,9: 66,74; C4: 77,83; C5:
91,25 ; C11,12: 114,16; C13,14: 119,11 ; C10: 132,91; C15:
156,26.

4-(3-(3-(trifluoromethyl)phenyl)-3,3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol-3ayl)morpholine (3e)

 

99 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
17

8

7

O
19

16

N

9

F3C

18

12

6

N1

14

4

5

2

3

H

(m, 1H16a), 1,91-1,98 (m, 1H18b), 2,02-2,14 (m, 2H17), 2,242,31 (m,1H18a),2,38-2,51 (m,4H6,7) ,3,67 (t, J= 4,70 4H8,9),
4,82 (dd, J= 3,51;J=5,52 1H4), 7,30 (d, J= 7,78 1H11), 7,43 (t, J=
8,03 1H13), 7,84 (d, J=8,28 1H15) ,7,97 (s, 1H12).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 23,26; C16: 32,16 ;
C18: 33,33; C6,7: 46,38; C8,9: 66,84; C4:78,44; C5: 90,90 ; C12:
110,42; C11: 112,78 ; C13: 119,34 ; C19: 125,76; C15: 129,60;
C14: 131,28; C10: 139,72 .

N

N

10
11

15
13

3e 
 



4-(3-(4-fluorophenyl)-3,3a,4,5,6,6a--hexahydrocyclopenta[d][1,2,3]triazol-3ayl)morpholine(3f)
 
 
17

8

7

O
9

18

16

N

5

6

12

N1

14

F

15

4

2

3

H
N

N

10
11
13

3f 

appearance: Yellow solid caramel
yield: 90% in ether
m.p : 80-82°C
IR: 2960,80 (m) ; 1637,63(m) ; 1108,56(F) ; 1499,44(m) ;
880,31 (m) ; 535,77 (p) .
H NMR (300 MHZ, CDCl3), δ: 1,21-1,32 (m, 1H16b), 1,581,68 (m, 1H16a), 1,86-1,94 (m, 1H18b), 2,00-2,06 (m,
2H17),2,16-2,24 (m,1H18a),2,40-2,44 (m,4H6,7), 3,68
(t,J=4,9 4H8,9), 19 (s,3H19), 4,78 (dd, J= 3,40;J=5,47 1H4),
7.01 (dd, J= 4,70;J=4,90 2H11,12), 7.58 (dd, J= 4,70;J=4,90
2H13,14).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 23,60; C16:
32,66 ; C18: 33,81; C6,7: 46,83; C8,9: 67,25; C4:78,39; C5:
91,57 ; C11,12: 115,95; C13,14: 118,77 ; C10: 136,08; C15:
157,96.
1

 
Methyl6a-morpholino-1-phenyl-1,3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol-3acarboxylate(4a)
 
appearance: Solid chocolate brown



17

8

7 18

O
9

N
12

14

6

16
5

N1
10

19

4
2

3

N

11

15
13

4a 



appearance: Light yellow solid
yield : 88% in ether
m.p: 86-88°C
1
H NMR (300 MHZ, CDCl3), δ: 1,28-1,36 (m, 1H16b), 1,62-1,71

20

CO2Me

N

yield : 19% in ether
m.p : 108-110°C
1
H NMR (300 MHZ, CDCl3), δ: 1,60-1,68 (m, 1H16b), 1,85-1,91

(m,1H16a),2,03-2,07 (m,1H18b),2,16-2,20 (m,2H17) ,2,29-2,32
(m,1H18a),3,50 (t, J=4,52 4H6,7), 3,68 (t,J=4,52 4H8,9), 3,68
(s,3H20), 7,15 (d, J= 7,281H15), 7.35 (t, J= 7,52 2H13,14), 7,45
(d, J=8,78 2H11,12).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 22,26; C16: 31,39 ;
C18: 33,55; C19: 52,95; C6,7: 46,84; C8,9: 67,77; C4:79,00; C5:
93,07 ; C11,12: 115,68; C15: 125,47 ; C13,14: 130,54 ; C10:
142,38; C20: 168,24. 

Methyl6a-morpholino-1-(4-nitrophenyl)-1,3a,4,5,6,6a-hexahydrocyclopenta[d][1,2,3]triazol3a-carboxylate (4b)
 
100 

 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768
17

8

7 18

O

N

9

12

5

6

N1

14

O2N

16
19

4
3

2

20

CO2Me
N

1,65(m,1H16a),1,81-1,94(m,1H18b),1,82-2,16(m,2H17) ,2,482,50(m,1H18a),2,53-2,60(m,2H6),2,63-2,74(m,2H7),
,3,453,58(m,4H8,9), 3,88(s,3H20), 7,61(d,J=9,51 2H11,12), 8,24 (d,
J=9,51 2H13,14).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 23,24; C16:
31,65; C18: 33,27; C6,7: 46,74; C19: 54,64 C8,9: 68,83;
C4:79,81; C5:95,48 ; C12:120,18; C14: 125,19 ; C13: 128,52 ;
C11: 130,22; C15: 145,84; C10: 142,85; C20: 170,25.

N

10
11

15

13

4b 
 
 


Methyl1-(4-methoxyphenyl))-6a-mopholino-1,3a,4,5,6ª hexahydrocyclopenta[d]
[1,2,3]triazole-3a-carboxylate (4c)
 
appearance: brown oil

 

17

8

7 18

O
12

5

6

14

N1

20

4

2

3

N

11

15

21

CO2Me

N

10

19

MeO

16

N

9

13

4c 


appearance: yellow solid
yield : 45% in ether
m.p : 190-192°C
1
H NMR (300 MHZ, CDCl3), δ: 1,14-1,40(m, 1H16b), 1,61-

yield: 14% in ether
1
H NMR (300 MHZ, CDCl3), δ: 1,62-1,66 (m, 1H16b),

1,82-1,87
(m,1H16a),2,08-2,12
(m,1H18b),2,37-2,41
(m,2H17),2,48-2,50(m,1H18a),3,47-3,50(m,4H6,7),
3,80(t,J=3,79 4H8,9),3,87 (s,3H19),3,80 (s,3H21),
6,87(d,J=9,27 2H11,12), 7,35(d, J=9,27 2H13,14).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 22,73; C16:
30,45 ; C18: 32,29; C6,7: 46,54; C21: 52,95; C19: 55,7;
C8,9: 67,74; C4:77,87; C5: 92,25 ; C11,12: 114,25;
C13,14: 118,15 ; C10: 134,91; C15: 152,26; C20: 169,54. 

 

Methyl6a-morpholino-1-(3-(trifluoromethyl)phenyl)-1,3a,4,5,6,6a-hexahydrocyclopenta[d]
[1,2,3]triazol-carboxylate(4d)
 
17

8

7 18

O
9

19

F3C

N
12

5

6

14

N1
10
11

15
13

4d 
 
 

16
20

4

2

N

3

21

CO2Me

N

appearance: Yellow solid honey
yield: 51% in ether
m.p : 84-86°C
1
H NMR (300 MHZ, CDCl3), δ: 1,27-1,35(m, 1H16b), 1,81-

1,88(m,1H16a),2,05-2,10(m,1H18b),2,19-2,37(m,2H17)
,2,45-2,48(m,1H18a),2,50-2,71(m,4H6,7),3,443,49(m,4H8,9),3,86(s,3H21),7,36(d,J=7,781H11),
7,43(t,J=8,031H13), 7,69(d, J=8,28 1H15) ,7,75(s,1H12).
13
C NMR (300 MHZ, CDCl3) , δ ppm : C17: 22,78; C16:
32,28 ; C18:39,50; C6,7:48,57; C21:52,90; C8,9: 67,29;
C4:92,96; C5:94,78 ; C12:106,95; C15:113,35 ; C11: 118,93;
C19: 120,04; C13: 129,84 ;C14: 131,81; C10: 139,98; C20:
170,64

References
[1] a) Genin, M. J., Allwin, D. A., Anderson, D. J., Barbachyn, M. R., Emmert, D. E., Garmon, S. A.,
Graber, D. R., Grega, K. C., Hester, J.B., Hutchinson, D. K., Morris, J., Reischer, R. J., Ford, C.
W., Zurenko, G. E., Hamel, J. C., Schaadt, R. D., Stapert, D., Yagi, B. H., J. Med. Chem., 2000,
43, 953.
b) Molteni, G., Buttero, P.D., Tetrahedron, 2005, 61, 4983-4987.
[2] Siddiqhia, N., Ahsana, W., Alama, M. S., Alia, R., Jainb, S., Azada, B., Akhtara, J., International
Journal of Pharmaceutical Sciences Review and Reseach, 2011, Vol.8, 1, 161-169.
101 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

[3] Jehl, F., Antibiotiques, 2000, 2(4), 229.
[4] Witkowski, J. T. et al., J. Med. Chem., 1972, 15(11), 150; b) Y. S. Sanghvi, Y. S. et al, J. Med. Chem.,
1990, 33(1), 336.
[5] a) Huisgen, R., Angew. Chem. Int. Edn. 1963, 2, 565; b) for a detailed review, see: Lwowski, W.,
«1,3- dipolar cycloaddition chemistry», Ed. A. Padwa, Wiley-Interscience, New York, vol 1,
1984. Chapitre 5.
[6] Nolting.E., Michel. O., Ber., 1893, 26,86.
[7] Ranu.B.C, Sarkar.A et Chakraborty.R, J.Org.Chem, 1994, 59, 15,4114.
[8] Ouasti, F-Z., Memory of Magister, University of Oran, 2011.

 

102 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

PhytoChem & BioSub Journal

2013
Vol. 7 No. 3

ISSN 2170-1768

Comparative study on the vibrational IR spectra of N-aryl imidazoline-2(thio) one derivatives by various semi-empirical methods
A. LARBI 1, N. BELBOUKHARI 2, A. CHERITI 1 & A. ZANOUN 3
1)

Phytochemistry & Organic Synthesis Laboratory
Bioactive Molecules and Chiral Separation Laboratory
University of Bechar, Bechar 08000, Algeria
3)
ENSET , Oran BP.1523 Oran El Mnaouer, 31000, Oran, Algeria.
2)

Received: March O5, 2013; Accepted: June 29, 2013
Corresponding author Email belboukhari.nasser@yahoo.com
Copyright © 2013-POSL

Abstract- The vibrational IR spectra of several derivatives of imidazoline and their dipole moment
have been calculated using the MNDO , AM1 and PM3 methods , using Jaguar software and the
results are compared with those calculated from ab initio and density functional theory (DFT)
calculation. The ability of the semi empirical methods to predict the vibrational frequencies has been
probed.
Key Words: Imidazoline; IR spectra; DFT; MNDO; AM1; PM3

INTRODUCTION
Because of the importance of imidazole derivatives for living bodies, we carry on
working on these molecules both at experimental and theoretical levels. To determine the
physical and chemical properties of the molecule studied: Di Me -4,5 4 N-Ethyl N-Phenyl
Imidazoline-thione-2 (1) as well as its derivatives, we conducts the theoretical calculations
with the software of molecular simulation (Jaguar) by using some semi empirical methods
(AM1,PM3 and MNDO) and of the ab initio methods (HF and B3LYP). This calculation will
be validated by the results of the infrared spectroscopy. The result of the comparison will be
to find the theoretical method that interprets best the physical and chemical experimental
properties of the molecule studied ie that it gives the results with the weakest mistakes. Thus,
we can project , with the found method, the calculation for the no determined parameters by
the experimental (Table 1).
R1

S
H2
C N
H3C
H3C

N
CH3

103

R2
R3

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

MATERIALS AND METHODS
Theoretical calculations were carried out with both semi empirical (AM1, PM3,
MNDO) , ab initio (3-21 G) and DFT B3LYP (6-31 +G*) methods with Jaguar software. We
are going to study the vibrational parameters with a separated way. For every case, we are
going to study the molecule 1 then we pass to the set of the derivatives to verify the
substituting effect.

RESULTS AND DISCUSSION
The analysis done for the set of imidazoline in strong state by the initialization of the
tablets prepared from KBr gives the following values :
Table 1. Experimental results of Infra red spectra of imidazoline derivatives (KBr)
R1

R2 R3

1

H

H

H

2

Me

H

H

Frequence band RI (KBr) cm-1
3094,3022,2920,1602,1578,1506,1482,1419,1332, 1311,1275, 1254, 912,
801,765,714,696, 674, 573

3100,3016,2926,1578,1506,1479,1416,1323,1290,1251,1146, 1101, 1089,
912,795,777,720,684, 585
3148,3106,3022,1605,1575,1509,1473,1446,1422,1320,1278, 1254,1198, 1152,
H
1101, 1020, 912,771, 732, 684

3 OMe

H

4

OEt

H

H 2997,1605,1551,1425,1335,1245,1150,675, 575

5

H

Cl

H 3160,1605,1590,1488,1458,1437, 1323, 1257, 1233, 1077, 774, 705, 687 and 669

6

H

H

Cl

3154, 3112, 3028, 1578, 1500, 1476, 1419, 1320, 1284, 1269, 1251, 1089, 912,
873,756, 741, 573

To understand the physical and chemical properties and in order to interpret the specters of
imidazoline and its derivatives, the IR specters give very interesting information on the types
of the functions that exist within molecules and especially that constructs the functional
groups. The frequencies of vibrations calculated by PM3, MNDO, AM1 that are near and
belong to the zone of the characteristic vibrations of the molecule and these derivatives
(Experimental) are presented in the Tables 3,4 and 5.
Table 2. Comparison of the theoretical results IR with the experimental
EXP

573

678
696
714
765
801

PM3

B3LYP(DFT)

HF



42.58
485.88
571.92
583.22
611.30
629.38
665.17
679.55

51.43
509.47
578.05
622.11
623.85

34.13
502.56
552.66
618.5

1
2
3
4
5
6
7
8
9
10
11
12
13
14

782.51
821.94
849.63

668.11
687.88
714.86
765.76
783.66
823.12
842.43

104

683.45
706.03
774.26
806.91
817.57
846.54

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

912

1254
1275
1311
1332

1419
1482
1506
1578
1602
2920

3094

944.89
950.36
1223.03
1267.18
1292.94
1314.40
1332.76
1377.37
1402.61
1406.61
1429.81
1456.93
1472.54
1540.50
1559.96
1605.32
2896.87
2966.65
3011.91
3046.23
3047.67
3049.99
3067.20
3083.54
3138.32
3144.63
3177.77

937.23
980.71
957.49
1205.04
1239.21
1321.61
1337.96
1360.83
1403.71
1424.76
1462.33
1499.43
1504.93
1506.60
1547.63
1645.61
3041.01
3050..90
3088.29
3099.13
3117.83
3203.73
3229.68

896.13
917.17
988.25
1237.89
1250.25
1285.80
1317.33
1322.39
1349.43
1425.93
1469.46
1496.81
1505.43
1573.31
1599.53
1623.17

3205.57
3207.85
3301.72
3398.27

15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40

The Tables 2 summarize the frequencies of the strips of vibration feature of this molecule.
The calculated values are those of the maximal vibration that characterize the position of the
absorption strip in IR
We present first the theoretical calculation of the frequencies of vibration of the molecule 1,
with the AM1 methods, PM3 and MNDO, compared to the experimental results (Table 2).
The specters of vibration of the molecule 1 have been calculated using the AM1, PM3 and
MNDO methods. The results are compared with the frequencies of vibration feature of the
experimental specters of these molecules, so the best method pulled from this comparison will
be compared again to the ab initio and DFT methods.
Table 3. Comparison of the molecule 1 vibration frequencies calculations

1
2
3
4
5
6
7

AM1

PM3

40.57
500.03
541.33
615.84
625.11

42.58
485.88
571.92
583.22
611.30
629.38
665.17

656.96

MNDO
37.70
531.29
585.75
597.93
604.00
621.82
640.40

105

Experiment

573

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49

684.49

679.55

696.17

728.16
814.80
853.93

714.09

890.26
954.09

782.51
821.94
849.63
944.89
950.36

841.02
873.58
897.34
920.65
967.54

1203.58
1251.12
1273.35

1223.03
1267.18
1292.94

1327.70
1341.30
1376.58

1314.40
1332.76
1377.37

1223.93
1257.63
1270.92
1276.41
1340.33
1366.47
1381.80

1393.71
1425.61
1430.74
1435.27

1402.61
1406.61
1429.81
1456.93

1445.32

1472.54

1525.83
1548.43
1572.21
1588.01
1636.51

1540.50
1559.96
1605.32

2954.18

2896.87
2966.65
3011.91

3021.14
3048.17
3053.73
3067.46
3068.08
3139.32
3142.79
3179.41

3046.23
3047.67
3049.99
3067.20
3083.54
3138.32
3144.63
3177.77

1426.88
1429.94
1432.08
1443.01
1480.63
1491.96
1508.21
1554.01
1591.73
1602.10

678
696
714
765
801

912

1254
1275
1311
1332

1419

1482
1506
1578
1602

2920

3022

3202.43

3094

3342.45
3408.19

The comparison with the ab initio and DFT methods succeeds to the most precise method that
presents the most complete specter. The geometries of different bases are considered from the
standards values of bases. The frequencies of vibration are calculated for the whole set of the
frequencies of the imidazoline derivatives.
The values of vibration frequencies compared are only those that belong to the same scale
(same zone of vibration): according to the type and mode of vibration with the characteristic
values. Others comparison between the methods in the zones non mentioned in the
experimental specter.
106

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

Table 4. Results of the Molecule 1 vibration frequencies calculations
EXP

573

678
696
714
765
801

912

1254
1275
1311
1332

1419
1482
1506
1578
1602
2920

3094

PM3

B3LYP(DFT)

HF



42.58
485.88
571.92
583.22
611.30
629.38
665.17
679.55

51.43
509.47
578.05
622.11
623.85

34.13
502.5
6
552.6
6
618.5

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40

782.51
821.94
849.63
944.89
950.36
1223.03
1267.18
1292.94
1314.40
1332.76
1377.37
1402.61
1406.61
1429.81
1456.93
1472.54
1540.50
1559.96
1605.32
2896.87
2966.65
3011.91
3046.23
3047.67
3049.99
3067.20
3083.54
3138.32
3144.63
3177.77

668.11
687.88
714.86
765.76
783.66
823.12
842.43
937.23
980.71
957.49
1205.04
1239.21
1321.61
1337.96
1360.83
1403.71
1424.76
1462.33
1499.43
1504.93
1506.60
1547.63
1645.61
3041.01
3050..90
3088.29
3099.13
3117.83
3203.73
3229.68

683.4
5
706.0
3
774.2
6
806.9
1
817.5
7
846.5
4
896.1
3
917.1
7
988.2
5
1237.
89
1250.
25
1285.
80
1317.
33
1322.
39
1349.
43
1425.
93
1469.
46
1496.
81

107

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

1505.
43
1573.
31
1599.
53
1623.
17

3205.
57
3207.
85
3301.
72
3398.
27

According to the Table 1, the PM3 method gives good nearest results to the experimental in
relation to AM1 and PM3. According to works already done on the heterocyclic, the DFT
determines better the structure and the relative energies of the molecule. In the same way the
ab initio method has been introduced in large biologically active molecule domains that are
difficult and very long for the macromolecules that contain some heterocyclic . Therefore, it is
necessary to compare the results already calculated by the semi empirical method : PM3
(interpreting the physical and chemical parameters of this class of bioactive compound better).
In this study , we notice that the DFT method is nearer to the experimental result. The zone of
distortion DFT is better than the PM3 . The study of the IR matrix of this molecule shows the
existence of the vibrations N-H, even we not really finds them as covalent link chemical in the
molecule, it can be Van der Waals 's link as hydrogen links N-H are the weak links that vibrate
toward 3100-3250 cm 1. The results of DFT are bad
in the region 3000-3400 cm-1, on the other hand for the other regions present a considerable
approach with the experimental results and that the DFT also represents the totality of the specter
as the case of the PM3. For example the strips respective 1114,1044 cm-1 calculated by the DFT
and ab initio are of frequency that belongs to the vibration safe C-N by the molecules that
contains the oxygen because are characteristic of the C-O link which is an intense strip. In the
contrary the C-N and C-C strips are weak strips. The comparison of the PM3 method with the ab
initio methods shows that for the totality of the IR matrixes, the DFT is more believable, but
without excluding the PM3 in zones of calculations that are not presented agreement well with the
experimental in the strips of very elevated vibrations, these characteristic strips of all aromatic
organic compounds, So, the PM3 finds a very important place to be complement to the ab initio
method. Generally we can conclude from the results that the frequencies calculated by the PM3
method are the nearest to the experimental values except that the MNDO especially presents an
important agreement in the zone of the distortion vibrations 500-1000 cm-1 that concerns the
vibration of distortion = C-H that appear to 696 cm-1 in the experimental specter and to 696,17 cm1 in the MNDO spectra (Table 3) , the MNDO is rightly after the PM3 method concerning the
totality of the characteristic frequencies (37 for PM3 and 34 for MNDO). We notice in the MNDO
specters and AM1 the absence of the frequencies of vibration of groupings CH elongation, CH2
108

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

and CH3 in the zone 2800-3000 cm-1 which are determinist values of functional 5 groupings in
this molecule. In the zone 1000 to 1800 cm-1, the AM1 method gives nearer calculations for the
vibration of the aromatic connection C=C that appears to 1578 cm-1 ,1506 cm-1 in the experimental
spectra and to 1572,21 cm-1, 1525,83 cm-1 in the AM1 spectra (Table 3) and distortion of CH3 that
appears to 1419 cm-1 in the experimental spectra and to 1425,61 cm-1 in the spectra AM1(Table 3).
In the zone of the most elevated frequencies of vibration that characterizes the vibrations aromatic
CH between 3000-3100 cm-1 and the vibrations N-H (3100-3400cm-1) the calculation by the PM3
method gives good results that agree well with the experimental. For example the vibrations C-HS
aromatic appear to 3016 cm 1, 3100 cm 1 in the experimental specter and to 3044,27 cm 1,
3100,21 cm 1 in the PM3 specter (Table 3).In the case of OEt phenyl and OMe phenyl we finds
the C-O aromatic vibration and aliphatic C-O whrer the calculations done by the PM3 method are
nearer to the results of spectroscopic analysis. For example the C-O aromatic vibrations and
aliphatic appear to 1020 cm 1 and 1101 cm-1 in the experimental specter and to 1014,66 cm 1,
1097,33 cm 1, in the PM3 specter (Table 4) also the molecule 4 present a C-O vibration to 1089
cm-1 in the experimental specter and 1083,10 cm 1 in the PM3 specter (Table 4).
Table 5. Results of the Molecule 2 vibration frequencies calculations
Exp

684
720

MNDO
37.61
501.24
574.92
591.92
644.02
709.93

777
795

783.51
848.02

912

913.14
1025.80
1087.91
1097.29
1106.74

585

1089
1101
1146
1251
1290
1323
1416
1479

1506
1578
2926
3016
3100

1168.79

PM3
60.22
522.71
579.39
590.71
668.35
734.81
758.96
784.00
870.41
887.85
930.29
1001.13
1087.20
1094.42
1098.16
1114.42
1137.85

AM1
25.11
500.84
580.91
658.85
690.19
705.38
738.26
766.17
897.05

1224.88
1285.92
1335.28

1269.40
1294.17
1338.37

1416.33
1480.64
1487.43
1496.28
1506.78
1548.93
1593.19
1600.5

1407.33
1448.19

1122.09
1164.33
1215.93
1280.76
1289.98
1323.92
1330.70
1403.59
1461.03

1535.76

1495.48

1576.19
1603.90

1569.63
1605.65

3044.27
3100.21
3157.80

3194.29

916.97
1007.5
1078.87

δ (=CH arom)
δ (-CH2)
δ (=CH arom)
δ (C=C arom)
δ
(C4=C5
heterocycle)

υ(C=S)

δ (-CH2)
δ (-CH3)
υ(C=C aroma)
υ(C=C aroma)

109

υ(C=C heterocycle)
υ(CH2 , CH3)
υ(=C-H aroma)
υ(=C-H aroma)


1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

3202.59
3302.61
3400.55
3418.42

3208.32
3215.74

35
36
37
38

3209.50
3325.36
3330.30

We observe in the theoretical specters of the values of vibration frequency raised between 31003500 cm 1 that represents the harmonic weak vibrations and non harmonic that are not present in
the experimental specters It remains the vibration to weak absorption. For example the zone of
vibration 3362,49 to 3457,51 for the MNDO specter (Table 5) . The strips calculated between
1650-1900 don't represent really experimental vibration, but very weak vibrations of the C=C to
different substitutions . For example the absorption to 1674,05 cm 1 in the AM1 specter (Table 6).
The vibrations 500-900 represent the zone of distortion vibration in the plan and outside of the
plan. The vibrations inferior to 500 cm-1 due to the vibration of conformation (distortion) the
molecule due to the rotation around the C-N link between the C hybridizes sp2 of the aryl in a
gate of very determined rotation that gives to this molecule an atrophic chirality.
Here is a comparison therefore between the characteristic strips of valences for the set of the
derivatives imidazole for the experimental IR specters : According to the Tables 10 and 11 the
frequencies _ =C-H arom generally appears between 3000 and 3150 cm-1 the strip of
absorption of the double heterocyclic link appears toward 1600 cm-1. We can differentiate
between this six imidazoline derivatives by IR spectroscopy according to the characteristic
vibrations of the substituting so the additive effect on the other links. The presence of
vibration in the domain 1000 -1150 cm-1 for the molecules 4 and 5 show the vibrations of the
aliphatic C-O link and aromatic = C-O .(Table 7 and 8)

Table 6. Results of the Molecule 3 vibration frequencies calculations

684
732
771

MNDO
97.92
504.63
623.65
703.46
765.62

912

919.26

1020
1101

1011.06
1145.41

1152
1198

1171.62
1196.52
1233.67
1259.34

exp

1254
1278
1320
1422
1446
1473
1509

1318.59
1418.41
1420.47
1452.02
1471.94
1507.70

PM3
60.19
521.04
668.16
737.56
759.97
784.27
929.74
1006.57
1014.66
1097.33
1104.39
1137.63
1189.12
1224.37
1269.08
1293.23
1338.27

AM1
14.67
515.37
699.29
733.40
773.18

1407.49
1447.72

1421.96
1453.94
1484.56
1494.80

1537.10

δ (=CH arom)
δ (=CH arom)
δ (-CH2)

911.09
1007.00
1013.89
1120.79

δ
(C4=C5
heterocycle)
υ(CH2 , CH3)

1160.92
1198.41
1226.88

υ(=C-O)

1275.23
1325.60

110

υ(C=S)

υ(C=C aroma)


1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

1575
1605
3022

1540.11
1596.37

3106
3148

υ(C=C aroma)
1578.97

1588.80

3044.28
3100.38
3109.12
3145.90
3208.52
3216.02

3194.20
3219.16
3325.41
3330.40

3362.82
3430.70
3457.98

υ(C=C aroma)
υ(C=C )
υ(=C-H aroma)
υ(=C-H aroma)

23
24
25
26
27
28
29
30
31
32
33
34

Generally, we can conclude from the results that the calculated frequencies by the PM3
method are the nearest to the experimental values except that the MNDO method especially
presents an important agreement in the zone of the distortion vibrations 500-1000 cm-1 which
concerns the = C-H distortion vibration that appear to 696 cm-1 in the experimental specter
and to 696,17 cm-1 in the MNDO spectra (Table2) , the MNDO is rightly after the PM3
method concerning the totality of the characteristic frequencies (37 for PM3 and 34 for
MNDO). We notice in the MNDO and AM1 specters, the absence of the CH elongation
groupings vibration frequencies, CH2 and CH3 in the zone 2800-3000 cm-1 which are
determinist values of five functional groupings in this molecule. In the zone 1000 to 1800 cm1
, the AM1 method gives nearer calculations for the vibration of the aromatic bond C=C that
appears to 1578 cm-1 ,1506 cm-1 in the experimental spectra and to 1572,21 cm-1, 1525,83 cm1
in the AM1 spectra (Table 2) and distortion of CH3 that appears to 1419 cm-1 in the
experimental spectra and to 1425,61 cm-1 in the spectra AM1(Table 2). In the zone of the
most elevated vibration frequencies that characterizes the aromatic vibrations CH between
3000-3100 cm-1 and the vibrations N-H (3100-3400cm-1) the calculation by the PM3 method
gives good results that agree well with the experimental. For example the aromatic vibrations
C-HS appear to 3016 cm-1, 3100 cm-1 in the experimental specter and to 3044,27 cm-1,
3100,21 cm-1 in the PM3 specter (Table 3).
In the case of OEt phenyl and OMe phenyl, we finds the C-O aromatic vibration and aliphatic
C-O (CH3-O or And Oh) the calculations done by the PM3 method are nearer to the results of
spectroscopic analysis. For example the C-O aromatic vibrations and aliphatic appear to 1020
cm-1 and 1101 cm-1 in the experimental specter and to 1014,66 cm-1, 1097,33 cm-1, in the PM3
specter (Table 5) also the molecule 4 present a C-O vibration to 1089 cm-1 in the experimental
specter and 1083,10 cm-1 in the PM3 specter (Table 5). We observe in the theoretical specters
of the vibration frequency values raised between 3100-3500 cm-1 that represents the harmonic
weak vibrations and non harmonic which are not present in the experimental specters It
remains the vibration to weak absorption. For example the vibration zone 3362,49 to 3457,51
for the MNDO specter (Table 7). The strips calculated between 1650-1900 don't represent
really experimental vibration, but very weak C=C vibrations to different substitutions. For
example the absorption to 1674,05 cm-1 in the AM1 specter (Table 8), The vibrations 500-900
represent the vibration distortion zone in the plan and outside of the plan. The vibrations
inferior to 500 cm-1 due to the conformation vibration (distortion) caused by the rotation
around the C-N bond between the C hybridizes sp2 of the aryl in a gate of very determined
rotation that gives to this molecule an atrophic chirality. Here is a comparison between the
characteristic strips of valences for the Imidazole derivatives set which concern the
experimental IR specters
According to the Tables 9 and 10 the frequencies ν =C-H arom generally appears between
111

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

3000 and 3150 cm-1 the strip of absorption of the double heterocyclic bond appears toward
1600 cm-1.
We can differentiate between this six imidazolines derivatives by IR spectroscopy according
to the vibrations characteristic of the substituting so the additive effect on the other bonds.
The presence of vibration in the domain 1000 -1150 cm-1 for the molecules 4 and 5 show the
aliphatic C-O bond vibrations and aromatic = C-O one.
Table 7. Results of the Molecule 4 vibration frequencies calculations
exp

575

675

1150
1245
1335
1425
1551
1605
2997

MNDO

PM3

AM1



56.32
502.25
518.73
561.80
586.17
607.53
629.13
703.96
925.62
1001.29
1079.56

9.69
510.90

12.20
513.85

574.40
618.22

584.14
614.96

668.67
740.53
939.95
1005.67
1099.21
1144.52
1224.22
1269.39

659.98
745.05
932.64
1006.00
1101.76
1150.94

1
2
3
4
5
6
7
8
δ (=CH arom) δ (- 9
CH2)
10
11
12
13
14
υ(=C-O)
15
16
17
υ(C=S)
18
19
δ (-CH2) , δ (-CH3)
20
21
δ (-CH3)
22
23
υ(C=C arom)
24
25
υ(C=C)
26
υ(=C-H arom)
27
28
29
30
31
32
33

1145.56
1246.94
1328.35
1345.64
1420.62
1540.27
1596.76
1665.16

1338.22
1412.58
1447.98
1538.27
1580.13
1604.75

1224.70
1274.76
1334.77
1399.81
1450.86
1494.51
1589.97
1657.57

3015.07
3100.38
3202.06
3216.10

3194.19
3202.35
3322.27
3330.37

3356.89
3412.56
3457.96

Table 8. Results of the Molecule 5 vibration frequencies calculations
exp

MNDO

PM3

AM1

112



PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

669
687
705
774

1077

1233
1254
1323

1437
1458
1488

1590
1605

3160

33.84
527.13
639.29
689.11
700.90
809.51
966.29
1005.68
1077.19
1106.64
1223.20
1240.30
1276.96
1340.74
1426.71
1431.38
1442.39
1480.31
1491.92
1507.82
1592.62
1601.07

3202.85

64.40
530.31
661.39

55.01
539.55
658.82

715.55
776.30
909.14
1006.56
1070.22
1130.69

700.20
789.37
914.70
1008.64
1076.68
1201.33
1207.15
1252.10

1238.40
1272.27
1339.76
1401.33
1407.63
1444.86

1327.02
1330.59
1401.63
1461.19

1508.24
1535.55

1494.05
1553.97

1605.83

1595.54
1674.29

3044.52
3108.85
3142.47
3208.57
3216.35

3343.49
3401.12
3414.61

3194.07

δ (=C-H)
δ (-CH2)
δ (=C-H)

υ(C=S)
δ (-CH2)

δ (-CH2)
δ (-CH3)
υ(C=C arom)

υ(C=C arom)
υ(C=C)

υ(=C-H arom)

3217.41
3325.20
3330.08

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33

Table 9. Results of the Molecule 6 vibration frequencies calculations
exp

573
741
756
837
912
1089

1251
1269
1284
1320

MNDO
127.51
504.80
567.23
592.15
676.15
802.38
809.16
931.03
1021.32
1067.39
1118.59
1246.10
1288.46
1316.31

PM3
64.59
501.95
565.63
580.39
709.59
782.77
837.94
931.11
1006.59
1083.10
1094.75
1100.35
1237.60
1271.67
1295.87
1339.77

AM1
57.76
553.59
561.51
588.29
738.10
779.02
840.27
919.22
1008.79
1076.86
1110.53
1258.09
1275.74
1290.24
1316.82

113

δ (-CH2)
δ (=C-H arom)
δ (C=C)
δ (C=C)

υ(C=S)

δ (-CH3)


1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

1419
1476
1500
1578

1325.52
1419.64
1471.60
1488.67
1515.66
1543.51
1618.88

1407.73
1446.88
1498.63
1535.61
1606.24

1401.04
1459.59
1496.81
1598.74
1674.05

3044.51
3112.91
3142.07

3028
3112
3154

3209.93
3216.39

3194.07
3217.31
3325.21
3330.08

3362.49
3430.90
3457.51

17
18
δ (-CH2)
δ
(-CH3),υ(C=C 19
20
arom)
21
υ(C=C arom)
22
23
υ(C=C)
24
25
26
υ(=C-H arom)
27
υ(=C-H arom)
28
29
30
31
32
33

Table 10. Comparison between methods and characteristic bands
Méthodes
ν c=c

ν

CH2

et

CH3

ν c=c arom

Molécule1

Molécule2

Molécule3

Molécule4

Molécule5

AM1
PM3
MNDO
HF
B3LYP
Experim
ν AM1
PM3
MNDO
HF
B3LYP

1588.01
1605.32
1602.10
1599.53
1645.61
1602
2954.18
2966.65

1569.63
1576.19
1593.19

1588.80
1578.97
1596.37

1589.97
1604.75
1596.76

1595.54
1605.83
1601.07

1598.74
1606.24
1543.51

1578
-

1605

1605

1605

1578

Experim
AM1

2920
1525.83
1572.21
1540.50
1559.96
1508.21
1591.73
1505.43
1506.60
1506
1578

2926
1461.03
1495.48
1448.19
1535.76
1480.64
1506.78

1484.56,
1494.80
1573.10

1589.97

1494.05

1538.27

1508.24

1471.94
1507.70

1540.27

1480.31
1592.62

1459.59
1496.81
1446.88
1498.63
1471.60
1488.67

1479
1506

1473
1509
1575
1160.92
1137.63
1171.62

1551

1590
1488

1500
1476

PM3
MNDO
HF
B3LYP
Experim
ν =c-o

AM1
PM3
MNDO
HF

Molécule6

3041.01

114

1150.94
1144.52
1145.56

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768

Experim

1152

1150

Tableau 11. The characteristic valence bands:
Mol1
ν c=c
ν CH2 et ν CH3
ν c=c arom
ν =c-H arom
ν c=s
ν =c-o
ν CH3-0 ou CH2-0

1602
2920
1482
1506
1578
3022
3094
1254

Mol2
1578
2926
1479
1506
3016
3100
1251

Mol3

Mol4

Mol5

Mol6

1605

1605

1605

1578

1473
1509
1575
3022
3106
1254
1152
1020

1551

1590
1488

1500
1476

2997

3160

1245
1150

1257

3028
3112
1251

Conclusion
However, the ab intio and DFT methods have given us the better results of IR spectra calculations,
the ability of the semi empirical methods and especially PM3 method to predict the vibrational
frequencies has been probed.

REFERENCES
[1] V. Subramanian, K. Venkatesh. Comparative study on the vibrational IR spectra of cytosine and
thiocytosine by various semi-empirical quantum mechanical methods. India . Chemical Physics
Letters 264 (1997) 92-100
[2] A. Koch , S. Thomas , E.Kleinpeter. A . Ab initio study, semiempirical calculation and NMR
spectroscopy of keto-enol tautomerism of triazolopyrimidines. Germany. Journal of Molecular
structure (Theochem) 401 (1997) 1-14
[3] C.I.Williams,M.A.Withehead. Semi empirical study of isocyanate geometries, and β-lactam
formation through alkene-isocyanate cyclo-addition reactions.Canada: Quebec. Journ. Molecul.
Struct. 491 (1999) 93-101
[4] R.M.Srivastava , W.M.Faustino. Semi-empirical (PM3 and AM1) and ab initio molecular orbital
calculations of 1,2,4-oxadiazoles, 4,5-dihydro-1,2,4-oxadiazoles and 4,4-di-n-butyl-2phenylbenzo-1,3-oxazine. Brazil. Journal of Molecular Structure (Theochem) 640(2003) 49-56
[5] Milan Remko, Th. Van Duijnen, Marcel Swart2. Theoretical Study of Molecular Structure,
Tautomerism, and Geometrical Isomerism of N-Methyl- and N-Phenyl-Substituted Cyclic
Imidazolines, Oxazolines, and Thiazolines. Structural Chemistry, Vol. 14, No. 3, June 2003
[6] Pyridinium, imidazolium and quinucludinium oximes : synthesis, interaction with native and
phosphylated cholinesterases and antidotes against organophosphorus compounds. J Med Chem
Def Vol. 2, June 2004
[7] B.Célariès,C.Amourette,C.Lion
,G. Rima. Nouveaux phosphorothioates dérivés de la
naphthylméthylimidazoline et de la naphthyléthylimidazoline: application en radioprotection
chimique. France : Toulouse . Juillet 2004.
[8] Howard B.Broughton . Studies of Imidazole and Pyrazole Protonation using Electrostatically
Trained Neural Networks, Department of Chemistry, Imperial College, London, MO 63130-4899.
Radioprotection 2005, Vol. 40, n° 1, p 57- 71
[9] Hitoshi Kusamaa, Hironori Arakawab, Hideki Sugiharaa . Density functional study of imidazole–
iodine interaction and its implication in dye-sensitized solar cell. Journal de la photochimie et du
Photobiologie (2005) 197-204 chimie 171

115

PhytoChem & BioSub Journal Vol. 7(3) 2013
ISSN 2170-1768 

PhytoChem & BioSub Journal

2013 
Vol. 7  No. 3  

ISSN 2170‐1768 

Antimicrobial activity of essential oils of Bubonium Graveolens (Forssk.)
and Anvillea Radiata (Coss.)
Djahida AICI 1, Abdelkrim CHERITI 1, Younes BOURMITA1, Nasser BELBOUKHARI 2
1
2

Phytochemistry & Organic Synthesis Laboratory
Bioactive Molecules & Chiral Separation Laboratory
Bechar University, 08000 Bechar, Algeria

 
Received: Presented at Young Chem & BioChem Days, April, 2013
Corresponding author Email aici.djahida@yahoo.fr
Copyright © 2013-POSL
 

Abstract- Essential oils are a group of secondary metabolites identified in several families of aromatic
plants. These natural products are distinguished by their chemical characteristics and their interesting
biological activities (inflammatory, antioxidant, antibacterial, insecticides,...). It is in this context
articulates our work aims to study the antimicrobial activity of essential oils of two endemic medicinal
plants of the Southwest Algerian: Bubonium graveolens Forssk (Tafs), Anvillea radiata Coss (Nogd),
testing their antibacterial effect on four bacterial strains: Escherichia coli, Pseudomonas aeruginosa,
Bacillus steorothermophilus and Staphylococcus aureus. At the concentrations studied, both species
manifested significant antibacterial property with a zone of inhibition more than 11 mm noted for
Pseudomonas aeruginosa by the effect of the essential oil of Bubonium graveolens.
Key words: Essential oil, Bubonium Graveolens, Anvillea Radiata , Biological activity, Bacterial strains.

 

 
 
1. Introduction
The aromatogramme is a method for measuring the in vitro antibacterial activity of
essential oils. This is the equivalent of a susceptibility where antibiotics are replaced by
essential oils. 1 The use of essential oils in medicine was never abandoned despite the
discovery of organic synthesis process and the birth of the pharmaceutical industry.  They are
considered a reservoir of basic molecules that are irreplaceable. 2 The essential oils which
were utilized centuries ago in cosmetics usually show interesting biological features. The
Asteraceae family contains many medicinal and aromatic plants. 3
Bubonium graveolens (Forssk) and Anvillea radiata (Coss) belonging to the family
Asteraceae, is an endemic herbaceous medicinal aromatic plant mainly distributed in southwestern Algerian and south-eastern Morocco. 4
Anvillea radiata is used in the folk medicine as excellent heating, for the treatment of
dysentery, gastric–intestinal disorders and has been reported to have hypoglycemic activity. 5
Bubonium graveolens has been used in Sahara folk medicine as a stomachic, for treating
fever, gastrointestinal tract complaints, cephalic pains, bronchitis and as an intiinflammatory. 6
116 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

2. Materials and methods
2.1. Plant material
Aerial parts of B. graveolens and A. radiata were collected during flowering stage, from the
region between Bechar and Lahmar (over a distance of 30 km) in south-western Algeria
(April 2012).
2.2. Extraction of essential oil
Samples of flowers and leaves were dried in shade. Both flowers and leaves are subjected to a
steam distillation for 6 hours, in a montage developed by a pressure cooker, for increasing
quantity of extracted oil. The oil was dried over anhydrous sodium sulphate and stored at 4°C
until analysis.
2.3. Antimicrobial activity
2.3.1. Bacterial strains
For the determination of antibacterial activity of B. graveolens and A. radiata essential oils,
standard and isolated strains of the following Gram-negative bacteria: Escherichia coli ATCC
25922, Pseudomonas aeruginosa ATCC 27853; Gram-positive bacteria: Bacillus
steorothermophilus ATCC 12980, Staphylococcus aureus ATCC 25923, were used. The
microorganisms were obtained from the “Pasteur Institute of Algiers, Algeria”.
2.3.2. Preparation of dilutions
Due to the immiscibility of essential oils in water, different dilutions were prepared using
DMSO eluent. 7
2.3.3. Screening for antibacterial activity :
Antimicrobial activity was tested by the agar-well diffusion method. All bacterial cultures
were first grown on Mueller Hinton agar at 37°C for 18–24 h prior to inoculation onto the
nutrient agar. One or several colonies of similar morphology of the respective bacteria were
transferred into API Suspension medium and adjusted to 0.5 McFarland turbidity standard (13 ×108 bacteria /ml) with a photometer (UV lamp type Spectrolin, Model ENF-260/ FE).
The inoculums of the respective bacteria were streaked onto Mueller Hinton agar plates using
a sterile swab. A sterile filter disc (diameter 6 mm, Whatman paper) was placed. The disc was
impregnated by four (04) different concentrations of the tested essential oils (4 µL/disc). The
treated Petri dishes were incubated at 37°C for 18–24 h.
Antimicrobial activity was evaluated by measuring the zone of growth inhibition around the
discs after 24 h of incubation at 37°C. The diameter of the zones of inhibition around each of
the discs was taken as measure of the antimicrobial activity. Each experiment was carried out
in quintuplicate and the mean diameter of the inhibition zone was recorded.
3. Statistical Analysis:
The conventional statistical methods were used to calculate averages and standard deviations.
All measurements were replicated five times, and data are presented as mean ± standard
deviation.
4. Results and discussion:
The antimicrobial activities of B. graveolens and A. radiata were evaluated by a paper disc
diffusion method against tested bacteria. The results showed that the essential oils were active
against the microorganisms assayed. Related to the inhibition of growth, significant
differences were detected among these cited oil types, since all of them showed an interesting
activity for all tested strains.
The Anvillea radiata essential oil showed antimicrobial activity against all microbial strains
tested. The concentration of 100 µg/ml showed a zone of inhibition against the bacterial strain
117 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

Bacillus steorothermophilus, then the concentration 50 mg / ml has an activity against
Pseudomonas aeruginosa.
The essential oil of Bubonium graveolens shows a large zone of inhibition, which appeared
with P. aeruginosa at the concentration of 25 µg/ml. This oil also has a large inhibition zone
against Escherichia coli at the same concentration.
Bubonium graveolens essential oil is effective against gram-positive bacteria, with remarkable
inhibitions zone for S. aureus and B. steorothermophilus. While Anvillea radiata essential oil
proved an average antibacterial activity against S. aureus strains.
The results of the antibacterial activity for the two essential oils studied are shown in Tables 1
and 2.
Table 1:Antibacterial activity of Anvillea radiata essential oils against bacterial strains.
Bacterial strains

Inhibition zone diameter (mm ± SD)
C1

C2

C3

C4

Gram-negative bacteria
E. coli ATCC 25922
P. aeruginosa ATCC 27853

7.8 ± 2.2
8.4 ± 1.6

8.2 ± 1.8
10.8 ± 1.2

9.6 ± 1.4
7.0 ± 3.0

8.5 ± 1.5
7.3 ± 1.7

Gram-positive bacteria
B. steorothermophilus ATCC 12980
S. aureus ATCC 25923

8.8 ± 1.2
7.6 ± 2.4

6.7 ± 0.3
7.4 ± 1.6

8.2 ± 2.8
6.5 ± 0.5

11.0 ± 1.0
6.9 ± 1.1

SD: standard deviation. C: significant concentration, C1: 25, C2: 50, C3: 75, C4:100.
Concentrations are expressed in µg/ml.

Table 2: Antibacterial activity of Bubonium graveolens essential oils against bacterial strains.
Bacterial strains

Inhibition zone diameter (mm ± SD)
C1

C2

C3

C4

Gram-negative bacteria
E. coli ATCC 25922
P. aeruginosa ATCC 27853

11.0 ± 1.0
11.8 ± 1.2

7.6 ± 2.4
7.5 ± 2.5

8.4 ± 0.6
11.4 ± 0.4

6.8 ± 1.2
10.8 ± 1.2

Gram-positive bacteria
B. steorothermophilus ATCC 12980
S. aureus ATCC 25923

8.9 ± 2.1
9.4 ± 2.6

6.7 ± 1.3
6.9 ± 1.1

9.3 ± 1.7
7.0 ± 3.0

9.0 ± 1.0
9.4 ± 1.6

SD: standard deviation, C: significant concentration, C1: 25, C2: 50, C3: 75, C4:100.
Concentrations are expressed in µg/ml.

5. Conclusion:
The antimicrobial activity of different oils was tested using the diffusion method and by
determining the inhibition zone. The results showed that all examined oil types had great
potential of antimicrobial activity against strains.
These first results we have obtained allow a systematic study of many essential oils on
pathogenic bacteria with increased resistance vis-à-vis conventional antibacterial agents,
including inpatient samples.
118 
 

PhytoChem & BioSub Journal Vol. 7(3) 2013
  ISSN 2170-1768

6. References:
1 J. Kaloustian, J. Chevalier, C. Mikail, M. Martino, L. Abou, M.-F. Vergnes, M.-F. Vergnes. Etude de
six huiles essentielles : composition chimique et activité antibactérienne. Phytothe´rapie (2008) 6:
160–164.
2 D. Ouraïni, A. Agoumi, M. Ismaïli-Alaoui, K. Alaoui,Y. Cherrah ,M.Amrani, M.-A. Belabbas. Étude
de l’activité des huiles essentielles de plantes aromatiques à propriétés antifongiques sur les
différentes étapes du développement des dermatophytes. Phytothérapie (2005), 4: 147-157.
3 Elhoussine Derwich, Zineb Benziane, Abdellatif Boukir. Chemical compositions and insectisidal
activity of essential oils of three plants Artimisia Sp: Artimisia herba-alba, Artimisia absinthium and
Artimisia pontica (Morocco). Electronic Journal of Environmental, Agricultural and Food Chemistry,
1202-1211.
4 Quezel P, Santa S. Nouvelle Flore de l’Algerie et des Regions Desertiques et Meridionales, vol. II.
CNRS : Paris, 1983.
5 B. El Hassany, F. El Hanbali, M. Akssira, F. Mellouki, A. Haidour, A.F. Barrero. Germacranolides
from Anvillea radiata. Fitoterapia 75 (2004), 573 – 576.
6 Abdelkrim Cheriti, Amel Saad, Nasser Belboukhari, Said Ghezali. The essential oil composition of
Bubonium graveolens (Forssk.) Maire from the Algerian Sahara. Flavour and Fragrance Journal.
(2007) 22: 286-288.
7 Kouamé Raphaël Oussou, Coffi Kanko, Nathalie Guessend, Séri Yolou, Gérard Koukoua, Mireille
Dosso, Yao Thomas N’Guessan, Gilles Figueredo, Jean-Claude Chalchat. Activités antibactériennes
des huiles essentielles de trois plantes aromatiques de Côte-d’Ivoire. C. R. Chimie, (2004), 1081–
1086.

119 
 

PhytoChem & BioSub Journal
Peer-reviewed research journal on Phytochemistry & Bioactives Substances

ISSN 2170 - 1768

ISSN 2170-1768

POSL

Edition LPSO
http://www.pcbsj.webs.com
Email: phytochem07@yahoo.fr



Télécharger le fichier (PDF)










Documents similaires


phytochem biosub journal vol 7 3 2013
phytochem biosub journal vol 8 n 1 2014
phytochem biosub journal vol 8 n 2 2014
phytochem biosub journal vol 8 n 3 2014
phytochem biosub journal vol 8 4 special 2014
03 0 pcbsj vol9 3 2015 cheriti editorial g

Sur le même sujet..