Content uploaded by Ibtisam Ahmed
Author content
All content in this area was uploaded by Ibtisam Ahmed on Feb 15, 2016
Content may be subject to copyright.
Available online www.jocpr.com
Journal of Chemical and Pharmaceutical Research, 2014, 6(7):151-156
Research Article
ISSN : 0975-7384
CODEN(USA) : JCPRC5
151
Commiphora myrrha and commiphora Africana essential oils
Suad A. Gadir
1
and Ibtisam M. Ahmed
2
1
Department of Chemistry, Faculty of Education, Alzaeim Alazhari University, Khartoum, Sudan
2
Faculty of Education, Faculty of Graduate Studies and Scientific Research Program Ph.D. Chemistry, Alzaeim
Alazhari University, Khartoum, Sudan
_____________________________________________________________________________________________
ABSTRACT
From generation to generation in all countries all around the world medicinal plants play an important role in our
live from ancient time till these days of wide drugs and pharmacological high technique industries , the studding of
biological and pharmacological activities of plant essential oils attracted the attention to the potential use of these
natural products from chemical and pharmacological investigation to their therapeutic aspects. In this paper two
resins commiphora Africana and commiphora myrrha were selected to discuss their essential oils for chemical
analysis and biological aspect the results of GCMS shows that the two resins are rich in sesqiuterpenes and
sesqiuterpene lactones compounds that possess anti-inflammatory and antitumor activity Antibacterial and
antifungal bioassay shows antibacterial and antifungal activity higher in the myrrha oil than the Africana oil while
antiviral bioassay shows higher antiviral activity in the Africana oil than myrrha oil
Key words: burseraceae, Essential oil, Sesquiterpenes, GCMS, brine shrimbs, NDV.
_____________________________________________________________________________________________
INTRODUCTION
The genus Commiphora (Burseraceae) includes over 150 species of trees and shrubs, distributed mostly in East
Africa, Arabia and India (1).The comminphora myrrha tree produces a type of aromatic resin known as myrrh (mur
higazy) ,commiphora Africana tree produce gum resin known in sudan as gafal resin . name Commiphora may be
based on the Greek words kommi gum and phero to bear (2) Biblically, Myrrh was given to Christ at his birth. It
was esteemed higher and cost more than gold. The Muslim physician al-Razi the greatest of all medieval
clinicians, used myrrh to treat ailments of the kidneys and bladder, to dissipate swellings in the stomach . (3)
Essential oils have been used for a wide variety of purposes including perfumes, cosmetics, aromatherapy ,
secondary metabolites .Terpenes and terpenoids are the most important constituents in essential oils terpenes are
distributed in the plant kingdom (4) pharmaceuticals composition reported some sesquiterpene lactones in treating
some diseases , helenalin is a anti-inflammatory drug and parthenolide a sesquiterpene lactones with anti cancer
effect (5 .6) .Gas chromatography mass spectroscopy were used to identify chemical components in essentialn oils
(7)
EXPERIMENTAL SECTION
2.1.Plant materials
Commiphora Africana resin were collected from the western sudan (Jabal mara ),and identified by Dr. Mohamed
Elmubark at sudan university of science and technology resins unit and commiphora myrrh resin which is known
and widely used in sudan and Arabic and African countries were bought from Omdurman market ,their essential oil
were extracted by steam distillation at the researches centre of medicinal and aromatic plants at Khartoum Sudan .
200 gm of each resin were grinded and the oil extracted by steam distillation .6.5 ml. of C.myrrha essential oil were
yielded while only 2ml. of C. Africana were yielded .
Suad A. Gadir
and Ibtisam M. Ahmed J. Chem. Pharm. Res., 2014, 6(7):151-156
______________________________________________________________________________
152
2.2.Thin layer chromatography:
Thin layer chromatography was carried out for commiphora Africana and commiphora myrrh essential oils, Pre-
coated silica gel plate used a stationary while Petroleum ether: Di-ethyl ether (9: 1) and Toluene : Ethyl acetate
(93: 7) were used as mobile phase 0.2 ml of the samples was dissolved in 105 ml of methanol and applied to the
bottom of the plated at 2 cm using capillary tube. Plate was kept to dry by air and then inserted in tank containing
the selected solvent system. After reached the height of 15 to 20 cm, plate was took out and allowed to air till
solvent was completely evaporated. The plate was inspected in day light, then examined under UV lamp and finally
sprayed with para anizaldehyde spray reagent and heated at 100 C for about five minutes. Rf values of separated
compounds which appeared under UV , after sprayed and heated retention factors were calculated as follows.
Rf= distant travelled by sample
distant travelled by solvent
Photo 2. 2. TLC plate of C.AF oil and C.MR oil
Table 2.2.1 Retention factor of commiphora Africana oil (petroleum ether and diethyl ether ( 9 : 1 )
Spot 1 2 3 4 5 6 7
Rf 0.2 0.3252 0.4172 0.5460 0.7362 0.8098 0.9754
2.2.2 Retention Factor of Commiphora Africana oil (Toluene and Ethyl acetate ( 93:7)
Spot 1 2 3 4 5 6 7
Rf 0.3333 0.5333 0.5666 0.7333 0.8000 0.9333 0.9867
Table 2.2.3 Retention Factor of Myrrha oil (petroleum ether and diethyl ether 9:1)
spot 1 2 3 4 5 6 7
Rf 0.16266 0.21687 0.5422 0.6458 0.7169 0.8132 0.8916
Table2.2.4 Retention factor of Myrrha oil (Toluene and Ethyl acetate93:7
Spot 1 2 3 4 5 6 7
Rf 0.3354 0.4051 0.0613 0.6709 0.7532 0.8798 0.9403
2.3.Gas chromatography mass spectroscopy
GC/MS is the method for identifying volatile compounds in complex mixtures. GCMS of these oils were done to
investigate chemical compounds in Commiphora .africana and Commiphora. myrrha oils . GCMS analysis were
done at the instrumentation lab of central petroleum laboratories Khartoum. the results shows they were rich in
terpenes specially sesquiterpene and sesquiterpene lactones and diterpenes which have wide interest for their
biological activities such as anti inflammatory and anti cancer effect
Suad A. Gadir
and Ibtisam M. Ahmed J. Chem. Pharm. Res., 2014, 6(7):151-156
______________________________________________________________________________
153
Figure 2.3.1 commiphora Africana essential oil chromatogram
Table 2.3.1 Chemical compounds in commiphora Africana oil detected by GCMS
NO NAME FORMULA RT MW AREA AREA%
1 Cyclobutane, 1,2-bis(1-methylethenyl)-, trans- C10H16 19.827 136 526436 0.6356
2 trans-4-Decene C10H20 23.575 140 558300 6.741
3 trans,trans-3,5-Heptadien-2-one C7H10O 27.027 110 315030 0.3804
4 Cyclopropane, octyl- C11H22 33.233 154 173500 20.95
5 Ethanone, 1-(1,3-dimethyl-3-cyclohexen-1-yl)- C10H16O 54.811 152 467205 0.5641
6 1,4-Methanoazulene, decahydro-4,8,8-trimethyl-9-methylene-, [1S-(1à,3aá,4à,8aá)]- C15H24 55.420 204 263600 31.83
7 Ethanone, 1-(1,4-dimethyl-3-cyclohexen-1-yl)- C10H16O 55.741 152 448518 0.5415
8 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, [S-(E,Z,E,E)]- C20H32 73.928 272 440500 5.319
9 Podocarpa-6,13-diene, 13-isopropyl- C20H32 74.203 272 908464 1.097
10 Kaur-16-ene, (8á,13á)- C20H32 74.453 272 495685 0.5985
11 Kaur-16-ene C20H32 74.536 272 884200 10.68
12 Phenanthrene, 7-ethenyl-1,2,3,4,4a,4b,5,6,7,9,10,10a-dodecahydro-1,1,4a,7-tetra
methyl-, [4aS-(4aà,4bá,7á,10aá)]- C20H32 74.837 272 977313 1.180
13 17-Norkaur-15-ene, 13-methyl-, (8á,13á)- C20H32 75.286 272 971531 1.173
14 Podocarpa-6,13-diene, 13-isopropyl- C20H32 75.505 272 228400 2.758
15 Isodihydrohistrionicotoxin 285a C19H27NO 75.775 285 111000 1.340
16 Bicyclo[9.3.1]pentadeca-3,7-dien-12-ol, 4,8,12,15,15-pentamethyl-, [1R-(1R*,3E,
7E,11R*,12R*)]- C20H34O 77.210 290 687298 0.8299
17 Cyclopentanemethanol, 5-methyl-2-[1-methylene-3-(5-isopropyl-2-methylcyclopent-
1-enyl)propyl]- C20H34O 77.389 290 107800 13.02
Chromatogram Plot
File: c:\varianws\data\sara elseikh\rcc\id1277411-13-2012.sms
Sample: ID12774 Operator: TAWFEEG
Scan Range: 1 - 4102 Time Range: 0.00 - 85.98 min. Date: 11/13/2012 11:54 AM
20 30 40 50 60 70 80 minutes
0
1
2
3
4
MCounts 35:450
1A
Apex: 19.827 min.
Area: 526436
Apex: 23.575 min.
Area: 5.583e+6
Apex: 27.027 min.
Area: 315030
Apex: 33.233 min.
Area: 1.735e+7
Apex: 54.811 min.
Area: 467205
Apex: 55.420 min.
Area: 2.636e+7
Apex: 55.741 min.
Area: 448518
Apex: 73.928 min.
Area: 4.405e+6
Apex: 74.203 min.
Area: 908464
Apex: 74.453 min.
Area: 495685
Apex: 74.536 min.
Area: 8.842e+6
Apex: 74.837 min.
Area: 977313
Apex: 75.286 min.
Area: 971531
Apex: 75.505 min.
Area: 2.284e+6
Apex: 75.775 min.
Area: 1.110e+6
Apex: 77.210 min.
Area: 687298
Apex: 77.389 min.
Area: 1.078e+7
Apex: 77.496 min.
Area: 310270
Seg 2, Ext EI, Time: 5.00-86.00
975 1443 1910 2356 2837 3283 3814 Scans
Suad A. Gadir
and Ibtisam M. Ahmed J. Chem. Pharm. Res., 2014, 6(7):151-156
______________________________________________________________________________
154
Figure 2.3.2 Commiphora Myrrha Oil GCMS chromatogram
Table table 2.3.2 Chemical compounds detected in Commiphora myrrh Essential oil by GCMS
NO NAME FORMULA RT MW AREA AREA%
1 Cyclohexene, 4-ethenyl-4-methyl-3-(1-methylethenyl)-1-(1-methylethyl)-, (3R-
trans)- C15H24 40.896 204 170171 0.3342
2 Longifolene-(V4) C15H24 44.462 204 2061000 4.048
3 Caryophyllene C15H24 46.195 204 151535 0.2976
4 1,4-Methanoazulene, decahydro-4,8,8-trimethyl-9-methylene-, [1S-(1à,3aá,
4à,8aá)]- C15H24 46.947 204 533347 1.047
5 1,6-Cyclodecadiene, 1-methyl-5-methylene-8-(1-methylethyl)-, [s-(E,E)]- C15H24 50.042 204 178100 0.3498
6 Naphthalene, 1,2,3,5,6,7,8,8a-octahydro-1,8a-dimethyl-7-(1-methylethenyl)-,
[1S-(1à,7à,8aà)]- C15H24 50.517 204 197277 0.3874
7 Benzofuran, 6-ethenyl-4,5,6,7-tetrahydro-3,6-dimethyl-5-isopropenyl-, trans- C15H20O 51.122 216 13560000 26.63
8 1H-Cycloprop[e]azulene, decahydro-1,1,7-trimethyl-4-methylene-, [1aR-(1aà,
4aá,7à,7aá,7bà C15H24 54.643 204 294150 0.5777
9 meso-Hydrobenzoin C14H14O2 55.861 214 175025 0.3437
10 1(2H)Phenanthrenone, 3,4,4a,9,10,10a-hexahydro-4a-methyl- C15H18O 58.868 214 25160000 49.41
11 Benzenemethanol, 3-methoxy-à-phenyl- C14H14O2 59.293 214 7021000 13.79
12 Cyclohexanemethanol, 4-ethenyl-à,à,4-trimethyl-3-(1-methylethenyl)-, [1R-
(1à,3à,4á)]- C15H26O 61.359 222 321120 0.6306
13 Testosterone C19H28O2 64.954 288 1097000 2.154
3.1 Antibacterial and antifungal bioassay
Commiphora myrrha and commiphora Africana oils were tested on their antibacterial and antifungal activity at
microbiology lab in medicinal and aromatic plants researches centre in Khartoum , staphauries and klebsilla bacteria
were selected to test the oils anti bacterial effect while Candida and Aspergillus fungi were selected to test the
antifungal activity bellow results reported the inhibition zone of bacteria or fungi growth measured in millimetres.
Chromatogram Plot
File: c:\varianws\data\sara elseikh\rcc\id1277511-14-2012.sms
Sample: ID12775 Operator: TAWFEEG
Scan Range: 1 - 4007 Time Range: 0.00 - 85.96 min. Date: 11/14/2012 8:12 AM
30 40 50 60 minutes
0.0
0.5
1.0
1.5
2.0
2.5
MCounts 35:450
Apex: 40.896 min.
Area: 170171
Apex: 44.462 min.
Area: 2.061e+6
Apex: 46.195 min.
Area: 151535
Apex: 46.947 min.
Area: 533347 Apex: 50.042 min.
Area: 178100
Apex: 50.517 min.
Area: 197277
Apex: 51.122 min.
Area: 1.356e+7
Apex: 54.643 min.
Area: 294150
Apex: 55.861 min.
Area: 175025
Apex: 58.868 min.
Area: 2.516e+7
Apex: 59.293 min.
Area: 7.021e+6
Apex: 61.359 min.
Area: 321120
Apex: 64.954 min.
Area: 1.097e+6
Seg 2, Ext EI, Time: 5.00-86.00
1417 1863 2327 2834 Scans
Suad A. Gadir
and Ibtisam M. Ahmed J. Chem. Pharm. Res., 2014, 6(7):151-156
______________________________________________________________________________
155
Table 3.1 Commiphora myrrha and commiphora Africana essential oil antibacterial and antifungal results of diameter of growth
inhibition zone in millilitres
Bacteria or fungi
S.a K.p A.n C.a
C.AF oil 16 15 17 19
C,MR oil 18 17 18 20
C.AF commiphora Africana . C.MR commiphora myrrha . S.a Staphylococcus aureus . K.p Klebsiella pneumoniae . A.n Aspergillus niger . C.a
Candida albica
Photo 3.1 inhibitition zone by Myrrha oil and Africana oil in Petri dish
3.2. Brine shrimp lethality bioassay
Brine shrimp lethality bioassay is a simple method to investigate the cytotoxcisisty Of extracts of medicinal plants
. Brine shrimps (Artemia salina) were hatched using sterile artificial seawater (prepared using sea salt 38 g in one
litre distilled water ) and let to 24 hours After hatching, ten active nauplii free from egg shells were collected from
brighter portion of the hatching chamber and used for the assay. three beaker prepared for each concentration as
follows , 0.02 ml of the oil dissolved in 2 ml ethanol to form 1000 PPM , then 0.2 ml from it were added to 1.8
ethanol to reach 100 PPM and from this 0.2 ml were added to 108 ethanol to have 10 PPM . these three
concentration kept to add them to the hatched nauplii , . Ten nauplii were drawn through a glass capillary and
placed in small beaker contain the prepared oil and artificial sea water were added to complete five millilitre and
maintained at room temperature for 24 h under the light and surviving larvae were counted. To calculate LC50 for
each oil .Caffeine LC50 306 ug/ml used as positive control and ethanol LC50 1000 ug/ml is negative control
Table 3.2.1 shrimps bioassay results of commiphora african oil
concentration 1000ug/ml 100 ug/ml 10 ug/ml Lc50 ug/ml
Percent death
100 90 76.67 60
Table 3.2.2 Shrimps bioassay of commiphora myrrha oil
concentration 1000ug/ml 100ug/ml 10ug/ml Lc50ug/ml
Percent death
86.67 73.33 63.33 68
3.3.Antiviral bioassay
Antiviral test were held at veterinary research institute virology section towards Newcastle virus (NDV) on chicken
embryo nine days old, according to Rajbahndari et al. method with some modifications , 0.1ml of NDV suspension
was treated with 0.1 ml of pure oil and 0.1ml of ethanol diluted oil 50% ,and the treated viruses were incubated at
4ºC for about 1 h. The treated viruses and the controls were then inoculated via the allantoic sac of 9 day old chick
embryos for NDV. Hank's balanced salt solution (negative control ) and the virus without treatment (positive
control ) were used as controls. Triplicate tests were carried out for each oil against the virus, the results were
compared to the sample without treatment
Antiviral screening of the two oil
Commiphora Africana Essential Oil : The pure and the ethanol oil shows high and clear antiviral activity
Suad A. Gadir
and Ibtisam M. Ahmed J. Chem. Pharm. Res., 2014, 6(7):151-156
______________________________________________________________________________
156
Commiphora Myrrha essential oil: The pure myrrha oil shows high antiviral activity while the ethanol oil shows
moderate antiviral activity
RESULTS AND DISCUSSION
Results of GCMS shows different terpene compounds in C.Af and C.Mr oils monoterpene, sesquiterpene,
sesquiterpene lactones and diterpene which have therapeutic aspects (8) , Caryophyllene C15H24 which found in
myrrha oil Anti-tumour antibacterial, anti- inflammatory .C19H27NO which found in Africana oil pentazocine
pain reliever. And , C20H32 refer to Kaurene may serve as a leading compound for the development of new
chemotherapeutic drugs to overcome resistance of tumours to chemotherapy. (9) were found in commiphora
africana oil
CONCLUSION
Commiphora myrrha and Commiphora Africana oils have antibacterial and antifungal activities and were rich in
compounds that play important role in therapy ,Commiphora Africana oil shows clear and high antiviral activity
towards NDV more investigation can lead to control this aggressive virus , low LC50 values of the two oils explains
their cytotoxicity and anti-tumour properties ,the main chemical compounds in the two oils were found as
sesquiterpenes and sesquiterpene lactones the antitumor properties of sesquiterpene lactones have attracted a good
interest ,more investigation is needed for these oils and their components can lead to interesting pharmaceuticals
natural products
Acknowledgements
The authors are grateful to the chemistry and microbiology labs at Institute Research Center of Medicinal and
Aromatic Plants Khartoum, Sudan and to the virology section at the Center for Veterinary Research Soba Khartoum
Sudan and to the instrumental lab at Central Petrolium laboratories Khartoum Sudan for their support of this work
REFERENCES
[1] Flora of Tropical East Africa, William Bertram Turrill, E. Milne-Redhead, Taylor & Francis, p 100 , 2003
[2] Poonia,Priyanka ,Mittal ,Sanjeev K.Gupta. Vivek Kumar International Journal of pharmacognosy and
Phytochemical Research , vol. 6 , p 347- 354 , 2014
[3] Dr. Sharif Kaf Al-Ghazal The Valuable Contributions of al-Razi in the History of Pharmcy , The Foundation for
Science and Technology and Civilisation,UK 2007
[4] Vidita V. Bhargava Shashank C. Patel , Kruti S. Desai international Journal of Herbal Medicine volume 1 pages
14-21 . 2013
[5] Michael F Neerman , International Journal of Aromatherapy vol. 13 pages 114-120. 2003
[6] V.cijo George , R.Ashok Kumar International journal of pharmacy vol. 3 pages 69-71, 2012
[7] Ivan LJ. Milovanovic1, Aleksandra . Journal of the Institute of Food Technology Volume 36, pages 75-79 ,
2009
[8] Qurshi; Alharbi MM; Ahmed MM; .Cancer Chemotherapy and Pharmacology, vol,33, :130–138 ,1993
[9] M asuo Kondoh, Ikue Suzuki, Masao Sato, Fumihiro Nagashima The journal of pharmacology and
experimental therapeutics vol. 311, P 115–122, 2004
