Antibacterial
activity of essential oil extracted from Coriandrum sativam (L.) and GC-MS analysis
Suganya S.1,
Bharathidasan R.1, Senthilkumar
G.1, Madhanraj P.2*
and Panneerselvam A.1
1PG and Research Department of Botany and Microbiology,
A.V.V.M Sri Pushpam College (Autonomous), Poondi, Thanjavur, Tamil Nadu.
2Dept of Microbiology, Thanthai
Hans Roever College of Arts and Science, Perambalur - 621 212, Tamil Nadu, India.
ABSTRACT:
Extraction of essential oils from locally available
plant Coriandrum sativam (L.) was carried out using steam
distillation followed by ether extraction. Dried and purified extracted oils
were screened for their antibacterial activity of both gram positive (Staphylococcus
aureus) and gram negative (Enterobacter
aerogenes, Klebsiella pneumoniae, Vibrio cholerae and Salmonella typhi) bacterial strains by Agar well diffusion
method. Coriandrum sativam shows antibacterial
activity against the above given five bacterial strains. GC-MS was done on Coriandrum sativam oil. These
results support that this plant oil can be used to cure bacterial infections
and may also have role as pharmaceuticals and preservatives.
KEY WORDS: Coriandrum sativam,
GC-MS, Essential oil, Antibacterial activity and Extraction.
INTRODUCTION:
Natural products have been a major source of new drugs
[19].Plants are used medicinally in different countries and are a source of
many potent and powerful drugs. Medicinal plants are used by 80% of the world
population as the only available medicines especially in developing countries
[7].Current research on natural molecules and products primarily focuses on
plants. Since they can be sourced more easily and be selected based on their
ethno-medicinal uses [2]. A wide range of medicinal plants parts is used to
extract as raw drugs and they possess varied medicinal properties while some of
these raw drugs are collected in smaller quantities by the local communities
and folk healers for local used many other raw drugs are collected in larger
quantities and traded in the market as the raw materials for many herbal
industries. Plant used for traditional medicine contains a wide range of
substances that can be used to treat chronic as well as infectious diseases
[18].
Plant oils and extracts have been used for a wide
variety of purposes for many thousands of years [8].These purposes vary from
the use of rosewood and cedar wood in perfumery, to flavouring
drinks with lime, fennel or juniper berry oil [9], and the application of
lemongrass oil for the preservation of
stored food crops [12]. In the antimicrobial activity of plant oils and xtracts has formed the basis of many applications including
raw and processed food preservation pharmaceuticals, alternative medicine and
natural therapies [17,11].
Essential oils of herbs and their components, which are
products from the secondary metabolism of plants have many applications in
ethno-medicine food, flavouring and preservation as
well as in the fragrance and pharmaceuticals industries[5]. The antimicrobial
properties of essential oils have been described [15], and because of the
growing demand on antimicrobials for preventing microbial food spoilage and
bacterial infections, there is an increasing interest in medicinal plants as an
alternative to synthetic preservatives and antibiotics [4]. Many essential oils
are already used in the food industry as flavouring
agents and some are known to exert
antimicrobial activity ,but the mechanism of action is often not entirely
understood. Coriandrum sativam (L.)
is a well known herb widely used as a spice, in folk medicine and in the
pharmacy and food industries [3].Coriander seed oil is one of the major
essential oils in the world market [10], and it is known to exert antimicrobial
activity however, its mechanism of action is still unclear.
MATERIALS AND METHODS:
Selection of Microorganisms:
Totally five human pathogenic bacteria were selected
for the present investigation. Among them, five bacterial strains such as, Staphylococcus aureus.
Enterobacter aerogenes, Klebseilla pnemoniae, Salmonella typhi and Vibrio cholerae . The human
pathogenic bacteria were originally obtained from Microbial Germ Plasm Culture Collection Unit (MGPCCU), Sri Gowri Biotech Research Academy, Thanjavur
and used for present investigation.
Preparation of Microbial
Inoculums:
The young microbial inoculum
culture were prepared and used during the research period. The nutrient broth
(NB) was prepared and poured into several tubes. Then these tubes were
sterilized. The pure microbial cultures were collected from the institute
(either solid or liquid medium) and included in the tubes by using inoculation
loops. After these tubes were incubated
at 37oC for 24-48hrs. After incubation the cultures were used
for the experiments.
Media Preparation:
Composition of Nutrient Agar
Medium:
Peptone - 5gm
Beef extract - 3gm
NaCl - 5
gm
Agar - 15gm
Distilled water - 1000ml
pH - 6.8
Preparation of Nutrient Agar Medium:
The ingredients (peptone– 5g; beef extract – 3g; Nacl
-5g) were weighed and taken in a conical flask contains 1000ml distilled water.
Then pH of the medium was adjusted to 6.8 using a pH meter by the addition of
either acid (or) alkali. The flask were
sterilized in an autoclave at 121ºC for 15 lbs pressure for 15 minutes and
allowed to cool.
Procedure for of oil extraction process:
About 750 g of dried fruits of coriander accurately weighed and
transferred to 1 litre distillation flask (Clevenger
Apparatus) together with 900ml of water .Added few pieces of porcelain to it in
order to avoid bumping during distillation. Distillation tank kept on the
heating mantle and set the distillation assembly. Graduated receiver filled
with water avoiding any air bubbles. The out let near the upper end of the
receiver was not tightened instead, loosely packed with cotton. Heating mantle
was switched on and continued distillation for four hours at a rate which keeps
the lower end of the condenser cool. The distillate allowed being collected in
the graduated receiver in which the aqueous portion of the distillate was
automatically separated and returned to the distillation flask. Measured the
volume of volatile oil which separated out as the upper layer in the graduated
tube and calculated the percentage v/w of isolated oil on a dry weight basis.
The volume of isolated volatile oil from the given sample of Coriander fruits
2.2ml.
Screening for Antibacterial Activity assay:
The antibacterial activities of the Coriander oil were analyzed by agar
- well diffusion method. The Coriander oil was tested against the selected
bacterial strains. The petriplates were washed and
placed in an autoclave for sterilization.
After sterilization, nutrient agar medium was poured into each sterile petriplates and allowed to solidify in a laminar air flow
chamber. After solidification, using a sterile cotton swabs, fresh bacterial
culture with known population count was spread over the plate by spread plate
technique. Then one well of 5mm size made in the agar plates with the help of
sterile cork borer, the wells were loaded with 200µl of oil. All the plates were
incubated at 37°C for 24-48hours. After
incubation, the plates were observed for formation of clear inhibition zone
around the well indicated the presence of antibacterial activity. The zone of inhibition was calculated by
measuring the diameters of the inhibition zone around the well.
Antibiotic sensitivity test on microbes (Positive control)
The antibiotic sensitivity test using standard antibiotics (ampicillin, Kanamycin and
tetracycline for bacteria) were analyzed by following the method of Bauer et
al., 1996.
GC-MS analysis:
The GC-MS analysis was carried out using a clarus
500 perkin –Elmer (Auto system XL).Gas chromatography
equipped and coupled to a mass detector turbo mass Gold –perkin
Elmer turbo mass 5.1 spectrometer with an Elite 1(100% Dimethyl
poly siloxane )
30m x0.25 mm IDX 1mm of capillary
column. The instrument was set to an initial temperature of 110˚C and
maintained at this temperature for 2 minutes. At the end of this period the
over temperature was rose up to 280˚C at the rate of an increase of
5˚C /min and maintained for 9 min. Infection part temperature was ensured
as 250˚C and Helium flow rates as 1ml/min. The ionization voltage was Foev. The samples were infected in split made as 10:1 mass
spectral scan range was set at 45-450(m/2) using computer searchers on a NIST Ver 2.1 Ms data library and comparing the spectrum obtained through GC-MS the compounds
present in the samples were identified.
RESULTS:
Antibacterial activity of Coriandrum
sativam essential
oil:
Antibacterial activities of Coriandrum
sativam oil was analyzed by agar well
diffusion method against human pathogenic bacterial strains such as, Staphylococcus
aureus, Enterobacter aerogenes, Klebsiella pneumoniae, Salmonella typhi and
Vibrio cholerae. Essential
oil extracted from dried fruits of Coriandrum sativam was
analysed for antibacterial activity.
The essential oil of Coriandrum sativam has highest antibacterial activity against the Staphylococcus
aureus (12mm), Enterobacter
aerogenes (12mm), Klebsiella
pneumoniae (10mm), Vibrio
cholerae (13mm) and Salmonella typhi (15mm)
given in table-1 and fig-1.
Antibiotic sensitivity test on bacteria:
The antibiotic sensitivity test using standard antibiotics viz., ampicillin, Kanamycin and
tetracycline were tested against pathogenic bacteria. The results of antibiotic
sensitivity test were presented in table-2.
GC-MS analysis of Coriandrum sativam essential
oil:
The essential oil of Coriandrum sativam was analysed by GC-MS
and 13 components were indentified (Table-3 and Fig-2). The major components in
the essential oil of Coriandrum sativam were
Bicycle(4.1.0),heptanes,3,7,7-trimethyl-(1a,6a,3a), (6.12%) propanoic
acid,2-methyl-3,7-dimethyl octadiennyl
ester,(E)-(6.62%),2- undecenal (7.57%), 2-Napthalenemethanol,
decahydro-a,a,4a-trimethyl-8-methylene- [2R-(2a,4aa,8aa)]- (9.87%).
Table -1: Antibacterial activity of Coriandrum
sativam essential oil
|
S. No.
|
Test
Organisms
(Bacterial
pathogens)
|
Zone of
inhibition (diameter in mm)
|
|
Essential
oil
|
|
1.
|
Enterobacter aerogenes
|
12
|
|
2.
|
Klebsiella pneumoniae,
|
10
|
|
3.
|
Salmonella
typhi.
|
15
|
|
4.
|
Staphylococcus
aureus
|
12
|
|
5.
|
Vibrio cholerae
|
13
|
Table -2: Antibiotic
sensitivity test on bacterial strains
|
S No.
|
Test
Organisms (Bacterial pathogens)
|
Zone of
inhibition (diameter in mm)
|
|
Ampicillin
|
Tetracycline
|
Kanamycin
|
|
1.
|
Enterobacter aerogenes
|
11
|
10
|
9
|
|
2.
|
Klebsiella pneumoniae,
|
8
|
9
|
10
|
|
3.
|
Salmonella
typhi.
|
7
|
8
|
9
|
|
4.
|
Staphylococcus
aureus
|
6
|
10
|
11
|
|
5.
|
Vibrio cholerae
|
9
|
11
|
10
|
Table -3:
GC-MS analysis of Coriandrum
sativam essential oil
Fig -1 Antibacterial activity of Coriandrum
sativam essential oil and standard antibiotics
Fig -2 GC-MS analysis of Coriandrum
sativam essential oil
DISCUSSION:
Microorganisms are the concealed enemies to the mankind. They are
small but cause a very profound damage in human body as well as other living
organism. The agents, which have the capacity to kill the microbes or arrest
the multiplication, are called the antimicrobial agents or drugs. There are a
lot of antimicrobial drugs of which some are discovered or established and some
are hidden in the nature. Hence, the last decade witnessed an increase in the investigations on plants
as a source of human disease management[1,16,13,20] and more natural
antimicrobials have driven scientist to investigate the effectiveness of
inhibitory compounds such as extracts from plants[14].There are several reports
for antibiotics resistance of human pathogens to available antibiotics [6,21]. Bimolecules of plant origin appear to be one of the
alternatives for the control of these antibiotic resistant human pathogens.
This study deals with five pathogenic bacterial strains in which four
bacteria Enterobacter aerogenes,
Klebsiella pneumoniae, Vibrio cholerae and Salmonella typhi are gram negative bacteria while Staphylococcus aureus
is gram positive bacteria. In the present work antibacterial activity of Coriandrum sativam essential
oil was found against five human pathogenic bacteria. All the five human
pathogenic bacterial strains were sensitive to essential oil of Coriandrum sativam. Coriandrum sativam essential
oil show highest inhibitory activity against
Salmonella typhi. The antibiotic
sensitivity test using standard antibiotics ampicillin,
Kanamycin and tetracycline were tested against
bacteria.
ACKNOWLEDGEMENT:
The authors are grateful to Sri
Gowri Biotech Research Academy, Thanjavur
(Dt),Tamil Nadu. For their permission to utilized the
laboratory facility.
REFERENCES:
1. Aiyclagabe OO.
Antibacterial activity of Jatropa multifida roots. Fitoterapia.
72; 2001: 544-546.
2. Arora DS, Kaur GJ. Antibacterial activity of some Indian medicinal
plant. Journal of Natural Medicine. 61; 2007: 313-317.
3. Bauer AW, Kirby WM, Sherris
JC. Antibiotic susceptibility testing by a standard single disc method. Am.
J. Clin. Pathol. 451;
1996: 493- 496.
4. Burdock GA and Carabin
IG. Safety assessment of Coriandrum sativum L. essential oil as a food ingredient. Food ChemToxicol. 47; 2009: 22-34.
5. Edris AE.
Pharmaceutical and therapeutic potentials of essential oils and their
individual volatile constituents. A Review Phytother.
21; 2007: 308-323.
6. Fabian D. Sabol
M., Domaracka K and Bujnakva
D. Essential oils –their antimicrobial activity against E.coli
and effect on intestinal cell viability. Toxicol.
20; 2006: 1435-1445.
7. Ganguly R., Mishra P., Sharma A. Microbes and infection. Indian
Journal of Microbiology. 41; 2001: 211-213.
8. Hashim H., Kamali EL, Mohamed Y. Antibacterial activity and phytochemical screening of ethanolic extracts obtained from
selected Sudanese medicinal plants. Current Research journal of Biological
Science. 2(2); 2010: 143-146.
9. Jones FA. Herbs–useful plants their role
in history and today. European Journal of
Gastroenterology and Hepatology. 8; 1996: 1227-1231.
10. Lawless J. The Illustrated Encyclopedia of
Essential oils, Shafttesbuy UK. Element Books Ltd.
1995
11. Lawrence BM, A planning scheme to evaluate
new aromatic plants for the flavour and fragrance
industries. In New Crops. Edited by Janick J
and Simon JE, New Yolk: wiley. 1993
12. Lis–Balchin M and
Deans SG. Bioactivity of selected plant essential oils against Listeria monocytogenes.
Journal of Applied Bacteriology. 82; 1997: 759-762.
13. Mishra AK and Dubey NK. Evaluation of some essential oils for their
toxicity against fungi causing deterioration of stored food commodities. Applied
and Environmental Microbiology. 60; 1994: 1101-1105.
14. Mounishwamy V., Davimans S., Gunasekaran R.
Antibacterial activity of Gossyptein isolated from Hibiscus
sabdariffa. The Antiseptic. 99; 2002:
81-82.
15. Nasar-Abbas SM, Halkman AK. Antimicrobial effect of water extract of sumac
(Rhuscoriaria L.) on the growth of some food borne
bacteria including pathogens. International Journal of Food Microbiology.
97; 2004: 63-69.
16. Pinto E., Pinavaz
C., Salgueiro L., Goncalves
M.J., Costadeoliveira S., Calvaleiro
C., Palmeira A., Rodrigues
A. and Martine 2-de-oliveira J. Antifungal activity of the essential oil of Thymus
pulegiodes on Candida, Aspergillus
and Dermatophyte species, J.Med.Microbiol.
55; 2006: 1367-1373.
17. Prashanth D., Asha MK, Amt. A. Antibacterial activity of Punica granatum. Fitoterappia. 72; 2001: 171-173.
18. Reynolds JEF. Martindale-the Extra
pharmacopoeia 31st edn London Royal
Pharmaceutical society of Great Britain. 1996
19. Uniyal SK, Sing
KN, Jamwal P, Lal B.
Traditional use of medicinal plants among the tribal communities of Chhota Bhangal. Western
Himalayan Journal of Ethonobiology and Ethnomedicine. 2; 2006: 1-14.
20. Vuorelaa P., Leinonenb M., Saikkuc P., Tammelaa P., Rauhad JP, Wennberge T., Vuorela H. Natural
products in the process of finding new drug candidates, Current Medicinal
Chemistry. 11; 2004: 1375-89.
21. Woldemichael GM, Watchter G., Sing MP. Antibacterial diterpenes
from Calceolaria pinifolia. Journal of
Natural Products. 66; 2003: 242-246.