ISSN 0974-3618
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0974-360X (Online)
RESEARCH ARTICLE
GC-MS/MS Study of Parthenium hysterophorus L(N.
Am) stem, Antimicrobial activity
M. Krishnaveni1 *, M. Kalaivani2, C. Ragina Banu2, G.
Krishna Kumari2
1Assistant
Professor, Department of Biochemistry, School of Bio-Sciences, Periyar
University, Salem
2M.Phil Students,
Department of Biochemistry, School of Bio-Sciences, Periyar University,
Salem
*Corresponding Author E-mail: logan.consolidated@gmail.com
ABSTRACT:
15 species of Parthenium are available under this genus. Parthenium hysterophorus belong to the family of Asteracea. Since, plant forms a fundamental raw material
for the development of therapeutic drug,
the phytocomponents of plants are vital tool for the treatment and cure. So,
the present work was decided to study the components in ethanol extract of Parthenium hysterophorus stem through GC-MS/MS and its antimicrobial
activity. The results of GC-MS/MS showed highest peak for Aciphyllene,
Stigmasteryltosylate, Benzophenone, Stigmasterol, 4-Nitrophenyl laurate.
Compounds such as Azulene,
1,2,3,5,6,7,8,8a-octahydro-1,4-dimethyl-7-(1-methylethenyl)-,
[1S-(1α,7α,8aβ)]-, Bicyclo[3.1.0]hexan-2-one,
1,5-bis(1,1-dimethylethyl)-3,3-dimethyl-,
1,2-Benzenediol, mono(methylcarbamate), 2,5-Dihydroxyacetophenone,
bis(trimethylsilyl) ether, 12-Tridecynoic acid, methyl ester, Phenol,
3,4-dimethoxy-, 4-Acetoxy-3-methoxystyrene, Benzoic acid,
4-methyl-2-trimethylsilyloxy-, trimethylsilyl ester, 12-Tridecynoic acid, methyl
ester, Tetradecanoic acid shows moderate peak area percent. The results of
antimicrobial activity shows that antimicrobial activity was observed with Escherichia coli, Aspergillus
niger, Candida albicans while, Staphylococcus aureus showed no zone
of inhibition.
KEY WORDS: Antibacterial, Antifungal, Parthenium, Phytocontituents,
GC-MS/MS.
INTRODUCTION:
Parthenium hysterophorus L is a herbaceous annual
plant, grows erect up to 2m height, have branched stem with trichomes. It forms
a good waste for composting. The stem contains fumeric acid, rich in histamine,
volatile oils, saponin, terpenoids, glycosides, amino acids, phenolic
derivatives, flavonoids etc. Therapeutic potential of plant includes
insecticidal, nematicidal, herbicidal and are natures gift to Parthenium hysterophorus. The
antimicrobial agents are available in both natural as well as in synthetic
form. The synthetic antimicrobials are not safe for regular use. So, it is
better to find an alternate source to kill the microbes. In order to find its
antimicrobial potency, the plant Parthenium
hysterophorus stem was screened for its phtocontituents trough
GC-MS/MS.
MATERIALS AND METHODS:
Sample
collection
Fresh
Parthenium hysterophorus stem was collected, shade dried, and powdered. 25grams of
powdered leaf sample was used for ethanol extraction. The extracted sample was
used for phytochemical analysis through GC-MS/MS, anti-microbial activity. The
plant was authenticated by Dr. A. Balasubramanian. The authentication number was AUT/PUS/069
dated 17/12/2014.
Analytical method
GC-MS/MS
was performed on a Scion 436-GC Bruker carrying Triple quadruple mass
spectrophotometer with fused silica capillary column BR-5MS (5% Diphenyl/ 95%
Dimethyl poly siloxane), 30m x 0.25mm ID x 0.25m df. The column oven temperature program was as
follows: 80°C hold for 2 min, Up to 160°C at the rate of 20°C/min-No hold, Up
to 280°C at the rate of 5°C / min-No hold, Up to 300°C at the rate of
20°C/min-10 min hold, Injector temperature 280°C, Total GC running time was 41
min. The inlet temperature was set at 280°C, source temperature 250°C;
ionization mode, ionization at 70-eV ionization energy; For single scan
analysis, the scan range was set from m/z 40 to 600; Solvent Delay: 0-3.5 min;
and the injection volume was 2μl.The GC-MS/MS was performed by Institute
of crop processing technology, Tanjavur.
Antimicrobial
assay
The
antimicrobial activity was assessed by means of Kirby- Bauer technique.1
RESULTS
AND DISCUSSION:
Table.1. Shows
the results of phytochemicals
analysed through GC-MS/MS.
The
results of components identified in ethanol extract of Parthenium
hysterophorus stem was shown in
Table.1. Totally 14 compounds were identified in Parthenium
hysterophorus stem. The compounds
identified are as follows: 1,2-Benzenediol, mono(methylcarbamate),
4-Acetoxy-3-methoxystyrene, Phenol, 3,4-dimethoxy-, Aciphyllene, Bicyclo[3.1.0]hexan-2-one,
1,5-bis(1,1-dimethylethyl)-3,3-dimethyl-, Benzophenone, Benzoic acid,
4-methyl-2-trimethylsilyloxy-, trimethylsilyl ester, 2,5-Dihydroxyacetophenone,
bis(trimethylsilyl) ether, Tetradecanoic acid, 12-Tridecynoic acid, methyl
ester, Stigmasteryl tosylate, Azulene,
1,2,3,5,6,7,8,8a-octahydro-1,4-dimethyl-7-(1-methylethenyl)-,
[1S-(1α,7α,8aβ)]-, Stigmasterol, 4-Nitrophenyl laurate. The
molecular formula and molecular weight are shown below: Compound 1: C8H9NO3,
167. Compound 2: C11H12O3, 192. Compound 3: C8H10O3,
154. Compound 4: C15H24, 204. Compound 5: C16H28O,
236. Compound 6: C13H10O, 182. Compound 7, 8: C14H24O3Si2,
296. Compound 9: C14H28O2, 228. Compound
10: C14H24O2, 224. Compound 11: C36H54O3S,
566. Compound 12: C15H24, 204. Compound 13: C29H48O,
412. Compound 14: C18H27NO4, 321. The peak
area percent of the identified compounds are: 4.16 for 1,2-Benzenediol, mono
(methylcarbamate), 2.85 for 4-Acetoxy-3-methoxystyrene, 3.66 for Phenol,
3,4-dimethoxy-, 17.60 for Aciphyllene, 4.62 for
Bicyclo[3.1.0]hexan-2-one, 1,5-bis(1,1-dimethylethyl)-3,3-dimethyl-,
12.86 for Benzophenone, 2.78 for Benzoic acid,
4-methyl-2-trimethylsilyloxy-, trimethylsilyl ester, 3.76 for 2,5-Dihydroxyacetophenone,
bis(trimethylsilyl) ether, 1.57 for Tetradecanoic acid, 3.69 for 12-Tridecynoic acid, methyl ester,
15.04 for Stigmasteryl tosylate, 6.61
for Azulene, 1,2,3,5,6,7,8,8a-octahydro-1,4-dimethyl-7-(1-methylethenyl)-,
[1S-(1α,7α,8aβ)]-, 11.45 for Stigmasterol, 9.35 for
4-Nitrophenyl laurate. The plant Parthenium hysterophorus was studied
for its air pollution tolerance index, antioxidant activities,2
phytochemicals,3 GC-MS/MS4
by Krishnaveni et.al.
Table.1 Components
identified in ethanol extract of Parthenium
hysterophorus stem.
|
S.No
|
RT
|
Name of the compound
|
Molecular Formulae
|
MW
|
Peak area %
|
|
1.
|
5.92
|
1,2-Benzenediol,
mono(methylcarbamate)
|
C8H9NO3
|
167
|
4.16
|
|
2.
|
6.88
|
4-Acetoxy-3-methoxystyrene
|
C11H12O3
|
192
|
2.85
|
|
3.
|
7.19
|
Phenol, 3,4-dimethoxy-
|
C8H10O3
|
154
|
3.66
|
|
4.
|
7.86
|
Aciphyllene
|
C15H24
|
204
|
17.60
|
|
5.
|
9.51
|
Bicyclo[3.1.0]hexan-2-one,
1,5-bis(1,1-dimethylethyl)-3,3-dimethyl-
|
C16H28O
|
236
|
4.62
|
|
6.
|
10.04
|
Benzophenone
|
C13H10O
|
182
|
12.86
|
|
7.
|
11.14
|
Benzoic acid, 4-methyl-2-trimethylsilyloxy-,
trimethylsilyl ester
|
C14H24O3Si2
|
296
|
2.78
|
|
8.
|
13.74
|
2,5-Dihydroxyacetophenone,
bis(trimethylsilyl) ether
|
C14H24O3Si2
|
296
|
3.76
|
|
9.
|
12.64
|
Tetradecanoic acid
|
C14H28O2
|
228
|
1.57
|
|
10.
|
14.97
|
12-Tridecynoic acid, methyl
ester
|
C14H24O2
|
224
|
3.69
|
|
11.
|
31.70
|
Stigmasteryl tosylate
|
C36H54O3S
|
566
|
15.04
|
|
12.
|
32.16
|
Azulene,
1,2,3,5,6,7,8,8a-octahydro-1,4-dimethyl-7-(1-methylethenyl)-,
[1S-(1α,7α,8aβ)]-
|
C15H24
|
204
|
6.61
|
|
13.
|
34.29
|
Stigmasterol
|
C29H48O
|
412
|
11.45
|
|
14.
|
36.13
|
4-Nitrophenyl laurate
|
C18H27NO4
|
321
|
9.35
|
Fig.1.
shows the chromatogram result of Partheium hysterophorus stem.
Antimicrobial assay
Table.2 Antimicrobial activity of ethanol extract of
Parthenium hysterophorus stem
|
Plant
used
|
Microbes
tested
|
Zone of inhibition
(mm)
|
|
Parthenium
hysterophorus
stem
|
Escherichia
coli
Staphylococcus
aureus
Aspergillus
niger
Candida
albicans
|
10
-
9
12
|
The antimicrobial
activity was tested for the bacteria and fungi selected. The bacteria selected was Escherichia
coli, Staphylococcus aureus and the fungi selected was Aspergillus
niger, Candida albicans. Among the bacteria and fungi selected, no activity
was observed for Staphylococcus
aureus. While, the zone of inhibition observed was 10mm for Escherichia
coli, 9mm for Aspergillus niger, 12mm for Candida albicans.
The ethanol extract of Partheium hysterophorus stem was found to be
active against Candida albicans, Escherichia coli. The difference
in the zone of inhibition might be due to the diffusing potency of the extract
against the particular organism.
CONCLUSION:
Knowledge
on plants and phytochemicals contained in it forms an essential study to
develop a new avenue in the field of
research. The results of GC-MS/MS study shows Aciphyllene, Stigmasteryl
tosylate, Benzophenone. The antimicrobial assay showed highest activity
for Candida albicans, Escherichia coli. Since, Benzophenone
is rich, isolation of this particular compound could be done and critically
evaluated for its sun screen potential in order to find a suitable
beautification agent from plant based source.
ACKNOWLEDGEMENT:
The
author wishes her thanks to Honorable Vice-chancellor Dr. C. Swaminathan Avl,
and Registrar Dr. K. Angamuthu Avl, Periyar University, Salem for their
administrative support and excellent infrastructure facilities provided and
also Co-ordinator, School of Bio-Sciences, Periyar University, Salem, Dr. A.
Balasubramanian, ABS Botanical garden, Salem for his help in authenticating
plants, Director Dr. K. Alagusundaram and Dr. S. Kumaravel, senior
scientist, Indian Institute of Crop
Processing Technology, Thanjavur for analyzing the samples. The author would
like express her gratitude to her dedicated teachers.
REFERENCES:
1. Bauer AW, Perry DM, Kirby WMM, Single
disc antibiotic sensitivity testing Staphylococci, AMA Arch Intern Med, 104,
1959, 208–216.
2. M.Krishnaveni. Air pollution tolerance
index and antioxidant activity of Parthenium hysterophorus,
Journal of Pharmacy Research, 2013; 7(4): 296-298,
3. Krishnaveni M, Dhanalakshmi R, Phytochemical analysis of Parthenium
hysterophorus L. leaf, World Journal of Pharmaceutical Research, 2014;
3(6):1066-1074.
4. Krishnaveni M, Kalaivani M, Krishna
kumari G, Ragina Banu C, GC-MS/MS analysis of
phytochemicals in Parthenium hysterophorus L (N.AM),
antimicrobial assay, World Journal of
Pharmacy and Pharmaceutical Sciences, 2015; 4(4): 1604-1608. .