GC-MS analysis of ethanolic plant extract of Trichopus Zeylanicus ssp. Travancoricus Burkill Ex K. Narayanan

Abstract

The aim of the present study is to analyze the bioactive phytochemical constituents present in the plant of Trichopus zeylanicus ssp. travancoricus Burkill ex K. Narayanan by using GC-MS. Methods: The dried and coarsely powdered plant materials were extracted using solvent ethanol for 5 hours by soxhlet extraction method. The presence of various phytochemical constituent was analyzed by GC-MS. Results: Totally 22 chemical constituents were identified. Among these twenty-two compounds detected from ethanolic extract,2-methyl-3-oxy-gamma-pyrone (1.87%), 2-methyl-3-hydroxypyrone (2.77%), Pyranone (4.88%), Coumaran (12.74%), Vanillin (1.24%), Isovanillicacid (1.96%), Chinasaure (Quinic acid) (9.15%), alpha-santonin (4.98%),digiprolactone (1.46%), Ascorbic acid 2,6-dihexadecanoate (0.43%), and phenolic derivates were identified as therapeutically active components. Conclusion: The obtained phytochemical constituents were identified as potentially active and can be used for the treatment of various diseases. Constituents supporting nutritive value were also identified.

Full text

~ 983 ~
Journal of Pharmacognosy and Phytochemistry 2020; 9(4): 983-986
E-ISSN: 2278-4136
P-ISSN: 2349-8234
www.phytojournal.com
JPP 2020; 9(4): 983-986
Received: 18-05-2020
Accepted: 20-06-2020
Athira Anilan
Department of Pharmacognosy
and Phytochemistry Research
Laboratory, Nehru College of
Pharmacy, Pampady,
Thiruvilwamala, Thrissur,
Kerala, India
Dr. Ragunathan Muthuswamy
Department of Pharmacognosy
and Phytochemistry Research
Laboratory, Nehru College of
Pharmacy, Pampady,
Thiruvilwamala, Thrissur,
Kerala, India
Corresponding Author:
Dr. Ragunathan Muthuswamy
Department of Pharmacognosy
and Phytochemistry Research
Laboratory, Nehru College of
Pharmacy, Pampady,
Thiruvilwamala, Thrissur,
Kerala, India
GC-MS analysis of ethanolic plant extract of
Trichopus Zeylanicus ssp. Travancoricus Burkill
Ex K. Narayanan
Athira Anilan and Dr. Ragunathan Muthuswamy
Abstract
The aim of the present study is to analyze the bioactive phytochemical constituents present in the plant of
Trichopus zeylanicus ssp. travancoricus Burkill ex K. Narayanan by using GC-MS.
Methods: The dried and coarsely powdered plant materials were extracted using solvent ethanol for 5
hours by soxhlet extraction method. The presence of various phytochemical constituent was analyzed by
GC-MS.
Results: Totally 22 chemical constituents were identified. Among these twenty-two compounds detected
from ethanolic extract,2-methyl-3-oxy-gamma-pyrone (1.87%), 2-methyl-3-hydroxypyrone (2.77%),
Pyranone (4.88%), Coumaran (12.74%), Vanillin (1.24%), Isovanillicacid (1.96%), Chinasaure (Quinic
acid) (9.15%), alpha-santonin (4.98%),digiprolactone (1.46%), Ascorbic acid 2,6-dihexadecanoate
(0.43%), and phenolic derivates were identified as therapeutically active components.
Conclusion: The obtained phytochemical constituents were identified as potentially active and can be
used for the treatment of various diseases. Constituents supporting nutritive value were also identified.
Keywords: Arogyapacha, coumaran, kani tribe, quinic acid
Introduction
Trichopus zeylanicus ssp. travancoricus Burkill ex K. Narayanan is a herb belonging to the
family Trichopodaceae. The plant is locally known as ‘Arogyapacha’ in Malayalam, ’Arogya’
means health and ‘ Pacha’ means green and hence the herb is literally called as green that
gives health. Three subspecies of Trichopus zeylanicus are identified and among these,
Trichopus zeylanicus ssp. travancoricus have reported as endemic to the Southern Western
Ghats in India with a restricted distribution in Agasthyamalai Biosphere Reserve. The’ Kani
tribe’ of Agasthyamalai has introduced various uses of this wild plant to the present medical
world. They also claim that one who consumes the fruits of Arogyapacha regularly, will
remains healthy, agile and disease resistant [1]. The powdered leaves of T. zeylanicus ssp.
travancoricus along with stem bark of Mangifera indica was used to treat venereal diseases [2].
The indigenous tribal community in Agastya hills traditionally uses this plant as an instant
energy booster that combat fatigue [3]. Apart from anti fatigue property, medicinal properties
such as anti-oxidant, anti-inflammatory, aphrodisiac [4], hepatoprotective [5], mast cell
stabilization [6], anti-hyperlipidemic, adaptogenic [7], cardioprotective [8], and anti-ulcer activity
[9] were also reported to this endemic herb. Literature survey revealed that various therapeutic
properties of Trichopus zeylanicus including anti-oxidant, anti-stress, anti-
microbial, ͕aphrodisiac, analgesic, anti-inflammatory etc., have been demonstrated using
various plant extracts both in in vitro and in vivo studies [3]. This study focuses to analyze the
constituents responsible for contributing various pharmacological activities and nutritive value
to the plant. The results of pharmacological activities and nutritive value of this plant
accentuates a point of view that helps in future research to explore its full pharmacological
potential and nutritive status. The present study focused to analyze the bioactive chemical
constituents present in the ethanolic extract of whole plant material of Trichopus zeylanicus.
Fig 1: Trichopus zeylanicus ssp. travancoricus

~ 984 ~
Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
Materials and Methods
1. Plant material collection and authentification
The plant Trichopus zeylanicus ssp. travancoricus( Fig.1.)
were collected from Rayirath Gardens, Pattikkad, Thrissur,
Kerala, India. The specimens collected from nursery were
identified with the standard literature and authenticated with
valid voucher specimens. The plant materials were
taxonomically identified by the Botanist, Dr. Udayan P.S,
Assistant professor and Head of P G Department of Botany
and Research, Sree Krishna College Guruvayoor. The
specimen of T. zeylanicus ssp. travancoricus voucher number
85 was submitted in the Herbarium.
2. Extraction
The whole plant material was shade dried, coarsely powdered
and subjected to soxhlet extraction by using solvent ethanol
for 5 hours. The extract was filtered through Whatman No. 1
filter paper and concentrated. The extract obtained was then
subjected to GC-MS analysis.
3. GC -MS Analysis
Gas chromatography Mass spectroscopy analysis of ethanolic
extract was performed using Shimadzu GC-MS Model
number: QP2010S equipped with Column - ELITE-5MS (30
meter length, 0.25 mm ID, and 0.25 µm thicknesses). Electron
ionization system was used; details of GC programme were
given in Table I. The oven temperature was programmed from
70.000C which is given in Table II. Helium gas was used as
the carrier gas. Details of GC-MS programme was given in
Table III. Programme specifications regarding Mass Spectra
were depicted in Table IV. GCMS Software: GCMS
Solutions, Libraries used: NIST 11& WILEY 8.
Table 1: GC programme (GC 2010)
GC-Parameters
Programme
Column oven temperature
70.000C
Injection temperature
260.000C
Injection mode
Split less
Sampling time
2.00 min
Flow control mode
Linear velocity
Pressure
61.5 kPa
Total flow
54.1mL/min
Column flow
1.00mL/min
Linear velocity
36.7 cm/sec
Purge flow
3.0mL/min
Split ratio
50.0
Table 2: Oven temperature programme
Rate
Temperature(0c)
Hold time(min)
-
70.0
2.00
10.00
200.0
5.00
5.00
280.0
15.00
Table 3: GC-MS programme (GCMS QP2010)
GC-MS Parameters
Programme
Ion source temperature
200.000c
Interface temperature
280.000c
Solvent cut time
6.50 min
Detector gain mode
Relative
Detector gain
1.01 kV+0.00 kV
Threshold
1000
Table IV: MS table
Mass spectroscopy parameters
Programme
Start time
6-7 min
End time
51.00 min
ACQ time
Scan
Event time
0.50 sec
Scan speed
1000
Start m/z
50.00
End m/z
500.00
Sample inlet unit
GC
4. Identification of compounds
The constituents in the extract were identified by comparing
their relative retention time and confirmation was done by
comparing the mass spectra with database from the Library of
NIST 11 and Wiley8. GC-MS Chromatogram obtained was
given in figure.2.
Results and Discussion
GC-MS analysis of ethanolic extract of Trichopus zeylanicus
were carried out and a group of 22 compounds were identified
which are depicted in Table V. This includes several phenolic
derivatives and other biologically and pharmacologically
active compounds. The compounds identified are -2-methyl-
3-oxy-gamma-pyrone (1.87%), 2-hydroxyacetylfuran
(4.37%), Erythrol (3.14%), 2-ethyl crotonaldehyde (9.16%),
2-methyl-3-hydroxy pyrone (2.77%), Glycerin (6.21%),
Pyranone (4.88%) 1, 2-benzenediol (3.21%), Coumaran
(12.74%), 5-Hydroxymethylfurfural (8.53%), 4-vinylguaiacol
(12.99% ),Phenol, 2, 6-dimethoxy- (2.95%), Vanillin (1.24%),
2,6-cresotaldehyde (3.83%), 4-tert-Butoxybenzoic Acid
(1.63%), Isovanillic acid (1.96%), -acetyl guaiacol (0.55%),
Chinasaure (9.15%), alpha-santonin (4.98%), gamma
hydroxyisoeugenol (1.96%), Digiprolactone (1.46%) -
Ascorbic acid 2, 6-dihexadecanoate (0.43%). Each of these
constituents is responsible for various pharmacological and
biological activities. Of these 22 compounds, phenolic
derivatives (4- vinylguaiacol 12.99%, phenol 2, 6 dimethoxy
2.95%, and acetyl guaiacol 0.55%) were found as the first
major compounds. Phenolic derivatives are well known for
their antioxidant activities. Coumaran (12.74%) were found as
the second major compound [10]. Coumarans (2, 3-
dihydrobenzofurans) have antitubercular, anti-HIV,
antioxidant, anti stress activity [11]. Chinasaure (Quinic acid
9.15%) were found to be third major therapeutically important
compound. Study suggested that the pharmacokinetic
properties of Quinic acid were more preferable to be used as a
potent drug candidate to combat prostate cancer [12]. Quinic
acid nutritionally supports the synthesis of tryptophan and
nicotamide in GI tract that in turn leads to the DNA repair
enhancement and NF-kB inhibition via increased
nicotinamide and tryptophan production. It also promotes
insulin secretion from pancreatic beta cells [13]. Now a days
pyrone and its derivatives are used for the treatment of an

~ 985 ~
Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
Anticancer, Antibiotics, Anticoagulants, and HIVprptease etc.
[14] Isovanillin posses antidiarrheal and anti motility effect on
GI tract [15]. Alpha santonin are sesquiterpene lactones and are
pharmacologically proved as Antipyretic, Anthelmintic,
Anticancer, and Analgesic.
Fig 2: GC-MS chromatogram of ethanolic extract of Trichopus zeylanicus ssp. travancoricus
Table 5: GC-MS analysis of ethanolic extract of Trichopus zeylanicus ssp. travancoricus
Peak#
R. Time
Area
Area %
Height
Height%
Name
Base m/z
1
6.933
1027896
1.87
110783
0.90
2-METHYL-3-OXY-GAMMA-PYRONE
126.10
2
7.083
2407378
4.37
235712
1.91
2-HYDROXYACETYLFURAN
95.00
3
7.268
1726530
3.14
458117
3.71
ERYTHROL
57.00
4
7.333
5039753
9.16
421784
3.41
2-ETHYL CROTONALDEHYDE
61.05
5
7.533
1523675
2.77
532583
4.31
2-METHYL-3-HYDROXYPYRONE
126.10
6
7.625
3420193
6.21
558796
4.52
Glycerin
61.05
7
8.006
2686098
4.88
913469
7.39
Pyranone
144.05
8
8.678
1764178
3.21
353294
2.86
1,2-BENZENEDIOL
110.05
9
8.923
7013530
12.74
1089506
8.81
Coumaran
120.10
10
9.174
4697227
8.53
1474539
11.93
5-Hydroxymethylfurfural
97.05
11
10.346
7148565
12.99
1879333
15.20
4-VINYLGUAIACOL
150.10
12
10.862
1621732
2.95
715220
5.78
PHENOL, 2,6-DIMETHOXY-
154.10
13
11.547
681135
1.24
256415
2.07
Vanillin
152.10
14
11.999
2108978
3.83
634986
5.14
2,6-CRESOTALDEHYDE
136.10
15
12.743
895719
1.63
118388
0.96
4-tert-Butoxybenzoic Acid
57.05
16
13.649
1077092
1.96
350559
2.84
Isovanillic acid
168.10
17
13.886
300568
0.55
171595
1.39
_ACETYLGUAIACOL
151.05
18
14.675
5036008
9.15
502602
4.07
CHINASAURE
60.05
19
14.738
2742771
4.98
894566
7.24
(-)-.ALPHA.-SANTONIN
182.10
20
15.695
1080247
1.96
414287
3.35
gamma.-Hydroxyisoeugenol
137.10
21
16.203
801232
1.46
199859
1.62
DIGIPROLACTONE
111.10
22
18.930
236629
0.43
77665
0.63
l-(+)-Ascorbic acid 2,6-dihexadecanoate
73.05
55037134
100.00
12364058
100.00
Conclusion
In the present study, twenty two chemical constituents have
been identified from the ethanolic extract of Trichopus.
Zeylanicus ssp. travancoricus. The obtained bioactive
compounds were identified as potentially active and can be
used for the treatment of various diseases. Constituents
supporting nutritive status were also identified. The presence
of various phytoconstituents helps in future research to
explore full pharmacological potential and nutritive value of
the plant.
Acknowledgement
We thank Department of Pharmacognosy and
Phytochemistry, Nehru college of pharmacy, Thiruvilwamala,
Thrissur, Kerala and Kerala Forest Research Institute, Peechi,
Kerala for helping us to carry out this research.
Conflict of Interest
The authors have no conflict of interest.
Abbreviation Used
GC-MS: Gas Chromatography- Mass Spectroscopy; Rt:
Retention time; NIST 11; National Institute Of Standard and
Technology 11.
References
1. Pushpangadan P, Rajasekharan S, Rathesh Kumar PK,
Jawahar CR, Nair VV, Lakshmi N et al. Arogyapacha
(Trichopus zeylanicus Gaertn.) The ginseng of Kani

~ 986 ~
Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
tribes of Agashtyar Hills (Kerala) for Ever Green.
Ancient Sci Life. 1988; 8(1):13-16.
2. Ayyanar M, Ignacimuthu S. Ethnobotanical survey of
medicinal plants commonly used by Kani tribals in
Tirunelveli hills of Western Ghats, India. J
Ethnopharmacology. 2011; 134(3):851-64.
3. Biju VC, Shidhi PR, Rajan VS, Anoop PK, Nair AS.
Therapeutic properties of Trichopus zeylanicus subsp.
travancoricus, a Rare, Endangered Medicinal Plant in
South India. A Review. Herb Med. 2019; (5):1-2.
4. Subramoniam A, Madhavachandran V, Rajasekharan S,
Pushpangadan P. Aphrodisiac property of Trichopus
zeylanicus extract in male mice. J. Ethnopharmacology.
1997; 57(1):21-7.
5. Subramoniam A, Evans DA, Rajasekharan S,
Pushpangadan P. Hepatoprotective activity of Trichopus
zeylanicus extract against paracetamol-induced hepatic
damage in rats. Indian. J Experimental Biol. 1998;
36(4):385-9.
6. Subramoniam A, Evans DA, Rajasekharan S,
Pushpangadan P. Inhibition of antigen-induced
degranulation of sensitized mast cells by Trichopus
zeylanicus in mice and rats. J Ethnopharm. 1999;
68(1):137-43.
7. Singh B, Chandan BK, Sharma N, Singh S, Khajuria A,
Gupta DK. Adaptogenic activity of glyco-peptido-lipid
fraction from the alcoholic extract of Trichopus
zeylanicus Gaerten. (Part II). Phytomedicine. 2005;
12(6):468-81.
8. Velavan S, Selvarani S, Adhithan A. Cardio protective
effect of Trichopus zeylanicus against myocardial
ischemia induced by isoproterenol in rats. Bangladesh J
Pharm. 2009; 4(2):88-91.
9. Rishikesh RS, Kumar SR, Ravindranath SB, Vaibhav
BV. Anti-ulcer potential of saponin fraction of Trichopus
zeylanicus on various experimental animal models. Inter
J Green Pharm. 2017; 11(1):11-6.
10. E.Ilya L, Kulikova EV. Van der Eycken L.
Voskressensky, Recent Advances in Phthalan and
Coumaran Chemistry. Chemistry Open. 2018; 7(11):914-
929.
11. Inbathamizh L, Padmini E. Quinic acid as a potent drug
candidate for prostate cancer. A comparative
pharmacokinetic approach. Asian journal of
pharmaceutical and clinical Research. 2013; 6(4):106-
112.
12. Pero RW, Lund H, Lenderson T. Antioxidant metabolism
induced by Quinic acid increased urinary excretion of
tryptophan and nicotinamide. Phytother Res. 2009;
23(3):335-346.
13. Janet Beula S, Dr. Suthakaran R, Rajaram Das, Laxmidar
Sahou, Muthu Mohammed J. A review on Pyrones,
Pyrone derivatives and their biological activity. World
Journal of Pharmacy and Pharmaceutical sciences. 2019;
8(2):442-450.
14. Sadraei H, Ghanadian M, Asghari G, Azali N.
Antidiarrheal activities of isovanillin, iso-acetovanillin
and Pycnocycla spinosa Decne ex.Boiss extract in mice.
Res Pharm Sci. 2014; 9(2):83-89.
15. Al-Harbi MM, Qureshi S, Ahmad MM, Raza M, Miana
GA, Shah AH. Studies on the anti-inflammatory,
antipyretic and analgesic activities of santonin. Japanies
Journal of Pharmacology. 1994; 64(30):135-139.