World Journal of Pharmaceutical Research

Vignesh et al.

World Journal of Pharmaceutical

Research
SJIF Impact
Factor 6.805

Volume 5, Issue 10, 499-517.

Research Article

ISSN 2277 7105

ANALYSIS OF PHYTOCHEMICAL CONSTITUENTS AND

ANTIMICROBIAL PROPERTIES OF ESSENTIAL OIL EXTRACTED
FROM THE LEAVES OF TRICHOPUS ZEYLANICUS SSP.
TRAVANCORICUS BURKILL EX K. NARAYANAN
1

*Vignesh Kumar Balasubramanian, 2 Ramasubbu Raju and 3 Sasikala N.

*Department of Biology, Gandhigram Rural Institute- Deemed University, Dindigul, Tamil

Nadu, India.
2

Assistant Professor, Department of Biology, Gandhigram Rural Institute-Deemed

University, Dindigul, Tamil Nadu, India.

Research Scholar, Department of Biology, Gandhigram Rural Institute- Deemed University,

Dindigul, Tamil Nadu, India.
ABSTRACT

Article Received on
26 July 2016,

Background:

Revised on 16 August 2016,

Accepted on 06 Sep. 2016

antimicrobial properties of essential oil extracted from the leaves of

DOI: 10.20959/wjpr201610-6978

Trichopus zeylanicus ssp. is clearly justified that this plant have a

Analysis

of

phytochemical

constituents

and

potentiality. Trichopus zeylanicus ssp. travancoricus Burkill ex K.

Narayanan is an endemic perennial and rhizomatous herb belongs to
*Corresponding Author
Vignesh Kumar

the

family

Dioscoreaceae growing in selected

forest areas of

Balasubramanian

Agasthyamalai, southern Western Ghats. The plant is locally known as

Department of Biology,

Arogyappachai and the herbal is normally used by the Kani tribe for

Gandhigram Rural Institute-

their good health and vitality. The herb is known to have unique

Deemed University,

medicinal properties and efficacy of this plant help as rejuvenating, age

Dindigul, Tamil Nadu,

India.

stabilizing, disease resistance building etc. The un-ripened fruits of

Trichopus are highly rejuvenates and are used by the Kani tribe to

ameliorate fatigue. The seeds are rich in saponins and the preparations from the leaves, stem
and seeds are used as a health tonic. Materials and Methods: To identify the chemical
constituents which make the plant as potent herbal, the essential oil was extracted from the
leaves. The oil from the leaves were isolated by steam distillation method through Clevenger
apparatus and analyzed by GC-MS to identify the bioactive compounds. Results: From this
leaf oil extract, about 12 bioactive compounds were identified viz. 5, 8-Epoxy-3H-2www.wjpr.net

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benzopyran, 4, 4a, 5, 8- tetrahydro-5, 8-dimethyl, 9-Acetylphenanthrene, 2, 13-Octadecadien1-ol, Hexadecanoic acid, methyl ester, 9-Oximino-2, 7-diethoxyfluorene, etc. Conclusions:
Whereas in dried leaves the isolated compounds were effectively tested against pathogenic
microorganisms and

also

compared

with standard antibiotics. Maximum antimicrobial

activity was observed at low concentration of essential oil. Main conclusions: Analysis of
phytochemical constituents and antimicrobial properties of essential oil extracted from the
leaves of Trichopus zeylanicus ssp. is clearly justified that this plant have a potentiality.
KEYWORDS: Trichopus zeylanicus ssp. travancoricus Burkill ex, Endemic Rhizomatous
herb, Agasthiyamalai hills, Arogyapachai, Kani tribe Jeevani.
1.0 INTRODUCTION
India, a mega diverse country reported with only 2.4% of the world's land area which
accounts for 20% of forest area, including over 45,000 species of plants. India has worlds
most biodiversity regions and the political boundaries of India encompass a wide range of
Eco

zones

via deserts,

high mountains,

highlands,

tropical and

temperate forests,

swamplands, plains, grasslands, as well as island archipelago. India has two hotspots viz.
Western Ghats and Himalayas. These hotspots harbor numerous endemic species of plants
and animals. From the two biodiversity rich centers, Western Ghats harbor more than 7388
species of flowering plants along with more than 1500 endemic species. Of these, 5584
species are indigenous, 377 are exotic naturalized and 1427 are cultivated or planted. Of the
5584 indigenous species, 2242 species are Indian endemics (found only in India) and 1261
are endemics to Western Ghats. Apart from the above, there are 586 taxa with subspecies and
variety status, bringing total taxa in the Western Ghats to 7974 (Nayar et al., 2014). The
Western Ghats are a chain of hills that run along the western edge of peninsular India. Their
proximity to the ocean and through orographic effect, they receive high rainfall. These
regions have wide range of forest from moist deciduous forest to rain forest. The region
shows high species diversity as well as high levels of endemism.
India has a rich culture of medicinal herbs and spices, which include about more than 2000
species and have a vast geographical area with high potential abilities for Ayurvedic, Unani
and Siddha are traditional medicines. Only very few researchers have been studied potential
bioactive compounds and pharmacological activity of medicinal herbs (Gupta et al., 2005).
Human beings have used plants for the treatment of diverse ailments for thousands of years
(Sofowara, 1982; Hill, 1989). According to the World Health Organization, most populations
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still rely on traditional medicines for their psychological and physical health requirements
(Rabe and Van Stoden, 2000), since they cannot afford the products of Western
pharmaceutical industries (Salie et al., 1996), together with their side effects and lack of
healthcare facilities (Griggs et al., 2001). Rural areas of many developing countries still rely
on traditional medicine for their primary health care needs and have found a place in day-today life. These medicines are relatively safer and cheaper than synthetic or modern medicine
(Iwu et al., 1999; Idu et al., 2007; Mann et al., 2008; Ammara et al., 2009). People living in
rural areas from their personal experience know that these traditional remedies are valuable
source of natural products to maintain human health, but they may not understand the science
behind these medicines, but knew that some medicinal plants are highly effective only when
used at therapeutic doses (Maheshwari et al., 1986; Van Wyk et al., 2000).
Herbal medicines are in great demand in both developed and developing countries as a source
of primary health care owing to their attributes having wide biological and medicinal
activities, high safety margins and lesser costs. Herbal molecules are safe and would
overcome the resistance produced by the pathogens as they exist in a combined form or in a
pooled form of more than one molecule in the protoplasm of the plant cell (Lai and Roy,
2004; Tapsell et al., 2006). Even with the advent of modern or allopathic medicine, Balick
and Cox (1996) have noted that a number of important modern drugs have been derived from
plants used by indigenous people. Traditional use of medicine is recognized as a way to learn
about potential future medicines. Researchers have identified number of compounds used in
mainstream medicine which were derived from ethno medicinal plant sources (Fabricant and
Farnsworth, 2001). Plants are used medicinally in different countries and are a source of
many potent and powerful drugs (Srivastava, et al., 1996; Mahesh and Sathish, 2008).
Trichopus zeylanicus ssp. travancoricus Burkill ex K. Narayanan is locally called
Arogyapachai or Ginseng of Kerala, a small perennial herb with rhizomatous stem belonging
to the family Dioscoriaceae. Scientists who have collected the plant species from the forest
areas of Agasthyamalai hills treated the species under the family Trichopodaceae. After
several nomenclatural confusions the species has included in Dioscoriaceae. The unripe fruits
are used by Kani tribes as a highly rejuvenating tonic comparable to Ginseng. The Kanis
claim to remain healthy, agile, young and resistant to various diseases or infections one
should consume the fresh fruits of the plant. The pharmacological investigation proved
antistress,

stamina

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boosting,

immunomodulatory,

hepatoprotective,

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aphrodisiac

and

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antioxidant properties associated with various parts of this plant. A critical survey of the
Ayurvedic classics suggests that the divine drug Varahi is very close to Arogyapachai. The
pharmacological studies showed that all parts of the plant possess medicinal properties like
inhibition of antigen induced de-granulation of sensitized peritoneal mast cells in rats.
2.0 MATERIALS AND METHODS Collection of Plant Materials
Trichopus zeylanicus ssp. travancoricus leaves were collected from Agasthyamalai Hills,
Kerala, India. The leaves were washed thoroughly with running tap water and finally with
distilled water. The plant materials were completely air-dried at room temperature for 3
weeks and then powdered by using electric blender to get a fine powder. The powdered
samples were kept in sealed air tight containers for the extraction of bioactive chemicals.
Trichopus zeylanicus ssp. travancoricus Burkill ex. K. Naryanan
Small, rhizomatous, glabrous, perennial herbs; rhizome 1-4 cm long, with slightly ascending
tips, seldom branched, covered with scales, older parts loosing scales and dying off
progressively; scales closely set, chaffy, lanceolate, acute or acuminate, 5-8 mm long; roots
many, fibrous; stem 3-8 from the axils of scales, petiole-like, each terminating in a solitary,
petioled leaf, terete, longitudinally striate or faintly ribbed, 5-15 cm long; petiole like the
stem, but dorsally grooved, 4-10 cm long; lamina highly variable in size, shape and venation,
broadly ovate, elliptic or linear-lanceolate, deeply cordate, truncate or cuneate at base, acute
or obtuse and apiculate or slightly emarginated at apex, 3-7 nerved from base with another
pair of nerves running along the whole leaf margin; fertile branch terminal on the stem, but
pushed towards one side by the petiole which usurps the line of the stem, short, sessile,
densely clothed with scaly, lanceolate bracts similar to the scales on the rhizome; flowers 1-6
on each branch, hermaphrodite; pedicel filiform, 2-4 cm long; perianth of 6 subequal lobes in
two series, campanulate, 3-10 mm long, constricted above the 'gynostemium', lobes
lanceolate, apiculate, broader and saccate at base, keeled on the back; stamens 6, filaments
short, flat, united with the top of the ovary and base of style, anthers 2- celled, connective
broad, projecting and meeting by their edges making a roof over a chamber into which
pollinating insects should enter, the whole structure forming what is called a 'gynostemium';
ovary inferior, 3-loculed with two superposed ovules in each chamber; style short, stout;
stigma 3, each 2-lobed, lobes fleshy, erect at first and reflexed later, always included in the
staminal chamber; fruit capsular, 3-lobed, 3-winged, ellipsoid or trapezoid, 1 -5 x 1 cm,
opening by irregular rupture of the fruit wall; seeds 2-6, oblong or ellipsoid, covered with

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dense brownish tomentum, ventrally flat and ruminate, to 4 x 2 mm.

Experimental Plant material
Fresh and dried leaves of T. zeylanicus ssp. travancoricus were collected from
Agasthiyamalai hills, Kerala. The plant material was identified by R. Ramasubbu, Assistant
Professor, Department of Biology, Gandhigram Rural Institute, Gandhigram and voucher
specimen was deposited in the Herbarium of Department of Biology, GRI-DU.
Oil isolation
Chopped Fresh leaves and dried powdered leaves were hydrodistilled separately using
Clevenger apparatus for 6 hr. the oils were collected dried over anhydrous sodium sulphate
and stored at 40 C until analyzed.
Analysis of the essential oil
GC-MS analysis were performed using a Perkin-Elmer GC clauses 500 system and Gas
Chromatograph interfaced to a mass spectrometer (GC-MS) equipped with a Elite-1, fused
silica capillary column (30 m 0.25 mm ID 1 df, composed of 100% Dimethyl poly
siloxane). For GC/MS detection, an electron ionization system with ionizing energy of 70 eV
was used. Helium gas (99.999%) was used as the carrier gas at constant flow rate 1 ml/min
and an injection volume of 2 l was employed (Split ratio of 10:1) injector temperature
250C; ion-source temperature 280C. The oven temperature was programmed from 110C
(isothermal for 2 min) with an increase of 10C/min to 2000C, then 5C/min to 280C,
ending with a 9 min isothermal at 280C. Mass spectra were taken at 70eV; a scan interval of
0.5 seconds and fragments from 45 to 450 Da. Total GC running time was 36 minutes.
Software adopted to handle mass spectra and chromatograms was a Turbo mass. Compounds
present in leaf extract of Trichopus were identified based on retention time, and their identity
was confirmed by comparing the mass spectra with the database from the Library of National
Institute of Standard and Technology.
Preparation of Inoculums
The microorganism used for the study was collected from the co-culture collection of
laboratory Department of Biology, GRI-DU Dindigul. Gram positive bacteria; Bacillus
cereus, Staphylococcus aureus and Micrococcus mucilaginosus, gram-negative bacteria;
Klebsiella pneumonia, Pseudomonas aeruginosa, Escherichia coli and Klebsiella terrigena,

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Fungi; Candida albicans, Candida glabrata and Candida sp. were used for the study. The
young bacterial and fungal strains were maintained in nutrient agar medium. The strains were
subculture of bimonthly and the cultured strains were allowed to grow for one week and
stored at 50 C for future analysis.
Effects of human pathogenic microorganism
Bacillus cereus
Bacillus cereus is widespread in the soil and the food industry in such foods as herbs, spices,
milk and vegetables. It is a spore-forming, Gram positive, facultative anaerobic bacterium
associated with food poisoning in humans. The most common is a diarrhea illness caused by
a heat-labile toxin, accompanied with abdominal pain.
Staphylococcus aureus
Staphylococcus aureus is a gram positive bacterium that causes disease in humans. It is the
leading cause of skin and soft tissue infections such as abscesses (boils), furuncles, cellulitis,
bloodstream infections, pneumonia or bone and joint infections.
Escherichia coli
Escherichia coli are gram negative bacteria that cause disease in humans. Its caused
following disease diarrhea illness. Infection with Escherichia coli is the leading cause of
travelers' diarrhea and a major cause of diarrhea disease in underdeveloped nations,
especially among children.
Candida albicans, Candida glabrata and Candida sp.
Candida albicans, Candida glabrata and Candida sp. are fungal pathogen responsible for
candidacies in human hosts.

Fatigue,

headache,

depression,

irritability, disorientation,

dizziness, inability to concentrate. In women, premenstrual syndrome (PMS), decreased

libido, recurrent vaginal yeast infection, vulvar itching and pain, prostate and bladder
infections.
Klebsiella pneumoniae and Klebsiella terrigena
Klebsiella pneumoniae and Klebsiella terrigena is a gram negative bacterium found in the
mouth, skin and intestinal tract, where it initially does not cause disease. K.pneumoniae can
progress into severe bacterial infections leading to pneumonia, bloodstream infections,
wound infection, urinary tract infections and meningitis.

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Micrococcus mucilaginosus
Micrococcus mucilaginosus is a gram positive bacteria it caused part of the normal human
oral flora and upper respiratory tract flora.
Culture Media Nutrient Agar
Nutrient Agar was used for the cultivation of less fastidious microorganisms and it was
enriched with blood or other biological fluids (Normal). The nutrient agar medium prepared
with the formulations as in the table 1.
Table 1. Nutrient Agar Composition
Peptic digest of animal tissue
Sodium chloride
Beef extract

5.0 gm/L
5.0 gm/L
1.5 gm/L

Yeast extract
Agar

1.5 gm/L
15 gm/L
pH

7.40.2

Bacterial growth medium

The bacterial activities of plant extract were carried out by using Muller Hinton Agar
Medium. The medium was prepared with the formulations as in the table 2.
Table 2. Bacterial Growth Medium
Beef infusion
Casein amino acids
Starch
Agar

3 g/L
17.5 g/L
1.5 g/L
17.0 g/L

pH

7.4

Fungal growth medium

Fungal activities of plant extract were carried out by using Potato Dextrose Agar Medium.
The medium was prepared with the formulations as in the table 3.
Table 3. Fungal Growth Medium
Potato
Dextrose
Agar
pH

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250 g/L
20 g/L
20 g/L
7.0

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Determination of antimicrobial activities Agar- well diffusion method

Agar well-diffusion method was followed to determine the antimicrobial activity. Nutrient
agar (NA) and Potato Dextrose Agar (PDA) plates were spreaded with the help of L- rod
inside the laminar air flow chamber with 8 hours old broth culture of respective bacteria and
fungi. Wells with 6 mm diameter and about 2 cm a part were made in a medium of Petri plate
each using sterile cork borer. Essential oil extracted from the leaves of Trichopus zeylanicus
was added with the help of sterile syringe into the wells and allowed to diffuse at room
temperature for 2hrs. Standard antibiotic, Ampicillin was (100 mg/ ml) selected to serve as
positive control. Control experiments comprising inoculums without plant extract were set up
and the plates were incubated at 37C for 18-24 h for bacterial pathogens and 28C for 48
hours for fungal pathogens. The diameter of the inhibition zone (mm) was measured by
Antibiotic scale (Himedia) and the activity index was also calculated for triplicates of each
experiment.
Disc diffusion Method
The essential oil extracts were subjected to antimicrobial assay using the agar disc diffusion
method. In this, different dilutions of extracts were prepared. Discs (6 mm diameter) of
whatman filter paper were used. All the Petri dishes, medium and paper discs were sterilized
using autoclave at 121C at 15 lbs for 15 minutes. Nutrient agar was poured into Petri plates
and allowed to cool in a laminar hood with the precaution to avoid any sort of atmospheric
contamination. After solidification, 24 h nutrient broth grown pathogenic cultures were
swabbed on the respective agar plates using sterilized cotton swabs. Different concentration
of extracts (200,150 and 100 g/ ml) was prepared reconstituting with methanol. The uniform
volumes of different concentrations of test and standard solutions were added to the cups in
the Petri dishes and the solutions were allowed to diffuse by leaving plates undisturbed for
one hour at room temperature. The Petri dishes were incubated at 371 0 C for 24 hrs and the
zones of inhibition were recorded in mm.
Determination of minimal inhibitory concentration (MIC) of the extract
The minimum inhibitory concentration (MIC) is defined as the lowest concentration of the
antimicrobial agent that will inhibit the visible growth of a microorganism after overnight
incubation. The wells were filled with 100 g/ml of the plant extracts were added to the wells
by serial two fold dilution from the suspension of essential oil extract stock solution. Each
well was inoculated with 100 l standard microbial suspension. The plates were covered,

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placed in plastic bags and incubated at 37C for 24 hrs. In this study, the MIC was the lowest
concentration of plant extracts that exhibited no growth of the organism in the wells by visual
reading. A loop full of broth was collected from those tubes which did not show any growth
and inoculated on sterile nutrient agar (for bacteria) and Potato dextrose agar (for fungi) by
streaking. Nutrient agar and Potato agar only were streaked with the test organisms
respectively to serve as control. Plates inoculated with bacteria were then incubated at 370 C
for 24 hours while those inoculated with fungi were incubated at room temperature (30
320 C) for 48 h.
Table 6. Standard microbial strains used for screening of antimicrobial activities of
Trichopus zeylanicus ssp. travancoricus
S. No. Name of the microbial strain Nature of organisms
1
Bacillus cereus
Gram-positive bacteria

Strain No.
GRIBIO1

2
3
4

Staphylococcus aureus
Micrococcus mucilaginosus
Klebsiella pneumonia

Gram-positive bacteria
Gram-positive bacteria
Gram-negative bacteria

GRIBIO5
GRIBIO8
GRIBIO2

Pseudomonas aeruginosa

Gram-negative bacteria

GRIBIO4

Escherichia coli

Gram-negative bacteria

GRIBIO6

7
8
9
10

Klebsiella terrigena
Candida albicans
Candida glabrata
Candida sp.

Gram-negative bacteria
Fungi
Fungi
Fungi

GRIBIO9
GRIBIO3
GRIBIO7
GRIBIO10

3.0 RESULTS
Phytochemical Analysis of Fresh and Dried Leaves Oil
Fresh and dried leaves of Trichopus zeylanicus ssp. travancoricus subjected into steam
distillation process for oil extraction through Clevenger apparatus. The temperature of the
apparatus slightly increased from 10-600 C. The maximum temperature of steam distillation
was fixed as for 4-5 hours. Essential oil extracted were diluted with acetone and carefully
removed from the apparatus and stored at specific temperature to avoid unwanted loss of
chemical constituents.

Hydro

distillation of the fresh leaves of T. zeylanicus ssp.

travancoricus afforded aromatic green oil with 1.25% (v/w) yield from fresh leaves and
0.07% (v/w) yield from dried leaves. The essential oil was collected and the extracted
essential oil was analyzed through GC-MS. Gas chromatography with mass spectrum was
used for the identification of secondary metabolites from leaves of T. zeylanicus ssp.
travancoricus. Bioactive compounds were eluted from the GC-MS at different interval time

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with Peak area percentage. Twelve different peaks from the compounds were detected
through GC-MS chromatogram which was shown in Fig. 1 along with retention time.
Whereas the Fig. 2 denotes the peaks with retention time also represent the compounds which
represent in the essential oil of fresh leaves. There were twelve compounds such as 5,8Epoxy-3H-2-benzopyran,4,4a,5,8-tetrahydro-5,8-dimethyl (RT 12.42), 9-Acetylphenanthrene
(RT 14.\2), 2,13-Octadecadien-1-ol (RT 16.18), Hexadecanoic acid, methyl ester (RT 17.22),
9-Oximino-2, 7-diethoxyfluorene (RT 17.93), 9-Octadecenoic acid methyl ester (RT 18.92),
Heptadecanoic acid, 16-methyl, methyl ester (RT 19.12), 9,15-Octadecadienoic acid, methyl
ester (RT 19.52), 5,8-Octadecadiyonic acid, methyl ester (RT 20.95), Ethanol, 2-(9octadecenyloxy) (RT 22.52), 18,19-Secoyohimban-19-oic acid, 16-methyl-, methyl ester (RT
25.35),

1H-Pyrazolo(3,4-b) quinoxaline, 1-phenyl-3-(1,2,3-trimethoxypropyl) (RT 28.23)

were identified from the fresh leaf oil of Trichopus zeylanicus ssp. travancoricus (Table 4
and Fig 1) and six compounds such as Hexadecanoic acid, methyl ester (RT 17.17), Methyl
10-Oxohexadecanoate (RT 17.88), 9-Octadecenoic acid, methyl ester (RT 18.95), (E)
Heptadecanoic acid (RT 19.18), 15-methyl, methyl ester, 6-Octadecadienoic acid, (Z) (RT
19.62) and E,E,Z-1,3,12-Nonadecatriene-5,14-diol(RT 19.73) were identified from the
essential oil of dried leaves (Table 5 and Fig 3). The Mass Spectroscopy of the fresh and
dried leaf oil of Trichopus zeylanicus ssp. travancoricus is given in the Fig 2 and Fig 4
respectively.
Antimicrobial Activity of Essential Oil of From Fresh Leaves
In this present study, antimicrobial activity was carried out from the essential oil of fresh
leaves oil of T. zeylanicus ssp. travancoricus by using agar-well diffusion method against the
various pathogens which are depicted in the Table 6. The table includes the gram positive
organisms are Bacillus cereus, Staphylococcus aurerus and Micrococcus mucilaginosis and
followed by gram negative (Klebsiella pneumonia, Pseudomonas aeruginosa, Escheria coli)
and fungi (Candida albicans, C.glabra and Candida sp). From the antimicrobial activity
observation, the essential oil from fresh leaves of T. Zeylanicus ssp. travancoricus is more
effective against the Gram negative bacteria. It exhibited a better antimicrobial activity
against the organisms such as Klebsiella pneumonia, Pseudomonas aeruginosa, Escherichia
coli, and Klebsiella terrigena followed by the fungi are Candida glabrata, Candida albicans
and Candida sp. The antimicrobial activity of T. Zeylanicus ssp. travancoricus against the
organisms such as Klebsiella pneumonia, Pseudomonas aeruginosa, Escherichia coli and
Klebsiella terrigena followed by the fungi are Candida glabrata, Candida albicans and

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Candida sp has been figured (Fig 5).

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Table 4. List of phytochemical constituents identified from fresh leaf oil of Trichopus
zeylanicus ssp. travancoricus through GC-MS
S. No
1.
2.
3.
4.

Compound Name
5,8-Epoxy-3H-2-benzopyran, 4, 4a,5,8-tetrahydro-5, 8-dimethyl
9-Acetylphenanthrene
2,13-Octadecadien-1-ol
Hexadecanoic acid, methyl ester

RT
12.42
14.2
16.18
17.22

5.
6.
7.

9-Oximino-2, 7-diethoxyfluorene
9-Octadecenoic acid methyl ester
Heptadecanoic acid, 16-methyl, methyl ester

17.93
18.92
19.12

8.
9.
10.
11.

9,15-Octadecadienoic acid, methyl ester

5,8-Octadecadiyonic acid, methyl ester
Ethanol, 2-(9-octadecenyl oxy)
18,19-Secoyohimban-19-oic acid, 16-methyl-, methyl ester

19.52
20.95
22.52
25.35

12.

1H-Pyrazolo(3,4-b)quinoxaline,1-phenyl-3-(1,2,3- trimethoxypropyl)

28.23

Table 5. List of phytochemicals constituents identified from dried leaf oil of Trichopus
zeylanicus ssp. travancoricus through GC-MS
S. No Name
1.
Hexadecanoic acid, methyl ester
2.
Methyl 10-Oxohexadecanoate
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RT
17.17
17.88
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3.
4.
5.
6.

9-Octadecenoic acid, methyl ester, (E)

Heptadecanoic acid, 15-methyl, methyl ester
6-Octadecadienoic acid, (Z)Petroselinic acid
E,E,Z-1,3,12-Nonadecatriene-5,14-diol

18.95
19.18
19.62
19.73

Table 6: Antimicrobial activity

Sl no

Name of the pathogen

Zone of inhibition by
sample
17.50.7071
210.4142
150
20.50.1213
17.50.7071
20.50.5355
180.707106781.4142

Zone of inhibition by
antibiotic (Gentamycin)
291.4142
32.50.7071
27.53.5355
29.50.7071
300
271.4142
311.4142

Zone of inhibition
by control
100
100
100
100
100
100
10.50.707106781

1
2
3
4
5
6
7

Bacillus cereus
Klebsiellapneumonia
Candida albicans
Pseudomonas aeruginosa
Staphylococcus aureus
Escherichia coli
Candida glabrata

Micrococcus mucilaginosus 17.53.535533906.1213

28.52.1213

12.53.535533906

9
10

Klebsiellaterrigena
Candida sp.

28.52.1213
294.2426

91.414213562
111.414213562

161.414213562
171.414213562.8284

4.0 DISCUSSION
Essential oils were extracted from fresh and dried leaves of T. zeylanicus ssp. travancoricus.
Among that, the Fig. 1 and Fig. 2 exhibited the highest peak in GC-MS chromatogram is 9Octadecenoic acid methyl ester commonly called as Methyl elaidate. It is insoluble in water
and it has many industrial uses such as adsorbent, absorbent, non pesticidal-agricultural
chemical, lubricants and water treatment product. In Fig. 1 second peak exhibited is
Heptadecanoic acid, 16-methyl, methyl ester, which have the ability to conjugate linoleic acid
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content in the human plasma. It is used to identify non-toxic small molecules and genes. It is
also

having property like anti human colon cancer,

antiasthmatics,

bronchodilators,

impotency, used as drug for dermatologic disorder and genital or sexual disorder
contraceptives,

antisporiatic,

non-central

analgesic,

anti-infective.

In

Fig.

6-

Octadecadienoic acid, (Z)Petroselinic acid exhibited as second peak used in cosmetics,

biodiesel engines and as well as lubricants, flavoring and fragrance agent, detergents, termites
spraying and antiperspirant. In Fig. 1 the third peak exhibited as Hexadecanoic acid, methyl
ester which is commonly called as Palmitic acid methyl ester and the compound also
presented in Fig. 2. Its having the activities such as antioxidant, hypocholesterolemic,
nematicide, pesticide, antiandrogenic, flavor, hemolytic, 5-Alpha reductase inhibitor. Other,
bioactive compounds extracted from fresh leaves of T. zeylanicus ssp. travancoriucs are 5, 8Epoxy-3H-2-benzopyran, 4, 4a, 5, 8-tetrahydro-5, 8-dimethyl is having a potential to inhibit
the human tumor cell line growth and it is also used for an antiviral and anticancer drug. 9Acetylphenanthrene compound used as drug for disorder of the nervous system and it is an
antiallergic agent, antineoplastic agent and bronchodilator. 9-Oximino-2, 7-diethoxyfluorene
is a compound that inhibits the capD of Bacillus anthracis. Ethanol, 2-(9-octadecenyl oxy) is
inhibiting the deposition of scale on surface and carrier for negatively charged drugs. It is
used in the preparation of progesterone. It has property like a pheromone dispenser and
antipsychotic. Methyl 10-Oxohexadecanoate used in the process for producing alcohol,
ceramides and molding compounds. It is also an anticancer drug.
From the above finding of this research has been clearly indicate that this plant Trichopus
zeylanicus ssp. Travancoricus is a potential medicinal plant was said by the previous
researchers such as Gupta et al., 2005, Srivastava, et al., 1996; Mahesh and Sathish, 2008 In
this present study, twelve different bioactive constituents have been identified from essential
oil extract from the leaves of T. zeylanicus ssp. travancoricus by Gas Chromatogram-Mass
spectrometry (GC-MS) analysis. The presence of various chemical compounds in the
essential oil justifies the use of the whole plant for various ailments by traditional
practitioners as said by Fabricant and Farnsworth, 2001. It also showed a positive
antimicrobial activity against Bacillus cereus, Staphylococcus aureus and Micrococcus
mucilaginosus. The leaf oil extract of Trichopus zeylanicus ssp. travancoricus exhibited a
common effect against the Gram positive bacteria. In all the Gram positive bacteria leaf oil
extract of Trichopus zeylanicus ssp. travancoricus shows a zone inhibition as 17.5mm. (AAcetone, B-Gentamycin (antibiotic), C- Trichopus zeylanicus ssp. Travancoricus oil extract

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sample).
CONCLUSION
Present research findings Trichopus zeylanicus ssp. travancoricus is an effective antibiotic
against all type of human pathogens due to the presence of various bioactive compounds
which are present in the essential oil. So, the plant is having the phyto- pharmaceutical
importance.
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