World Research Journal Biology of Biological Sciences

WRJBBS
Vol. 2(1), pp. 005-011, July, 2017. www.premierpublishers.org. ISSN: 3713-2135

Research Article

GC-MS Analysis of Ethanolic Extract of Alternanthera

Philoxeroides and Alternanthera Bettzickiana from India
U. Arul Pamila1 and S. Karpagam2
1
PhD Research Scholar, 2Assistant Professor, Department of Botany, Queen Mary's College (A), Chennai-600 004.

The present study is the comparative GC-MS analysis of an edible plant Alternanthera
philoxeroides (Mart.) Griseb and Alternanthera bettzickiana (Regel) G. Nicholson. The shade dried
aerial parts of plant powder A. philoxeroides and A. bettzickiana were extracted with ethanol. The
GC-MS analysis shows different peaks with low and high molecular weight determining the
presence of many bioactive compounds. The phytoconstituents in the ethanolic extract of
Alternanthera philoxeroides and Alternanthera bettzickiana have been screened by using GC-MS
analysis. A. bettzickiana ethanol extracts showed higher phytoconstituents when compared to
the ethanol extracts of A.philoxeroides. This study helps to explore the potential compounds and
the presence of these compounds may proceed to find out various therapeutic activities.

Keywords: GC-MS analysis, phytoconstituents, Alternanthera philoxeroides, Alternanthera bettzickiana, ethanol extract.

INTRODUCTION

Medicinal plants have occupied an important position in
the socio-cultural, development of rural people of India.
Plants leaves are considered one of the main sources of
biologically active compounds. It continues to play a
dominant role as sources of bioactive compounds in the
maintenance of human health. Plant are rich sources of
secondary metabolites with interesting biological activities
(Chaman and Verma 2006; Yousuf et al., 2014). In
developing countries, the practice of medicine still relies
heavily on plant and herbal extracts for the treatment of
human ailments. Dietary agents consist of a wide variety
of biologically active compounds that are ubiquitous in
plants, and many of them have been used as traditional
medicines (Aggarwal et al., 2003; Aggarwal and Shishodia
2004; Aggarwal and Shishodia 2006). The green plants
synthesize and accumulate a variety of biochemical
products, many of which are extractable and used as a
chemical feed stocks or as raw material for various
scientific investigations.
Plants are capable of synthesizing an overwhelming
variety of low-molecular weight organic compounds called
secondary metabolites, usually with unique and complex
structures. Many metabolites have been found to possess
interesting biological activities and find applications, such
GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India
as pharmaceuticals, insecticides, dyes, flavors and
fragrances. Plants used for traditional medicine contain a
wide range of substances that can be used to treat chronic
as well as infectious diseases (Duraipandiyan et al., 2006).
The modern methods describing the identification and
quantification of active constituents in plant material may
be useful for proper standardization and formulations.
Many species of Alternanthera traditionally are used in the
treatment of infections, as analgesic, anti-nociceptive,
antiviral, antibacterial, antifungal and diuretic agents
(Siqueira, 1987; Siqueria, 1984)
Alternanthera is one of the medicinally important genus
belonging to the family Amaranthaceae commonly known
as joyweeds, or Joseph's coat in English 'joyweeds or
Joseph's coat in English 'Ponnaganni Keerai and Seemai
Ponnaganni in Tamil. This genus consists of
approximately 80-200 species and is widespread genus
with cosmopolitan distribution (Pino et al., 2012;
ZafraStone et al., 2007)
*
Corresponding author: Arul Pamila, PhD Research
Scholar, Department of Botany, Queen Mary's College (A),
Chennai-600 004.India. Email: pamilastalin2004@gmail.com
Pamila and Karpagam 006
Figure 1: GC-MS analysis of phytochemicals identified from ethanolic extract of Alternanthera philoxeroides
Alternanthera philoxeroides is a perennial stoloniferous
herb found in both aquatic to terrestrial habitats. A.
philoxeroides is a perennial with prostrate, sprawling,
floating hollow stems, often in a dense tangled mass,
rooted in shallow water or growing from the shoreline,
occasionally free-floating (Long and Lakela., 1971;
Godfrey and Wooten., 1981). The hollow stems provide
considerable buoyancy of the mat (Buckingham., 1996).
Roots form at stem nodes. The preliminary phytochemical
analysis showed the presence of alkaloids, carbohydrates,
saponins, phenols, flavonoids, aminoacids, diterpenes,
tannin, terpenoids, protein, steroid, oxalate, coumarin and
quinone in the ethanol extract of A. philoxeroides isolated
from unpolluted site (Pamila and Karpagam., 2017b). A.
philoxeroides Griseb is being prescribed clinically in the
People's Republic of China for the treatment of viral
hepatitis, epidemic parotitis, hemorrhagic fever and
influenza (Chaterjee and Pakrashi., 1991). A.
GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India
philoxeroides has been suggested to be one of the 20 most
dangerous invasive alien species in China (Lu and Ding.,
2012)
Alternanthera bettzickiana (Regel) G. Nicholson is
commonly used as an ornamental edging plant. It is
particularly popular in China where it is cultivated in nearly
all the large cities. It is native to South America. Its
common name includes calico plant. The whole plant is
reported to be useful in purifying and nourishing blood and
is claimed to be a soft laxative, a galactagogue and an
antipyretic, in addition to its wound healing property
(Petrus et al., 2014). The preliminary phytochemical
analysis showed primary and secondary metabolites such
as alkaloids, carbohydrates, saponins, phenols,
flavonoids, diterpenes, tannin, terpenoids, steroid, oxalate,
anthocyanin, leucoanthocyanin, Xanthoprotein, coumarin
 World Res. J. Biol. Biol. Sci. 007
Table 1: Phytochemical compounds identified in ethanol extract of Alternanthera philoxeroides

PK# PEAK NAME Molecular formula Molecular weight Retention time % peak area
1. Ar-tumerone C15H20O 216.324 g/mol 13.351 8.14
2. Phenol, 5-(1,5-dimethyl-4-hexeny C15H22O 218 g/mol 14.272 2.77
3. Bicyclo [3.1.1] heptane, 2,6,6- C10H18 138.2499g/mol 15.141 2.04
trimethyl-
4. 2-pentadecanone, 6,10, 14- C18H36O 268.485 g/mol 15.200 1.23
trimethyl
5. n-Hexadecanoic acid C16H32O2 256.4241 g/mol 16.367 29.23
6. 3-Acetoxypentadecane C17H34O2 270.451 g/mol 16.641 2.10
7. 9,12-octadecadienoic acid (Z, Z)- C18H32O2 280.4455) g/mol 17.934 12.50
8. 9,12-octadecadienoic acid (Z, Z)- C18H32O2 280.4455 g/mol 17.979 13.61
9. Octadecanoic acid C18H36O2 284.4772 g/mol 18.179 3.45
10 Tris (tert-butyldimethylsilyloxy) C8H2O2Si 871.302 g/mol 26.105 7.27
arsane
11. Temazepam C16H13ClN2O2 300.742 g/mol 26.469 13.20
12. 5-Methyl-2phenylindolizine C15H13N 207.276 g/mol 26.871 4.47

Figure 2: GC-MS analysis of phytochemicals identified from ethanolic extract of Alternanthera bettzickiana

GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India
Pamila and Karpagam 008

Table 2 Phytochemical compounds identified in ethanol extract of Alternanthera bettzickiana

PK# Peak Names Molecular formula Molecular weight Retention time % peak area

1 Ar-tumerone C15H20O 216.32 g/mol 13.350 6.67

2. Tumerone C15H22O 218.33 g/mol 13.395 1.31
3. Curlone C15H22O 218.34 g/mol 13.752 1.43
4. Phenol, 5-(1,5-dimethyl-4-hexeny C15H22O 218.33 g/mol 14.272 2.14
5 Bicyclo[3.1.1]heptane, 2,6,6-triglycerides C10H18 138.25g/mol 15.141 5.24
6 Cyclohexanol, 1-ethynyl C8H12O 124.18g/mol 15.386 0.74
7 7-Heptadecyne, 1-chloro C17H31Cl 270.885 g/mol 15.564 1.86
8 n-Hexadecanoic acid C16H32O2 256.42g/mol 16.359 18.10
9 Dichloroacetic acid, heptadecyl C19H36Cl2O2 367.395g/mol 16.634 2.43
10 Phytol C20H40O 296.53g/mol 17.748 1.87
11 9,12-Octadecadienoic acid (Z,Z)- C18H32O2 280.45 g/mol 17.941 15.08
12 9,12,15-Octadecatrien-1-ol, (Z,Z... C18H32O2 264.44g/mol 17.993 15.05
13 Squalene C30H50 410.72g/mol 23.305 1.06
14 Vitamin E C29H50O2 430.71g/mol 25.318 4.29
15 Cyclotrisiloxane, hexamethyl- C6H18O3Si3 222.46g/mol 26.105 0.27
16 Arsenous acid, tris(trimethylsil C9H27AsO3Si3 342.48 g/mol 26.469 8.76
17 Hexestrol, di-TMS C24H38O2 Si2 414.73 g/mol 26.871 2.27
18 Farnesol isomer a C15H26O 222.37 g/mol 30.607 4.80
19 Cyclotrisiloxane, hexamethyl C6H18O3Si3 222.46 g/mol 32.071 2.46
20 Cyclotrisiloxane, hexamethyl C6H18O3Si3 222.46 g/mol 33.148 4.18

and glycosides in the ethanol extract of A bettzickiana
(Pamila and Karpagam., 2017a). GC-MS is one of the
paramount technique to identify the phytoconstituents
present in the plants. Hence, the objective of the present
study is to identify the phytochemical constituents with the
aid of GC-MS technique, which may provide an insight in
its use in traditional medicine.
MATERIALS AND METHODS
Collection and authentication of plant materials
A. philoxeroides and A. bettzickiana specimens were
collected from Pechiparai of Kanyakumari District, Tamil
Nadu, India. The collected plants were identified in the
Department of Botany, Queen Mary's College and
confirmed by Prof. P. Jayaraman, Director, Plant Anatomy
Research Centre (PARC) Chennai.
Plant material
Fresh plants were washed thoroughly three to four times
with running tap water then finally with sterile water
followed by shade drying at room temperature for 20-30
days and powdered by using an electric blender and stored
in airtight container.
Preparation of extract
The 10g of sample was taken and soaked for 24h in 30ml
of ethanol. The extract was filtered using Whatman filter
paper No. 1, evaporated to dryness and re-dissolved in
DMSO (Dimethyl Sulphoxide). The extracts were
preserved in airtight container and kept at 4-50C for further
use.
Gas Chromatography-Mass spectrometry (GC-MS)
analysis
The GC-MS was performed by using PerkinElmer Clarus
500 Model and the software used is Turbomass ver 5.2.
The fused silica column was packed with Elite -5MS (5%
Phenyl 95%dimethylpolysiloxane,30m x 250m). The
oven temperature was set up from 50C with an increase
of 8 C/minto 220 C for 5 min and 7C /min to 280 C for
15 mins. Helium gas (99.999%) was used as the carrier
gas at constant flow rate of 1 ml/min. An aliquot of 2l of
sample was injected into the column with the injector
temperature at 280C and the Split ratio of 10:1. The
ionizing energy of 70 eV was used and the
electronionization is involved. The mass range is 40-
600amu. The inlet line temperature was 200 C and
source temperature was 150 C. Total GC running time
was 50 minutes. The compounds were identified referring
to NIST 2005 library.
Identification of compounds
Interpretation of mass spectrum of GC-MS was conducted
using the database of National Institute Standard and
Technique (NIST Version-Year 2005) having more than
62,000 patterns. The relative percentage amount of each
component was calculated by comparing its average peak
area to the total areas. The spectrum of the unknown
component was compared with the spectrum of the known
component stored in the NIST data library (version 2005).

GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India
 World Res. J. Biol. Biol. Sci. 009

Table 3 Biological activity of phytoconstituents identified in the ethanol extracts of the plant of A. philoxeroides

PK# Name of the compounds Biological Activity

1. Ar-tumerone Antiviral.
2. Phenol, 5-(1,5-dimethyl-4-hexeny Antioxidant, Antibacterial/Antiseptic and disinfectant aromatic alcohol., Topical
Anesthetic
3. Bicyclo [3.1.1] heptane, 2,6,6-trimethyl- No Activity reported.
4. 2-pentadecanone, 6,10, 14-trimethyl Cancer-preventive
5. n-Hexadecanoic acid Antioxidant, hypocholesterolemic, nematicide, pesticide, antiandrogenic,
flavour, hemolytic, 5-alpha reductase inhibitor
6. 3-Acetoxypentadecane Antinephrotoxic and antioxidant activities
7. 9,12-octadecadienoic acid (Z, Z)- Anti-inflammatory Hypocholesterolemic Cancer preventive
HepatoprotectiveNematicideInsectifuge, Antihistaminic AntieczemicAntiacne, 5-
Alpha reductase inhibitor AntiandrogenicAntiarthriticAnticoronary
8. 9,12-octadecadienoic acid (Z, Z)- Anti-inflammatory Hypocholesterolemic Cancer preventive
HepatoprotectiveNematicideInsectifuge, Antihistaminic AntieczemicAntiacne, 5-
Alpha reductase inhibitor AntiandrogenicAntiarthriticAnticoronary
9. Octadecanoic acid Cancer preventive Insectifuge/Cosmetic, Flavor, Hypocholesterolemic, Lubricant,
Perfumery, Propecic, Suppository
10 Tris (tert-butyldimethylsilyloxy) arsane No Activity reported.
11. Temazepam Therapy of insomnia, anti-anxiety agent, anticonvulsant activity, motor
coordination, and memory loss.
12. 5-Methyl-2phenylindolizine No Activity reported.

Table 4 Biological Activity of phytoconstituents identified in the ethanol extracts of the plant of A. bettzickiana

PK# Name of the compound Biological Activities

1 Ar-tumerone Antiviral.
2. Tumerone Antiviral.
3. Curlone No Activity reported.
4. Phenol, 5-(1,5-dimethyl-4-hexeny Antioxidant, Antibacterial/Antiseptic and disinfectant aromatic alcohol., Topical
Anesthetic
5 Bicyclo[3.1.1]heptane, 2,6,6-tri No Activity reported
6 Cyclohexanol, 1-ethynyl No Activity reported.
7 7-Heptadecyne, 1-chloro No Activity reported.
8 n-Hexadecanoic acid Antioxidant, HypocholesterolemicNematicide, Pesticide,Lubricant,
Antiandrogenic, Flavor, Hemolytic, 5-Alpha reductase inhibitor
9 Dichloroacetic acid, heptadecyl No Activity reported.
10 Phytol Cancer-Preventive, Antimicrobial Anti-inflammatory Anti cancer Diuretic.
11 9,12-Octadecadienoic acid (Z,Z)- Anti-inflammatory Hypocholesterolemic Cancer preventive
HepatoprotectiveNematicideInsectifuge, Antihistaminic AntieczemicAntiacne, 5-
Alpha reductase inhibitor AntiandrogenicAntiarthriticAnticoronary
12 9,12,15-Octadecatrien-1-ol, (Z,Z... Fatty acid/Antioxidant, Antibacterial
13 Squalene Antibacterial,Antioxidant,Antitumor, Cancer preventive, Immunostimulant, Chemo
preventive, Lipoxygenase-inhibitor,Pesticide Diuretic
14 Vitamin E Antiageing, Analgesic,Antidiabatic, Antiinflammatory, Antioxidant, Antidermatitic,
Antileukemic, Antitumor, Anticancer, Hepatoprotective, Hypocholesterolemic,
Antiulcerogenic, Vasodilator, Antispasmodic, Antibronchitic,
Anticoronary,Antialzheimeran, Antidermatitic, , Immunostimulant
15 Cyclotrisiloxane, hexamethyl- Antimicrobial potential, antimicrobial, antioxidant.
16 Arsenous acid, tris(trimethylsillyl) Ester No Activity reported.
17 Hexestrol, di-TMS Hormonal antineoplastic agent
18 Farnesol isomer a Antineoplastic, antibacterial
19 Cyclotrisiloxane, hexamethyl Antimicrobial potential, antimicrobial, antioxidant.
20 Cyclotrisiloxane, hexamethyl Antimicrobial potential, antimicrobial, antioxidant.
Source: Dr. Duke's phytochemical and ethnobotanical databases [Online database]

The name, molecular weight, molecular formula and RESULTS AND DISCUSSION
structure of the components of the test material were
determined. The components present in the ethanol extract of aerial
parts of Alternanthera philoxeroides and Alternanthera
GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India
Pamila and Karpagam 010
bettzickiana were identified by GC-MS analyzed (Figure 1
and 2). The 12 and 20 phytoconstituents were detected in
ethanol extract of aerial parts of Alternanthera
philoxeroides and Alternanthera bettzickiana. The peak
names with their molecular formula, molecular weight
(MW), retention time (RT) and peak area are exhibited in
Tables 1 and 2. The name of phytoconstituents and their
biological activities are showed in Tables 3 and 4.
GC-MS is one of the most precise methods to identify
various secondary metabolites present in the plant extract
(Deshpande and Kadam., 2013; Al-Huqail et al., 2015;
Payum., 2016). Diversity of medicinal plants and herbs
containing various phytochemicals with biological activity
can be of valuable therapeutic key. Different
phytochemicals have been found to have a broad range of
activities, which may help in protection against chronic
diseases (Liu., 2003). In the present study, the GC-MS
analysis of the ethanolic extract of plants of A
philoxeroides and A. bettzickiana aerial parts showed the
presence of many bioactive compounds. Five compounds
are commonly present in both plants, they aren-
Hexadecanoic acid, 9,12-Octadecadienoic acid (Z,Z), Ar-
tumerone, Bicyclo [3.1.1] heptane, 2,6,6-trimethyl, and
Phenol, 5-(1,5-dimethyl-4-hexeny.Vidhya etal.,(2015)
reported that the GC-MS analysis identified the presence
of N N-Dimethyltryptamine, N-Dimethylaminomethyl-
isopropyl benzyl phosphine, N-Hexadecanoic acid and
Cyclotrisiloxane Hexane methyl in the leaf of A.
bettzickiana. Huang et al., (2017) reported that ethyl
propionate was identified as the most abundant
component of the root aqueous extracts of A.
philoxeroides. In best of our knowledge, there is no
documental evidence of gas chromatography and mass
spectrum analysis to identify the chemical compounds
from the ethanol extract of aerial parts of A. philoxeroides.
Hexadecanoic acid and vitamin E may have the role in
antioxidant and anti-inflammatory effects (Kalpana Devi et
al., 2012). Among the identified phytochemicals,
Squalene has antioxidant activity and has been found that
squalene possesses chemo-preventive activity against the
colon carcinogenesis (Rao et al., 1998). The presence of
phytol compounds attributes to the antimicrobial, anti-
inflammatory and anticancer property of the plant leaves
(Cho et al., 2010; Munakata., 1983). Phenol compounds
have antioxidant, antibacterial/antiseptic, disinfectant
properties. The aromatic alcohol also is used in topical
applications and is promising as anesthesia.
CONCLUSION
In the present study, A. bettzickiana ethanol extracts
showed higher phytoconstituents when compared to the
ethanol extracts of A.philoxeroides. The presence of these
phytoconstituents justifies the use of these plants for
GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India
various ailments by traditional practitioners. It would be
worthwhile to further isolate the compounds and determine
their specific activity and also to understand the synergistic
effect of compounds for therapeutic roles.
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GC-MS Analysis of Ethanolic Extract of Alternanthera Philoxeroides and Alternanthera Bettzickiana from India