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GC-MS ANALYSIS OF BIOACTIVE COMPOUNDS OF
ACHYRANTHES ASPERA
A. Pauline Fatima Mary1*
and Dr. R. Sagaya Giri1
*1
Department of Botany, K. N. Govt. Arts for Women (Autonomous),
Thanjavur-613 007, Tamilnadu, India.
ABSTRACT
Achyranthes aspera L. (Amaranthaceae) is an important medicinal
herb found as a weed throughout India. It has long been used in
different systems of medicine in the treatment of cancer, leprosy,
asthma, wound, insect and snake bite, dandruff, hepatitis, renal
disorders, dermatological disorders, gynecological disorders, malaria,
fever, cough, diabetes, dysentery, toothache etc.,. The powdered leaves
were subjected to phytochemical screening and were found to contain
alkaloids, flavonoids, Phenols, Saponins, Tannins, Phytosterols and
Terpenoids. To determine the bioactive components of ethanolic leaf
extracts of A. aspera by GC-MS analysis were carried out. Twenty five
compounds were identified. Some important compounds are; 9-Octadecenamide, (Z)
(21.15%), Squalene (19.04%), Phytol (7.06%), Phenol, 4,6-di (1,1dimethylethyl)
2-methyl- (6.50%), à–Amyrin (4.33%), Cyclopropane, nonyl- (3.37%), Dibutyl phthalate
(3.19%) etc.,. Some of the identified compounds have been reported to possess various
biological activities such as Antimicrobial, Antioxidant, Antiseptic, Pesticide, Fungicide,
Diuretic, Anti-inflammatory and Anticancer.
KEYWORDS: Achyranthes aspera, Antifungal activity, Bioactive compoounds,
Phytochemical.
INTRODUCTION
Nature has provided a complete store-house of remedies to cure all ailments of mankind. For
a long period of time, plants have been a valuable source of natural products for maintaining
human health, especially in the last decade, with more intensive studies for natural therapies.
World Journal of Pharmaceutical Research SJIF Impact Factor 7.523
Volume 7, Issue 1, 1045-1056. Research Article ISSN 2277– 7105
*Corresponding Author
Dr. A. Pauline Fatima Mary
Department of Botany, K. N.
Govt. Arts for Women
(Autonomous), Thanjavur-
613 007. Tamilnadu, India.
Article Received on
13 November 2017,
Revised on 3 Dec. 2017,
Accepted on 24 Dec. 2017
DOI: 10.20959/wjpr20181-10540
8533
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Mary and Giri. World Journal of Pharmaceutical Research
A number of plants have been documented for their medicinal potential, which is in use by
the traditional healers, herbal folklorists and in Indian system of medicine namely Siddha,
Ayurveda and Unani.[1]
The use of herbal medicine has become increasingly worldwide
popular and medicinal plants are believed to be an important source of new chemical
substances with potential therapeutic effects. WHO has considered phytotherapy in its health
programme; because these drugs are safe, cost effective and most importantly people have
faith in them. The demand for crude drugs has undergone a considerable change in recent
years due to aggressive marketing of the crude drugs. Standardized extracts from them or
pure phyto-pharmaceuticals need to be studied extensively for their quality, purity, potency,
safety and efficacy.[2]
The novel molecules from plant sources have been instrumental in development of
structurally modified compounds, which assist a lot in the development of modern
therapeutic system.[3]
Phytochemicals are responsible for medicinal activity of plants and
these biochemicals are naturally ccurring in the plants that have defence mechanism and
protect from various diseases[4]
The phytochemical are very important in medicine and
constitute most of the valuable drugs.[5]
This biochemicals are often referred to as secondary
metabolites which is useful to traditional medicine system are identified by GC-MS
technique.[6]
In recent years Gas Chromatography – Mass Spectrum (GC-MS) studies have
been increasingly applied for the analysis of medicinal plants as this technique has proved to
be a valuable method for the analysis of essential oil, alcohols, acids, esters, alkaloids,
steroids, amino and nitro compounds.[7]
Achyranthes aspera L. (Amaranthaceae) is an important medicinal herb found as a weed
throughout India, tropical Asia and other parts of the world. It has long been used in different
systems of medicine in the treatment of cancer, leprosy, asthma, fistula, piles, arthritis,
wound, insect and snake bite, diabetes, malaria, pneumonia fever, dandruff, hepatitis, renal
disorders, dermatological disorders, gynecological disorders, malaria, fever, cough, diabetes,
dysentery, toothache etc. The aim of this study was to investigate the Achyranthes aspera
leaves for its phytochemicals by qualitative test and to study the phytochemical components
by GCMS.
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MATERIAL AND METHODS
Sample Authentication
Achyranthes aspera was identified and authenticated by Botanist, Dr. Soosai Raj, M.Sc.,
Ph.D., Department of Botany, St. Joseph’s college, Trichy, Tamil Nadu, India. A Voucher
specimen has been deposited at the Rapinat Herbarium, St. Joseph’s College, Tiruchirappalli,
Tamil Nadu, India. (Figure-1).
Botanical Description
Achyranthes aspera Linn
Systematic position
Class : Dicotyledons
Sub Class : Monochlamydeae
Serious : Curvembryeae
Family : Amaranthaceae
Genus : Achyranthes
Species : aspera
Achyranthes aspera Plants are small, much branched, monoecious perennial sub shrub up to
0.8–1×0.8 m. Rootstock stout, woody. Stems somewhat succulent at first, ribbed, becoming
basally woody with age, densely covered in velutinous, appressed hairs. Leaves opposite,
densely clustered toward branch tips 40–50×25–30 mm, spreading to decurved, mostly
broadly ovate, ovate-orbicular or elliptic; apex blunt to abruptly sub acute, sometimes very
shortly apiculate; base attenuate; lamina somewhat fleshy, purple-grey, veins often purple,
abaxial and adaxial surfaces silkycanescent, margins crenulate to crenate. Petioles 5–10mm
long, pink, fleshy, velutinous, basal abscission zone present. Inflorescence a terminal erect
spike, 150– 200mm long; peduncle 15mm long, fleshy, white villous; spike rachis fleshy,
white-villous to purple villous; flowers bisexual, retrorse, sessile, 180–200 per spike, these
spaced initially at 10-mm intervals along rachis, diminishing rapidly to <1-mm intervals
toward inflorescence apex.
Collection of Plant materials
Fresh leaves of Achyranthes aspera were collected (Figure-2) from vaduganputhupatti,
Thanjavur Dt., Tamilnadu. The leaves were washed in clean water and air dried in room
temperature. The dried plant materials were milled to a fine powder using grinder and stored
in the dark at room temperature in airtight containers.
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Preparation of plant extracts
50 g of dried powder of plant was extracted with ethanol by continuous hot percolation, using
soxhlet apparatus (Figure-3) and concentrated up to 50 ml in desiccator under reduced
pressure. The concentrated extract were lyophilized and used for further studies.
Qualitative analysis of phytochemical compounds
Phytochemical analysis of the all plant extracts was undertaken using standard qualitative
methods as described by various authors. The plant extracts were screened for the presence of
biologically active compounds such as alkaloids, flavonoids, carbohydrates, phytosterols,
proteins, phenolics, tannins and saponins.
Quantitative analysis of Phytochemical compounds by using GCMS
The ethanolic extracts of the selected plant samples were subjected to GC-MS studies for
identification of phytochemical compounds. The GC-MS analysis was carried out using a
Clarus 500 Perkin- Elmer (Auto System XL) Gas Chromatograph equipped and coupled to a
mass detector Turbo mass gold – Perking Elmer Turbomas 5.2 spectrometer with an Elite-1
(100% Dimethyl ply siloxane), 300 m x 0.25 mm x 1 μm df capillary column. The instrument
was set to an initial temperature of 110°C and maintained at this temperature for 2 min. At
the end of this period, the oven temperature was raised upto 280°C, at the rate of an increase
of 5°C/min and maintained for 9 min. Injection port temperature was ensured as 250°C and
Helium flow rate as 1 ml/min. The ionization voltage was 70 eV. The samples were injected
in split mode as 10:1. Mass Spectral scan range was set at 45-450 (mhz). The chemical
constituents were identified by GC-MS. The fragmentation patterns of mass spectra were
compared with those stored in the spectrometer database using National Institute of Standards
and Technology Mass Spectral database (NIST-MS). The percentage of each component was
calculated from relative peak area of each component in the chromatogram.
Identification of Compounds
Interpretation of mass spectrum of GC-MS was conducted using the database of National
Institute Standard and Technology (NIST) having more than 62,000 patterns. The spectrum
of the known component was compared with the spectrum of the unknown components
stored in the NIST library. The Compound name, Retention time, Molecular formula and
Nature of the compounds and Area % of the test materials were ascertained.
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RESULTS AND DISCUSSION
Ethanolic extracts of leaves of A. aspera showed the presence of alkaloids, flavonoids,
carbohydrates, proteins, phenols, saponins and terpenoids whereas phytosterol and tannins
were absent (Table-1and Figure-4). Priya[8]
screened the phytochemical of A. aspera,
methanol extract showed the presence of carbohydrates, phenolic compounds, oil and fats,
saponins, flavonoids, alkaloids and tannins, whereas, aqueous extract contained phenolic
compounds, saponins, flavonoids and tannins as major phytochemical groups.
Danial[9]
reported the presence of polysaccharides, ecdysterone (harmone), achyranthine,
betaine (Alkaliods), vanillic acid, syringic acid, p-coumaric acid (phenolic acids), saponin A,
saponin B (saponins), protein and carbohydrates in A. aspera. Presence of phenolic
compounds in the plant suggests the potential use of A. aspera as a source of antioxidant
compounds[10]
.
The GC-MS spectral studies revealed the presence of 25 compounds from the leaf ethanol
extract of A. aspera. The GCMS chromatogram of ethanolic leaf extract of Achyranthes
aspera are shown in the figure-5. The compound name, retention time (RT), molecular
formula, compound nature and area % were presented in Table 2. The major compounds were
cyclopropane, nonyl- (3.37) 1-Hexadecanol (3.11%), Phenol, 4, 6-di (1,1-dimethylethyl)-2-
methyl- (6.5%), Squalene (19.04%), Dibutyl phthalate (3.19%), Phytol (7.06%), 9-
Octadecenamide, (Z)- (21.15%) and á-Amyrin (4.33%). The details of the identified
phytoconstituents and its therapeutic activity were given in the Table 3. The structure of the
identified compounds among were given in the Figure-6. Saurabh Srivastav et al., (2011)11
revealed that wide numbers of phytochemical constituents have been isolated from
Achyranthes aspera which possesses activities like antiperiodic, diuretic, purgative, laxative,
antiasthmatic, hepatoprotective, anti-allergic and various other important medicinal
properties.
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Figure 1: Sample Authentication.
Figure-2: Collection of Achyranthes aspera leaves.
Figure-3: Preparation of Plant Extract.
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Figure-4: Preliminary phytochemical analysis.
Table 1: Analysis the phytochemicals of Achyranthes aspera by qualitative test.
Figure 5: GC-MS Chromatogram of Ethanolic leaf extracts of Achyranthes aspera.
Phytochemicals Result
Alkaloids +
Flavonoids +
Carbohydrates +
Protein +
Phenols +
Saponins +
Tannins -
Phytosterols -
Terpenoids +
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Table 2: Components detected in Ethanolic leaf extracts of Achyranthes aspera.
S. No Compound Name RT Molecular
Formula Compound Nature
Area
%
1 Cyclotetrasiloxane, octamethyl- 5.99 C8H24O4Si4 OrganoSilicone
compound 2.15
2 3-Carene 6.29 C10H16 Monoterpene 1.14
3 Cyclopropane, nonyl- 9.06 C12H24 Gaseous
Hydrocarbons 3.37
4 1-Hexadecanol 11.86 C16H34O Terpene alcohol 3.11
5 Caryophyllene 12.4 C15H24 Sesquiterpene 0.8
6 Precocene I 12.93 C12H14O2 Essential oils 1.99
7 Cycloheptasiloxane,
tetradecamethyl- 13.17 C14H42O7Si7
Organo Silicone
compound 0.83
8 1-Hexadecanol 14.37 C16H34O Terpene alcohol 2.42
9 Phenol, 4,6-di (1,1dimethylethyl)
2-methyl 15.28 C15H24O Phenolic compound 6.5
10 Trichloroacetic acid, tetradecyl
ester 16.61 C16H29Cl3O2 Acetic acid compound 0.97
11 3-Eicosyne 17.08 C20H38 Organic compound 2.15
12 Squalene 17.94 C30H50 Triterpene 19.0
4
13 Dibutyl phthalate 18.42 C16H22O4 Plastilizer compound 3.19
14 Hexadecanoic acid, ethyl ester 18.63 C18H36O2 Palmitic acid eater 1.72
15 Phytol 19.78 C20H40O Diterpene 7.06
16 9,12,15 Octadecatrienoic acid,
ethyl ester,(Z,Z,Z) 20.28 C20H34O2 Linolenic acid ester 1.59
17 9-Octadecenamide,(Z) 22.07 C18H35NO Amide 21.1
5
18 Hexasiloxane,1,1,3,3,5,5,7,7,9,9,
11,11dodecamethyl 22.86 C12H38O5Si6
Organo Silicone
compound 0.55
19 Eicosane, 7-hexyl 23.09 C26H54 Aliphatic hydro
compounds 1.28
20 Diisooctyl phthalate 23.7 C24H38O4 Plastilizer compound 1.53
21 Docosanoic acid, 1,2,3-
propanetriyl ester 24.12 C69H134O6 Fatty acid 2.25
22 à –Amyrin 24.79 C30H50O Triterpene 4.33
23 9-Hexacosene 25.13 C26H52 Hydrocarbon 2.2
24 Heptacosane, 1-chloro 25.44 C27H55Cl Hydrocarbon 1.77
25
Glycine,N-[(3à,5á,7à,12à) 24-
oxo-3,7,12-tris [(trimethylsilyl
,methyl ester
25.95 C36H69NO6
Si3 Organic compound 0.77
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Table 3: Bioactive components and their activity in Ethanolic leaf extracts of A. aspera
by GCMS study.
S. No Compound Name ***Activity
1 Cyclotetrasiloxane, octamethyl-
Antimicrobial, Antiseptic, Hair
Conditioning Agent, Skin- Conditioning
Agent-Emollient; Solvent.
2 3-Carene Allergenic, Fungicide, Irritant,
Pesticide.
3 Cyclopropane, nonyl- Anethetic agent.
4 1-Hexadecanol Antimicrobial , Anti-inflammatory.
5 Caryophyllene
Anti-inflammatory, Antibiotic,
Antooxidant, Anticarcinogenic agent,
local anaesthetic activities.
6 Precocene I Antifeedant, juvabional, Pesticide.
7 Cycloheptasiloxane, tetradecamethyl-
Antimicrobial, Antiseptic, Hair
Conditioning Agent, Skin- Conditioning
Agent-Emollient; Solvent.
8 1-Hexadecanol Antimicrobial, Anti-inflammatory.
9 Phenol, 4,6-di (1,1dimethylethyl) 2-
methyl
Antimicobial, Anesthtic, Antooxidant,
Antiseptic, Cancer preventive, Pesicide,
Fungicide.
10 Trichloroacetic acid, tetradecyl ester Precipatation agent, An Ingredient
agent, Herbicide.
11 3-Eicosyne Antimicrobial.
12 Squalene Antimicrobial, Antioxidant, Antitumor,
Cancer-Preventive, Pesticide.
13 Dibutyl phthalate Antimicrobial, Solvent, Plastilixer,
Pesticide, Repellent.
14 Hexadecanoic acid, ethyl ester Antioxidant, Nematicide, Pesicide.
15 Phytol Antimicrobial, Anticancer, Diuretic,
Anti -inflammatory.
16 9,12,15 Octadecatrienoic acid, ethyl
ester,(Z,Z,Z)
Analgesic, Antipyretic, Anticonvulsant,
Antiseptic.
17 9-Octadecenamide,(Z) Antimicrobial.
18 Hexasiloxane,1,1,3,3,5,5,7,7,9,9,11,11do
decamethyl
Antimicrobial, Antiseptic, Hair
Conditioning Agent, Skin- Conditioning
Agent-Emollient; Solvent.
19 Eicosane, 7-hexyl No Activity reported.
20 Diisooctyl phthalate Antimicrobial, Solvent, Plastilixer,
Pesticide, Repellent.
21 Docosanoic acid, 1,2,3-propanetriyl ester Antimicrobial.
22 à-Amyrin Antimicrobial, Antitumor, antiviral,
Anti- inflammatory.
23 9-Hexacosene Antiinflammatory
24 Heptacosane, 1-chloro Anticancer drug
25 Glycine,N-[(3à,5á,7à,12à) 24-oxo-3,7,12-
tris [(trimethylsilyl ,methyl ester Antibacterial.
***Source: Dr. Duke’s phytochemical and ethno botanical databases (online database).
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1. 3-Carene 2. Cycloheptasiloxane, tetradecamethyl
3.1-Hexadecanol 4. Phenol,4,6-di(1,1-dimethylethymethyl
5. Squalene 6. Dibutyl phthalate
7. Phytol 8. 9-Octadecenamide,(Z)-
9. á-Amyrin
Figure-6: Structure of Bioactive compounds
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CONCLUSION
In the present study 25 compounds from the ethanol leaves extract of Achyranthes aspera
were identified by Gas Chromatography Mass Spectrometry (GC-MS) analysis. The
biological activities of each of the identified phytocomponents used for antimicrobial,
antioxidant, anti-inflammatory and pesticide activities. The research findings have shown that
the leaves extract of A. aspera is extensively rich in secondary metabolites. The plant leaves
has a high potential for a vast number of bioactive compounds which justified its use for
various ailments by traditional practitioners.
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