Sharma (2018)
Sharma (2018)
Sharma (2018)
TM
ISSN: 2321-3272 (Print), ISSN: 2230-7605 (Online)
IJPBSTM | Volume 8 | Issue 4 | OCT-DEC | 2018 | 889-903
Research Article | Biological Sciences | Open Access | MCI Approved|
|UGC Approved Journal |
KEY WORDS
GCMS; Fatty acids; sesquiterpenoid; Spilanthes ciliata
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made into powder in a warring blender to yield S. ciliata 3. RESULTS AND DISCUSSION
floral powder. A picture of the plant used in the current study is
2.2 Preparation of hexane fraction presented (Figure 1). The plant has several features
200g of floral powder was extracted with methanol (2L, common to its close relatives Spilanthes paniculata and
100%) by incubatiing for 72h at 28±2oC temperature. Spilanthes calva [8]. Several species cultivated under
The methanol solution was then filtered through the name of Spilanthes were differentially identified.
Whatman No.1 filter paper and the filtrate obtained was For example based on phenotypic details and
rotary evaporated to obtain the crude extract. The description, S. ciliata has been used as synonym for S.
crude extract was then subjected to normal phase acmella, S. mauritiana and S.calva [8]. Similarly, S.
chromatography using silica (200-400) gel column with oleracea is used as a synonym for A. oleracea and S.
hexane as mobile phase. The hexane extract was acmella [9]. We propose a systematic study of all the
evaporated, and a yellow oil was obtained. An aliquot of species under Spilanthes/Acmella at molecular and
the oil was mixed with fresh hexane and subjected to phytochemical level is needed for fine tuning the
GCMS. species confirmation. Nevertheless, the crop of the
2.3 GCMS Analysis cultivated Spilanthes species is mostly used as
The mass analysis of the extracted oil was performed toothache plants and the determination of
using Perkin Elmer GCMS (Model: Clarus 680). GC used phytoconstituents of the plant and their biological
in the analysis employed a fused silica column (30m x properties is a subject of extensive research in many
0.25mm, ID: 250μm) packed with Elite-5MS (5% labs around the globe [10,11].
biphenyl 95% dimethylpolysiloxane) and the
components were separated using helium as carrier gas
set at a constant flow of 1 ml/min. The injector
temperature was set at 260°C during the
chromatographic run. 1μL of the sample was injected
into the instrument and oven temperature varied from
60°C (2 min) to 300°C with increment set at 10°C/min
and a hold of 6 min at 300°C. The mass detector
parameters include transfer line and ion source
temperature both set at 240°C, ionization mode
electron impact at 70 eV and a scan time 0.2 sec with a
scan interval of 0.1 sec. The spectrums of the
components were compared with the database of
spectrum of known compounds available in
the GCMS NIST (2008) library.
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Figure 1: Spilanthes ciliata crop at flowering stage growing in the experimental garden of Department of
Biotechnology, Pondicherry University
Figure 2: GC chromatogram showing peaks for different compounds present in hexane fraction of flower extract
of S. ciliata
A picture of chromatogram obtained after GC analysis is terpenoids were present in the extract. Barring
presented (Figure 2). An observation of the eicosanoic acid all other compounds are new report for
chromatogram reveals presence of many distinct peaks S. ciliata. 11-tridece-1-ol constituted the most abundant
indicating presence of several compounds in the hexane (18.72%) component in the extract. This was followed
fraction of the flower extract. Using GCMS coupled with by 2R-acetoxymethyl-1,3,3-trimethyl-4T-(3-methyl-2-
NIST library search a total of 15 different compounds buten-1-yl)-1T-cyclohexanol, Pentadecanoic acid, 1-
could be identified. The fragmentation pattern of the hexacosanol, Nonadecane, Hexatriacontane,
individual compounds along with their deduced 4,4,6a,6b,8a,11,11,14b-Octamethyl-
structure is presented (Figure 3). The compounds 1,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-
identified by GCMS followed by library search with octadecahydro-2H-picen-3-one, Sulfurous acid, 2-
name, molecular formula, molecular weight and their propyl tridecyl ester, 3-O-Acetyl-6-methoxy-
potential biological properties is presented (Table 1). cycloartenol and 2,4,4-Trimethyl-3-hydroxymethyl-5A-
Among the compounds identified there were a number (3-methyl-but-2-enyl)-cyclohexene while others were in
of fatty acids, esters, fatty alcohols, higher alkanes and trace amount. This study revealed 14 new compounds
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in S. ciliata of which 12 were oxygenated. Earlier in S. appear to be associated with antineoplastic, anti-
acmella, there were 20 compounds identified in the inflammatory, anti-microbial, anti-cancerous and other
essential oil with only terpen-4-ol being oxygenated biological properties (Table 1). The sesquiterpenoid (2R-
[12]. Later on, in S. acmella 44 compounds and in A. acetoxymethyl-1,3,3-trimethyl-4t-(3-methyl-2-buten-1-
oleracea 71 compounds were identified [8,9]. In another yl)-1tcyclohexanol) found in the oil is a potential
study the hexane fraction of A. oleracea obtained anticancer agent [17]. Another compound identified in
following a similar procedure as ours was found to the oil sample namely 2,4,4-Trimethyl-3-
contain several fatty acids as is reported here [13]. But hydroxymethyl-5A-(3-methyl-but-2-enyl)-cyclohexene
there were differences in number and type of satisfies the rule of five and indicates favorable
compounds identified in the present and other studies bioavailability as determined by molinspiration
cited above which might be attributed to the difference calculation and is predicted to be a substrate for
in isolation procedures followed. We infer that applying Cytochrome P450 2J (CYP2J) enzyme which are expressed
different procedures for isolation and fractionation in heart tissues [18,19]. Moreover, similar esters have
would facilitate identification of diverse array of also been also reported in members of Malvaceae [20].
compounds with some of them becoming potential lead Thus, our study paves the way for large scale isolation
for developing new drugs. Often hydro-distillation of these compounds and subjecting them to validation
method [7,8, 14-16] or gradient elution [13] have been for clinical application.
used. The new compounds identified in our study
Table 1: List of compounds identified from flower extract of S. ciliata and their associated biological properties
Area Chemical Molecular Molecular Bioactivity
S.No RT Compound Name Reference
(%) Nature Formula Weight Properties
Pentafluoropropi
1 18.53 2.720 onic acid, Ester C21H37O2F5 416 NR NA
octadecyl ester
Peroxisome
proliferator
activated
receptor
2 19.4 1.850 Eicosanoic acid Fatty acid C20H40O2 312 delta, alpha 21
and gamma
(PPAR-
d/a/g)
agonist
Peroxisome
proliferator
activated
Pentadecanoic
3 19.5 8.368 Fatty acid C15H30O2 242 receptor 22
acid
delta
(PPARd)
agonist
Neuromusc
ular
4 19.84 8.320 1-hexacosanol Fatty alcohol C26H54O 382 23
regeneratio
n
Bleaching
agent,
5 19.94 2.446 Lauroyl peroxide Peroxide C24H46O4 398 Polymerizati 24
on catalyst,
Plasticizer
Tetratetracontan Higher Anti-
6 20.58 2.849 C44H90 618 25
e alkane microbial
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Figure 3: GCMS fragmentation pattern and deduced chemical structure of the identified compounds
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Received:04.08.18, Accepted: 07.09.18, Published:01.10.2018
*Corresponding Author:
Neelakantan Arumugam*
Email: [email protected]
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