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Document heading doi: 2012 by the Asian Pacific Journal of Tropical Biomedicine. All rights reserved.
Article history: Objective: To characterize the phytochemical constituents of Eupatorium triplinerve using GC
Received 14 August 2012 -MS. Methods: Ten grams of the powdered sample was subjected to column chromatography
Received in revised form 23 August 2012 over silica gel (100-200 mesh) and eluted with n-hexane, chloroform, ethanol and methanol
Accepted 5 October 2012
respectively. n-Hexane and Chloroform did not elute much of the compounds. The methanol
Available online 28 December 2012
fraction of the Eupharbatum triplinerve was taken for GC-MS analysis. The analysis was carried
out on a GC Clarus 500 GC system with a column packed with Elite - 1 (10% dimethyl poly
Keywords: siloxane, 30 0.25 mm ID 1 EM df), the compounds are separated using with Helium as carrier
GC-MS analysis gas at a constant flow 1ml/min. sample extract (2 L) injected into the instrument was detected
Eupatorium triplinerve by Turbo gold mass detector (Perkin Elmer) with the aid of the Turbo mass 5.1 software. Results:
Phytocomponents The GC MS analysis provided peaks of eleven different phytochemical compounds namely
Hexadecanoic acid hexadecanoic acid (14.65%), 2,6,10-trimethyl,14-ethylene-14-pentadecne (9.84%), Bicyclo[4.1.0]
heptane, 7-butyl- (2.38%), Decanoic acid, 8-methyl-, methyl ester (3.86%), 1-undecanol (7.82%),
1-hexyl-1-nitrocyclohexane (2.09%), 1,14-tetradecanediol (6.78%), Octadecanoic acid, 2-hydroxy-
1,3-propanediyl ester (19.18%) and 2-hydroxy- 3-[(9E) -9-octadecenoyloxy] propyl (9E)-9-
octadecenoate (8.79%). Conclusions: The bioactive compounds in the methanolic extract of
Eupatorium triplinerve have been screened using this analysis. Isolation of individual components
would however, help to find new drugs.
2.1 Collection and preparation of plant material volume of 0.5 EI was employed (split ratio of 10:1 injector
temperature 250 ; ion-source temperature 280. The oven
Fresh plants of E. triplinerve were collected from the temperature was programmed from 110 (isothermal for 2
natural habitats of Tiruchirappalli, Tamil Nadu, India. The min). with an increase of 10 /min, to 200 then 5/min to
samples were washed thoroughly in running tap water to 280, ending with a 9 min isothermal at 280. Mass spectra
remove soil particles and other adhered debris and finally were taken at 70 eV; a scan interval of 0.5s and fragments
washed with sterile distilled water. The whole plants were from 40 to 550 Da.
shade dried and ground into fine powder. The powdered
materials were stored in air tight polythene bags until use. 2.3.Identification of components
2.2. Plant sample extraction Interpretation on mass spectrum GC-MS was conducted
using the database of N ational I nstitute S tandard and
Plant sample extraction and Column chromatography Technology (NIST) having more than 62,000 patterns. The
Ten grams of powdered sample was extracted with 50 spectrum of the unknown component was compared with
mL methanol overnight and filtered through ash less filter the spectrum of the known components stored in the NIST
paper with sodium sulphate (2 g). The crude extract was library. The name, molecular weight and structure of the
subjected to column chromatography over silica gel (100-200 components of the test materials were ascertained.
mesh) and eluted with n-hexane, chloroform, ethanol and
methanol respectively. n-Hexane and Chloroform did not 3. Results
elute much of the compounds. The methanol fraction of the 500,000 Chromatogram
Eupharbatum triplinerve was taken for GC-MS analysis.
Gas Chromatography- Mass Spectrum Analysis (GC-MS)
GC-MS analysis was carried out on a GC Clarus 500 Perlin
Elmer system comprising a AOC-20i autosampler and gas
chromatograph interfaced to a mass spectrophotometer (GC -
MS) instrument employing the following conditions: column
17.151
20.148
21.619
15.084
15.752
18.383
19.986
16.960
Elite - 1 fused silica capillary column (30 0.25 mm ID 1
16.201
16.701
EM df, composed of 100% Dimethyl polysiloxane), operating TIC*1.00
100
100 78 60 78
43 60
41
41
HO
HO O 256
129
O 85
129 27 98 213
115 157 171 185
27 85 185 143 227
228
98 115 171
143 157
20 40 60 80 100 120 140 160 180 200 220 240
20 40 60 80 100 120 140 160 180 200 220 240 Figure 2 F.Hexadecanoic acid
Figure 2 A.Tetradecanoic acid
100 O
55 83
69 H
97
41 N O
100
68
111
82 95
167
39
43 125
123 168
41
109 20 40 60 80 100 120 140 160 180 200 220 240 260
137
179 208
278 Figure 2 G.1-hexyl-1-nitrocyclohexane
27 151 193 263
20 40 60 80 100 120 140 160 180 200 220 240 260 100
55
Figure 2 B.2,6,10-trimethyl,14-ethylene-14-pentadecne 41 69
82
HO
OH
95
100
82 109
27 123
27 55
96
10 20 30 40 50 60 70 80 90 100 110 120 130
Figure 2 H.1,14-tetradecanediol
109
152
124
154 100
43 57
o 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360
41
57
o Figure 2 I.Octadecanoic acid, 2-hydroxy-1,3-propanediyl ester
256 100
55
41
69
40 60 80 100 120 140 160 180 200 220 240 260 280
83 OH
o o
Figure 2 D. Decanoic acid, 8-methyl-, methyl ester
97
o o
129 264
27 112
151
100 55 69
41 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
Figure 2 J.2-hydroxy-3-[(9E) -9-octadecenoyloxy]propyl(9E)-9-\
70
octadecenoate
97 OH
and 2,6,10,-trimethyl,14-ethylen-14-pentadecne, while [4] Garg SC, Nakhare S. Studies on the essential oil from the flowers
1-hexyl-1-nitrocyclohexane and 1,14-tetradecanediol other of Eupatorium triplinerve. Indian Perfumer 1993; 37(4): 318 - 323.
compounds show antimicrobial and anti-inflammatory [5] G hani, A . 1998 . M edicinal plants of B angladesh: C hemical
activities. constituents and uses. 1st edn. Asiatic Society of Bangladesh. pp.
T here is growing awareness in correlating the 174.
phytochemical components and their biological activities [6] Fernie AR, Trethewey RN, Krotzky AJ, Willmitzer L. Innovation -
[6,7,8]. E. triplinerve is a plant used in Ayurvedic medicine Metabolite profiling: from diagnostics to system biology. Nat Rev
however there are no reports on the thorough phytochemical Mol Cell Biol. 2004; 5: 763 - 769.
analysis of the plant. We report the presence of some of the [7] Sumner LW, Mendes P, Dixon RA. Plant metabolomics: largescale
important components resolved by GC-MS analysis and phytochemistry in the functional genomics era. Phytochem 2003;
their biological activities. Thus this type of GC-MS analysis 62(6): 817 - 836.
is the first step towards understanding the nature of active [8] Robertson DG. Metabonomics in toxicology: A review. Toxicol Sci
principles in this medicinal plant and this type of study will 2005;85:809 - 822.
be helpful for further detailed study.