Essential Oil Composition of the Fruits of

Galium tricornutum
Abdul Khaliq Jan, 1Mohammad Rasul Jan, 2Mohammad
1

Shakirullah, 3Mohammad Raza Shah and 1Mohammad Ali

Sheringal High Altitude Research Centre University of

1

Malakand, NWFP, Pakistan

2
Institute of Chemical Sciences University of Peshawar,
Pakistan.
3
H.E.J. Research Institute of Chemistry, University of
Karachi, Pakistan
Corresponding author Email: abdukhaliq@gmail.com
Phone +92(944) 885479, Cell 0300-5856017, Fax 0944-
885805
 Introduction

• In recent times, the number of traditional healers

claiming the efficacies of various Galium species for a
variety of purposes, mainly as a diuretic and in the
treatment of some stomach, gout and epilepsy diseases.
• Iridoid in Galium have anti-inflammatory, cardiovascular
activities.
• Cleavers are believed to have mild diuretic effects, which
may reduce stone formation by increasing urinary flow
and volume.
• The word Galium has been derived from the Greek term
”gala” meaning milk, because the species of Galium
coagulate the milk, due the presence of an enzyme.
• For this reason, it is known as “Yogurt herb” in
Turkish and is famous as jeshay at district Dir of
Pakistan.
• Galium species are distributed in Pakistan from
plains to mountains and are used in folklore
remedies.
• The essential oils have been reported in Galium
aparine L. and Galium odoratum L.
• The fruits of Galium tricornutum (Khoug jeshay)
have been chosen for essential oil analysis.
 Experimental
Plant material
• Galium tricornutum (Khoug Jeshay), whole plant,
was collected in village Bunr of District Dir,
Pakistan, (March 2006).
• The Plant materials were identified by Ali Hazrat,
plant taxonomist and research officer at
Sheringal Research Centre.
• Voucher specimens (SHPC-AKJ-40) have been
deposited at the Herbarium of the Sheringal
High Altitude Research Centre, University of
Malakand.
 Isolation of oils
• The ground fruits (200g) of Galium tricornutum
(Rubiaceae) were subjected to steam distillation
using a Clevenger type apparatus for 3 h.
• The oil obtained was extracted with diethyl ether
to obtain the water free oil.
• The oil obtained by normal steam distillation
from Galium tricornutum was analyzed by
GC/MS analysis.
• The yield of oil on dry weight basis was 0.4%.
 GC/MS analysis
• The oil was analyzed on a Hewlett-Packard GCD
system. Innowax FSC column (60 m x 0.25 mm, with
0.25 µm film thickness).
• Helium was used as a carrier gas.
• The GC oven temperature was kept at 60˚C for 10
minutes and programmed to 220˚C at a rate of 4˚C/min
and then kept constant at 220˚C for 10 mins to 240˚C at
a rate of 1˚C/min.
• The Injector temperature was 250˚C. Split flow was
adjusted at 1 ml/min.
• The mass range was recorded from m/z 35 to 425 and
the MS were recorded at 70 eV.
Identification of Compounds
• The components of essential oils were identified
by comparison of their mass spectra with those
of the Baser Library of Essential Oil
Constituents, Wiley GC/MS Library, TBAM
Library of Essential Oil Constituents, Mass
Finder Library and confirmed by comparison of
their retention indices.
• Relative percentage amount were calculated
from TIC by the computer.
• The composition of oil is given in the Table I.
 Results and Discussion
• The oil was characterized with the help of
GC/MS and it was found that the oil composition
of the fruits of Galium tricornutum was differ from
the oils, reported from the leaves of most closely
related genera,Galium aparine and Galium
odoratum .
• From the Relative retention indices (RI), it was
found that oil isolated from the fruits of Galium
tricornutum consist of hexadecanoic acid,
hexadecanoic acid, 4H-1-benzopyran-4-one, 2-
(3,4-dihydroxyphenyl)-6,8-di-a`-D-
glucopyranosyl-5, 7-dihydroxy-, oleic acid,
eicosyl ester etc. (Table I).
• The maximum RI peak appeared at
1534,representing the maximum amount of oleic
acid, eicosyl ester(12.7%).
• The detection of 4H-benzopyrane-4-0ne,2-(3,4-
dihydroxyl phenyl)-6,8-di-`a-D-glucopyranosyl-
5,7-dihydroxy showed the presence of essential
oil in the fruit of Galium tricornutum.
• The detail of the composition of oil yields are
given in Table I.
Table I: Composition of Essential Oil from the fruit of Galium tricornutum

Compound RI % Compound RI %
16-octadecanoic acid, methyl ester 1276 0.4 4H-1-benzopyran-4-one, 2-(3,4- 1790 3.6
dihydroxyphenyl)-6,8-di-a`-D-
glucopyranosyl-5, 7-dihydroxy-
cyclopropanebutanoic acid, 2-[[2-[[2-[(2- 1280 1.2 myrtenol 1800 2.3
paentylcyclopropyl) methyl] cyclopropyl] methyl]-,
methyl ester
octadecanoic acid, 17-hydroxy-, methyl ester geraniol 1860 0.5

hexadecanoic acid 1372 5.6 hexznoic acid 1876 0.7

hexadecanoic acid, 1-(hydroxymethyl)-1,2-ethanediyl 1378 5.2 isomethyl butyrate 1912 1.6
ester
9-octadecanoic acid (Z)-, tetradecyl ester 1435 2 β-ionone 1960 0.2
8-octadecanoic acid, methyl ester, (E)- 1438 1.9 octanoic acid 2090 0.7
octadecanoic acid, 4-hydroxy-, methyl ester 1444 1.6 isothymol(2-isopropyl-4-methyl 2118 0.5
phenol
1510 1.9 nonanoic acid 2192 0.3

1518 5.2 thymol 2205 0.9

9-octadecanoic acid (Z)-, 2-hydroxy-1- 1520 5.5 carvacrol 2230 0.2

(hydroxymethyl) ethyl ester
oleic acid, eicosyl ester 1534 12.7 intermedeol 2248 0.9
9-octadecanoic acid (Z)-, tetradecyl ester 1539 12.3 decanoic acid 2290 0.2
 Summary
• The aim of this study was to evaluate the chemical composition of
the oil from the fruits of Galium tricornutum found in northern
areas of Pakistan, and is locally know as Khog Jeshay.
• The oil obtained (yellowish colored) was extracted with diethyl
ether to obtain the water free oil.
• The oil obtained by normal steam distillation was analyzed by
GC/MS analysis.
• The main components of the hydrodistilled oil were found to be
oleic acid, eicosyl ester (12.7%), 9-octadecanoic acid (Z)-,
tetradecyl ester(12.3%),hexadecanoic acid (5.6%), hexadecanoic
acid, 1-(hydroxymethyl)-1,2-ethanediyl ester (5.2%), 9-
octadecanoic acid (Z)-, 9-octadecanoic acid (Z)-, 2-hydroxy-1-
(hydroxymethyl) ethyl ester(3.7%) , myrtenol,(2.3%) and
tetradecyl ester(2%).
• The presence of 4H-benzopyrane-4-0ne,2-(3,4-dihydroxyl
phenyl)-6,8-di-`a-D-glucopyranosyl-5,7-dihydroxy(3.6%),being an
aromatic indicates that the plant under investigation may be the
source of essential oil.
• To the best of our knowledge it is the first report of the oil
composition of G.tricornutum.