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Gopalakrishnan S and Kalaiarasi T. et al. / I nternational J ournal of Biological & Pharmaceutical Research. 2013; 4(7): 523-527.

e- ISSN 0976 - 3651
Print ISSN 2229 - 7480

International Journal of Biological
&
Pharmaceutical Research
Journal homepage: www.ijbpr.com


DETERMINATION OF BIOLOGICALLY ACTIVE CONSTITUENTS
OF THE FRUITS OF CUCUMI S SATI VUS LINN.
USING GC- MS ANALYSIS

Gopalakrishnan S
1,*
and Kalaiarasi T
2


1*
Department of Chemistry, Noorul Islam University, Kumaracoil-629180, K.K.District, Tamil Nadu, India.
2
Department of Pharmaceutical Chemistry, Manonmaniam Sundaranar University, Tirunelveli-627012, Tamil Nadu, India.

ABSTRACT
The plant Cucumis sativus (Fam.Cucurbitaceae) commonly known as Mullu vellari in Tamil, Sakusa in Sanskrit,
Kheera in Hindi and Cucumber in English is reported to possess a number of medicinal values. The present investigation
was carried out to determine the possible bioactive components of the ethanolic extract of the fruits of Cucumis sativus using
GC-MS analysis. Seven compounds were identified by GC-MS analysis. The major phytoconstituents were 2,3-Dihydro-3,5-
dihydroxy-6-methyl-4H-pyran-4-one (10.86 %), 5-(Hydroxyl methyl)-2-furancarboxaldehyde (64.69 %), 4-Hydroxy-3-methyl-
2-butenyl-acetate (3.08 %), 2-(2-Methylcyclohexylidene)-hydrazinecarboxamide (11.54 %),n-Hexadecanoic acid (2.95 %), 1,2-
Benzenedicarboxylic acid-diisooctyl ester (6.88 %) which are possessing many biological activities. Hence this study creates a
platform to screen many bioactive components to treat liver disorder.

Key Words: Cucumis sativus, GC-MS analysis, Bioactive components, Liver disorder.

INTRODUCTION
Herbal medicines are safe than synthetic
medicines because the phytochemicals in the plant extract
target the biochemical pathway (Zaidan et al., 2005). The
herbs are constantly being screened for their biological and
pharmacological activities such as anti-diabetic, anti-
oxidant, anti-microbial, laxative, and anti-cancer activities
(Evans and Trease, 1997; Anbuselvi et al., 2012; Makky et
al., 2012; Kavitha et al., 2011; Meurer-Grimes et al., 1996;
Koduru et al., 2006). The herbs are having numerous
bioactive components which are identified (at less than
1ng) by using GC or LC-MS.
The cucumber plant is an annual climber which
grows to a height of 15-30 cm and has large leaves that
form a canopy over the fruit. The cucumber is the edible

Corresponding Author

S. Gopalakrishnan
Email: sgkmsu@yahoo.co.in
fruit of the cucumber plant Cucumis sativus, which belongs
to the gourd family Cucurbitaceae. Cucumis sativus fruit is
shown to possess various activities such as anti-
hyperglycemic activity (Roman-Ramos et al., 1995),
inhibitory effects on protein kinase C (PKC) activity
Sudheesh et al., 2007), anti-oxidant activity (Stratil et al.,
2006; Chu YF et al., 2002; Pellegrini et al., 2003),
amylolytic activity (Repka and Fischerova, 1999), anti-
cancer activity (Villasenor et al., 2002), anti-clastogenic
activity (Edenharder et al., 1998), and anti-mutagenicity
activity (Edenharder et al., 1994; Zhao et al., 1992). The
juice is used in many beauty products (Katsambas and
Lotti 2003). C. sativus is amongst the constituents of
cosmetics marketed as treatments for skin inflammations
and other skin disorders, and as skin protectants (Aburjai
and Natsheh, 2003). Taking into consideration of the
medicinal importance of this plant, the ethanol extract of
fruits of Cucumis sativus was analyzed for the first time
using GC-MS. This work will be helpful to identify the
compounds of therapeutic value. GC-MS is
IJBPR
524
Gopalakrishnan S and Kalaiarasi T. et al. / I nternational J ournal of Biological & Pharmaceutical Research. 2013; 4(7): 523-527.

one of the best techniques to identify the bioactive
constituents of long chain branched chain hydrocarbons,
alcohols, acids, ester etc.

MATERIALS AND METHODS
Collection of plant materials
The fruits of Cucumis sativus was collected in the
month of July from Alangulam, Tirunelveli District, Tamil
Nadu and identified by Prof. P. Jayaraman, Plant Anatomy
Research Centre, West Thambaram, Chennai- 600 045,
Tamil Nadu, India.
A voucher specimen (MSU/PHAR/HER141) has
been preserved in the Herbarium of the Department of
Pharmaceutical Chemistry, Manonmaniam Sundaranar
University, Tirunelveli -627 012, Tamil Nadu, India.

Preparation of extracts
The collected fruits were cut into pieces, shade-
dried at room temperature and powdered. The dried fruit
powder (500 g) was successively extracted using
petroleum ether (40- 60C), benzene, chloroform, ethanol
and water by using Soxhlet apparatus. The last trace of
solvent was removed under reduced pressure distillation
and then vacuum dried. The dried crude ethanolic extract
of the fruits has been used for the GC-MS analysis.

GC-MS ANALYSIS
Preparation of extract
2 l of the ethanolic extract of the fruits of
Cucumis sativus was employed for GC-MS analysis
(Merlin et al., 2009).

I nstruments and chromatographic conditions
GC-MS analysis of the ethanolic extract was
carried out on a GC-MS Clarus 500 Perkin Elmer system
comprising a AOC-20i autosampler and gas
chromatograph interfaced to a mass spectrometer (GC-MS)
instrument employing the following conditions: column
Elite-1 fused silica capillary column (30 mm x 0.2 5mm ID
x 1Mdf, composed of 100 % Dimethyl poly siloxane),
operating in electron impact mode at 70 eV; helium (99.
999 %) was used as carrier gas at a constant flow of
1ml/min and an injection volume of 0.5l was employed
(split ratio of 10:1); injector temperature 250C. The oven
temperature was programmed from 110C (isothermal for
2 min), with an increase of 10C/min, to 200C, then 5C /
min to 280C, ending with a 9 min isothermal at 280C.
Mass spectra were taken at 70 eV; a scan interval of 0.5
seconds and fragments from 40 to 550 Da.

I dentification of phytochemical constituents
Interpretation on mass spectra of GC-MS was
conducted using the database of National Institute of
Standards and Technology (NIST). The mass spectra of the
unknown components were compared with those of the
known components stored in the NIST library. The name,
molecular weight and structure of the phytochemical
constituents of the ethanolic extract were ascertained
(Nezhadali et al., 2010; Sathyaprabha et al., 2011).

RESULTS AND DISCUSSION
The biologically active constituents present in the
ethanolic extract of the fruits of Cucumis sativus were
identified by GC-MS analysis (Fig. 1) and the mass spectra
of the six major constituents were obtained (Fig. 2 to Fig.
7). The mass spectra of the constituents were compared
with those of the known spectra which are stored in the
NIST library. The retention time (RT), molecular formula,
molecular weight and peak area (%) are presented in Table
1.

Table 1. Phytoconstituents identified in the ethanolic extract of the fruits of Cucumis sativus by GC-MS analysis.
No. RT Name of the compound
Molecular
formula
Molecular
weight
Peak
Area %
Compound
Nature
**Activity
1 3.61
2,3-Dihydro-3,5-dihydroxy-6-
methyl-4H-pyran-4-one
C
6
H
8
O
4

144 10.86
Flavonoid
fraction
Antimicrobial Anti-
inflammatory
2 4.59
5-(Hydroxymethyl)-
2-furancarboxaldehyde
C
6
H
6
O
3

126 64.69
Aldehyde
compound
Antimicrobial Preservative
3 7.50
4-Hydroxy-3-methyl-2-
butenyl- acetate
C
7
H
12
O
3

144 3.08
Acetate
compound
Antimicrobial
4 10.60
2-(2-Methylcyclohexylidene)-
hydrazinecarboxamide
C
8
H
15
N
3
O
169 11.54
Amino
compound
Antimicrobial
5 13.32 n-Hexadecanoic acid
C
16
H
32
O
2

256 2.95
Palmitic
acid
Antioxidant, Hypo
cholesterolemic
Nematicide, Pesticide,
Lubricant, Antiandrogenic,
Flavor, Hemolytic 5-Alpha
reductase inhibitor
6 24.24
1,2-Benzenedicarboxylic
acid-diisooctyl ester
C
24
H
38
O
4

390 6.88
Plasticizer
compound
Antimicrobial Anti-
fouling
**Source: Dr.Duke's phytochemical and ethnobotanical databases [Online database].
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Gopalakrishnan S and Kalaiarasi T. et al. / I nternational J ournal of Biological & Pharmaceutical Research. 2013; 4(7): 523-527.

Fig 1. GC-MS Chromatogram of ethanol extract of the fruits of Cucumis sativus


Fig 2. Mass spectrum of 2, 3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (RT: 3.61)


Fig 3. Mass spectrum of 5-(Hydroxymethyl)-2-furancarboxaldehyde (RT: 4.59)



Fig 4. Mass spectrum of 4-Hydroxy-3-methyl-2-butenyl-acetate (RT: 7.50)
(mainlib) Acetate, 4-hydroxy-3-methyl-2-butenyl-
20 30 40 50 60 70 80 90 100 110 120 130 140 150

0

50

100
27
31
41
43
55
61
71
84
126
O
HO O

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Gopalakrishnan S and Kalaiarasi T. et al. / I nternational J ournal of Biological & Pharmaceutical Research. 2013; 4(7): 523-527.

Fig 5. Mass spectrum of 2-(2-Methylcyclohexylidene)-hydrazinecarboxamide (RT: 10.60)
(mainlib) Hydrazinecarboxamide, 2-(2-methylcyclohexylidene)-
20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180

0

50

100
26
41
55
61
67
81
95
98
110
126
141
152
169
N
NH
H
2
N O


Fig 6. Mass spectrum of n-Hexadecanoic acid (RT: 13.32)


Fig 7. Mass spectrum of 1, 2-Benzenedicarboxylic acid, diisooctyl ester (RT: 24.24)


In the present study, six phytochemical
constituents have been identified from the ethanol extract
of the fruits of Cucumis sativus by Gas Chromatography-
Mass spectrometry (GC-MS) analysis. All the identified
phytoconstituents, 2,3-Dihydro-3,5-dihydroxy-6-methyl-
4H-pyran-4-one, 5-(Hydroxylmethyl)-2-furan carboxal
dehyde, 4-Hydroxy-3-methyl-2-butenyl-acetate, 2-(2-
Methylcyclohexylidene)-hydrazinecarboxamide, 1,2-
Benzene dicarboxylic acid-diisooctyl ester are reported to
possess antimicrobial activity.n-Hexadecanoic acid has
antioxidant, hypocholesterolemic, nematicide, pesticide,
lubricant, antiandrogenic, flavor, hemolytic 5- reductase
inhibitor activities. Out of these six bioactive constituents
2-Furancarboxaldehyde-5-(hydroxyl methyl) (64.69 %) is
present in high quantity and n-Hexadecanoic acid (2.95 %)
is present in less quantity, and the other constituents are
present in considerable amounts. Thus, this type of GC-MS
analysis is the first step towards understanding the nature
of active principles in this medicinal plant and this type of
study will be helpful for further detailed phytochemical
study.
The investigation concluded that the stronger
extraction capacity of ethanol could have produced number
of active constituents responsible for many biological
activities.

CONCLUSION
The six phytochemical constituents identified by
GC-MS of the ethanolic extract of Cucumis sativus are
medicinally valuable and possess various pharmaceutical
applications. GC-MS analysis of the ethanolic extract
showed the presence of Hemolytic 5- reductase inhibitor
like n-Hexadecanoic acid which may be responsible for the
liver disorder curing effect. The identified
phytoconstituents needs further research on toxicological
aspects to develop safe drug.
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Gopalakrishnan S and Kalaiarasi T. et al. / I nternational J ournal of Biological & Pharmaceutical Research. 2013; 4(7): 523-527.

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