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GC-MS Characterization of Volatile Odorous Compounds in Allium Cepa

Conference Paper · October 2014

DOI: 10.13140/2.1.3278.7523

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GC-MS Characterization of Volatile Odorous Compounds
in Allium Cepa

N.C.J. Packia Lekshmi1, S. Viveka2, M.B. Viswanathan3, G. Manivannan4

and T. Mini Shobi5

Department of Biotechnology, Sathyabama University, Solinganallur, Chennai 600119, Tamil Nadu, India
1

2
Department of Biotechnology, Udaya School of Engineering, Vellamodi, Kanniyakumari 629204,
Tamil Nadu, India
3, 5
Department of Plant Science, Bharathidhasan University, Tiruchirappalli 620024, Tamil Nadu, India
4
Department of Microbiology, NMSSVN College, Nagamalai, Madurai 625019, Tamil Nadu, India
e-mail: [email protected]

Abstract:
An analytical technique for the determination of volatile phytochemical compounds by GC-MS in onion,
Allium cepa L., was reported here. Three different cultivars of onion bulb (O1, O2 and O3) were extracted
with petroleum ether, and the volatile compounds were analyzed by GC-MS. Terpenes such as humulaneand
phytosterols and aldehydes are found common in all the three cultivars. Thiophenes and disulphides were
identified as major compounds in O1 and O3 respectively. Caryophyllene, a terpene was observed in both
O1 and O2. Squalene, allicin and sulphide compounds were identified specifically in O3. The study further
reported that the composition of volatile compounds in the cultivars is varied and these differences may be
due to the variations in geographic, climatic, soil fertility and cultivation practices. The intensity of flavour
is governed by hereditary factors and environmental conditions under which the onions grow.
Keywords: Phytochemicals, GC-MS, onion, Terpenes, Disulphides.

INTRODUCTION
Allium is the prime and most important genus in the family of Alliaceae with about 450 species. Technical
investigation on these plants was started in the middle of 19th century. Allium species are rich sources of
phytonutrients [1]. Onions occupy a major portion in food market. Cultivars and typologies of onion can
diverge from a chemical point of analysis and variations can be important in their classification [2].
Bulbs of Allium species are broadly used as food flavouring agents and as remedial agents because of its
valuable compounds such as allicin, alliin, flavonoid and their derivatives [3-7]. The oily substance which
has the characteristic onion odour from bulb of Allium cepa was di- and trisulphide [8] and thioaldehydes
[9]. These odoriferous volatile lipid compounds were analyzed by several research groups [10]. The nature
and composition of volatile components of onion be still contentious. Geographic, climatic and differences
in varieties may also play a vital part in the proportions of volatile compounds of onion [10].
The volatile lipid soluble organo sulphur compounds were analysed using GC-MS [11]. This method is
used to measure secondary metabolites formed during enzymatic/metabolic pathways such as disulphides
and trisulphides. The analytical strategy (GC-MS) used to analyse the volatile compounds, and selected
compounds were structurally analysed by MS transposing the method to GC-MS. In this paper, we evaluated
the organic volatile compounds which are responsible for biological activity, flavour and odour in this
particular medicinally valuable onion by GC-MS.
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MATERIALS AND METHODS

Extraction of organo-sulphur compounds (using non-polar solvent of petroleum ether)
The plant samples were collected from different cultivation localities such as Alankulam (O1) and Surandai
(O2) in Tirunelveli District and Vilathikulam (O3) in Tuticorin district of Tamil Nadu. The extraction of
organo-volatile compounds was accomplished using Soxhlet apparatus in which the polar barrier was broken
up and allowed the solvent to reach the non-polar compounds in order to achieve their extraction. Often, the
sample was dried, ground and prepared for the extraction.
GC-MS analysis
The filtered plant extracts were subjected to GC-MS analyses such as petroleum ether, chloroform, ethyl
acetate and methanol. GC-MS was carried out on an HP 5890 GC system coupled to a Quadrupole Mass
Detector. Helium was used as a carrier gas in a constant flow mode at 1ml/min. The initial temperature of
the column was 70°C, which was gradually increased by 10°C up to 280°C. The instrument was set to an
initial temperature of 70°C and maintained at this temperature for 2 min. At the end of this period, the Oven
temperature was raised up to 280°C, at the increased rate 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 70eV.
Separation was achieved by RTS-volatile column about 30 m long. Quadrupole Mass Detector was employed
to detect compounds when they were vented from the column. Temperature of the detector was 300°C. Using
data library such as WILEY8.LIB and NIST 08 database, the mass spectra obtained through GC–MS, were
analysed, and the volatile compounds of the plant samples were identified.

RESULTS AND DISCUSSION

The volatile organic compounds from petroleum ether extracts of onion cultivars were identified by GC-MS
analysis since this method was found to be a great value in the identification of organic volatile compounds
(Table 1).

Table 1: List of volatile compounds identified by GC-MS

S.No. R.Time Area Area% Compound name Present in

1 8.642 2697349 3.55 Undecane O1, O2 and O3
2 50.242 6081063 8.01 Gamma-Sitosterol O1, O2 and O3
3 19.605 829867 1.09 Phenol O1, O2 and O3
4 32.194 128551 0.17 Octadecanoic acid (Oleic acid) O1, O2 and O3
5 49.143 4996524 6.58 Ergost-8-en-3-ol O1, O2 and O3
6 49.606 9601417 12.65 9,19-Cycloergost-24(28(-en-3-ol O1, O2 and O3
7 48.008 172143 0.23 Cholesta-4,6-dien-3-ol O1, O2 and O3
8 48.609 2825006 3.72 17,(1,5-Dimethyl-hexyl)-10 O1, O2 and O3
9 48.457 341303 1.83 Cholesterol (Phytosterol) O1, O2 and O3
10 17.225 89109 1.13 Humulane (Sesquiterpenes) O1, O2 and O3
11 24.118 251871 0.33 Myristic acid O1, O2 and O3
12 50.247 106771 0.57 Fucosterol (Phytosterol) O1, O2 and O3
13 49.143 4996524 6.58 Ergost-8-en-3-ol, 14-methyl-, (3 beta,5 alpha.)- O1, O2 and O3
14 32.060 327236 1.76 Fumaric acid O1, O2 and O3
15 36.736 85464 0.46 Oxalic acid O1, O2 and O3
16 9.655 593153 0.78 2-Mercapto-3,4-dimethyl-2,3-dihydrothiophene O1
17 17.904 2533702 3.34 3(2H)-Furanone O1
18 19.007 224765 0.30 Thiophene O1
19 19.358 914661 1.20 Ethanone O1
20 22.177 543096 0.72 1,2-Dimethoxy-4-(2-methoxyethenyl)benzene O1

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21 24.118 251871 0.33 Tetradecanal O1

22 29.625 110637 0.15 1,2-Benzenedicarboxylic acid O1
23 33.411 207295 0.27 Tricosane O1
24 35.110 384977 0.51 Octadecane O1
25 36.742 332279 0.44 Tetratriacontane O1
26 17.223 776709 4.17 Caryophyllene (Sesquiterpene) O1 and O2
27 35.432 1260074 1.66 Octanamide O1 and O2
28 28.400 1955725 2.58 7,9-Di-tert-butyl-1-oxaspiro(4,5)deca-6,9-diene-2,8-dione O1 and O2
29 32.501 157762 0.21 Decanamide (Capramide) O1 and O2
30 29.388 187318 0.25 n-Hexadecanoic acid O1 and O2
31 50.851 14138860 18.63 9,19-Cyclolanost-24-en-3-ol O1 and O3
32 48.181 169854 0.22 Tetratetracontane O1 and O3
33 24.214 117164 0.63 Pentadecanal- O2
34 31.636 100535 0.54 1-Hexadecyne O2
35 29.249 1036092 5.56 Dibutyl phthalate O2 and O3
36 28.466 101132 0.54 Heptadecanoic acid O2 and O3
37 8.179 4447752 0.18 Diallyl disulphide (Sulfides) O3
38 8.17 311631 0.63 Allicin (sulphides) O3
39 44.108 100051 0.52 Squalene (Triterpene) O3

Most of the identified compounds were present in all the three cultivars. The important ones among
them were gamma-sitosterol, ergos-8-en-3-ol, cholesta-4,6-dien-3-ol, cholesterol, humulene, myristic acid,
fucosterol, fumaric and oxalic acids. Thiophene, an active compound specifically identified in O1. Similarly,
the volatile compounds, disulphides and thiophenes were identified in yellow onions by Jarvenpaa et al. [12]
in their solid phase micro-extraction combined GC-MS study and by Boelens et al. [13]. Caryophyllene, a
terpenoid was identified in both O1 and O2. Liu et al. [14] reported the presence of aromatic compounds
caryophyllene, thiophene and other terpenoids by GC-MS analysis (Figs.1-3).

Figure 1: GC-MS chromatogram of O1

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Figure 2: GC-MS chromatogram of O2

Figure 3: GC-MS chromatogram of O3

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The cultivar O3 showed the presence of volatile sulphide compounds –allicin, diallyl disulfide and a
terpeneof squalene. Keusgen [15] identified the volatile compounds of allicin and sulphides. Most of the
disulphides and thiol groups in onion were also determined by Marie Lokke et al. [16]. In this study, we
identified the precursor molecules for the synthesis of the volatile sulphide and terpenoids in all the three
cultivars. Besides, we also determined the presence of ethanol, isoamyl acetate, isobutyl alcohol, propyl
alcohol, palmitic acid, stearic acid, and lanosterol in the onion cultivars. Similar results were reported by
Ohta and Osajima [17]. They detected the typical sulphur containing aldehyde compounds, di- and trisulphide
in onion (Fig. 4).

Figure 4: Chemical structure of volatile compounds

The volatile compounds of onion were studied by many researchers [18-20]. The potential antibiotic
and flavour properties of these volatile compounds in onion received greater attention [21]. However, there
is a disparity in the identified compounds among the three cultivars. These variations may be due to the
variations in geographic, climatic, soil fertility and cultivation practices. The intensity of flavour is governed
by hereditary factors and environmental conditions under which the onions grow. The ability to intake sulfate
and the efficiency of producing flavour precursors differs among the onion cultivars [1]. Increased flavour
intensity in onion was contributed by improved sulphate fertility, dry growing conditions and increased
growing temperatures.

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CONCLUSION
In the present study, an effective and easy method for the analysis of volatile compounds in red onion using
GC-MS is reported here. This method is very sensitive and reproducible to identify the phytocompounds
with accuracy and precision. Since this medicinal plant, onion, possesses a lot of phytochemicals, it can be
a superior and easily accessible material for nutraceuticals.

ACKNOWLEDGEMENT
The authors are thankful for the Jawaharlal Nehru University, New Delhi, for providing GC-MS analytical
data.

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