Antioxidants 2013, 2, 384-397; doi:10.3390/antiox2040384
OPEN ACCESS

antioxidants
ISSN 2076-3921
www.mdpi.com/journal/antioxidants
Article

Chemical Composition and Antioxidant Activity of Essential
Oils from Cinnamodendron dinisii Schwacke and
Siparuna guianensis Aublet
Milene Aparecida Andrade 1, Maria das Graças Cardoso 1,*, Juliana de Andrade 1,
Lucilene Fernandes Silva 1, Maria Luisa Teixeira 1, Juliana Maria Valério Resende 2,
Ana Cristina da Silva Figueiredo 3 and José Gonçalves Barroso 3
1

2

3

Department of Chemistry, University of Lavras, Lavras, MG 37200-000, Brazil;
E-Mails: [email protected] (M.A.A.); [email protected] (J.A.);
[email protected] (L.F.S.); [email protected] (M.L.T.)
Department of Food Science, University of Lavras, Lavras, MG 37200-000, Brazil;
E-Mail: [email protected]
Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, Instituto de Biotecnologia e
Bioengenharia, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016 Lisboa, Portugal;
E-Mails: [email protected] (A.C.S.F.); [email protected] (J.G.B.)

* Author to whom correspondence should be addressed; E-Mail: [email protected];
Tel.: +55-35-3829-1202; Fax: +55-35-3829-1271.
Received: 31 August 2013; in revised form: 8 October 2013 / Accepted: 28 October 2013 /
Published: 26 November 2013

Abstract: The objectives of this study were to chemically characterize and evaluate the
antioxidant activity of essential oils Cinnamodendron dinisii Schwacke (pepper) and
Siparuna guianensis Aublet (negramina). The essential oil was isolated by hydrodistillation
using a Clevenger modified apparatus, and the identification and quantification of
constituents, through GC/MS and GC-FID analysis. The antioxidant activity was evaluated
using -carotene/linoleic acid system and the DPPH radical sequestering method. In
chromatographic analysis, the majority constituents found in the essential oil of C. dinisii
were bicyclic monoterpenes, -pinene (35.41%), -pinene (17.81%), sabinene (12.01%)
and sesquiterpene bicyclogermacrene (7.59%). In the essential oil of the fresh leaves of
Siparuna guianensis Aublet, acyclic monoterpene, -myrcene (13.14%), and
sesquiterpenes, germacrene-D (8.68%) and bicyclogermacrene (16.71%) were identified.
The antioxidant activity was low by the -carotene/linoleic acid test and was not evidenced
by the DPPH test, for both oils evaluated.

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Keywords: volatile oils; chemical composition; free radicals

1. Introduction
The use of medicinal plants by humans dates back thousands of years due to their medicinal and
nutritional properties. Many natural compounds extracted from plants have important biological
activities. Among these compounds, we highlight the essential oils, which are increasingly attracting
the attention of various segments of industry due to their multiple functions, especially antioxidant and
antimicrobial activities.
Essential oils are defined by ISO as the products obtained from different parts of plants through
distillation by steam distillation, hydro distillation and pressing the citrus fruit pericarp techniques.
They are complex mixtures of volatile and lipophilic substances, formed by the secondary metabolism
of plants characterized by a pleasant odor most of them presented [1,2].
Essential oils are marketed by various companies as raw material for various products with
applications in perfumery, cosmetics, foods, and as adjuncts in medicines, among others. There are
approximately 300 essential oils of commercial importance in the world. In the food industry, essential
oils, besides imparting aroma and flavor to food, have important antioxidant activity, a property that
further encourages its use [3].
In foods, lipid peroxidation is responsible for the development of unpleasant flavors and odors,
making them unfit for consumption, and causes other changes that may affect the nutritional quality
due to degradation of fat soluble vitamins and essential fatty acids, as well affecting the integrity and
safety of food. In biological systems, lipid peroxidation on cell membrane unsaturated lipids causes
membrane damage, disrupting the metabolite exchange mechanisms, and may cause cell death,
becoming largely responsible for early aging and cardiovascular disease, cataracts, immune system
decline and brain dysfunction, among others [4,5].
To prevent lipid peroxidation of foods, synthetic antioxidants commonly used in the industry are
BHA, BHT, TBHQ and PG. However, studies assessing the toxicology of these compounds have
demonstrated their carcinogenic potential in animals. Given the evidence of problems that can be
caused by the consumption of synthetic antioxidants, research has emerged with the goal of finding
natural products with antioxidant potential, which are an alternative to substitute the synthetic
compounds or even promote an association between them, order to reduce their amount in food. One
advantage of natural antioxidants versus the synthetic is that legislation regarding them is more
flexible, but the natural antioxidants, in most cases, have lower antioxidant capacities than those of
synthetic compounds [4,6].
The antioxidant activity of volatile oils, even very pronounced, can hardly be attributed to the
components alone since due to the complex chemical composition of these natural compounds, which
may contain molecules with different functional groups, the magnitude of the antioxidant activity
shown by them can be related to the effect caused by the interaction of all constituents present in the
essential oil, from those present in a greater proportion to those present in minor amounts. This
interaction may produce a synergistic effect, when the interaction enhances the effect of the oil, or

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antagonistic, when the interaction negatively affects the antioxidant activity of the oil in study, which
makes it very important to investigate the antioxidant properties of essential oils without considering
only its major components [7].
The Cinnamodendron dinisii Schwacke species is a tree, 10­20 feet high, of the family Canellaceae,
commonly known as "pepper" or "cure-all", because the bark of the trunk, with a spicy flavor like true
pepper, possesses medicinal properties and is slightly numbing [8,9].
The species Siparuna guianensis Aublet is used as a medical resource medicinal among the
population of Mato Grosso, where the decoction of its leaves is used to combat sinusitis symptoms,
fever, rheumatism, migraine, flu, body aches and "malina". It is popularly known as "negramina" and
belongs to the family Siparunaceae [10].
Given the growing importance of essential oils in the world market, their biological potential and
existing species diversity as yet unexplored, further studies are needed that make their use viable. The
objectives of this study were to chemically characterize and evaluate the antioxidant activity of
essential oils of C. dinisii and S. guianensis.
2. Experimental Section
2.1. Plant Material
Leaves Cinnamodendron dinisii and Siparuna guianensis were collected in February 2011, in the
morning, in the Medicinal Plant Garden of UFLA and on the UFLA campus respectively. The
C. dinisii collection site has the following coordinates: -21°13'49.0476" latitude, -44°58'27.4764"
longitude and 933 m of altitude, and the S. guianensis collection site: -21°13'41.9952" latitude,
-44°58'9.0048" longitude and 951 m of altitude.
The species were duly identified and a voucher specimen of each species is registered in with the
ESAL Herbarium, located in the Biology Department of UFLA, with the following registration
numbers: 26,285 (Cinnamodendron dinisii Schwacke) and 26,623 (Siparuna guianensis Aublet).
The collected material was sent to the Laboratory of Organic Chemistry--Essential Oils of the
Chemistry Department, Federal University of Lavras (UFLA) Lavras/MG, in which mature
healthy leaves were selected with no injuries and blemishes caused by pathogens, insects or sunburn.
The plant material was properly packaged and maintained under refrigeration (7 °C) until extraction
of essential oils.
2.2. Essential Oil Extraction
The essential oil extractions were carried out in the Laboratory of Organic Chemistry--Essential
Oils, Federal University of Lavras. The extraction method was hydro distillation using a modified
Clevenger apparatus [11].
Three repetitions of the extraction from each plant were conducted using 300 g of fresh, chopped
leaves for each repetition, distilling for 2 h. After this time, the oil was separated from the hydrolyte by
centrifugation at 1100 g for 5 min, using a bench top centrifuge with horizontal crosspiece (Fanem
Baby® I Model 206 BL). The oil was removed with the aid of a Pasteur pipette, placed in a glass
bottle, stored under refrigeration (4 °C), and protected from light [12].

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2.3. Identification and Quantification of the Essential Oil Constituents
The identification of the constituents of the essential oils was performed by gas-liquid
chromatography coupled to mass spectrometry (GC-MS), using an Autosystem XL equipped with a
DB-1 fused silica column (30 m × 0.25 mm ID, film thickness 0.25 m; J & W Scientific Inc., Rancho
Cordova, CA, USA) connected to a Perkin-Elmer Turbomass. The oven temperature was programmed
from 45 to 175 °C in increments of 3 °C/min and subsequently at 15 °C/min to 300 °C. On reaching
300 °C, the temperature was kept isothermal for 10 min, temperature of the transfer line, 280 °C;
temperature of the ionization chamber 220 °C, helium carrier gas, adjusted to a linear velocity of
30 cm/s; split-flow ratio 1:40, ionization energy 70 eV, ionization current, 60 A; mass range, 40­300 u
(atomic mass unit), scan time 1 s. The compounds were identified by comparison of their retention
indices, relative to those of n-alkanes C9-C21 and by comparison with a library of mass spectra
developed in the laboratory of the Centre for Plant Biotechnology, Faculdade de Ciências da
Universidade de Lisboa--Portugal [13].
The content of each constituent was determined by gas chromatography (GC-FID) in a Perkin
Elmer 8700 gas chromatograph equipped with two Flame Ionization Detectors (FID), a data processing
system and an injector, in which two columns of different polarity were installed: DB-1 fused silica,
with immobilized methyl silicone phase (30 m × 0.25 mm ID, film thickness 0.25 m; J & W Scientific
Inc.) and DB-17HT fused silica (30 m × 0.25 mm ID, film thickness 0.25 mm; J & W Scientific Inc.).
The oven temperature was programmed from 45 °C to 175 °C in increments of 3 °C/min and
subsequently at 15 °C/min to 300 °C. On reaching 300 °C, the temperature was kept isothermal for
10 min. Injector and detector temperature, 290 °C and 280 °C, respectively. Hydrogen was used as
carrier gas adjusted to a linear velocity of 30 cm/s. Split-flow ratio of 1:50. The percentage of oil
constituents was determined by integration of peak areas without using correction factors. The values
shown correspond to the average value of two injections [13].
2.4. Determination of Antioxidant Activity
2.4.1. Reducing Method of Stable Radical DPPH
A DPPH methanolic solution was prepared at a concentration of 40 µg mL-1. For the evaluation,
2.7 mL of the stock DPPH solution were added in a test tube, followed by the addition of 0.3 mL of
each dilution of the oil in methanol (300, 250, 200, 150, 100, 50; 25 g mL-1). In parallel, the control
was prepared containing all reagents except the essential oil. After 60 min, readings were taken using a
spectrophotometer (Shimadzu UV-160 1PC) at a wavelength of 517 nm [14].
The antioxidant activity was calculated as percentage inhibition of DPPH, using the following equation:
%I = 100 - (DPPHs/DPPHw/s)/100

(1)

where %I is the inhibition percentage of the DPPH radical, DPPHs corresponds to the absorbance of
DPPH with the sample and DPPHw/s the absorbance of DPPH without sample (blank).
For comparison, activities of BHT, ascorbic acid and thymol standards were evaluated.

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2.4.2. Assay of -Carotene/Linoleic Acid Oxidation System
In a round bottom flask, 60 µL of linoleic acid, 600 mg of Tween 20, 6 mg of -carotene and 30 mL
of chloroform were added. All the chloroform was removed using a rotary evaporator with a water
bath at 50 °C (Büchi Rotavapor R 114). Subsequently, the residue was dissolved in 150 mL of distilled
water saturated with oxygen under vigorous stirring. In test tubes, 2.7 mL of this solution were added to 0.3
mL of each oil (300, 250, 200, 150, 100, 50, 25 µg/mL), the control comprised only of methanol. The
absorbance was measured immediately in a spectrophotometer (Shimadzu UV-160 1PC) at 470 nm [15].
After reading the initial absorbance, the tubes were incubated in a water bath at 50 °C for the
oxidation reaction, a second reading being conducted after 60 min of incubation. All readings were
performed in triplicate.
According to Wang et al. [16] the antioxidant activity was expressed as percentage of inhibition
after 60 min of incubation using the following equation:
%AA = 100 × (Drc - Drs)/Drc

(2)

where %AA is the inhibition percentage of -carotene degradation, Drc is the degradation ratio of the
control [(ln(a/b)/60], Drs the degradation ratio in the presence of the sample [(ln(a/b)/60], "a"
corresponds to absorbance at time zero and "b" corresponds to absorbance at 60 min.
For comparison, activities of BHT, ascorbic acid and thymol standards were evaluated.
2.4.3. Statistical Analysis
For both tests, a completely randomized design (CRD) was used with seven concentrations and
three replicates for each sample (or standard). The SISVAR [17] statistical program was used.
Data were subjected to analysis of variance and the averages obtained were subjected to regression to
5% probability.
The adjusted equations were used to calculate the IC50 and graphs were plotted with the %I values
of the DPPH assay, or %AA for the -carotene/linoleic acid system versus the concentrations analyzed
using the software Origin 6.0.
3. Results and Discussion
3.1. Identification and Quantification of the Constituents of Essential Oils
We identified 39 constituents present in the essential oil of C. dinisii, which are shown in Table 1. It
can be observed that the essential oil presents almost entirely monoterpene hydrocarbons (76.20%),
presenting as majority components bicyclic monoterpenes, -pinene (35.41%), -pinene (17.81%),
sabinene (12.01%) and sesquiterpene bicyclogermacrene (7.59%). We noted the presence of a
drimane-type sesquiterpene, drimenol (0.20%), present in the composition of many species of Canellaceae.
In the essential oil of the fresh leaves of S. guianensis, 41 constituents were identified (Table 1),
which are composed mainly of sesquiterpene hydrocarbons (41.50%), oxygenated sesquiterpenes
(19.40%) and monoterpene hydrocarbons (17.90%), respectively. As majority compounds, acyclic
monoterpene, -myrcene (13.14%) and sesquiterpenes germacrene-D (8.68%) and bicyclogermacrene
(16.71%) were identified.

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Table 1. Chemical composition of the essential oil of fresh leaves of Cinnamodendron
dinisii and Siparuna guianensis.
RIc *
921
924
930
938
958
963
975
995
1000
1002
1003
1004
1009
1005
1017
1027
1035
1037
1064
1066
1074
1098
1099
1106
1110
1110
1114
1121
1134
1148
1153
1159
1168
1265
1275
1332
1334
1345
1375
1379
1388

Compound
Tricyclene
-Thujene
-Pinene
Camphene
Sabinene
-Pinene
-Myrcene
-Phellandrene
-3-Carene
-Terpinene
p-Cymene
1,8-Cineole
Limonene
-Phellandrene
cis--Ocimene
trans--Ocimene
-Terpinene
trans-Sabinene hydrate
Terpinolene
cis-Sabinene hydrate
Linalool
-Campholenal
trans-p-2-Menthen-1-ol
trans-Pinocarveol
allo-Ocimene
cis-p-2-Menthen-1-ol
trans-Verbenol
Pinocarvone
Borneol
Terpinen-4-ol
Myrtenal
-Terpineol
Myrtenol
Bornyl Acetate
2-Undecanone
-Elemene
-Terpinyl Acetate
-Cubene
-copaene
-Bourbonene
-Elemene

C. dinisii
t ***
1.06
35.41
0.71
12.01
17.81
1.46
t
0.24
1.21
4.37
1.54
t
1.99
1.82
0.75
0.15
0.17
0.15
0.65
t
0.09
0.11
0.03
t
0.03
t
0.36
2.50
t
0.11
0.05
0.10
0.32
0.32
0.11
0.21

%I **
S. guianensis
t
t
1.83
0.04
t
0.86
13.14
0.03
0.72
t
1.23
0.06
0.03
t
t
t
1.69
0.58
0.04
0.27
0.31
2.08

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Table 1. Cont.

1385
1400
1414
1426
1428
1428
1447
1456
1455
1469
1474
1476
1487
1493
1494
1500
1500
1505
1505
1549
1533
1551
1561
1566
1569
1600
1616
1620
1626
1656
1764
-

-Cubebene
-Gurjunene
-Caryophyllene
-Elemene
-Copaene
Aromandrene
-Humulene
allo-Aromadendrene
trans--Farnesene
trans-Cadina-1(6)-4-diene
Germacrene-D
-Selinene
Bicyclogermacrene
Curzerene
-Muurolene
-Muurolene
(trans,trans) -Farnesene
-Cadinene
trans-Calamene
-Cadinene
trans-Nerolidol
Germacrene-B
Spathulenol
-Caryophyllene Oxide
Globulol
Viridiflorol
Humulene epoxide II
1-epi-Cubenol
T-Cadinol
-Eudesmol
-Cadinol
-Bisabolol
Drimenol
Atractylone
Total identified (%)
Monoterpene hydrocarbons
Oxygenated monoterpenes
sesquiterpene hydrocarbons
Oxygenated sesquiterpenes
Others

1.88
0.23
t
0.20
t
7.59
0.09
0.14
0.05
1.88
0.42
0.32
0.16
0.20
99.00
76.20
9.00
10.80
3.00
1.00

0.18
t
1.12
0.05
0.04
0.04
2.07
0.05
t
8.68
0.20
16.71
2.15
t
1.17
2.13
0.29
1.04
2.34
4.16
0.45
0.40
3.00
0.63
0.15
4.14
1.02
1.95
3.53
t
80.48
17.90
0.00
41.50
19.40
1.70

* RIc = retention index calculated; ** % = concentration in percentage; *** t = traces.

Some research was carried out to evaluate the chemical composition of essential oils obtained from
different plant parts of the same species (or species belonging to the same family) evaluated in this
study, as can be seen in Table 2.

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Table 2. Chemical composition of the essential oils obtained from different plant parts of
species of the Canellaceae family and of the Siparuna guianensis specie found by
others authors.
Authors

Plant

Part of
plant used

Place

Chemical composition
(main compounds)

Torres
et al. [18]
Adams
and
Zanoni
[19]
Tucker
et al. [20]

Capsicodendron
dinisii

Bark

Guarapuava
(PR)--Brazil

Monoterpenes--(68.5% of limonene).

Cinnamodendron
ekamani

Wood

Hispaniola--
Caribbean
island

1,8-cineole, -humulene, -caryophyllene,
4-terpineol, germacrene-D, -elemene,
-pinene and -terpineol.

Cinnamosma
fragrans

Commercial
essential oil

Madagascar

1,8-cineole and sabinene.

Setzer [21]

Canella winterana

Leaves

Amiguet
et al. [22]

Pleodendron
costaricense

Leaves and
bark

Valentini
et al. [23]

Siparuna
guianensis

Leaves

Antônio
et al. [24]
Rebouças
[25]
Montanari
[26]
Fischer
et al. [27]

Siparuna
guianensis
Siparuna
guianensis
Siparuna
guianensis
Siparuna
guianensis

Zoghbi
et al. [28]

Siparuna
guianensis

Leaves
Leaves
Leaves
Leaves and
fruits

Leaves

Islands of
Abaco--
Bahamas
Parrita (Costa
Rica)
Cerrado in
Mato
Grosso--
Brazil
Panama
Rio Branco
(AC)--Brazil
Tocantins
(MG)--Brazil
Brazilian
cerrado

Different
places of the
Amazon

Myrcene, -caryophyllene,
cis- and trans--ocimene.
-pinene, -pinene, -myrcene, -thujene
and -caryophyllene.
Sesquiterpenes and sesquiterpene
hydrocarbon.
Curzerene, derivatives their degradation and
myristicin.
-cadinene, bergamotene and
-caryophyllene.
-terpinolene and -bisabolol.
Leaves: decanoic acid and 2-undecanone.
Fruit: undecanone-2, -pinene and limonene.
Moju (PA): epi--bisabolol and spathulenol.
Rio Branco (AC): spathulenol,
selin-11-en-4-ol, -eudesmol and elemol.
Belém (PA): germacrone, germacrene-D,
bicyclogermacrene, germacrene-B and
atractilone.

Torres et al. [18] evaluated the chemical composition of the volatile components present in the bark
of Capsicodendron dinisii Schwancke, botanical synonym for C. dinisii, and verified the presence of
23 compounds (90% of the total oil composition), as 86.8% monoterpenes and limonene was found as
the majority compound (68.5%). Similar results were observed in this study as to the class of
compounds present in the highest percentage since 76.20% monoterpenes was found in the oil under
study, but it was different regarding the majority component, limonene, that in the present work
presented a percentage of 1.54%.

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One can see how in the present study, the other studies described that evaluated the chemical
composition of the essential oils of species present in four of the five genera of the family Canellaceae
(Cinnamodendron, Canella, Cinnamosma, Warburgia and Pleodendron), presented, as a common
characteristic, that all components are of terpenic origin, being mostly monoterpenes and sesquiterpenes.
Studies by Adams and Zanoni [19] evaluating the chemical composition of the essential oil
obtained from wood of Cinnamodendron ekamani, a species of the same family as C. dinisii, verified
the presence of 1,8-cineole (35.9%), -humulene (9.1%), -caryophyllene (6.5%), 4-terpineol (5.0%),
germacrene-D (4.9%), -elemene (4.8%), -pinene (3.6%) and -terpineol (3.0%). Subsequently
Tucker et al. [20] observed the presence of 1,8-cineole (51.0%) and sabinene (10.6%) in the chemical
composition of a commercial essential oil from Madagascar, named Mandravasarotra (Cinnamosma
fragrans Baill., Canellaceae).
The essential oil of the leaves of Canella winterana (L.) Gaertn. (Canellaceae), from the islands of
Abaco (Bahamas), presented a total of 19 compounds (100% of the total composition), the majority
components being myrcene (32.4%), -caryophyllene (18.8%) cis- and trans--ocimene (15.9%
and 14.0%, respectively) [21]. Preliminary analysis of the essential oils of leaves and bark of
Pleodendron costaricense indicated that the composition of the essential oils of the two parts of the
plant analyzed were very similar, showing high levels of -pinene, -pinene, -myrcene, -thujene and
-caryophyllene with a lesser amount of linalool. The main difference between the volatile
compositions of the two parts of the plant was the content of -caryophyllene, which was the second
most abundant component in the leaves, but almost absent in the bark [22].
The results obtained for the chemical composition of S. guianensis corroborate the results found by
Valentini et al. [23], which identified and quantified the majority compounds of the volatile chemicals
extracted from leaves of S. guianensis in an area of cerrado in Mato Grosso, for twelve months. They
found that in relation to the variability of the classes of compounds in question, 70% of the identified
compounds were sesquiterpenes and sesquiterpene hydrocarbons that appeared in the rainy season and
dry-rainy season transition, with increased production in February 2008, the same month oil was
obtained in the present study.
Antônio et al. [24] found, in the leaves of S. guianensis Panama, curzerene (25.64%), derivatives
their degradation (42.31%) and myristicin (7.93%). In the same period, Rebouças [25] studied the
essential oil of S. guianensis, plants collected in Rio Branco, Acre, and observed that the
majority components of the essential oil were -cadinene (21.8%), bergamotene (14.2%) and
-caryophyllene (15.1%).
Subsequently, Montanari [26] noted that in the essential oil of plants collected in the city of
Tocantins (MG), two constituents, the monoterpene -terpinolene and the sesquiterpene alcohol
-bisabolol, together, accounted for about 80% of the oil composition throughout the year, results that
differ from those found in our work where only the -bisabolol was found, but in smaller quantities.
Essential oils from leaves and fruits of S. guianensis collected in the southeastern Brazilian cerrado
presented as major constituents in the leaf oil, decanoic acid (46.6%) and 2-undecanone (31.7%), in the
fruit oil mainly undecanone-2 (32.5%), -pinene (19.6%) and limonene (13.6%) were found, results
that differ from those found in this present study [27].
The essential oils of S. guianensis obtained from plants collected in different places of the Amazon
showed differences in their constitution. In the essential oil sample collected in Moju (PA) were the

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majority constituents epi--bisabolol (25.1%) and spathulenol (15.7%), the essential oil sample
collected in Rio Branco (AC) presented as majority compounds spathulenol (22.0%), selin-11-en-4-ol
(19.4%), -eudesmol (10.0%) and elemol (10.0%), while in the oil sample collected Belém (PA) the
presence was verified of germacrone (23.2%), germacrene-D (10.9%), bicyclogermacrene (8.6%),
germacrene-B (8.0%) and atractilone (31.4%) as majority compounds [28]. The chemical composition
of essential oils evaluated in our study showed similarities with the oil of the plant collected in Belém
(PA) regarding the majority presence of germacrene-D and bicyclogermacrene.
Differences in content and chemical composition of essential oils extracted from a species often
occur, since the production of secondary metabolites, including essential oils, is strongly influenced by
the environment in which the producing organism is inserted, and the factors responsible for such
variations are, seasonality, circadian rhythm, age and plant development, as well as the different plant
organs, temperature, water availability, nutrients, altitude, atmospheric composition and attack of
pathogens and herbivores [29,30].
3.2. Antioxidant Activity
By the -carotene/linoleic acid method, the essential oils studied showed little antioxidant activity,
but did not provide IC50 values (>300 g mL-1) in the concentration ranges tested. Among standards
tested, BHT (IC50 > 25 g mL-1) was the most efficient, followed by thymol (IC50 105.82 g mL-1)
and ascorbic acid (IC50 118.15 g mL-1), respectively (Table 3).
Employing the DPPH technique, antioxidant activity was not observed for the essential oils studied,
but among the standards assessed, ascorbic acid (IC50 44.36 g mL-1) was more efficient than BHT
(IC50 48.84 g mL-1) followed by thymol (IC50 > 300.00 g mL-1) (Table 3).
Table 3. Antioxidant activity of essential oils of S. guianensis and C. dinisii and of the
standards thymol, BHT and ascorbic acid by the -carotene/linoleic acid test and the DPPH
radical sequestration method.
Methods
Components
C. dinisii
S. guianensis
BHT
Thymol
Ascorbic acid

-carotene/linoleic acid
IC50 * (µg mL-1)
>300.00
>300.00
300.00
44.36

* IC50 = Inhibition Concentration of 50%; ** NI = no inhibition in the concentration ranges tested.

According to the results, it can be said that the essential oils rich in terpenes showed better
values for the antioxidant activity in the -carotene/linoleic acid system oxidation assay, as the
-carotene/linoleic acid method can be especially useful for investigations of lipophilic antioxidants
and is appropriate for the investigation of antioxidant activity of essential oils. On the other hand, if
polar compounds, such as ascorbic acid, were only tested by it, they would be considered weak
antioxidants [31]. This explains the lower efficiency of ascorbic acid, compared with thymol.

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According to Ruberto and Baratta [32], in lipid systems, phenolic compounds are effective
antioxidants, thus, thymol and carvacrol molecules are indeed responsible for the antioxidant activity
of many essential oils that contain them and a scant antioxidant activity is given to monoterpene and
sesquiterpene hydrocarbons. Only three monocyclic components, terpinolene, -terpinene and
-terpinene, and to a lesser degree, sabinene (a bicyclic), show considerable activity. The presence of
these strongly activated methylene group molecules, according to the authors, is probably the reason
for this behavior. These results, may explain the low antioxidant activity of essential oils of C. dinisii,
being composed of 87% of monoterpene and sesquiterpene hydrocarbons as well as S. guianensis,
composed of 58.40% of this group of substances.
According to Mata et al. [33], the absence of antioxidant activity observed for the terpene
compounds in the DPPH reduction can be explained by the fact that they are not capable of donating a
hydrogen atom and the low solubility provided by them in the reaction medium of the assay, because
this test utilizes methanol or ethanol as solvent. Thus, the fact that the essential oils of our study do not
show significant antioxidant activity can be explained, since both oils are composed almost entirely of
monoterpene and sesquiterpene hydrocarbons. For Viuda-Martos et al. [34], the cited factors can be
considered as the main limitation of this assay for measuring the antioxidant activity of lipophilic
samples, like many essential oils.
Despite the essential oils tested in this study not showing significant antioxidant activity, many
essential oils have shown antioxidant potential. As an example there is the research conducted by
Guimarães [35], who investigated the antioxidant activity of essential oils of Lippia sidoides, Alomia
fastigiata, Ocotea odorifera, Mikania glauca and Cordia verbenacea, and their majority constituents,
by the methods of the -carotene/linoleic acid oxidation system, the formation of thiobarbituric acid
reactive species (TBARS) and the reduction of the stable DPPH radical, and found that the essential oil
of L. sidoides showed higher antioxidant activity, presenting the lowest IC50 values in all trials, and the
antioxidant activity presented by the essential oil of L. sidoides was attributed to its majority
constituent carvacrol, which also showed high antioxidant activity when assessed in isolation. It was
also observed that there is influence of the methodology on the antioxidant activity presented by
different essential oils and compounds evaluated, demonstrating the importance of the methodology
for the determination of activity. Lima et al. [36] observed that the essential oils of Myristica fragrans
and Salvia microphylla showed antioxidant potential by the -carotene/linoleic acid test, with IC50
values of 976 and 770 µg mL-1, respectively.
4. Conclusions
We concluded that the main components found in the essential oil of the fresh leaves of C. dinisii
were -pinene (35.41%), -pinene (17.81%), sabinene (12.1%) and bicyclogermacrene (7.59%)
and the essential oil of the fresh leaves of S. guianensis were identified, -myrcene (13.14%),
germacrene-D (8.68%) and bicyclogermacrene (16.71%) and antioxidant activities of the essential oils
were evaluated by testing low acid -carotene/linoleic and were not evidenced by DPPH to test both
oils evaluated.

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Acknowledgments
To the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), the Fundação de
Apoio à Pesquisa e Extensão de Minas Gerais (FAPEMIG), to CAPES (Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior) and to Pest OE/EQB/LA0023/2011, for financial aid
and grants provided.
Conflicts of Interest
The authors declare no conflicts of interest.
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