International Journal of Food Properties

ISSN: 1094-2912 (Print) 1532-2386 (Online) Journal homepage: https://www.tandfonline.com/loi/ljfp20

Biological activities of volatile extracts from two

varieties of Habanero pepper (Capsicum chinense
Jacq.)

Odri Sosa-Moguel, Jorge A. Pino, Guadalupe Ayora-Talavera, Enrique Sauri-

Duch & Luis Cuevas-Glory

To cite this article: Odri Sosa-Moguel, Jorge A. Pino, Guadalupe Ayora-Talavera, Enrique Sauri-
Duch & Luis Cuevas-Glory (2017) Biological activities of volatile extracts from two varieties of
Habanero pepper (Capsicum�chinense Jacq.), International Journal of Food Properties, 20:sup3,
S3042-S3051, DOI: 10.1080/10942912.2017.1397694

To link to this article: https://doi.org/10.1080/10942912.2017.1397694

© 2017 Taylor & Francis Group, LLC

Published online: 08 Jan 2018.

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INTERNATIONAL JOURNAL OF FOOD PROPERTIES
2017, VOL. 20, NO. S3, S3042–S3051
https://doi.org/10.1080/10942912.2017.1397694

Biological activities of volatile extracts from two varieties of

Habanero pepper (Capsicum chinense Jacq.)
Odri Sosa-Moguela, Jorge A. Pinob, Guadalupe Ayora-Talaverac, Enrique Sauri-Ducha,
and Luis Cuevas-Glorya
a
Departamento de Ingeniería Química y Bioquímica, Instituto Tecnológico de Mérida, Mérida, Yucatán, México;
b
Instituto de Investigaciones para la Industria Alimenticia, Havana, Cuba; cDepartamento de Biomedicina de
Enfermedades Infecciosas, Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Mérida, Yucatán, México

ABSTRACT ARTICLE HISTORY

Biological activity of Habanero pepper is principally due to capsaicinoids, Received 7 July 2017
flavonoids, carotenoids and peptides. However, the volatile fraction of this Accepted 24 October 2017
fruit has not been analysed earlier. Isolation of the volatile compounds from
KEYWORDS
two varieties of Habanero pepper was performed by simultaneous distilla-
Volatile extract; Capsicum
tion-extraction method. Gas chromatography-mass spectrometry analysis chinense; antioxidant;
revealed 109 and 110 compounds in Mayapan and Jaguar varieties, respec- antibacterial; GC-MS
tively. The main compounds found were hexyl 3-methylbutanoate, 3,3-
dimethylcyclohexanol, hexyl 3-methylbutanoate, (Z)-3-hexenyl 3-methylbu-
tanoate and heptyl 3-methylbutanoate. Highest antioxidant activity was
achieved from extracts of the Jaguar variety by means of the ABTS•+
method. Minimum inhibitory concentration analysis showed that V. cholerae
was the most sensitive bacteria to the volatile extracts from both varieties.

Introduction
Pepper fruits (Capsicum spp.) are a significant source of bioactive compounds such as vitamins C
and A, carotenoids, phenolic compounds, terpenoids, steroids and alkaloids known for their health
promoting effect against degenerative diseases.[1] The genus Capsicum comprises more than 200
varieties; the fruits vary widely in size, shape, flavour and sensory heat. The five main species of
Capsicum cited in the literature are as follows: C. annuum, C. baccatum, C. chinense, C. frutescens
and C. pubescens.[2] The Yucatan is one of the main producers of C. chinense Jacq. In Mexico, also
known as Yucatan Habanero pepper. Due to its designation of origin acquired in 2010 as well as to
its cultural, culinary and economic value, this is one of the main horticultural crops in the area with
high potential for industrialisation and exportation.[3]
Habanero pepper is a highly aromatic fruit and is the hottest chili pepper in the world. The typical
aroma is one of its most attractive properties, representing a quality parameter for the consumer.[4,5]
Previous works have found 102 and 53 volatile compounds (GC-MS) in Habanero pepper from the
Yucatan by utilising simultaneous steam distillation-solvent extraction and headspace solid-phase
microextraction, respectively.[3,4] Depending on the ripeness of each species and the isolation method,
the aroma of pepper fruits was varied due to differences in the volatile compounds profiles.[6] The
volatile fraction is generally constituted by low molecular weight compounds and a class of lipophilic
secondary metabolites with high vapour pressure. These constituents have been shown to exhibit
antimicrobial[7–9], antioxidant[10], anticancer[11], antidiabetic, and antithrombotic properties.[12] Several
scientific studies have reported biological activity of the C. chinense as antimicrobial[13], anti-
inflammatory[14], antioxidant[15] and anticancer.[16] This effect is mainly due to the presence of

CONTACT Odri Sosa-Moguel odrimar@hotmail.com Departamento de Ingeniería Química y Bioquímica, Instituto

Tecnológico de Mérida, Mérida, Yucatán, México
© 2017 Taylor & Francis Group, LLC
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES S3043

capsaicinoids, flavonoids, and carotenoids.[17,18] However, the volatile fraction from this fruit has never
been analysed in regards to the active function.
The aim of this study was to analyse the chemical composition of the volatile fraction of two
varieties (Mayapan and Jaguar) of Habanero pepper and to investigate other properties of volatile
extracts, such as antioxidant and antibacterial activity in vitro, in order to focus on the benefits of
this fruit as a functional food in the human diet.

Material and methods

Chemicals and reagents
Pure chemicals were obtained from Aldrich (St. Louis, MO, USA) as follows: n-alkane solution (C8-
C32), solution, a mixture of C5-C7, methyl nonanoate, chemical standards, dichloromethane, 2,2ʹ-
azinobis-3-ethylbenzothiazoline-6-sulphonate (ABTS), potassium persulphate, 1,1-diphenyl-2-picryl-
hydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), p-iodo-nitro-
tetrazolium violet (INT) and dimethylsulfoxide (DMSO).

Samples
Fresh Habanero pepper fruits on mature stage (Table 1) were provided by the National Institute for
Forestry Agriculture and Livestock (INIFAP) from the Yucatan, Mexico. Jaguar and Mayapan
varieties of Habanero pepper have been registered as CHL-008–101109 and CHL-009–170908
respectively by National Service Seed Inspection and Certification (SNICS).

Extraction of volatiles
Fresh fruits (200 g) were cut and homogenise with distilled water (600 mL) in a commercial blender
for 1 min. The resultant puree was immediately transferred into 2 L-flask of an apparatus for
simultaneous steam distillation–solvent extraction (SDE) and 500 µL (35.7 mg/mL) of methyl
nonanoate as internal standard was added. Dichloromethane (40 mL) was utilised as the extracting
solvent and the condenser was kept to 5°C. The SDE system was operated for 1 h as reported
previously.[19] The volatile extract was dried over anhydrous sodium sulphate and concentrated on a
Kuderna-Danish evapourator until nearly 1 mL was obtained. The final volume of 0.2 mL was
obtained by means of a gentle nitrogen stream. Extractions were done in triplicate.

Gas chromatography-mass spectrometry

GC-MS analyses were performed on a Perkin Elmer Clarus 500 (Shelton, CT, USA) gas chromato-
graph coupled to a MSD (Shelton, CT, USA). Injection was on splitless mode (ratio 1:10) at 250°C.
Separation was performed on an AT-5 MS column (30 m x 0.25 mm x 0.50 μm film thickness)
(Alltech, IL, USA). The column temperature was programmed as follows: 50 °C for 2 min then
increased to 250°C at a rate of 4°C/min, held at 250°C for 10 min. The gas carrier (helium at 99.99%

Table 1. Characteristics of the two varieties of Habanero pepper.

Name of the cultivar Mayapan Jaguar
Average fruit size (cm) 4.2 x 3.0 4.0 x 2.8
Soluble solids (ºBrix) 4.7 4.9
Color at maturity Orange Orange
L* 74.2 73.2
a* 24.0 21.7
b* 49.2 47.41
S3044 O. SOSA-MOGUEL ET AL.

of purity) flow rate was 1 mL/min and detector temperature was at 250°C. Mass spectrometer
parameters were as follows: electron impact ionisation mode at 70 eV; acquisition range, m/z
35–400; interface and source temperatures were 250°C and 250°C, respectively.
The retention times of n-alkanes mixture were used to calculate lineal retention indexes (LRI) for
all identified compounds and for reference standards. Identification of volatile compounds was
performed comparing their LRI and mass spectra with the authentic standards from Sigma-Aldrich.
Tentative identification of compounds, for which it was not possible to find reference compounds,
was performed by comparison of their mass spectra with those from various libraries (NIST 02,
Wiley 275, Palisade 600 and our specific library for volatile compounds, Flavourlib) and experi-
mental LRIs with those reported in the literature.[20] Concentrations were expressed as mg methyl
nonanoate equivalents kg of fresh weight, response factors being taken as 1.0 for all compounds with
reference to the internal standard and a recovery factor of 70% was considered.

Antioxidant activity
ABTS•+ radical-scavenging assay
The ABTS•+ assay was based on the method described[21] with slight modifications. The ABTS radical
cation (ABTS•+ radical dot) was generated by mixing a 7 mm ABTS solution in a 2.45 mm solution of
potassium persulphate. The ABTS•+ solution was diluted in ethanol until it reached an absorbance of
0.70 ± 0.02 at 740 nm. Different concentrations (0 to 40 mg/mL) were made by diluting the volatile
extract in pure ethanol. A 50 µL aliquot of the volatile extract solution was added in 150 µL of the radical
ABTS•+ solution. Absorbance was measured at the time of 8 min in a microtiter plate reader (Thermo
Science, Finland) at 740 nm. Measurements were performed in triplicate. A control experiment was
conducted by using a solution without the test material. Free radical scavenging activity of each solution
was calculated as percentage of inhibition, according to the following equation:
% Inhibition ¼½ðAo A1 Þ=Ao  x 100
Where Ao: Absorbance of control, A1: Absorbance of sample.

DPPH radical-scavenging assay

The free radical scavenging activity of the extracts of Habanero pepper was measured by using the
modified method described early[22] as follows: volumes of 50 μL of the samples (0–40 mg/mL) were
mixed with 150 μL of DPPH solution (0.1 mM in ethanol) in a 96-well microtiter plate. After
incubation for 30 min in the dark at room temperature, the reduction of the absorbance was
measured at 540 nm in a microtiter plate reader (Thermo Science, Finland). All measurements
were carried out in triplicate. A pure DPPH solution was used as control.

Antibacterial activity
Bacterial cultures
The antimicrobial activity of the volatile extract was tested against different microorganisms,
including Gram-positive bacteria: Staphylococcus aureus (ATCC 25923) and Enterococcus faecalis
(ATCC 29212); Gram negatives: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC
27853) and Vibrio cholera (ATCC 14033). LB (Luria-Bertani) agar was utilised for cultivating
bacteria and LB broth medium for MIC test. Inoculum was prepared by making a dilution of the
bacterial suspension in sterile NaCl solution until absorbance of 0.1 at 620 nm (1 × 108 cfu/mL) and
then (100 μL) a further mixing with 9.9 mL NaCl solution (1 × 106 cfu/mL).

Determination of minimum inhibitory concentrations

A two-fold broth microdilution method was utilised to determine the MIC of the volatile extracts.[23] The
volatile extracts were dissolved in DMSO (5%) and in LB broth medium, in order to prepare a stock
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES S3045

solution (20 mg/mL). LB broth medium (100 µL) was poured into each of the 96 wells of a microplate and
then 100 µL of the stock solution (volatile extracts) was added into the first well, then a serial dilution was
carried out by dispensing previous solution into the following well and so on, in order to obtain
concentrations from 10 to 0.02 mg/mL (wells 1 to 10). A bacterial suspension of 100 µL was inoculated
into the wells. The same test was performed simultaneously for growth control (LB + DMSO) and
negative control (LB + DMSO + microorganism). Ampicillin (1 mg/mL) was utilised as a positive
control. The plate was incubated at 37°C for 24 h and immediately after, 40 µL of INT solution (0.5 mg/
mL) was added to each well. The plates were then incubated for another 60 min to ensure adequate color
development. The MIC was determined as the lowest concentration at which it did not reveal any visual
growth of microorganisms.

Statistical analysis
The data were analysed statistically by means of the Student’s t-test and analysis of variance for
individual parameters was performed by Duncan’s test on the basis of mean values to find the
significance at p ≤ 0.05. Statgraphics 5.0 software was utilised for statistical analysis.

Results and discussion

Volatile components analysis
GC-MS profile of the volatile constituents of the Habanero pepper varieties Mayapan and Jaguar demon-
strated the presence of a wide range of compounds, including esters, followed by terpenes, long-chain
hydrocarbons, alcohols, aldehydes, ketones and acids. In total, 109 and 110 constituents were identified and
quantified from Mayapan and Jaguar varieties, respectively (Table 2). This finding was similar to those
previously reported for other varieties of C. chinense, such as Creole (102 components) from the Yucatan[4]
and Cachucha (134 components) from Cuba.[24] The results of this work demonstrated that esters were the
predominant compounds in both varieties (29.4 and 30% Mayapan and Jaguar, respectively). The
abundance of esters in C. chinense has been determined in the volatile fraction through different extraction
methods, such as 35–58% in HS-SPME[5,25–27] and 30–43% in SDE.[4] The most important esters, in terms
of their concentration, were hexyl pentanoate, hexyl 3-methylbutanoate and heptyl 3-methylbutanoate.
Citronellyl 3-methylbutanoate is reported here for the first time in Habanero peppers from the Yucatan.
The second most important chemical group in both varieties were the terpenes (19%), particularly
mono- and sesquiterpene hydrocarbons. The sesquiterpenes γ-himachalene (20.14 and 23.24 mg/mL
from Mayapan and Jaguar varieties, respectively), α-copaene (3.0 and 4.28 mg/mL), β-cubebene (6.32
and 1.35 mg/mL) and (Z)-β-farnesene (3.40 and 2.96 mg/mL) were quantitatively predominant, in
contrast to Pino et al.[4,19], who found β-caryophyllene and germacrene D as the most abundant
sesquiterpenes in a creole variety of Habanero pepper from the Yucatan.
Moreover, a higher amount of cubenenes was determined in fruits of C. chinense from Ecuador.[26]
According to a previous study[25] himachalene is one of the most characteristic compounds of the
more pungent genotypes of Habanero pepper. Other compounds such as aliphatic alcohols, aldehydes,
ketones and acids were qualitatively minor classes in Mayapan and Jaguar varieties. However, 3,3-
dimethylcyclohexanol (83.74 and 38.69 mg/mL from Mayapan and Jaguar varieties, respectively) was
quantitatively the major alcohol present in both varieties.
Branched hydrocarbons such as 2-methyl-1-tetradecene (6.57 and 7.70 mg/mL from Mayapan and
Jaguar varieties, respectively) and 2-methyltetradecane (9.15 and 10.47 mg/mL from Mayapan and Jaguar
varieties, respectively) were predominant. The methyl-branched hydrocarbons are thought to be related to
capsaicin biosynthesis.[25] The main components in both volatile extracts from Mayapan and Jaguar
varieties were hexyl 3-methylbutanoate (84.08 and 140.71 mg/mL from Mayapan and Jaguar varieties,
respectively), hexyl pentanoate (90.09 and 34.87 mg/mL), 3,3-dimethylcyclohexanol (83.74 and 38.69 mg/
mL) and heptyl 3-methylbutanoate (54.69 and 16.75 mg/mL).
S3046 O. SOSA-MOGUEL ET AL.

Table 2. Volatile compounds (mg/kg) in Habanero pepper cultivated varieties.

Compound LRIa Identificationb Mayapan Jaguar Significante
Acids
3-Methylbutanoic acid 841 A 0.52 0.77 *
Hexanoic acid 1020 B ndc 0.03
Alcohols
2-Methylpropan-1-ol 621 A 0.42 0.67 *
Butan-1-ol 663 A trd tr
3-Penten-1-ol 686 B 0.24 0.25
3-Methylbutan-1-ol 733 B 0.38 0.38
Pentan-1-ol 764 A 0.26 0.07 *
(Z)-2-Penten-1-ol 767 B 0.30 0.26
4-Methylpentan-1-ol 833 A 5.73 5.30
(Z)-3-Hexen-1-ol 857 A 0.49 2.08 *
(E)-2-Hexen-1-ol 861 B 2.93 1.89 *
Hexan-1-ol 870 A 2.80 0.82 *
Octan-1-ol 1069 A tr 0.02
2,3-Dimethylcyclohexanol 1389 C 10.19 4.28 *
3,3-Dimethylcyclohexanol 1392 B 83.74 38.69 *
Tetradecan-1-ol 1811 B 0.92 0.38 *
Hexadecan-1-ol 1876 A 1.36 0.62 *
Octadecan-1-ol 2086 A 0.21 0.09 *
Aldehydes
Acetaldehyde 408 A tr tr
2-Propenal 498 A 0.08 0.30 *
Butanal 593 A tr tr
3-Methylbutanal 654 B 0.07 0.08
(Z)-2-Pentenal 754 B 0.19 0.25
Hexanal 802 A 0.36 0.61 *
(E)-2-Hexenal 853 A 7.13 5.81 *
Heptanal 904 B 0.01 tr
Benzaldehyde 958 A tr tr
Phenylacetaldehyde 1045 B 0.37 0.21
(E)-2-Nonenal 1164 C 0.07 tr
Tetradecanal 1615 B 1.78 1.37 *
Pentadecanal 1707 B 7.35 3.86 *
(E)-11-Hexadecenal 1759 B 3.10 2.23 *
Hexadecanal 1831 B 0.42 0.14 *
Ketones
2,3-Butanedione 595 B tr tr
2-Butanone 603 A tr tr
3-Methyl-2-butanone 657 B 0.45 0.24 *
3-Pentenone 694 B 0.05 0.04
3-Hydroxy-2-butanone 705 B tr tr
2-Methyl-3-pentanone 748 B 0.04 0.02
3-Methyl-2-pentanone 750 B 0.02 tr
2-Heptanone 889 A 0.05 0.06
(E)-3-Hepten-2-one 942 C 0.25 0.18 *
Oxacyclotetradecan-2-one 1650 B tr tr
Esters
Isobutyl 2-methylbutanoate 1004 B 0.01 0.02
Isobutyl 3-methylbutanoate 1006 A 0.14 0.21
Hexyl acetate 1010 A 0.10 0.06
Isopentyl 3-methylbutanoate 1103 A 3.46 2.39 *
2-Methylbutyl 3-methylbutanoate 1107 A tr 0.15
Pentyl 2-methylbutanoate 1142 A 0.20 0.13
(Z)-3-Hexenyl 2-methylpropanoate 1145 A 0.01 0.09 *
Pentyl 3-methylbutanoate 1148 A 1.76 2.99 *
Hexyl 2-methylpropanoate 1151 B 0.37 0.63 *
Isoprenyl pentanoate 1152 B 0.25 0.44 *
Methyl salicylate 1193 A tr tr
Hexyl 2-methylbutanoate 1237 A 10.05 6.33 *
Hexyl 3-methylbutanoate 1240 A 125.35 140.71 *
Heptyl 2-methylpropanoate 1243 A 0.35 tr
(Z)-3-Hexenyl 2-methylbutanoate 1293 A 0.24 0.24
(Continued )
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES S3047

Table 2. (Continued).
Compound LRIa Identificationb Mayapan Jaguar Significante
(Z)-3-Hexenyl 3-methylbutanoate 1295 A 2.75 1.80 *
Hexyl pentanoate 1298 B 90.05 34.87 *
(E)-2-Hexenyl pentanoate 1299 B tr 2.67
Heptyl butanoate 1300 A nd tr
Hexyl tiglate 1328 C tr 0.48
Octyl 2-methylpropanoate 1329 B 0.17 0.67 *
Heptyl 2-methylbutanoate 1332 B 6.23 1.97 *
Heptyl 3-methylbutanoate 1338 B 54.69 16.75 *
Octyl butanoate 1379 A tr 0.22
(Z)-3-Hexenyl hexanoate 1385 A 1.08 0.40 *
Heptyl hexanoate 1448 B 3.38 2.96
Citronellyl 3-methylbutanoate 1558 B 3.08 1.06
Hexyl benzoate 1577 A tr tr
Benzyl benzoate 1762 A tr tr
Ether
2-pentylfuran 992 A 0.03 tr
Hydrocarbons
Cyclohexene 667 B 2.87 1.71 *
2-Methyltetradecene 1445 B 6.57 7.70 *
2-Methyltetradecane 1467 B 9.15 10.47 *
3-Methylpentadecane 1533 B 0.14 2.12 *
1-Hexadecane 1600 A tr tr
1-Tricosane 2300 A 0.23 0.24
1-Tetracosane 2400 A 0.12 0.06 *
1-Pentacosane 2500 A 0.44 0.11 *
Terpenes
α-Pinene 934 A 0.03 0.50 *
p-Cymenene 1023 A tr 0.08 *
Limonene 1029 A 0.24 0.45 *
(Z)-β-Ocimene 1046 A 0.03 0.03
γ-Terpinene 1061 A tr 0.20
Terpinolene 1079 A 0.01 tr
Linalool 1100 A tr tr
α-Terpineol 1192 A tr tr
α-Copaene 1380 B 3.00 4.28 *
β-Cubebene 1390 B 6.32 1.35 *
(E)-β-Caryophyllene 1421 B tr tr
(E)-α-Ionone 1426 A tr tr
(E)-β-Farnesene 1458 A 3.40 2.96 *
β-Chamigrene 1475 B 3.84 1.77 *
γ-Himachalene 1481 B 20.14 23.24 *
(E)-β-Ionone 1485 A tr tr
α-Murolene 1502 B tr 0.05
β-Himachalene 1504 B 1.00 0.51 *
δ-Cadinene 1515 A 2.40 0.46 *
Cubenol 1621 B 0.43 0.16 *
α-Cadinol 1632 B tr tr
Others
Pyridine 760 A 0.03 0.02
Toluene 765 B 0.02 0.03
p-Xilene 866 B 0.08 0.09
Guaiacol 1092 B tr tr
a
LRI, retention indices obtained using a 5% phenyl/95% dimethylpolysiloxane capillary column.
b
The reliability of the identification proposal is indicated by the following: A, mass spectrum and retention index agreed with
standards; B, mass spectrum and retention index agreed with database or literature; C, mass spectrum agreed with mass spectral
database.
c
nd, not detected.
d
tr, compound at trace level (<0.01 mg/kg).
e
Different letters in the same row indicate significant difference by Duncan’s test at p ≤ 0.05.
S3048 O. SOSA-MOGUEL ET AL.

Total yield of volatiles, determined by an internal standard addition, was 539.12 and 357.89 mg/
mL for Mayapan and Jaguar varieties, respectively. Similar concentration was reported (110.66–
302.53 mg/mL) for Cachucha peppers from Cuba.[24] Lesser concentrations were determined (103–
228 mg/mL) in commercial fruits[4] and (1.37–11.84 mg/mL) in experimental varieties of Habanero
peppers from the Yucatan.[19] According to the species and level of ripeness, the composition varies
due to differences in volatile compound profile.[5,19]

Antioxidant activity
Figure 1 illustrates how the extracts reduce the radicals DPPH and ABTS•+ in a dose-dependent
manner. A strong inhibition was observed at 40 mg/mL. However, the values of ABTS•+ were

70
60.8
60 54.9
50.1
ABTS•+ Inhibition (%)

50 43.4

40 34.3
30.4
30
18.2 18.5
20

10

0
10 20 30 40
Extract concentration (mg/mL)

35
29.6
30
25.2
DPPH Inhibition (%)

25

20 17.2 16.6

15 12.2
9.8
10 8.6

4.4
5

0
10 20 30 40
Extract concentration (mg/mL)

Mayapan Jaguar

Figure 1. Inhibition effect of ABTS•+ and DPPH in volatile extracts of two Habanero pepper varieties.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES S3049

significantly higher than the DPPH values. Certain factors, such as stereo selectivity of the radicals or
the solubility of the extract in different testing systems, may affect the capacity of the extracts to react
and quench different radicals.[28] On the other hand, inhibition of the extract from the variety Jaguar
was significantly higher than the extract of Mayapan variety by the DPPH method. This could be due
to mechanisms of the reaction of DPPH with the structural conformations of antioxidant com-
pounds. According to Prior et al.[29] small molecules have facilitated access to the radical site. The
volatile extract from Jaguar variety exhibited a higher antioxidant activity in both radicals than
Mayapan variety. This is probably due to compounds such as (Z)-3-hexen-1-ol, α-pinene, p-cymene,
limonene and γ-terpinene[10,30], whose concentration is greater than in Mayapan variety.
Antioxidant activity of ethanolic extracts, hexane extracts and methanolic extracts of C. chinense
has been previously reported.[17] However, there are no studies concerning antioxidant activity from
volatile extracts. Phenolic compounds, carotenoids, ascorbic acid and capsaicin are principally
responsible for antioxidant activity in Capsicum species.[15] However, due to their high boiling
points, these organic compounds are not present in the volatile extracts, which explains in part
the difference in activity found. According to the results elucidated in the present study, antioxidant
activity of volatile fraction of the varieties Mayapan and Jaguar of Habanero peppers depends mainly
on concentration of the volatile compounds present in each one.

Antibacterial activity
The results (Table 3) present effective antimicrobial activity against all microorganisms tested, except
for E. coli and E. faecalis (Mayapan volatile extract). The strongest antimicrobial activity was
observed with the extract from Jaguar variety. Thus, growing inhibition of S. aureus and P.
aeruginosa with Jaguar volatile extract (5 mg/mL) requires less concentration than required with
the Mayapan volatile extract (10 mg/mL). Differences may be associated to a higher content of (Z)-3-
hexen-1-ol, α-pinene, p-cimene and γ-terpinene in the volatile extract from Jaguar variety.
According to other authors[7,30] these components have shown significant antimicrobial activity on
S. aureus and P. aeruginosa. Other compounds such as α-terpineol[7], linalool[8] and (E)-β-
caryophyllene[31] (some minor constituents in the volatile extracts) are also known to have effective
antimicrobial properties. E. faecalis showed inhibition only to extracts of the Jaguar variety (10 mg/
mL), which can be attributed to antibacterial properties of high concentrations of γ-terpinene[7,32],
α-pinene[7,8], (Z)-3-hexen-1-ol[8] and p-cymene.[7,32] On the other hand, Elaissi et al.[33] suggested
that Gram-positive bacteria E. faecalis is more sensitive to p-cymene. The lowest MIC found against
V. cholerae, using extracts from Jaguar and Mayapan varieties, were 2.5 and 5 mg/mL, respectively. It
is known that antimicrobial activity of essential oils and their components are more pronounced
against Gram-positive than against Gram-negative bacteria, since they possess an outer membrane
surrounding the cell wall, which inhibits diffusion of hydrophobic compounds through the lipopo-
lysaccharide cover.[9] However, cyclic terpenes such as α-pinene, limonene, p-cymene and γ-

Table 3. Minimum inhibitory concentration (MIC) of the volatile extracts from Habanero
pepper varieties.
MIC (mg/mL)
Microorganisms Mayapan Jaguar
Gram-positive
Staphylococcus aureus 10.0 5.0
Enterococcus faecalis na 10.0
Gram-negative
Escherichia coli na na
Vibrio cholerae 5.0 2.5
Pseudomonas aeruginosa 10.0 5.0
na: not active
S3050 O. SOSA-MOGUEL ET AL.

terpinene may become accumulated in the lipid bilayer and distort the lipid-protein interaction;
thus, the membrane can lose integrity and increase permeability to protons and ions.

Conclusion
A total of 109 and 110 compounds were identified in the volatile extracts of Mayapan and Jaguar
Habanero peppers. Both varieties exhibited the same prevalence of volatile compounds. Nevertheless,
the differences between both varieties, by means of the volatile compounds identified by GC-MS,
were mainly quantitative rather than qualitative. The volatile extract from the Jaguar variety showed
higher antioxidant and antibacterial activity than those from the Mayapan variety. According to the
findings of this study, antioxidant and antibacterial activity of volatile extracts of Mayapan and
Jaguar Habanero pepper are most probably due to the presence of minor components rather than
compounds present in larger quantities.

Funding
The authors want to thank to National Council of Science and Technology (CONACYT) for the financial support
provided to carry out this work.

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