Journal of Medicinal Plants Research Vol. 6(9), pp.

1758-1763, 9 March, 2012

Available online at http://www.academicjournals.org/JMPR
DOI: 10.5897/JMPR11.1576
ISSN 1996-0875 ©2012 Academic Journals

Full Length Research Paper

Antioxidant activity and bioactive compounds of

Chiltepin (Capsicum annuum var. glabriusculum) and
Habanero (Capsicum chinense): A comparative study
Alfonso Rodríguez-Maturino1, Aura Valenzuela-Solorio1, Rosalba Troncoso-Rojas2, Daniel
González-Mendoza1*, Onécimo Grimaldo-Juarez1, Mónica Aviles-Marin1 and
Lourdes Cervantes-Diaz1
1
Instituto de Ciencias Agrícolas -Universidad Autónoma de Baja California, Carretera Delta S/N, Ejido Nuevo León
21705, Baja California, México.
2
Centro de Investigación en Alimentación y Desarrollo, A.C., Dirección de Tecnología de Alimentos de Origen Vegetal,
Carretera a La Victoria km. 0.6, Hermosillo, Sonora 83000, México.
Accepted 20 December, 2011

Phenolic compounds, carotenoids content (red and yellow), extractable colour (total pigments) and
antioxidant activity in fruits from Chiltepin and Habanero pepper were investigated. The Habanero
pepper had significantly higher total phenolic content (5.92 ± 0.47 mg/g GAE) than the Chiltepin pepper
(4.85 ± 0.23 mg/g GAE). On the other hand, the highest content of total antioxidant activity was found in
Chiltepin chili (11.10 ± 1.6 mg of GAE /g of dry weight). In contrast, Habanero chili showed the lowest
values of total antioxidant activity. In this way, the Habanero chilli has the lowest content (14 mg/g dry
weight) of carotenoids (yellow isochromic fraction) while the Chiltepin chilli had higher levels of yellow
isochromic fraction (36 mg/g dry weigh). Finally, the Chiltepin showed the highest value of extractable
colour (80 ASTA units) than Habanero (40 ASTA units). Therefore Chiltepin and Habanero peppers are a
good prospect for developing varieties with higher levels of bioactive compounds.

Key words: Chiltepin pepper, Habanero pepper, phenolic compounds, carotenoids content, extractable colour,
antioxidant activity.

INTRODUCTION

Reactive oxygen and nitrogen species, ROS/RNS are
essential to energy supply, detoxification, chemical
signaling and immune function. They are continuously
produced in the human body and they are controlled by
endogenous enzymes (superoxide dismutase,
glutathione peroxidase, and catalase). When there is an
over-production of these species, an exposure to external
oxidant substances or a failure in the defense
mechanisms, damage to valuable biomolecules (DNA,
lipids, proteins) may occur (Aruoma, 1998). On the other
hand, several studies have shown that consumption of
fruit, vegetables, and derived food products have health
benefits against chronic diseases including cardiovascular
*Corresponding author. E-mail: daniasaf@gmail.com. Tel: +52
686 5230079. Fax: +52 686 5230217.
disease and neurodegenerative disorders. In this way,
the health-promoting properties of fruits are due to the
presence of bioactive compounds (e. g. antioxidants and
phenolics compounds), dietary fiber, and nonessential
phytochemicals presents in these food products (Gil et
al., 2002; Gonzalez-Mendoza et al., 2010). With respect
to phenolics in vegetables, these compounds are present
in free and bound forms. Bound phenolics, mainly in the
form of β- glycosides, may survive human stomach and
small intestine digestion and reach the colon intact,
where they are released and exert their bioactivity
(Sosulski et al., 1982; Oboh and Rocha, 2007). Between
the plants of the Solanaceae family, Chiltepin pepper
(Capsicum annuum L. var. glabriusculum) is found and its
fruits are consumed mainly in the north-western part of
Mexico. On the other hand, consumption of fruit of
Habanero pepper (Capsicum chinense Jacq.) is limited
mainly to the south\western region of Mexico. In recent

studies, Forero et al. (2009) showed that Chiltepin pepper
has higher amounts of volatile components (hexyl
isopentanoate, hexyl 2-methylbutanoate, limonene, hexyl
isohexanoate, (E)-2-hexenal, isopentyl isopentanoate
and (Z)-3-hexenyl isopentanoate) in green stage than red
fruits. In this way, Montoya-Ballesteros et al. (2010)
observed that Chiltepin fruits green and red stages of
maturity showed differences in the color, pungency,
capsaicin and dihydrocapsaicin content. On the other
hand, Antonious et al. (2006, 2009) reported that fruits of
Habanero peppers are known to be excellent sources of
different phytochemicals, including vitamins A and C,
phenolic compounds, flavonoids, and carotenoids
essentially. Additionally, several studies has been
estimated that hot peppers are the second most
consumed vegetable by the Mexican population after
tomatoes with a consumption of approximately 7 to 9
kg/person per year (Ornelas-Paz et al., 2008; Alvarez-
Parrilla et al., 2011). Therefore, is important to
characterize the content of bioactive compounds from
both peppers fruits (Chiltepin and Habanero) would also
be helpful in increasing the awareness of the consumers
regarding the level of beneficial phytochemicals present
in these nutritious plants.
For this reason, the aim of the present study was to
determine the compounds such as phenolic compounds,
extractable colour (total pigments) and antioxidant activity
present in the fruits from Chiltepin and Habanero pepper.
This research will permit a comparison of some basic
attributes of fruit quality between the genotypes
evaluated. As well as provide a basis on which any future
selections may be evaluated and compared.
Additionally, any information on the bioactive
compounds of fruit of these chilis will provide a
knowledge base that may be of some benefit to the
developing fruit culture and processing industry in
Sonora, Mexico.
MATERIALS AND METHODS
Collection of fruits from plant materials
Fresh fruits of selected Chiltepin (C. annuum L. var. glabriusculum)
and Habanero (C. chinense) respectively, were purchased from
regional markets of Mexicali, Mexico. The fruits of each pepper
were washed, and the edible portion of fresh samples was
dried using a cabinet-type convective dryer at 65°C by 24 h.
Total phenolics content
Total phenolic content in Chiltepin and Habanero chili´s fruit was
determined with Folin-Ciocalteu reagent according to the method of
Slinkard and Singleton (1977) and modified by Khandaker et al.
(2008), using gallic acid as a standard phenolic compound. In brief,
300 mg of biomass dried were placed in an eppendorf tube, with 1.5
mL of methanol (80%), grinded at 4 °C and centrifuged at 14000 × g
for 15 min. Reaction mixture consisted of mixing 30 µl of the extract
added with 90 µl of Na2CO3 and 150 µl of Folin-Ciocalteau reagent in
a 96-well microplate.
Rodríguez-Maturino et al. 1759
After 30 min, absorbance readings (725 nm) were taken in a KC-4
spectrophotometer (Biotek 2 ® Instruments, Inc. Winooski, Vt.). Total
phenolic content of tissue was expressed as mg gallic acid
equivalents per gram of dry weight (mg GAE/g DW) through the
calibration curve with gallic acid, ranging from 0 to 300 µg/ml.
Total antioxidant activity
This assay is based on the reduction of Mo(VI) to Mo(V) by the
extract and subsequent formation of green phosphate/Mo(V)
complex at acid pH (Prieto et al., 1999). The extract (0.1 ml) was
mixed with 3 ml of reagent solution (0.6 M sulphuric acid, 28 mM
sodium phosphate and 4 mM ammonium molybdate). The tubes
were incubated at 95ºC for 90 min. The mixture was cooled to room
temperature, and then the absorbance of the solution was
measured at 725 nm (Thermo Scientific Genesys 20) against blank.
The antioxidant capacity was expressed as mg gallic acid
equivalent per gram dry weight (mg GAE/g DW).
Extractable colour measurements
The extractable colour was measured according to ASTA 20
method. An amount of 0.3 g of Chiltepin and Habanero powder
respectively was put into a graduated 100 ml flask. Acetone was
added to the mark; the mixture was shaken and kept in the dark for
6 h. An aliquot of the transparent decanted extract was taken. The
absorbance of the solution was measured using a Thermo Scientific
Genesys 20 spectrophotometer, set at 460 nm and calibrated with
an acetone blank. ASTA 20 colour units were calculated from
following equation:
ASTA 20 = Absorbance 16.4 I/ Weight of sample (g)
where I is a correction factor for the apparatus, which is calculated
from the absorbance of a standard solution of potassium
dichromate, ammonium sulphate and cobalt sulphate.
Red and yellow carotenoid measurements
The carotenoids of red (R = capsanthin and capsorrubin) and
yellow (Y = b-carotene, b-criptoxanthin,zeaxanthin) isochromic
fractions of total carotenoid were measured according to Hornero-
Mendez and Minguez-Mosquera (2001). Carotenoids from Chiltepin
and Habanero were extracted from dried samples by placing 300
mg of powder in 30 ml acetone and left under constant agitation for
1 h in the dark. The homogenate was centrifuged at 6000 rpm for
five minutes. The supernatant was separated and the absorbances
were read at 472 (yellow) and 508 (red) nm using a Thermo
Scientific Genesys 20 spectrophotometer. The red and yellow
carotenoid measurements were expressed as mg/100 g dry weight.
Statistical analysis
The significant differences between Chiltepin and Habanero fruits a
sample was analyzed using the Kruskal–Wallis test (Statistical
Package version 5.5, Statsoft, USA).
RESULTS
Total phenolics content
In this study, significant variations were observed in
content of total phenolic compounds from different
species of selected Capsicum (Figure 1a).
1760 J. Med. Plants Res.

Figure 1. Total phenolic content (a) and total antioxidant activity (b) from
Chiltepin and Habanero chilis. (Boxplots, boxes show mean, bars show mean
± SE; n = 3).

The results showed that total phenolic content of
Chiltepin (4.85 ± 0.23 mg/ g GAE) was significantly minor
compared to Habanero fruits (5.92 ± 0.47 mg/g GAE).
Total antioxidant activity
antioxidant activities (p<0.05) with respect to Habanero
chili (8.20 ± 0.6 mg of GAE /g of dry weight) that showed
low values of total antioxidant activity (Figure 1b). In this
way, the order of total antioxidant activity of peppers
extracts can be seen as Chiltepin> Habanero.

On the other hand, in the total antioxidant activity Red and yellow carotenoid measurements
evaluation the results revealed , that Chiltepin chili (11.10
± 1.6 mg of GAE /g of dry weight) had significant The carotenoids are widely distributed in nature, where

Figure 2. Values of carotenoids (red isochromic fraction and yellow isochromic fraction)
of Chiltepin and Habanero chili, respectively. Means ± standard error; n = 3.
they have an important role in the plants. In the present
study, the results of the analyses show that Chiltepin
chili´s have higher levels of carotenoids, particularly in
terms of the yellow isochromic fraction which includes the
provitamin A carotenoids (b-carotene and b-criptoxanthin)
and on minor concentrations red isochromic fraction
which includes capsanthin and capsorrubin than those of
Habanero chili (Figure 2). The non-parametric ANOVA
Kruskal–Wallis test showed significant differences in
ASTA units concentrations among them (p<0.01).
Extractable colour measurements
Alternatively, the extractable colour analysis (ASTA 20
units) was used as measure of total pigment content. In
the present study, the Chiltepin chili was characterized by
accumulated higher concentration of ASTA units with
respect to Habanero chili (Figure 3). The non-parametric
ANOVA Kruskal–Wallis test showed significant
differences in ASTA units concentrations among them (P
= 0.014).
DISCUSSION
The peppers (Capsicum sp.) are known for their richness
in micronutrients and bioactive compounds (e. g. phenolic
compounds), thus their consumption has been distinctly
recognized as being important factor for good health. In
Rodríguez-Maturino et al. 1761
the present study, the results observed in Chiltepin
peppers are comparable with total phenolic content
observed in others hot peppers (Antonoius et al., 2006).
In this sense, our results showed that phenolics content
of red Chiltepin was higher than phenolic content present
in C. annuum var. aviculare (Tepin) and C. chinense,
(Oboh and Ochoa 2007 and Nuñez-Ramirez et al.,2011
respectively). These might be attributed to the increased
extractability of phenolic compounds due to the disruption
of plant cell walls during heat treatment (e.g., 65°C),
which might cause phenolic compounds to be released
more easily than in fresh materials (Scalzo et al., 2004;
Ornelas-Paz et al., 2008). The total antioxidant activity
differences observed between Chiltepin and Habanero
chilis could possibly be due to the diversity and
complexity of antioxidants compounds present in
Chiltepin and Habanero fruits. In this sense, several
studies show that difference exists in total antioxidant
activity in pepper, may be attributed to the different
causes, like fertilization, fruit ripening and temperature
(Menichini et al., 2009; Nuñez-Ramirez, 2011; Alvarez-
Parrilla et al., 2011). On the other hand, our results did
not showed correlation between bioactive compounds
(total phenolic compounds) and antioxidant capacity of
Chiltepin and Habanero fruits evaluated, respectively. In
this sense, one possibility is that the antioxidant capacity
could be affected by the presence of carotenoids. The
carotenoids are widely distributed in nature, where they
have an important role in the plants. These compounds
can act as antioxidants, with functions that include
1762 J. Med. Plants Res.
Figure 3. Change in extractable colour (ASTA units) of fruits from Chiltepin and
Habanero chilis (Boxplots, boxes show mean, bars show mean ± SE; n = 3).
protection of membranes against damage by free radicals
and retardation of ageing processes (Bulda et al., 2008).
In this way, the low content of red isochromic fraction
carotenoids observed in Chiltepin and Habanero fruits
could be due to the presence of a mutation (deletion) in
the gene which codifies for the enzyme capsanthin-
capsorrubin synthase (CCS) prevents the synthesis of
the red pigments (Rodríguez-Burruezo et al., 2009). In
consequence, the Chiltepin and Habanero fruits only
contain yellow and orange carotenoids. These results are
important because carotenoids present in peppers
particularly the yellow/orange, b-cryptoxanthin, and b-
carotene, are vitamin A precursors, with the high
antioxidant activity (Wall et al., 2001). Additionally, in this
study we found that both species of Capsicum showed
high concentrations of ASTA units in their fruits with
respect to reported for others Capsicum species
(Mendez-Trujillo et al., 2005; Eissa et al., 2007).
Therefore, Chiltepin and Habanero peppers are a good
prospect for developing varieties with higher levels of
bioactive compounds (e.g., yellow and orange
carotenoids). Finally, these results provide useful and
important information for researchers in order to increase
the antioxidant capacity and functional value of Chiltepin
and Habanero chilis for the food and nutraceutical
industries.
Conclusion
In the present study, Chiltepin and Habanero chili appear
to be a good and safe source of antioxidants. Chiltepin
fruits could be used for direct consumption or as extracts
to increase the nutritional value of different foods and
diets. Future studies include identification of the
remaining antioxidant constituents in the semi purified
aqueous fractions and study of the antifungal effects of
these aqueous extracts.
ACKNOWLEDGEMENTS
This study was supported by the 15ª Convocatoria
Interna de Apoyo a Proyectos de Investigación de la
Universidad Autónoma de Baja California (UABC) and
SUBNARGEM (subsistema nacional de recursos
genéticos microbianos).
REFERENCES
Alvarez-Parrilla E, de la Rosa LA, Amarowicz R, Shahidi F (2011).
Antioxidant activity of fresh and processed Jalapeño and Serrano
peppers. J. Agric. Food Chem., 12: 163-173.
Antonious GF, Kochhar TS, Jarret RL (2009). Pungency in Capsicum
chinense: variation among countries of origin. J. Environ. Sci. Health
B., 44: 179-184.
Antonious GF, Kochhar TS, Jarret RL, Snyder JC (2006). Antioxidants
in hot pepper: Variation among accessions. J. Environ. Sci. Health
Part B, 41: 1237-1243.
Aruoma OI (1998). Free radicals, oxidative stress, and antioxidants in
human health and disease. J. Am. Oil. Chem. Soc., 75: 199-212.
Bulda O, Rassadina V, Alekseichuk H, Laman N (2008).
Spectrophotometric measurement of carotenes, xanthophylls, and
chlorophylls in extracts from plant seeds. Russ. J. Plant Physiol.,
55(4): 544-551.
Eissa AH, Mostafa BE, Hussein AS (2007). Capsaicin content and
quality characteristics in different local pepper varieties (Capsicum
annum) and acid-brine pasteurized purre. J. Food Technol., 5: 246-
255.

Forero MD, Quijano EC, Pino AJ (2009). Volatile compounds of Chile
pepper (Capsicum annuum L. var. glabriusculum) at two ripening
stages. Flavour. Fragr. J., 24: 25-30.
Gil MI, Tomas-Barberan FA, Hess-Pierce B, Kader AA (2002).
Antioxidant capacities, phenolic compounds, carotenoids, and vitamin
C contents of nectarine, peach, and plum cultivars from California. J.
Agric. Food Chem., 50: 4976-4982.
González-Mendoza D, Grimaldo-Juárez O, Soto-Ortiz R, Escoboza-
Garcia F, Santiaguillo Hernández JF (2010). Evaluation of total
phenolics, anthocyanins and antioxidant capacity in purple tomatillo
(Physalis ixocarpa) genotypes. Afr. J. Biotechnol., 9: 5173-5176.
Hornero-Méndez D, Mínguez-Mosquera MI (2001). Rapid
spectrophotometric determination of red and yellow isochromic
carotenoid fractions in paprika and red pepper oleoresins. J. Agric.
Food Chem., 49: 3584-3588.
Khandaker L, Ali MB, Oba S (2008). Total polyphenol and antioxidant
activity of red amaranth (Amaranthus tricolor L.) as affected by
different sunlight level. J. Jpn. Soc. Hortic. Sci., 77: 395-401.
Méndez-Trujillo V, González-Mendoza D, Gutiérrez-Miceli FA (2005).
Carotenoid content and color of new cultivars inextractable chile
pepper. Chapingo Mag. Hortic. Ser., 11: 215-218.
Menichini F, Tundis R, Bonesi M, Loizzo MR, Conforti F, Statti G, De
Cindio B, Houghton PJ, Menichini F (2009). The influence of fruit
ripening on the phytochemical content and biological activity of
Capsicum chinnense Jaqc. cv habanero. Food Chem., 114: 553-560.
Montoya-Ballesteros LC, Gardea-Bejar A, Ayala-Chavez GM, Martinez-
Nuñez YY, Robles-Ozuna LE (2010). Capsaicinoids and color in
chilpetin (Capsicum annuum var. aviculare). Processing effect on
sauces and pickles. Mex. J. Chem. Eng., 9: 197-207.
Núñez-Ramírez F, González-Mendoza D, Grimaldo-Juárez O, Díaz LC
(2011). Nitrogen fertilization effect on antioxidants compounds in
fruits of habanero chili pepper (Capsicum chinense). Int. J. Agric.
Biol., 13: 827-830.
Rodríguez-Maturino et al. 1763
Oboh G, Rocha JBT (2007). Polyphenols in red pepper (Capsicum
annuum var. aviculare (Tepin) and their protective effect on some
pro-oxidants induced lipid peroxidation in brain and liver. Eur. Food
Res. Technol., 225: 239-247.
Ornelas-Paz JJ, Yahia EM, Gardea-Bejar AA (2008). Changes in
external and internal color during postharvest ripening of ‘Manila’ and
‘Ataulfo’ mango fruit and relationship with carotenoid content
determined by liquid chromatography-APcI+ time of flight mass
spectrometry. Postharvest Biol. Technol., 50: 145-152.
Prieto P, Pineda M, Aguilar M (1999). Spectrophotometric quantification
of antioxidant capacity through the formation of a
phosphomolybdenum complex: specific application of vitamin E.
Anal. Biochem., 269: 337-341.
Rodriguez-Burruezo, Prohens J, Raigon MD, Nuez F (2009). Variation
for bioactive compounds in aji (Capsicum baccatum L.) and rocoto
(C. pubescens R. & P.) and implications for breeding. Euphytica, l70:
l69-l8I.
Scalzo RL, Iannoccari T, Summa C, Morelli R, Rapisarda P (2004).
Effect of thermal treatments on antioxidant and antiradical activity of
blood orange juice. Food Chem., 85: 41-47.
Slinkard K, Singleton VL (1977). Total phenol analysis: automation and
comparison with manual methods. Am. J. Enol. Viticult., 28: 49-55.
Sosulski F, Krygier K, Hogge L (1982). Free esterified and insoluble-
bound phenolic acids.: Composition of phenolic acids in cereal and
potato flours. J. Agric. Food Chem., 30: 337-340.
Wall MM, Wadell CA, Bosland PW (2001). Variation in B- carotene and
total carotenoid content in fruits of Capsicum. Hort. Sci., 36: 746-749.