Food Chemistry 141 (2013) 2606–2613

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

The effect of domestic processing on the content and bioaccessibility

of carotenoids from chili peppers (Capsicum species)
Alessandro Pugliese a,b, Monica Rosa Loizzo a,⇑, Rosa Tundis a, Yvonne O’Callaghan b, Karen Galvin b,
Francesco Menichini a, Nora O’Brien b
a
Department of Pharmacy, Health and Nutrition Sciences, University of Calabria, 87036 Rende (CS), Italy
b
School of Food and Nutritional Sciences, University College Cork, Cork, Ireland

a r t i c l e i n f o a b s t r a c t

Article history: The content and bioaccessibility of carotenoids from different chili peppers were analysed and the effects
Received 7 November 2012 of typical domestic processing were investigated. Peppers were analysed before and after cooking by con-
Received in revised form 14 April 2013 ventional boiling (10 min in 100 °C water) and also following a freezing period of four months in a
Accepted 14 May 2013
domestic freezer (20 °C). The content and bioaccessibility of the eight carotenoids quantified varied,
Available online 23 May 2013
depending on cultivar, species, colour and processing. Provitamin A carotenoids (b-carotene and b-cryp-
toxanthin) and capsanthin were present at highest concentrations in the samples before and after pro-
Keywords:
cessing. In general, yellow and orange peppers were the best sources of lutein, zeaxanthin and
Carotenoids
Capsicum species
neoxanthin. Xanthophyll carotenoids were more efficiently transferred to the micelles and, therefore,
Bioaccessibility were also more bioavailable. Processing decreased the carotenoid content in certain samples; however,
Domestic processing the micellar content was generally not lower for processed peppers; therefore the bioaccessibility of
carotenoids from processed peppers is enhanced relative to unprocessed peppers.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction
Chili peppers are an indispensable spice, used as a basic ingre-
dient in a great variety of cuisines all over the world. They are also
used as flavourings, colorants and add tang and taste to otherwise
insipid foods. Peppers have been the subject of study, mainly as a
source of capsaicin, which produces different pungency levels
(Oboh, Puntel, & Rocha, 2007), as well as carotenoids and phenolic
compounds, which are used as natural pigments and antioxidant
agents (Suna, Youl Ha, & Park, 2008). The concentrations and com-
position of carotenoids are responsible for the diverse and attrac-
tive colours observed in peppers (Howard, 2001). Chlorophyll
contributes to the green colour (Marin, Ferreres, Tomas-Barberan,
& Gil, 2004), a- and b-carotene, zeaxanthin, lutein and b-crypto-
xanthin to the yellow–orange colour (Howard, 2001), whereas
red-pigmented chili peppers are the result from the presence of
capsanthin and capsorubin (Matsufuji, Nakamura, Chino, & Takeda,
1998).
The colour of each Capsicum variety in the full-ripe stage de-
pends on its capacity for synthesising carotenoids and even for
retaining chlorophyll pigments. In fact, variety is an important fac-
⇑ Corresponding author. Tel.: +39 0984493071; fax: +39 0984493107.
E-mail addresses: monica_rosa.loizzo@unical.it, mr.loizzo@unical.it
Loizzo).
tor, determining both the composition and content of plant pig-
ments (Guil-Guerrero & Rebolloso-Fuentes, 2009).
The genus Capsicum is classified into the family of Solanaceae
and is comprised of 25 wild and five domesticated species (C. ann-
uum L., C. frutescens L., C. chinense Jacq, C. baccatum Jacq, and C.
pubescens L.), which include more than 200 varieties (Conforti,
Statti, & Menichini, 2007), only a few of which have been studied
in detail.
The purported health benefits of peppers have been attributed,
in part, to their carotenoid content. In fact these fat-soluble com-
pounds have been shown to protect against certain cancers, pre-
vent gastric ulcers, stimulate the immune system, protect against
cardio-vascular diseases and age-related macular degeneration
and cataracts (Krinsky & Johnson, 2005).
In the Mediterranean diet, chili peppers are largely eaten in raw,
cooked, or processed forms. Typical processing methods include
freezing of the pepper, in order to store the fruit for a longer period.
These processes may influence the content and bioaccessibility of
the carotenoids contained in food.
Bioaccessibility is defined as the amount of an ingested nutrient
that is available for absorption in the gut after digestion (Hedren,
Diaz, & Svanberg, 2002). It is possible to determine the bioaccessi-
bility of a carotenoid by measuring the quantity transferred to the
micelle fraction following a simulated in vitro digestion procedure
(M.R. (Jiwan, Duane, O’Sullivan, O’Brien, & Aherne, 2010). The micelle
fraction is isolated by ultracentrifugation of the digested samples.

0308-8146/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2013.05.046
 A. Pugliese et al. / Food Chemistry 141 (2013) 2606–2613
This method is suitable for the routine screening of plant foods for
carotenoid bioaccessibility and may be used as an indicator of po-
tential bioavailability (Garrett, Failla, & Sarama, 2000). Previous
studies have reported the effect of processing on the carotenoid
content of vegetables (Nunn, Giraud, Parkhurst, & Hamouz, 2006;
Ryan, O’Connell, O’Sullivan, Aherne, & O’Brien, 2008). However,
information on the effects of processing on content and bioaccessi-
bility of carotenoids from peppers is limited.
The objective of the present study was to investigate the effect
of different processing treatments (freezing and boiling) on the
composition and bioaccessibility of carotenoids from seven varie-
ties of coloured (orange, red and yellow) peppers belonging to dif-
ferent species (C. annum, C. chinense, C. baccatum, C. chacoense).
2. Materials and methods
2.1. Chemicals, reagents and instruments
All reagents, including Hank’s Balanced Salts solution (HBSS),
lutein, capsanthin, zeaxanthin, b-carotene, b-apo-80 -carotenal and
cholesterol esterase were purchased from Sigma–Aldrich Chemical
Co. (Dublin, Ireland). b-Cryptoxanthin, violaxanthin, neoxanthin
and antheraxanthin were purchased from LGC Prochem (Middle-
sex, UK). All solvents employed were of HPLC grade. An Ultra-Tur-
rax T25 homogeniser, (IKA-Labortechnik, Staufen, Germany),
Sorvall TC6 centrifuge (DuPont Instruments, Herts, UK), HPLC sys-
tem (Finnigan SpectraSYSTEM; Thermo Scientific, Philadelphia, PA,
USA) and YMC C30 reversed phase column (TMC Europe GmbH,
Dinslaken, Germany) were used.
2.2. Pepper sample preparation
The seven commercial pepper cultivars used in this study were
harvested from a local farm (Miceli s.r.l., Scalea, Cosenza, Italy) in
October 2011 at maturity stage and at full fruit size from a random
sample of 20 plants in order to obtain a set of peppers representa-
tive of all the cultivars. Fruits from cultivar Cayenna (C. annuum)
are yellow at the mature ripe stage and are characterised by a
length of 9 cm and a width of 1 cm; fruits from cultivar Campana
(C. baccatum) are red at the mature ripe stage and are characterised
by a length of 5 cm and a width of 5 cm; fruits from cultivar Chaco
(C. chacoense) are red at the mature ripe stage and are character-
ised by a length of 1 cm and a width of 1 cm; fruits from cultivar
Aji Angelo (C. baccatum) are red at the mature ripe stage and are
characterised by a length of 12 cm and a width of 4 cm; fruits from
cultivar Taballo (C. annuum) are pale orange at the mature ripe
stage and are characterised by a length of 3 cm and a width of
1 cm; fruits from cultivar Hierro (C. annuum) are deep red at the
mature ripe stage and are characterised by a length of 5 cm and
a width of 1.5 cm; fruits from cultivar Capezzolo di Scimmia (C.
chinense) are orange at the mature ripe stage and are characterised
by a length of 1 cm and a width of 1 cm. All peppers received sim-
ilar water and fertiliser treatments.
Peppers were examined for integrity and absence of dust and
insect contamination and they were chopped uniformly to ensure
replication of conditions. All manipulations of the pepper fruits
were conducted under subdued (yellow) light to minimise photo-
decomposition of the carotenoids. The peppers were analysed
fresh, and following two different processing mechanisms, namely
boiling and freezing. For boiling, peppers were added to 500 ml of
boiling water and were cooked at a steady rolling boil for 10 min.
For freezing, fresh peppers were stored at 20 °C for 4 months in
order to replicate the common traditional method of storage. Fol-
lowing processing, 2 g of each pepper were weighed and the sam-
ples were subjected to an in vitro digestion procedure. Commercial
2607
vegetables are generally blanched before freezing as this process
reduces the microbial load and inactivates enzymes which may
have a detrimental effect on the nutritional content (Cano, 1996);
however, in domestic processing, peppers are typically frozen
without blanching.
2.3. In vitro digestion procedure
The in vitro digestion model was performed by the method of
O’Sullivan, Jiwan, Daly, and O’Brien (2010). The samples were
homogenised (Janke and Kunkel, Ultra-Turrax T25, IKA-Labortech-
nik, Staufen, Germany) at 13,500 rpm for approximately 20 s in
5 ml of HBSS. A portion of the homogenate (1 ml) was frozen at
80 °C after overlaying the headspace with nitrogen gas, in order
to quantify the carotenoid content in the peppers prior to diges-
tion. Digestion was carried out on the remaining homogenate
(4 ml). The samples were adjusted to pH 2, using HCl, and porcine
pepsin (0.04 g/ml) was added, followed by incubation at 37 °C in an
orbital shaking water bath for 1 h. After gastric digestion, pH was
increased to 5.3, followed by the addition of glycodeoxycholate
(0.8 mM), taurodeoxycholate (0.45 mM), taurocholate (0.75 mM)
and pancreatin (0.08 g/ml). Cholesterol esterase (1 unit/ml) was
added, in order to hydrolyse carotenoid esters or cleave different
xanthophyll esters. The pH of each sample was increased to 7.4,
followed by incubation at 37 °C for 2.5 h in an orbital shaking
water bath. Each digestate was then centrifuged at 53,000 rpm
for 95 min to isolate the micellar (aqueous) fraction which was col-
lected and frozen at 80 °C prior to further analysis.
2.4. Carotenoid extraction and saponification
All aliquots of samples, collected before and after digestion,
were thawed and vortexed. A recovery standard, b-apo-80 -carote-
nol, was added to all samples, which were extracted twice with
hexane/ethanol/acetone (50:25:25, v/v/v). Samples were centri-
fuged at 3000 rpm for 5 min, and the top layers were removed,
pooled, and dried under nitrogen gas. To enable better chromato-
graphic separation and detection of the carotenoids, the samples
were saponified, using a procedure previously described by Olives
Barba, Hutado, Sanchez Mata, and Fernadez Ruiz (2006). Briefly,
0.1 M ethanolic pyrogallol and 40% (w/v) potassium hydroxide
were added to the extracts. After vortexing, distilled deionised
water and hexane/dichloromethane (5:1, v/v) were added to the
samples which were then vortexed and centrifuged at 3000 rpm
for 5 min. The resultant supernatants were removed and the sol-
vent was evaporated. The samples were frozen at 80 °C under a
layer of N2 gas prior to further analysis.
2.5. HPLC analysis
Samples were reconstituted in 200 ll of methanol:ethyl acetate
(4:1, v/v), and the carotenoid content of the samples was quanti-
fied by a reverse-phase HPLC procedure. The HPLC method used
was based on the method of Taylor, Brackenbridge, Vivier, and
Oberhoster (2006). The HPLC system consisted of a P2000 pump
connected to an AS3000 auto-injector and a UV6000LP photodiode
array (PDA) detector. Separation of carotenoids was achieved using
a YMC C30 reversed phase column (250  4.6 mm, 5 lm particle
size) and column temperature was maintained at 30 °C by an inter-
nal column oven. Carotenoids were separated by gradient elution.
Mobile phase A was composed of methanol:water (97:3 v/v), con-
taining 10 mM ammonium acetate, 4.5 mM butylated hydroxytol-
uene, and 3.6 mM triethylamine, and mobile phase B was TBME
(tert butyl methyl ether 100%), containing 4.5 mM butylated
hydroxytoluene, and 3.6 mM triethylamine. The gradient was as
follows: 95% A for 35 min, 62% A in 36 min, 32% A in 39 min,
2608 A. Pugliese et al. / Food Chemistry 141 (2013) 2606–2613

Table 1 efficiency, based on recovery of b-Apo-80 -carotenal, and quantified

Peppers samples. after correction for initial weight and dilution factors.
Sample Cultivar Species Colour
P1 Yellow Cayenne C. annuum Yellow
P2 Campana C. baccatum Red 2.6. Statistical analysis
P3 Aji Angelo C. baccatum Red
P4 Chaco C. chacoense Red
Bioaccessibility is defined as the fraction of a nutrient available
P5 Taballo C. anuum Orange
P6 Hierro C. annuum Red for absorption into mucosa of the gut during human digestion. Bio-
P7 Capezzolo di scimmia C. chinense Orange accessibility is quantified by calculating the amount of a caroten-
oid present in the micelles as a percentage of the amount of
carotenoid in the original, undigested food. All experiments were
carried out in triplicate. Data were expressed as means ± SEM. Re-
sults among the different cultivars of chili peppers and the differ-
returned to 95% A in 45 min and re-equilibrated for 10 min. Carote- ent processing methods were analysed by two-way analysis of
noids were detected in 55 min with a flow rate of 1 ml/min at variance test (ANOVA), followed by a multicomparison Bonferroni
450 nm. Data were collected and analysed using ChromQuest post hoc test, using GraphPad Prism version 5.0 for Windows
software (version 4.2, Thermo Fisher Scientific). Violaxanthin, neo- (GraphPad Software, San Diego, CA, USA). The carotenoid content
xanthin, antheraxanthin, lutein, capsanthin, zeaxanthin, b-crypto- of each of the pepper varieties (P1–P7: Table 1) is expressed as
xanthin, and b-carotene levels in the samples were extrapolated lg/100 g of their fresh, frozen or cooked pepper weight, as appro-
from carotenoid standard curves after correction for extraction priate (Table 2).

Table 2
Carotenoid content of fresh (lg/100 g fresh pepper), frozen (lg/100 g frozen pepper) and cooked (lg/100 g cooked pepper) peppers.

Capsanthin Zeaxanthin Antheraxanthin Violaxanthin Neoxanthin Lutein b-Cryptoxanthin b-Carotene

Fresh
P1 0.0 ± 0.0 40.4 ± 14.1 20.1 ± 9.0 151 ± 48.7 34.6 ± 9.2 266 ± 45.0 9.8 ± 5.1 24.0 ± 11.7
(2,3,4,6,7) (2,5) (3,4,5,6) (5,7) (7) (2,3,4,5,6,7) (2,3,4,5,6) (2,3,4,5,6)
P2 230 ± 43.8 70.7 ± 5.9 116 ± 21.4 172 ± 35.6 30.7 ± 8.7 20.5 ± 7.0 139 ± 18.6 1258 ± 33.0
(1,3,4,5,6) (1,5) (3,4,5) (5,7) (7) (1,5) (1,3,4,5,6,7) (1,4,5,6,7)
P3 592 ± 64.4 95.6 ± 10.6 283 ± 38.7 214 ± 55.1 35.1 ± 11.4 59.2 ± 11.4 335 ± 27.7 1150 ± 132
(1,2,4,5,6,7) (5,7) (1,2,7) (5,7) (7) (1,5) (1,2,4,6,7) (1,4,5,6,7)
P4 848 ± 27.1 58.8 ± 0.9 225 ± 11.3 194 ± 38.1 32.1 ± 4.2 31.0 ± 5.2 238 ± 52.2 682 ± 50.3
(1,2,3,5,7) (5) (1,2,7) (6,7) (7) (1,5) (1,2,3,6,7) (1,2,3,5,6,7)
P5 0.0 ± 0.0 279 ± 43.8 249 ± 46.0 1119 ± 70.7 54.4 ± 11.9 614 ± 10.5 277 ± 28.8 548 ± 51.8
(2,3,4,6) (1,2,3,4,6,7) (1,2,7) (1,2,3,6,7) (7) (1,2,3,4,6,7) (1,2,6,7) (1,2,3,4,6,7)
P6 786 ± 45.7 56.4 ± 5.8 186 ± 29.2 249 ± 45.1 34.2 ± 6.8 17.8 ± 7.3 490 ± 15.1 1524 ± 42.3
(1,2,3,5,7) (5) (1,7) (4,5,7) (7) (1,5) (1,2,3,4,5,7) (1,2,3,4,5,7)
P7 104 ± 10.4 0.0 ± 0.0 13.7 ± 1.5 9.4 ± 2.5 0.0 ± 0.0 0.0 ± 0.0 9.0 ± 1.5 10.4 ± 0.3
(1,3,4,6) (3,5) (3,4,5,6) (1,2,3,4,5,6) (1,2,3,4,5,6) (1,5) (2,3,4,5,6) (2,3,4,5,6)
Frozen
P1 0.0 ± 0.0 240 ± 56.9⁄⁄ 45.7 ± 12.1 274 ± 10.1 47.8 ± 9.7 539 ± 9.8⁄⁄ 4.0 ± 0.3 39.7 ± 10.6
(2,3,4,6) (3,4,6,7) (2,4,5,6,7) (2,4,7) (2,3,4,5,6,7) (2,3,5,6,7)
P2 359 ± 57.5 89.5 ± 22.9 73.3 ± 32.3 91.1 ± 19.3 18.2 ± 7.4 10.3 ± 1.9 50.1 ± 10.0⁄ 363 ± 107⁄⁄
(1,5) (1,3,5) (1,5,6) (1,5) (1,6,7)
P3 376 ± 37.9⁄⁄ 34.9 ± 7.0 153 ± 54.7⁄ 209 ± 49.8 30.7 ± 12.8 21.8 ± 9.2 64.1 ± 22.7⁄⁄ 370 ± 72.4⁄⁄
(1,5) (1,5) (7) (2,4,5,7) (5,7) (1,5) (1,6,7)
P4 349 ± 30.0⁄⁄ 17.6 ± 3.8 73.0 ± 3.6⁄⁄ 62.5 ± 9.7⁄ 9.5 ± 1.6 4.8 ± 0.4 22.9 ± 3.2⁄⁄ 164 ± 46.1⁄⁄
(1,5) (1,5,6) (1,3,5,6) (1,5,6,7) (1,5) (6)
P5 0.0 ± 0.0 168 ± 63.5⁄ 176 ± 73.9 1065 ± 24.9 61.0 ± 11.2 370 ± 67.7⁄⁄ 19.1 ± 12.2⁄⁄ 459 ± 65.0
(2,3,4,6,7) (3,4,7) (1,2,3,4,6,7) (2,3,4,7) (1,2,3,4,6,7) (1,6,7)
P6 422 ± 62.2⁄⁄ 26.7 ± 6.4 88.2 ± 9.1 119 ± 28.1 49.6 ± 12.8 19.3 ± 1.5 48.4 ± 9.8⁄⁄ 1255 ± 88.1⁄
(1,5,7) (1,4) (1,4,5) (2,4,7) (1,5) (1,2,3,4,5,7)
P7 268 ± 49.2 30.8 ± 5.7 25.1 ± 12.9 26.3 ± 2.9 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 27.4 ± 15.8
(5,6) (1,5) (3) (1,3,5) (1,3,4,5,6) (1,5) (2,3,5,6)
Boiled
P1 11.7 ± 4.3 60.6 ± 21.3 8.5 ± 1.5 0.0 ± 0.0⁄ 21.8 ± 9.4 183 ± 37.2 2.0 ± 0.2 7.1 ± 1.8
(3,4,6) (3,4,5,6) (3,6) (2,3,4,6,7) (3,6) (2,3,4,6)
P2 115 ± 6.6 35.4 ± 9.0 54.7 ± 12.5 50.6 ± 17.3 11.6 ± 09 0.0 ± 0.0 37.5 ± 15.5⁄ 268 ± 66.7⁄⁄
(3,4,6) (3,4,6) (3,6) (1,5) (6) (1,6,7)
P3 813 ± 74.2⁄⁄ 85.8 ± 12.9 208 ± 17.1 188 ± 25.0 25.3 ± 2.1 2.8 ± 2.8 89.9 ± 39.3⁄⁄ 303 ± 108.7⁄⁄
(1,2,5,6,7) (7) (1,2,5,7) (1,2,5,7) (1,5) (1,6,7) (1,6,7)
P4 792 ± 73.5 42.2 ± 6.5 175 ± 46.6 105 ± 22.4 0.0 ± 0.0⁄ 0.0 ± 0.0 39.6 ± 13.2⁄⁄ 293 ± 65.8⁄⁄
(1,2,5,6,7) (1,2,5,7) (6) (1,5) (1,6,7)
P5 0.0 ± 0.0 71.2 ± 15.9⁄⁄ 26.5 ± 7.7⁄⁄ 0.0 ± 0.0⁄ 18.2 ± 2.4⁄ 161 ± 40.1⁄⁄ 30.2 ± 2.6⁄⁄ 177 ± 14.0⁄⁄
(3,4,6) (1,3,4,6,7) (3,6) (2,3,4,6,7) (6) (6)
P6 1016 ± 71.6⁄⁄ 34.3 ± 7.9 217 ± 49.1 223 ± 39.2 29.9 ± 6.6 0.0 ± 0.0 170 ± 46.5⁄⁄ 1544 ± 73.7
(1,2,3,4,5,7) (1,2,5,7) (1,2,5) (4,7) (1,5) (1,2,3,5,7) (1,2,3,4,5)
P7 66.7 ± 9.7 0.0 ± 0.0 12.2 ± 1.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.5 ± 0.5 22.8 ± 1.7
(3,4,6) (3) (3,4,5,6) (3,6) (6) (1,5) (3,6) (2,3,4,6)

Analysis of carotenoid content in the peppers is described under Materials and methods. Data are the means and SE of three independent experiments. Statistical analysis of
each carotenoid was by two-way ANOVA and Bonferroni post hoc test. Asterisks denote a significant difference (⁄⁄P < 0.01) (⁄P < 0.05) of the processed method against the
fresh control; numbers within the same column denote a significant difference (P < 0.05) and correspond to the number of the peppers next to the letter P.
 A. Pugliese et al. / Food Chemistry 141 (2013) 2606–2613 2609

3. Results
3.1. Carotenoid contents in chili peppers
The carotenoid contents of the pepper varieties (Table 1) are
listed in Tables 2 and 3. Capsanthin was one of the predominant
carotenoids in red peppers, with contents ranging from 230 to
848 lg/100 g pepper. Sample P7 also contained capsanthin at an
appreciable level (104 lg/100 g). There was no capsanthin detect-
able in raw or frozen yellow pepper (P1) or in the raw or frozen or-
ange pepper (P5). Similarly, no capsanthin was detectable in boiled
P5. However, in the boiled P1 samples, a small quantity (11.7 lg/
100 g) of capsanthin was detected. Following freezing, a significant
(P < 0.01) decrease was observed in the capsanthin content of pep-
per samples P3, P4 and P6. The capsanthin content of the remain-
ing samples was not significantly altered in comparison with the
corresponding raw samples. Boiling of the peppers resulted in a
significant increase (P < 0.01) in the capsanthin contents of sam-
ples P3 and P6.
Zeaxanthin was not present in the fresh and boiled orange pep-
per P7. In general, processing did not significantly alter the zeaxan-
thin content of the peppers. However, a significant increase was
observed in frozen P1 samples compared with raw P1, while both
freezing and boiling caused a significant decrease in the zeaxanthin
content of pepper sample P5. Similarly, the antheraxanthin content
of the peppers was not significantly affected by processing overall,
with the exception of the freezing of samples P3 and P4 and the
boiling of sample P5 which resulted in significant decreases
(P < 0.05, P < 0.01 and P < 0.01, respectively) in the antheraxanthin
contents.
The content of violaxanthin was highest in sample P5 (1119 lg/
100 g). A significant decrease (P < 0.05) was observed following
boiling in sample P4. A loss of violaxanthin, to levels which could
not be detected, was evidenced after boiling of the peppers. Neo-
xanthin was the carotenoid present at lowest concentrations in
most of the samples tested, with values ranging from 0.0 lg/
100 g (P7) to 54.4 lg/100 g (P5). The freezing process did not sig-
nificantly alter the neoxanthin content of any of the pepper sam-

Table 3
Carotenoid contents of micelles from fresh (lg/100 g fresh pepper), frozen (lg/100 g frozen pepper) and cooked (lg/100 g cooked pepper) peppers.

Capsanthin Zeaxanthin Antheraxanthin Violaxanthin Lutein b-Cryptoxanthin b-Carotene

Fresh
P1 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 50.4 ± 1.1 0.0 ± 0.0 0.0 ± 0.0
(2,3,4,6) (2,3,4,5,6,7) (2,3,4,6,7) (3) (2,3,4,5,6,7) (3,6) (3,6)
P2 169 ± 20.0 42.7 ± 7.0 41.5 ± 9.3 0.0 ± 0.0 0.0 ± 0.0 15.1 ± 2.2 28.7 ± 3.6
(1,3,4,5) (1,5,6,7) (1,3,5,7) (3) (1,5) (3,6) (3,6)
P3 400 ± 57.4 50.3 ± 14.3 74.7 ± 15.9 18.1 ± 5.0 0.0 ± 0.0 45.4 ± 11.9 79.5 ± 14.2
(1,2,4,5,6,7) 17 (1,2,4,5,6,7) (1,2,4,5,6,7) (2,5) (1,2,4,7) (1,2,4,5,7)
P4 278 ± 78.8 34.4 ± 11.6 74.4 ± 32.3 0.0 ± 0.0 0.0 ± 0.0 15.7 ± 5.5 28.7 ± 5.8
(1,2,3,5,7) (1,5,7) (1,3,5,7) (3) (2,5) (3) (3,6)
P5 0.0 ± 0.0 77.6 ± 32.9 0.0 ± 0.0 0.0 ± 0.0 177 ± 19.3 19.8 ± 11.2 17.2 ± 5.9
(2,3,4,6) (1,2,4,6,7) (2,3,4,6) (3) (1,2,3,4) (3,6)
P6 257 ± 22.8 38.4 ± 6.5 54.0 ± 21.3 0.0 ± 0.0 0.0± 0.0 28.7 ± 5.1 108 ± 7.9
(1,3,5,7) (1,2,5,7) (1,3,5,7) (3) (6,7) (1,2) (1,2,4,5,7)
P7 67.0 ± 17.9 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 4.3 ± 1.5 4.0 ± 0.4
(3,4,6) (1,2,3,4,5,6) (2,3,4,6) (3) (1,5) (3) (3,6)
Frozen
P1 0.0 ± 0.0 52.0 ± 9.6⁄⁄ 0.0 ± 0.0 0.0 ± 0.0 86.6 ± 15.2 0.0 ± 0.0 2.6 ± 0.3
(2,3,4,6) (4,6) (2,3) (2,3,6) (2,3,4,5,6,7) (3,6) (3,6)
P2 181 ± 24.5 25.1 ± 5.3 47.1 ± 6.3 18.2 ± 5.3⁄⁄ 0.0 ± 0.0 8.9 ± 1.5 27.1 ± 12.6
(1,5,7) (1,5,7) (1,4,5,7) (1,5) (3) (3,6)
P3 244 ± 10.7⁄⁄ 28.7 ± 6.6 92.2 ± 28.6 20.4 ± 4.0 0.0 ± 0.0 35.5 ± 9.9 93.9 ± 41.4
(1,4,5,7) (5) (1,5,7) (1,4,5,7) (1,5) (1,2,4,5,7) (1,2,4,5,7)
P4 114 ± 29.7⁄⁄ 14.0 ± 3.0 32.3 ± 5.2⁄⁄ 7.1 ± 3.7 0.0 ± 0.0 8.5 ± 2.2 19.2 ± 4.0
(1,3,5) (1,5) (2,3) (1,5) (3,6) (3,6)
P5 0.0 ± 0.0 74.0 ± 14,3 0.0 ± 0.0 0.0 ± 0.0 53.9 ± 2.3 8.5 ± 1.0 18.6 ± 2.5
(2,3,4,6) (3,4,6,7) (2,3) (2,3) (1,2,3,4,6,7) (3) (3,6)
P6 139 ± 66.4 16.9 ± 7.0 33.5 ± 8.0⁄ 14.0 ± 4.2⁄⁄ 0.0 ± 0.0 40.9 ± 26.4 114 ± 17.6
(1,5) (1,5) (1,5,7) (1,5) (1,4,5,7) (1,2,4,5,7)
P7 23.8 ± 5.6 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3.0 ± 0.9
(2,3) (5) (2,3) (2,3,6) (1,5) (3,6) (3,6)
Boiled
P1 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 43.6 ± 9.0 0.0 ± 0.0 0.0 ± 0.0
(3,6) (2) (2,3) (2,3,4,5,6,7) (6)
P2 92.6 ± 3.2 62.3 ± 2.9 51.4 ± 2.6 0.0 ± 0.0 0.0 ± 0.0 14.4 ± 1.3 30.4 ± 6.2
(1,3,4,5,6,7) (1,4,5,6,7) (1) (6)
P3 194 ± 32.5⁄⁄ 0.0 ± 0.0⁄⁄ 41.5 ± 10.3 0.0 ± 0.0⁄⁄ 0.0 ± 0.0 23.8 ± 3.8 38.3 ± 10.8
(1,4,5,7) (2) (1,4,5,6,7) (1) (6)
P4 69.1 ± 37.1⁄⁄ 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 6.4 ± 4.0 12.0 ± 1.1
(3) (2) (2,3) (1)
P5 0.0 ± 0.0 0.0 ± 0.0⁄⁄ 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 1.4 ± 1.4 18.5 ± 3.0
(3,6) (2) (2,3) (1) (6)
P6 126 ± 2.7⁄ 0.0 ± 0.0⁄ 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3.9 ± 2.6 72.9 ± 8.0
(1,5) (2) (2,3) (1) (1,2,3,5,7)
P7 33.1 ± 9.2 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3.3 ± 3.3 16.8 ± 3.3
(3,6) (2) (2,3) (1) (6)

Analysis of carotenoid content in the micelles is described under Materials and methods. Data are the mean and SE of three independent experiments. Neoxanthin is absent in
all analyzed samples and data are not reported. Statistical analysis of each carotenoid was by two-way ANOVA and Bonferroni post hoc test. Asterisks denote a significant
difference (⁄⁄P < 0.01) (⁄P < 0.05) of the processed method against the fresh control; numbers within the same column denote a significant difference (P < 0.05) and
correspond to the number of the peppers next to the letter P.
2610 A. Pugliese et al. / Food Chemistry 141 (2013) 2606–2613

ples; however, cooking significantly reduced (P < 0.05) its content
in both P4 and P5. Yellow peppers P1 and orange peppers P5 had
the highest content of lutein, but this compound was not present
in the orange peppers P7. The content of lutein in the red peppers
was low, ranging from 17.8 lg/100 g (P6) to 59.2 lg/100 g (P3).
Freezing significantly increased (P < 0.05) the lutein content of
the yellow pepper (P1). The lutein content of pepper sample P5
was significantly decreased (P < 0.01) following freezing and
boiling.
Both freezing and boiling processes significantly decreased
(P < 0.05, P < 0.01, respectively) the b-cryptoxanthin content of
the majority of the pepper samples (P2–P6). The remaining sam-
ples, P1 and P7, had a low content of b-cryptoxanthin in fresh pep-
pers, less than 10 lg/100 g. b-Carotene was present at high
concentrations in pepper samples P2-P6 and each of the red pep-
pers contained more b-carotene than did the yellow and orange
peppers. Freezing significantly reduced the b-carotene content of
each of the red peppers (P2–P6) and boiling significantly reduced
the b-carotene content of pepper samples P2–P5. b-Carotene, b-
cryptoxanthin, capsanthin and violaxanthin were generally the
predominant carotenoids in fresh chili peppers. With the exception
of b-carotene and b-cryptoxanthin, carotenoid contents were not
affected by processing in the majority of pepper samples. Where
changes in carotenoid contents were observed, they were usually
decreased. However, increases were found in the capsanthin con-
tent of samples P3 and P6 following cooking, the zeaxanthin con-
tent of P1, following freezing, and in the lutein content of
samples P1 and P5, following freezing and boiling, respectively.
3.2. Micelle content and bioaccessibility of carotenoids from chili
peppers
The bioaccessibility of capsanthin from fresh peppers ranged
from 0% in samples P1 (yellow) and P5 (orange) to 76% in P2 (Ta-
ble 4). The content of capsanthin in the micelles was significantly
lower (P < 0.01) for samples P3 and P4, following freezing, and
P3, P4 and P6, following boiling, in comparison with the micellar
content derived from raw peppers (Table 3). No zeaxanthin was

Table 4
Bioaccessibility of carotenoids from fresh, frozen and cooked peppers.

Capsanthin Zeaxanthin Antheraxanthin Violaxanthin Lutein b-Cryptoxanthin b-Carotene

Fresh
P1 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 20.7 ± 6.5 0.0 ± 0.0 0.0 ± 0.0
(2,3,4,6,7) (2,9) (2,3,4) (2,3,4,5,67) (7)
P2 76.0 ± 6.3 59.6 ± 5.2 37.5 ± 9.0 0.0 ± 0.0 0.0 ± 0.0 11.3 ± 2.4 2.3 ± 0.3
(1,4,5,6) (1,5,7) (1,3) (1,5) (7)
P3 69.7 ± 13.3 53.9 ± 17.9 27.1 ± 6.6 11.1 ± 5.3 0.0 ± 0.0 14.3 ± 4.6 7.4 ± 2.0
(1,4,5,6,7) (1,2,4,5,6,7) (1,5) (7)
P4 32.2 ± 8.0 58.7 ± 20.0 33.7 ± 15.3 0.0 ± 0.0 0.0 ± 0.0 8.8 ± 5.3 4.2 ± 0.6
(1,2,3,5,7) (1,3) (1,5) (7)
P5 0.0 ± 0.0 27.3 ± 11.3 0.0 ± 0.0 0.0 ± 0.0 28.9 ± 3.6 6.5 ± 3.6 3.3 ± 1.4
(2,3,4,6,7) (2) (3) (1,2,3,4,6,7) (7)
P6 33.3 ± 4.7 68.5 ± 11.6 35.2 ± 19.9 0.0 ± 0.0 0.0 ± 0.0 5.9 ± 1.1 7.1 ± 0.5
(1,2,3,5,7) (3) (1,5) (7)
P7 69.3 ± 26.2 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 53.3 ± 20.0 38.5 ± 3.4
(1,3,4,5,6) (2) (3) (1,5) (1,2,3,4,5,6)
Frozen
P1 0.0 ± 0.0 26.3 ± 9.7 0.0 ± 0.0 0.0 ± 0.0 16.0 ± 2.6 0.0 ± 0.0 8.2 ± 3.3
(2,3,4,6) (2,3,4,6) (3,4) (2,3,4,5,6) (5,6)
P2 53.3 ± 11.4 29.7 ± 6.2 87.3 ± 28.4⁄⁄ 19.2 ± 2.2⁄⁄ 0.0 ± 0.0 18.4 ± 3.1 6.9 ± 1.3
(1,5,7) (1,4,5,6,7) (3,4,5,6,7) (1,5) (6) (3)
P3 66.2 ± 7.1 81.2 ± 2.4 62.4 ± 3.5 9.9 ± 0.4⁄⁄ 0.0 ± 0.0 58.0 ± 3.8 23.2 ± 6.5
(1,5,6) (7) (1,5,7) (1,2,5,7) (1,5) (2,5)
P4 32.1 ± 7.0 79.6 ± 2.7 44.8 ± 8.5 9.9 ± 5.3 0.0 ± 0.0 36.6 ± 7.8 16.0 ± 8.3
(1,5) (7) (1,2,5,7) (1,2,5,6) (1,5)
P5 0.0 ± 0.0 89.8 ± 62.1 0.0 ± 0.0 0.0 ± 0.0 15.8 ± 3.4⁄⁄ 97.2 ± 41.5 4.3 ± 1.0
(2,3,4,6) (7) (2,3,4) (2,3,4,6) (1,2,3,4,6) (1,7) (3)
P6 29.4 ± 11.9 73.7 ± 36.0 37.4 ± 6.7 11.4 ± 00⁄⁄ 0.0 ± 0.0 98.7 ± 71.5 9.0 ± 0.8
(1,3,5) (1,2) (2,5,7) (1,5) (1,2,7)
P7 8.8 ± 0.0⁄⁄ 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 17.4 ± 6.5⁄
(2) (3,4,5) (2,3,4) (2,3,4,6) (1) (5,6)
Boiled
P1 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 25.5 ± 5.8 0.0 ± 0.0 0.0 ± 0.0
(2,7) (2) (2) (2,3,4,5,6,7) (3,7)
P2 81.0 ± 2.4 202 ± 54.1⁄⁄ 103 ± 20.0⁄⁄ 0.0 ± 0.0 0.0 ± 0.0 51.3 ± 15.9 13.2 ±4.0
(1,3,4,5,6) (1,3,4,5,6,7) (2,3,4,5,6,7) (1) (7)
P3 23.7 ± 2.8⁄⁄ 0.0 ± 0.0 20.8 ± 5.9 0.0 ± 0.0 ⁄⁄
0.0 ± 0.0 68.3 ± 50.7 16.7 ± 6.2
(2,4,7) (2) (2) (1) (1,7)
P4 9.7 ± 5.4 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 31.6 ± 24.9 4.4 ± 0.8
(2,3) (2) (2) (1) (7)
P5 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0⁄⁄ 5.4 ± 5.4 10.8 ± 2.4
(2,7) (2) (2,6) (1,7) (7)
P6 12.5 ± 1.1 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3.5 ± 2.6 4.7 ± 0.5
(2,7) (2) (2,5,7) (1) (7)
P7 47.8 ± 6.3 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0⁄⁄ 73.3 ± 2.6⁄⁄
(1,2,3,5,6) (2) (2,6) (1,5) (1,2,3,4,5,6)

Bioaccessibility (%) is defined as the proportion of carotenoids present in the micelles compared with that contained in the original food. Data are the means and SE of three
independent experiments. Neoxanthin is absent in all analyzed samples and data are not reported. Statistical analysis of each carotenoid was by two-way ANOVA and
Bonferroni post hoc test. Asterisks denote a significant difference (⁄⁄P < 0.01) (⁄P < 0.05) of the processed method against the fresh control; numbers within the same column
denote a significant difference (P < 0.05) and correspond to the number of the peppers next to the letter P.
 A. Pugliese et al. / Food Chemistry 141 (2013) 2606–2613
detectable in the micelles of raw samples P1 and P7. The bioacces-
sibility of zeaxanthin ranged from 0% (P1 and P7) to 68.5% (P6) for
the raw pepper samples. Boiling significantly reduced the micellar
content, and hence bioaccessibility, of zeaxanthin from all of the
samples apart from P2.
Antheraxanthin was not detected in the micelles of the yellow
and orange cultivars. In the red peppers, the bioaccessibility ranged
from 27.1% to 37.5% before processing. The micelle content was
significantly reduced in samples P4 (P < 0.01) and P6 (P < 0.05) fol-
lowing freezing and no antheraxanthin was detectable in these
samples following cooking (Table 3).
Violaxanthin was not detected in the micelles from any of the
fresh samples with the exception of peppers P3 (bioaccessibility
of 11.1%). The violaxanthin content of P3 micelles was not affected
by freezing. However, no violaxanthin was found in P3 micelles of
the boiled peppers. Carotenoids are enclosed in plant cell walls and
cell organelle structures which may be disrupted during food pro-
cessing, including freezing, thereby increasing their bioaccessibili-
ty (Van Buggenhout, Alminger, Lemmens, & Colle, 2010). Therefore,
although not detectable in the micelles of the raw peppers, the mi-
celles from frozen pepper samples P2 and P6 contained violaxan-
thin at concentrations of 18.2 and 14.0 lg/100 g, which
corresponded to bioaccessibilities of 19.2% and 11.4%, respectively.
Neoxanthin was not found in the micelles of any of the samples.
Lutein was only detected in the micelles of P1 and P5 with bioac-
cessibilities of 20.7% and 28.9%, respectively. Freezing and boiling
did not significantly affect the lutein content of the micelles from
sample P1. For sample P5, freezing of the micellar fraction signifi-
cantly decreased the lutein content. This carotenoid was not de-
tected in the boiled fraction (Table 3).
b-Cryptoxanthin was not found in the micelles of fresh, frozen
and boiled P1 peppers and it was very low in the P7 sample. How-
ever, as the content of b-cryptoxanthin in the fresh P7 samples was
also low, the percent bioaccessibility appears high (53.3%) and is
greater than the bioaccessibility for any of the remaining samples,
which ranged from 5.9% to 14.3%. The micelle content of b-crypto-
xanthin was not significantly different for the frozen or cooked
peppers (compared with the fresh samples). Although the b-cryp-
toxanthin contents of the micelles are broadly similar for all sam-
ples, there is a large range in the percent bioaccessibility, which
reflects variation in the b-cryptoxanthin content of the undigested
peppers.
Although b-carotene was one of the predominant carotenoids in
most of the samples, the content in the micelles was low, corre-
sponding to a bioaccessibility ranging from 2.3% to 7.4% for the
red peppers. Similarly to b-cryptoxanthin, the bioaccessibility of
b-carotene from sample P7 was high (38.6%) but the actual micelle
content was low (4.0 lg/100 g). Freezing and cooking did not affect
the b-carotene contents of their micelles.
4. Discussion
Studies have associated the health benefits of consuming fruits
and vegetables rich in carotenoids with a reduced risk of human
chronic diseases (Rao & Rao, 2007), but it is known that the absorp-
tion of these compounds may be highly variable (Faulks & Southon,
2005). It is possible to determine the proportion of a nutrient
which is available for intestinal absorption by studying the transfer
of the compound from the food matrix to the mixed micelles. This
transfer is defined as bioaccessibility and represents the fraction of
a nutrient accessible for intestinal absorption (Van Buggenhout
et al., 2010). Carotenoids in chili peppers have been studied for
decades; studies have generally been performed in C. annuum cul-
tivars and, to a lesser extent, in the related cultivars C. chinense, C.
baccatum and C. cachoense.
2611
In the present study, capsanthin, b-carotene and b-cryptoxan-
thin were found to be very abundant in all red genotypes of C. ann-
uum, C. baccatum and C. cachoense species; capsanthin was also
present at significant concentrations in the orange cultivar P7 (C.
chinense), contrary to the other yellow (P1) and orange (P5)
peppers.
Lutein has been reported to be the main carotenoid in the un-
ripe fruit of both red and yellow/orange genotypes of C. annuum
(Matus, Deli, & Szabolcs, 1991). However, the content of lutein de-
creases in red-fruited accessions during ripening, while it remains
in yellow/orange accessions (Hornero-Méndez, Go9 mez-Ladro9 n de
Guevara, & Mínguez-Mosquera, 2000). In support of these studies,
we found a higher quantity of lutein in yellow and orange samples
than in red samples. Interestingly, lutein was not found in the or-
ange C. chinense sample.
The content and bioaccessibility of carotenoids from plant foods
is determined by the physiological, genetic, and biochemical attri-
butes of a plant (Maiani, Casto9 n, Catasta, & Toti, 2009), including
species, part of the plant analysed, degree of maturity at harvest,
post-harvest handling, geographical origin, and seasonality (Chen
& Chen, 1992). Xanthophylls have been reported to be more
efficiently transferred than carotenes (Chitchumroonchokchai,
Schwartz, & Failla, 2004) as the lipophilicities of b-carotene and
b-cryptoxanthin result in a lower solubility of these carotenoids
in the micelles (van Het Hof, West, Weststrate, & Hautvast,
2000). Transfer efficiency varies among foods (van Het Hof et al.,
2000) and, furthermore, the transfer of carotenes to the micelles
can be impaired by the presence of other carotenoids (both
carotenes and xanthophylls) (Reboul, Richelle, Perrot, &
Desmoulins-Malezet, 2006).
Granado-Lorencio, Olmedilla-Alonso, Herrero-Barbudo, and
Blanco-Navarro (2007) reported that carotenoid micellarisation
can vary from one carotenoid to another in a given food, as well
as for each carotenoid in the different foods. O’Sullivan, Jiwan,
Daly, and O’Brien (2010) found that the bioaccessibility of carote-
noids from peppers ranged from 6.2% to 100%, depending on the
carotenoid and the food tested. In the present study, fresh red pep-
pers contained high amounts of the lipophilic provitamin A carote-
noids (b-carotene and b-cryptoxanthin) but these carotenoids were
significantly lower (P < 0.05) in the yellow and orange pepper sam-
ples (Table 2). Following in vitro digestion, we found that b-caro-
tene and b-cryptoxanthin showed lower% bioaccessibility, than
did the xanthophylls analysed, which had greater values of bioac-
cessibility, particularly capsanthin and zeaxanthin and, to a lesser
extent, antheraxanthin and lutein. In general, the bioaccessibility
of carotenoids from the red pepper samples was similar to that
previously reported by Hervert-Hernández, Sáyago-Ayerdi, and
Goñi (2010) and O’Sullivan, Jiwan, Daly, and O’Brien (2010).
Few studies have investigated the bioaccessibility of capsan-
thin, violaxanthin or neoxanthin in chili peppers. Following diges-
tion, the amount of carotenoids, neoxanthin and violaxanthin,
detected in the micelles, ranged from non-detectable to very low
and consequently the bioaccessibility was close to zero for the
majority of pepper samples, with the exception of the frozen red
pepper which showed a violaxanthin bioaccessibility of 9.9–
19.2% (Table 4).
Similarly, capsanthin was not found in the micelles of yellow
(P1) and orange (P5) pepper and hence the bioaccessibility was
0%. Interestingly, capsanthin was present at high concentrations
in the raw food and micelles of the orange pepper (P7) and the bio-
accessibility was 69.3% for fresh and 47.8% for cooked peppers.
Similar to the carotenoid content, carotenoid bioavailability
may also be influenced by a number of factors, such as processing,
cooking, fat content and/or the presence of fibres (Garrett et al.,
2000; Hedren et al., 2002; Rock, Lovalvo, Emenhiser, & Ruffin,
1998). Carotenoid release from the food matrix can be enhanced
2612 A. Pugliese et al. / Food Chemistry 141 (2013) 2606–2613
by cooking; however, cooking and processing have also been found
to have detrimental effects on carotenoid bioaccessibility from
vegetables (Bugianesi, Salucci, Leonardi, & Ferracane, 2004; Nunn
et al., 2006). Leaching of nutrients may occur during boiling; how-
ever, Liu, Perera, and Suresh (2007) did not detect carotenoids in
the water used to boil vegetables. Carotenoids have been shown
to leach into oil, due to their fat-soluble nature (Kao, Chiu, Tsou,
& Chiang, 2012). Processing can cause isomerisation and degrada-
tion of all-trans-b-carotene (Deming, Teixeira, & Erdman, 2002). In
our study we found that the influence of processing methods var-
ied with each cultivar and also with the processing method em-
ployed. b-Carotene and b-cryptoxanthin were most affected by
processing and both freezing and boiling resulted in significant de-
creases in these two carotenoids in several of the pepper samples.
The stability of specific carotenoids to thermal treatment has pre-
viously been shown to vary and Aparicio-Ruiz, Mínguez-Mosquera,
and Gandul-Rojas (2011) found that lutein was more stable to
heating than were either b-carotene or b-cryptoxanthin in olive
oil subjected to heat.
However, there was no significant difference in the micelle con-
tent of b-carotene and b-cryptoxanthin among the fresh, frozen
and cooked samples. Therefore, an increase in bioaccessibility fol-
lowing processing may compensate for the lower contents of these
carotenoids in the processed peppers. Ryan, O’Connell, O’Sullivan,
Aherne, and O’Brien (2008) found that b-carotene micellarisation
of red pepper strips was significantly increased following boiling
(10 min in boiling water).
Oruna-Concha, González-Castro, Lo9 pez-Hernández, and Simal-
Lozano (1997) found that the quantities of b-carotene and lutein
in frozen Padro9 n peppers were more or less constant over a
12 month period. Several additional studies have reported that, fol-
lowing freezing, carotenoid losses varied from 5% to 48%, while
there was one study which observed an increase in b-carotene con-
tent of corn following freezing (Scott & Eldridge, 2005). Our results
show that freezing decreases the content of carotenoids in peppers,
particularly b-carotene and b-cryptoxanthin in red peppers
(P < 0.01 and P < 0.05) but, to a lesser extent, also the other carote-
noids tested. However, despite the decrease in the carotenoid con-
tent following freezing and boiling, the micellar content was not
significantly lower for the processed peppers in comparison with
the fresh peppers and consequently the bioaccessibility of carote-
noids from processed peppers is often similar or slightly higher
than bioaccessibility of carotenoids from unprocessed peppers.
This is particularly evident for b-cryptoxanthin, b-carotene, and
for zeaxanthin.
5. Conclusion
This study was conducted to investigate the effects of different
processing methods (domestic freezing or boiling) on the caroten-
oid composition of seven varieties of coloured peppers. Red, orange
and yellow peppers were confirmed to be good sources of carote-
noids. Gaining knowledge on the effects of cooking and processing
on carotenoid-containing foods facilitates the optimisation of
carotenoid bioavailability for food manufacturers and helps con-
sumers obtain adequate intakes of carotenoids. Overall, with a
few exceptions, the carotenoid contents of the micelles were sim-
ilar, for both fresh and processed peppers. Therefore, the process-
ing conditions examined in this study do not negatively affect
the bioaccessibility of carotenoids from peppers.
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