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Food Chemistry
Red Chicory (Cichorium intybus) Extract Rich in
Anthocyanins: Chemical Stability, Antioxidant
Activity, and Antiproliferative Activity In Vitro
Aline Alves Migliorini, Camila Sztoltz Piroski, Taiana Gomes Daniel, Thiago Mendanha Cruz, Graziela Bragueto Escher,
Mariana Ara´
ujo Vieira do Carmo, Luciana Azevedo , Mariza Boscacci Marques, Daniel Granato , and Neiva Deliberali Rosso
Abstract: Red chicory leaves are appreciated sensorially and their constituents contain bioactive properties. The
objectives of this study were as follows: to use an experimental design to extract anthocyanins from red chicory in
aqueous solution at pH 2.5; to determine the stability of the extracts in relation to temperature and pH; and to evaluate
the antioxidant activity and in vitro cytotoxic effect of the lyophilized and purified extracts. The best extraction conditions
for the bioactive compounds from red chicory were a temperature of 64.2 °C for 25 min; the anthocyanin content
was 73.53 ±0.13 mg per 100 g fresh weight basis sample. The EC50 (Half maximal effective concentration) value for
the antioxidant activity assay in relation to DPPH (2,2-diphenyl-1-picrylhydrazyl) with optimized extract was 0.363,
which corresponds to a concentration of 39.171 µmol/L of anthocyanins. The activation energy for the degradation
reaction of the anthocyanins from the red chicory extract was 84.88 kJ/mol. The optimized extract, which was rich
in anthocyanins, showed chemical and biological antioxidant activity (protection against erythrocyte hemolysis) and
inhibited lipid peroxidation in vitro.TheCichorium intybus L. extracts interfered on the levels of reactive oxygen species
generation and the crude extract did not present procarcinogenic effect.
Keywords: Cichorium intybus L., free radicals, functional foods, thermal stability
Practical Application: Red chicory is basically consumed as a part of traditional dishes worldwide. Here, we developed a
process to extract and purify the anthocyanins from Cichorium intybus leaves and test the extracts in terms of the chemical
composition, thermal stability, antioxidant activity, and antiproliferative effects. The anthocyanin-rich extract presented
antioxidant activity in chemical and biological assays and low cytotoxicity and cytoprotective effects in relation to HepG2,
HCT8, and Caco-2 cell lines. Additionally, the red chicory extract protected human erythrocytes against hemolysis. This
extract may be used as a natural colorant/antioxidant in foods.
Introduction
Agricultural by-products are abundant, renewable sources of
natural compounds. Raw materials from plants are sources of di-
etary fiber, carotenoids, tocopherols, and polyphenols. Among
vegetables, red chicory (Cichorium intybus L.) has been the focus of
research regarding its content of phenolic compounds and antho-
cyanins. Red chicory leaves are appreciated sensorially and are used
in various culinary preparations (Innocenti et al., 2005). Sixty-
four chemical compounds were detected in the leaves of C. inty-
JFDS-2018-1631 Submitted 11/8/2018, Accepted 2/15/2019. Authors Miglior-
ini, Piroski, Escher, Granato, Daniel, and Rosso are with Graduation Program in Food
Science and Technology, State Univ. of Ponta Grossa, Av. Carlos Cavalcanti, 4748,
84030-900, Ponta Grossa, Brazil. Authors Cruz and Marques are with Department
of Chemistry, State Univ. of Ponta Grossa, Av. Carlos Cavalcanti, 4748, 84030-900,
Ponta Grossa, Brazil. Authors Carmo and Azevedo are with Department of Biolog-
ical Sciences, Federal University of Alfenas, Rua Gabriel Monteiro da Silva, 714,
37130-000, Alfenas, Brazil. Direct inquiries to author Daniel Granato (E-mail:
dgranato@uepg.br) and author Neiva D. Rosso (ndrosso@uepg.br).
Disclaimer: Dr. Bradley Bolling served as Scientific and Associate Editor over-
seeing single-blinded review of this manuscript. It is the policy of JFS to
blind Editorial Board members from the peer-review process of their own
submissions, just as all authors are blinded.
Part of a series of papers presented at the Dalian Food Nutrition and Health
Symposium 2018.
bus (var. “Treviso,” “Treviso Belgium,” and “Verona”), including
derivatives of hydroxycinnamic acid, mono and dicaffeoylquinic
acids, and three derivatives of tartaric acid. Thirty-one flavonols,
two flavone glucosides, and 10 anthocyanins were also identified
(Carazzone, Mascherpa, Gazzani, & Papetti, 2013). Reif, Ar-
rigoni, Sch¨
arer, Nystr¨
om, and Hurrell (2013) detected consid-
erable concentrations of lutein (1.76 to 6.98 mg/100 g) and β-
carotene (1.05 to 4.16 mg/100 g) in C. intybus var. foliosum. The
hydroalcoholic extraction of leaves of C. intybus L. detected the
presence of flavonoids, phenolic acids, tannins, saponins, and rel-
evant amounts of Mg and Zn (Abbas et al., 2015).
Va r i e t i e s o f C. intybus have been used in folk medicine to
treat liver disorders. The major anthocyanin identified in Inty-
bus Balou, Indigo, Manchini, Leonardo, and Erfano varieties of
chicory was cyanidin-3-O-(6-malonyl-glucopyranoside); the level
was over 95% and the highest content was detected in the Indigo
variety. The aqueous extract of Indigo, Balou, Leonardo, Man-
chini, and Erfano varieties inhibited lipid peroxidation in vitro,
and the anthocyanins isolated from this species presented the best
inhibition of lipid peroxidation and cyclooxygenase (Mulabagal,
Wang, Ngouajio, & Nair, 2009).
Red chicory extracts presented antioxidant, cytoprotective, and
antiproliferative activities in Caco-2 intestinal cell models. Red
chicory extract had a modulating effect on the oxidative stress
induced by 4-tert-OP (4-tert-octylphenol) and hepatotoxicity.
C2019 Institute of Food Technologists R
990 Journal of Food Science rVol. 84, Iss. 5, 2019 doi: 10.1111/1750-3841.14506
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