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Broa, an Ethnic Maize Bread, As a Source of Phenolic Compounds

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Broa, an Ethnic Maize Bread, As a Source of Phenolic Compounds antioxidants Article Broa, an Ethnic Maize Bread, as a Source of Phenolic Compounds Andreia Bento-Silva 1,2,3 , Noélia Duarte 4 , Elsa Mecha 2 , Maria Belo 2, Ana Teresa Serra 5 , Maria Carlota Vaz Patto 2 and Maria Rosário Bronze 2,4,5,* 1 FCT NOVA, Faculdade de Ciências e Tecnologia, Campus da Caparica, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal; [email protected] 2 ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras, Portugal; [email protected] (E.M.); [email protected] (M.B.); [email protected] (M.C.V.P.) 3 FFULisboa, Faculdade de Farmácia da Universidade de Lisboa, Av. das Forças Armadas, 1649-019 Lisboa, Portugal 4 iMed.ULisboa, Instituto de Investigação do Medicamento, Faculdade de Farmácia, Universidade de Lisboa, Avenida Prof. Gama Pinto, 1649-003 Lisboa, Portugal; [email protected] 5 iBET, Instituto de Biologia Experimental e Tecnológica, Avenida da República, Quinta-do-Marquês, Estação Agronómica Nacional, Apartado 12, 2780-157 Oeiras, Portugal; [email protected] * Correspondence: [email protected] Abstract: Maize is an important source of phenolic compounds, specially hydroxycinnamic acids, which are widely known for their antioxidant activity and associated health benefits. However, these effects depend on their bioaccessibility, which is influenced by the different techniques used for food processing. Several traditional products can be obtained from maize and, in Portugal, it is used for the production of an ethnic bread called broa. In order to evaluate the effect of processing Citation: Bento-Silva, A.; Duarte, N.; Mecha, E.; Belo, M.; Serra, A.T.; Vaz on maize phenolic composition, one commercial hybrid and five open-pollinated maize flours and Patto, M.C.; Bronze, M.R. Broa, an broas were studied. The total phenolic content and antioxidant activity were evaluated by the Folin- Ethnic Maize Bread, as a Source of Ciocalteu and ORAC assays, respectively. The major phenolics, namely ferulic and p-coumaric acids Phenolic Compounds. Antioxidants (in their soluble-free, soluble-conjugated and insoluble forms), insoluble ferulic acid dimers and 2021, 10, 672. https://doi.org/ soluble hydroxycinnamic acid amides were quantitated. Results show that the total phenolic content, 10.3390/antiox10050672 antioxidant activity and hydroxycinnamic acids resisted traditional processing conditions used in the production of broas. The content in soluble-free phenolics increased after processing, meaning Academic Editors: Monica that their bioaccessibility improved. Portuguese traditional broas, produced with open-pollinated Rosa Loizzo and Rosa Tundis maize varieties, can be considered an interesting dietary source of antioxidant compounds due to the higher content in hydroxycinnamic acids and derivatives. Received: 27 March 2021 Accepted: 24 April 2021 Keywords: maize; broa; ferulic acid; p-coumaric acid; dehydrodiferulic acids; hydroxycinnamic Published: 26 April 2021 acid amides Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- 1. Introduction iations. Maize (Zea mays) is considered a staple cereal in many countries, where it is used to prepare different types of food products [1]. In Portugal, maize is the main ingredient (50–100%) of an ethnic bread known as broa. Rye, wheat, (0–50%) or a combination of the flours are also commonly added to the recipe [2]. Broas are traditionally prepared Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. using open-pollinated maize varieties. However, Portuguese maize landraces are at risk of This article is an open access article disappearing, due to the progressive adoption of more productive hybrid varieties, which distributed under the terms and originate broas less appreciated by consumers [2]. conditions of the Creative Commons Epidemiological studies have demonstrated that the consumption of phenol-rich Attribution (CC BY) license (https:// foods, as wholegrain cereals, in a regular diet is inversely associated with the risk of de- creativecommons.org/licenses/by/ veloping chronic diseases, such as oncologic and cardiovascular diseases, and metabolic 4.0/). Antioxidants 2021, 10, 672. https://doi.org/10.3390/antiox10050672 https://www.mdpi.com/journal/antioxidants Antioxidants 2021, 10, 672 2 of 21 syndrome [3]. Compared to other cereals, maize contains a high level of phenolic com- pounds, specially hydroxycinnamic acids, which can be found in their soluble or insoluble forms [4]. In particular, the ferulic acid (FA) content is at least ten-fold higher than in other cereal grains, which makes maize one of the most interesting sources of FA in the human diet [4]. Soluble phenolic compounds can be present in their free form or conju- gated with smaller molecules, such as simple sugars and amines, as hydroxycinnamic acid amides [5–9]. Insoluble phenolic compounds are mostly (>94%) bound to arabinoxy- lans [4,5,10], and include dehydrodiferulic, dehydrotriferulic and dehydrotetraferulic acids [4,5] and their derivatives, such as the recently described insoluble hydroxycinnamic acid amides [11]. Evidence from human intervention trials on the protective effects of phenol-rich foods, such as wholegrains, has been inconsistent, mainly due to differences in the food composition and in the bioaccessibility and bioavailability of phenolic compounds [3]. Their bioaccessibility is influenced by the processing techniques involved in the preparation of food products [4,12]. Soluble compounds are usually available for absorption by a simple diffusion mechanism [4,13]. Conversely, insoluble compounds have a very low bioaccessibility [4,13] and need to be liberated from the food matrix during small intestinal digestion or colonic fermentation, in order to be absorbed and become bioavailable [12]. In a previous study, several phenolic compounds, mainly hydroxycinnamic acids and hydroxycinnamic acid amides, were identified in maize flours and broas [11]. Since the total content and bioaccessibility of phenolic compounds is influenced by the techniques used for food processing [3], it is mandatory to evaluate the phenolic composition of the final food product. The understanding of the influence of processing also allows the selection of processing conditions that are capable of preserving and increasing the bioaccessibility of phenolic compounds. The study reported herein aimed at elucidating the changes occurring in the soluble-free, soluble-conjugated and insoluble phenolic compositions caused by the processing of raw flours to broas, and the potential implications on their bioaccessibility, bioavailability and bioactivity. 2. Materials and Methods 2.1. Reagents The reagents and commercial standards used for chromatographic analysis were described elsewhere [11]. Additionally, Folin-Ciocalteu reagent, sodium carbonate, 20,20- azobis-(2-amidinopropane) dihydrochloride (AAPH), Trolox and fluorescein sodium salt were obtained from Sigma-Aldrich, St. Louis, MO, USA. 2.2. Maize Flour and Broas Preparation Five traditional Portuguese open-pollinated maize varieties (Broa-213, Pigarro, Castro Verde, Verdeal de Aperrela, Fandango) (Table S1), and their corresponding broas were studied. All maize samples were obtained from controlled trials conducted at ESAC (Escola Superior Agrária de Coimbra) and flours were obtained after milling the grain in an artisan water-mill with millstones (Moinhos do Inferno, Viseu, Falling Number 3100). A commercial hybrid maize flour, commonly used for broa production (Nacional Type 175) was also included in the study for comparison. This flour was acquired already milled and is described elsewhere [2]. Broas were prepared in a bakery following a traditional recipe [14]. The ingredients included 70% maize flour, 20% commercial rye flour (Concordia type 70, Portugal) and 10% commercial wheat flour (National type 65, Portugal). Before extraction of phenolic compounds, broas were milled using a grinding mill (IKA MF 10.2, Königswinter, Germany) with a 1.5 mm sieve. The commercial rye and wheat flours used for broa production were also analyzed. 2.3. Preparation of Phenolic Fractions Cereal flours (maize, rye and wheat) and broas (4 g) were submitted to a conventional extraction procedure [11] with 20 mL of EtOH/H2O (50%, v/v), to obtain an ethanolic Antioxidants 2021, 10, 672 3 of 21 solution containing the soluble phenolic compounds and a solid residue comprising the insoluble compounds. Alkaline hydrolyses were applied to both the ethanolic solution and residue. Three phenolic fractions were obtained: the soluble (SF), the soluble-hydrolyzed (SHF) and the insoluble (IF) fractions (Figure1). The abbreviations used throughout the paper, including tables and figures, are listed in Table1. Table 1. List of abbreviations. Abbreviation Definition Abbreviation Definition 55DFA 5-50-Dehydrodiferulic acid Fan Fandango 85DFA 8-50-Dehydrodiferulic acid GAE Gallic acid equivalents 8O4DFA 8-O-40-Dehydrodiferulic acid HCAAs Hydroxycinnamic acid amides AA Antioxidant activity HPLC High performance liquid chromatography AAi Insoluble antioxidant activity IF Insoluble fraction AAs Soluble antioxidant activity LOD Limit of detection B213 Broa 213 LOQ Limit of quantification CFP Coumaroyl feruloyl putrescine n/a Not applicable Com Commercial ORAC Oxygen radical absorbance capacity CV Castro Verde PC Phenolic content DAD Diode array detector PCi Insoluble phenolic content DCS Dicoumaroyl spermidine PCs Soluble phenolic content
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