Phenolic Content and Profile and Antioxidant
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molecules Article (Poly)phenolic Content and Profile and Antioxidant Capacity of Whole-Grain Cookies are Better Estimated by Simulated Digestion than Chemical Extraction 1,2 3 1, 4 4 Francesca Danesi , Luca Calani , Veronica Valli y, Letizia Bresciani , Daniele Del Rio and Alessandra Bordoni 1,2,* 1 Department of Agricultural and Food Sciences (DISTAL), University of Bologna, Piazza Goidanich 60, 47521 Cesena, Italy; [email protected] (F.D.); [email protected] (V.V.) 2 Interdepartmental Center for Agri-food Industrial Research (CIRI Agrifood), University of Bologna, Via Quinto Bucci 336, 47521 Cesena, Italy 3 Department of Food and Drug, University of Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy; [email protected] 4 Department of Veterinary Science, University of Parma, Medical School, Building C, Via Volturno 39, 43125 Parma, Italy; [email protected] (L.B.); [email protected] (D.D.R.) * Correspondence: [email protected]; Tel.: +39-0547-338955 Current address: Sociedad Española de Colorantes Naturales y Afines (SECNA), Polígono 33, Parcela 254, y El Muladar, 46370 Chiva, Valencia, Spain. Academic Editor: Francesco Cacciola Received: 28 May 2020; Accepted: 14 June 2020; Published: 17 June 2020 Abstract: It is widely recognized that the biological effects of phytochemicals cannot be attributed to the native compounds present in foods but rather to their metabolites endogenously released after intake. Bioavailability depends on bioaccessibility, which is the amount of the food constituent that is released from the matrix in the gastrointestinal tract. The use of chemical extraction to evaluate the content and profile of phytochemicals does not mirror the physiological situation in vivo, and their bioaccessibility should be considered while assessing their nutritional significance in human health. The current study was designed to compare the (poly)phenolic profile and content and antioxidant capacity of whole-grain (WG) cookies using chemical extraction and a more physiological approach based on simulated digestion. Three types of organic WG cookies (made with durum, Italian khorasan, or KAMUT® khorasan wheat) were considered, either fermented by Saccharomyces Cerevisiae or sourdough. Although the flour type and the fermentation process influenced the release of phytochemicals from the cookie matrix, in almost all samples, the simulated digestion appeared the most efficient procedure. Our results indicate that the use of chemical extraction for evaluation of the phytochemicals content and antioxidant capacity of food could lead to underestimation and underline the need for more physiological extraction methods. Keywords: whole-grain wheat; bioaccessibility; antioxidants; (poly)phenols; durum wheat; KAMUT® khorasan wheat; simulated digestion; sourdough fermentation 1. Introduction Phytochemicals are bioactive non-nutrient components of fruits, vegetables, grains, and other plant foods that may have biological significance and provide desirable health benefits beyond basic nutrition, reducing the risk of major chronic diseases [1]. (Poly)phenolic compounds (PCs) represent one of the major groups of phytochemicals. It is widely recognized that most of the biological effects of PCs cannot be attributed to the native compounds present in foods but rather to their metabolites of various origins gaining contact with the Molecules 2020, 25, 2792; doi:10.3390/molecules25122792 www.mdpi.com/journal/molecules Molecules 2020, 25, 2792 2 of 11 Molecules 2020, 25, x FOR PEER REVIEW 2 internal compartments after food intake [2]. Therefore, the health benefits of dietary PCs depend not onlythe on internal their concentration compartments in after the food intake but mainly [2]. Therefore, on their absorption,the health benefits metabolism, of dietary and PCs availability depend in thenot target only tissue on [their3]. Bioavailability, concentration i.e.,in thethe proportionfood but mainly of a food on constituent their absorption, that is absorbed metabolism, and utilizedand in theavailability normal metabolismin the target [4tissue], depends [3]. Bioavailability, on bioaccessibility, i.e., the which proportion is the amount of a food of constituent the food constituent that is absorbed and utilized in the normal metabolism [4], depends on bioaccessibility, which is the amount that is released from the solid food matrix in the gastrointestinal tract [5]. The bioaccessibility of PCs is of the food constituent that is released from the solid food matrix in the gastrointestinal tract [5]. The greatly influenced by their interactions with the food matrix. In turn, these interactions depend on the bioaccessibility of PCs is greatly influenced by their interactions with the food matrix. In turn, these physicochemical properties of the food matrix (e.g., pH, temperature, texture), on the presence of other interactions depend on the physicochemical properties of the food matrix (e.g., pH, temperature, components that can improve or antagonize accessibility [6], and on the technologies used for food texture), on the presence of other components that can improve or antagonize accessibility [6], and processing [7,8]. on the technologies used for food processing [7,8]. TheThe majority majority of of studies studies examining examining thethe PC content or or the the antioxidant antioxidant activity activity of offoods foods do do not not considerconsider bioaccessibility bioaccessibility andand rely on on non non-physiological-physiological extraction extraction conditions conditions [9] that [9] often that ignore often ignorePCs PCsremaining remaining in the in theresidues residues of the of chemical the chemicallyly extracted extracted material. material. RecentRecent studiesstudies evidenced evidenced that these that thesenon non-extractable-extractable bioactive bioactive molecules molecules represent represent a major a major part partof total of totaldietary dietary PCs, PCs,and andexhibit exhibit a a substantialsubstantial biological activity activity [10] [10].. Therefore, Therefore, the the concentration concentration and and profile profile of ofPCs PCs in infood food after after chemicalchemica extractionl extraction do do not not mirror mirror the thephysiological physiological situation inin vivo vivo [11][11,], and and could could give give misleading misleading resultsresults while while assessing assessing their their nutritional nutritional significance significance inin human health. health. InIn the the current current study,we study, compared we compared two different two different approaches approaches to evaluate to evaluate the PC profilethe PC and profile antioxidant and capacityantioxidant of organic capacity whole-grain of organic (WG) whole cookies:-grain (WG) a standard cookies: approach a standard based approach on chemical based extraction on chemical and a moreextraction physiological and a approachmore physiological based on in approach vitro-simulated based digestion.on in vitro Simulated-simulated digestion digestion. was Simulated followed by ultrafiltration,digestion was allowing followed the by selection ultrafiltration of compounds, allowing that the were selection small enough of compounds to be potentially that were absorbable small throughenough the to intestinalbe potentially mucosa absorbable (<3 kDa through digested the fraction, intestinal as mucosa bioaccessible (<3 kDa fraction) digested [12 fraction,]. WGs as are concentratedbioaccessible sources fraction) of phytochemicals,[12]. WGs are concentrated mainly PCs sources [13], and of phytochemicals, cookies were chosen mainly as PCs the model[13], and food sincecookies they were are a chosen popular as foodstuff, the model consumed food since bythey a wideare a popular range of foodstuff, populations consumed [14]. by a wide range of populations [14]. Three different types of organic WG cookies were considered:( i) WG durum wheat cookies (DUR); Three different types of organic WG cookies were considered: i) WG durum wheat cookies (ii) WG Italian khorasan wheat cookies (KHO); and iii) WG KAMUT® khorasan wheat cookies (KAM). (DUR); ii) WG Italian khorasan wheat cookies (KHO); and iii) WG KAMUT® khorasan wheat cookies To be sold under the trademark KAMUT®, khorasan wheat must meet specific standards that so far (KAM). To be sold under the trademark KAMUT® , khorasan wheat must meet specific standards that are consistently satisfied only in the growing region of North America [15,16]. In this study, we used so far are consistently satisfied only in the growing region of North America [15,16]. In this study, we ® Italianused khorasan Italian khorasan wheat andwheat KAMUT and KAMUTkhorasan® kho wheatrasan wheat to evaluate to evaluate the impact the impact of environmental of environmental factors onfactors the content on the and content bioaccessibility and bioaccessibility of PCs. of PCs. ToTo verify verify the the possible possible influence influence ofof productionproduction processes [17] [17],, the the WG WG cookies cookies were were prepared prepared by by standardstandard fermentation fermentation using usingSaccharomyces Saccharomyces cerevisiae cerevisiae(DURS ,(DUR KHOS,, KAMKHOSS), orKAM sourdoughS) or sourdough fermentation usingfermentationS. cerevisiae usingand S. cerevisiae lactic acid and bacterialactic acid (LAB) bacteria (DUR (LAB)L, (DUR KHOLL, KHO, KAML, KAML).L The). The study study design design is schematicallyis schematically reported reported in Figure in Figure1. 1. FigureFigure 1. Schematic1. Schematic