R 2.00 g/100 g > ¨ urgens, Johnson, & , and Susana I.P. Casal (Kotze, J 2019 Institute of Food Technologists Further reproduction without permission is prohibited C Metadata, citation and similar papers at core.ac.uk at papers similar and citation Metadata, L.) (Salem, Msaada, Hamdaoui, Limam, Acacia cyclops ´ opez, Almajano, Rodrigo, & Huguet, 2005; ,JorgeA.Saraiva ) During Flowering Carthamus tinctorius e´ A. Pereira Hoffmann, 2010), and rose species (Dafny-Yelin et al., 2005; carotenoids and flavonoids (Gamsjaegerto et our al., knowledge, only 2011). onedifferences According work associated has with investigated compositional (Skowyra petal et al., colors 2014). (red, However, noat one yellow, has and their characterized different violet) pansies flowering( stages, as already studied& for Marzouk, safflower 2011), although they can have awintergreen mild, taste, fresh depending flavor, orflower on a tastes the more prominent part strongerto of than salads flower or the (thecakes, used petals whole sorbets, as and alone). a icedas They garnish drinks), a as and are sweet well treat. to added multipurpose as, However, in embellish crystallized medicinal past, desserts and pansies agents, (frosted eaten expectorant, have with also been some emetic, used laxative,sedative, as antioxidant, depurative, and alterative, antiseptic propertiesSome (Tang et anti-inflammatory, al., of 2010). thosephenolic diuretic, biological compounds activitiesSchulz, and Heiselmayere, can & Musso, carotenoids be 2011; RopCalvo, (Gamsjaeger, et attributed Gallego, al., Azman, 2012; Baranska, to Skowyra, &Until Almajano, their 2014; now, few Vukics et studiesof al., on 2008). pansies the have physicochemicalactivity been characterization (Carazo, reported, L withVukics et some al., 2008), data mineral on composition (Rop antioxidant et al., 2012), and ,Jos -carotene/g dry weight) and monomeric anthocyanins (303 to β g µ ¸a, Portugal. ´ ario de Santi- through the analysis of the nutritional and nutraceutical features of pansies wittrockiana , Paula Baptista wittrockiana), × × ´ onia, 5300-253, Braganc Vol. 84, Iss. 3, 2019 doi: 10.1111/1750-3841.14482 r Viola ) from Violaceae family, represents The market of edible flowers is increasing, although little information in nutritional view is Viola , Elsa Ramalhosa wittrockiana × Edible flowers consumption and use are an increasing food trend worldwide, although information concerning Antioxidant activity, bioactive compounds, fatty acids, flowering stages, nutritional composition, pansies ¸a, Campus de Santa Apol Viola g Cy-3 glu/g dry weight); however, white and yellow pansies showed an increase in the values of total reducing µ

Journal of Food Science

to completely open flower stage.at Our distinct results stages underline of theimprove the development nutritional nutritional and differences properties between their of pansies potential foods. with health different benefits, colors suggesting that they can be used as ingredient to 402 capacity (total phenols), hydrolysable tannins, flavonoids, monomeric anthocyanins, and antioxidant activity from the bud highest contents of total carotenoids (873 to 1300 fresh weight), while red pansiesacid had the profiles, highest linoleic content acid ofthe carbohydrates was palmitic (8.0 always g/100 and predominant g linolenicin fresh (ranging weight). white acids. between Regarding and the During 18.7 fatty yellow and flowering, pansies, 51.0 there whereas g/100 in was red g an pansies fatty increase the acids), in values followed protein, did by fat, not and change. Red linolenic pansies acid were contents characterized by the with different colors (white, yellow,flower and composition. When red) completely and open, white flowering and stages. yellow pansies Both had flower the type highest contents and of flowering protein stage ( influenced the popularization of pansies ( their nutritional composition and nutraceutical value is still scarce. Thus, the aim of this study was to contribute to the study underline the nutritionaltheir differences potential between health pansies benefits,foods. with suggesting different that colors they at can distinct be stages used of as development ingredient and to improve the nutritional properties of available. So, the presentprospective source study for the was food conducted industry, as to well contribute as a to promising the product for popularization human nutrition. of The edible results flowers of the as present a new and Practical application: Keywords: Abstract: Pansies ( The consumption and use of edible flowers have increased in re- JFDS-2018-1785 Submitted 11/8/2018, Accepted 1/30/2019. Authors Fer- reira, 228, 4050–313, Porto, Portugal.(E-mail: Direct [email protected]). inquiries to authors, Elsa Ramalhosa, Pereira are with Mountain Researchnic Centre Inst. (CIMO), School of of Braganc Authors Agriculture, Polytech- Fernandes and Casalmatology and are Hydrology, with Faculty LAQV@REQUIMTE/Laboratory of of Pharmacy, Bro- Porto Univ., Rua Jorge Viterbo Fer- nandes and Saraiva(QOPNA), are Chemistry with Dept., Organicago, Univ. 3810-193, Chemistry, of Aveiro, Natural Aveiro, Portugal. Products Campus Authors and Universit Fernandes, Agrifood Ramalhosa, Baptista, and of rainbow pastel colors,same often flower (Lim, with 2014). two The or whole three flower and colors buds on are the edible, sufficiently exploited fromview. the nutritional and health pointsone of of the most popular edible flowers. Petals come in a myriad increased compared to the pastJurikova, (Loizzo Neugebauerova, et & al., Vabkova, 2016; 2012).ble Rop, Even flowers Mlcek, can though be edi- considered food sources, they have not been cent years. Supermarkets are beginning tochefs sell flowers to and gourmet use themmore, on the their number dishes, drinks, of and scientific desserts. papers Further- regarding this topic has 490 Introduction

Luana Fernandes Nutritional and Nutraceutical Composition of Pansies ( CORE

Provided by Biblioteca Digital do IPB do Digital Biblioteca by Provided Food Chemistry eemaue ihadgtlclpr( o10m;Powerfix, Aquino-Bola mm; by 150 to (0 caliper digital UK). a Leeds, width with and balance measured length, digital dimensions, were a Axial on Germany). weighed Balingen, were (Kern, color and stage flowering each a nlzdi ifrn onst suerepeatability. assure epidermis to petal points different of in structure analyzed (Nikon was camera The a Florida). with Tallahassee, equipped SMZ-U, (Leitz UK) microscope Chippenham, light a 12, on laborlux sec- observed and longitudinal water and in transversal mounted of random tions, sections into cut Thin then structure. were petals microscopic their regarding analyzed temperature room at kept light. and from Denmark), protected powder, Lynge, pre- homogenous Coolsafe, flowers to (Scanvac, in ground later lyophilization performed by on were served performed others were were while flowers analyses flowers, Some the fresh laboratory. harvest, the After ap- to of from plants. transported stage collected, different were and flowers 50 color color stemless proximately each each of For of g 1). 500 plants half-opened development, (Figure and (bud, selected flower), randomly stages open were flowering completely three and at flower, Braganc harvested of were Inst. ers School Polytechnic the of Agriculture, greenhouse of the from obtained were red) and yellow, hscceia analysis Physicochemical petals pansies of analysis Microscopic Samples Methods and Material industries. supplements or food and the consumer for the for applications relevant different are affect latter that The stages traits. flowering nutritional distinct their at and colors different and on of information open, pansies provides work half this (bud, Thus, stages open). completely flowering flower (white, different colors at different red) with and pansies yellow, of power) ac- reducing scavenging and radical tivity DPPH and capacity, reducing antho- total cyanins, monomeric acids, tannins, fatty hydrolysable individual flavonoids, composition, the carotenoids, nutritional in weight, knowledge and and physicochemical color the (a activity their increase water in (dimensions, to properties and biological was petals food of So, research the structure nutrition. this for microscopic human of for source product prospective aim promising and the a new as well a as as industry, flowers edible of & flowering. Vyas, during changes Sood, protein activity, antioxidant and 2010; phenolics, volatiles, Stampar, on focusing 2006), & Nagar, Osterc, Veberic, Schmitzer, . . . pansies of composition Nutritional ee Nvsn,Lbwf-w ahn Switzerland). Lachen, LabSwift-aw, (Novasina, meter ie ih5 Lo itle ae,bie o r lee,and filtered, Amorim, hr, HI8417, (Hanna 1 Portugal). potentiometer for a boiled with water, measured distilled pH wasthe of sample mL each 50 of with g mixed 1 Briefly, modifications. some with 920.149, Chav Hwsmaue codn otemto described method the to according measured was pH egt iesos oo,p,ada and pH, color, dimensions, Weight, were flowers open completely of color each of petals Three ( Pansies hssuywscnutdt otiuet h popularization the to contribute to conducted was study This ae ciiy(a activity Water zSri,adVerdalet-Guzm and ez-Servia, ´ Viola × o,Urrutia-Hern nos, ˜ w wittrockiana a eemndi otbewtractivity water portable a in determined was ) ftredfeetclr (white, colors different three of ) n(03 n OCmethod AOAC and (2013) an ´ ne,DlCastillo-Lozan, Del andez, ´ a(otgl.Flow- (Portugal). ¸a w . w ensamplesof o s e l p m a s n Te ,p,flowers’ pH, ), urtoa composition Nutritional urcuia composition Nutraceutical nrywscluae codn oE.(1): Eq. to according calculated was difference. Energy mass 985.29 by No. calculated were method Carbohydrates official 2003). AOAC enzymatic- (AOAC, an on by based method determined was gravimetric analysis fiber acid Dietary fatty below. the detailed for as preserved were samples These apparatus. (2,6-di- BHT of 0.01% tert weight with known ether petroleum a with extracting sample by powdered determined to were according Lipids (2012). 6.25, of factor L conversion macro- Sotelo, a the by with estimated method, was samples Kjeldahl the of content Protein ashes. 550 to sample at 105 the cination drying at by the weight determined following weight constant was fresh analyzed g content were g/100 Moisture in (fw). sample expressed and each (1990), of procedures AOAC fiber) dietary and naSlc AE(0 m out (100 carried FAME was GC Select separation by a acid on ioniza- Fatty determined (GC–FID). flame were detection with acids tion Netherlands) Fatty model, CP-9001 (2011). (Chrompack, according 12966-2 2M, cold ISO –20 hydroxide by at obtained to potassium were stored methanolic esters was methyl with and acid hydrolysis Fatty BHT analysis. had acid fraction fatty lipid the tion, n eeto ntefl cnmd iha with mode scan full the in detection and source Agilent (MS an detector on 5977B 230 MSD column an similar with peaks a 7890A, using chromatograph chromatographic GC–MS the by confirmed of was identification The Netherlands). System B.V., Middelburg, Data International Chromatography (Chrompack 2.5 Maitre Version program, of CP the pressure by 250 a performed were were detector at and gas injector 260 the carrier and of as temperatures The used kPa. was 190 Helium column. USA) bobnea 5 madcmaigterslst a to 8.8 µ results to the (0.22 comparing curve with and calibration the nm mL reading 450 by 100 at determined to absorbance water, was completed content residual carotenoid and any Total with filtered hexane. eliminate mixed solution to added was remaining sulphate was the phase sodium water anhydrous distilled acetone-free g of The 5 mL acetone. 200 eliminate and to funnel separation mL combined were a 20 extracts of in with Both gram v/v). twice One (1:1, (2018). extracted solution al. acetone:hexane was et sample Fernandes powdered by used freeze-dried method the to Library. ing Spectral Mass 14 NIST the and standards analytical using ihsih oictos reedidpwes( )o each of (2014) g) al. et (1 Li powders by Freeze-dried described modifications. method slight the on with capacity. based reducing was formed total and tannins, g h urtoa opsto mitr,ah a,carbohydrates, fat, ash, (moisture, composition nutritional The at acids. Fatty Carotenoids. ooei nhcais oa aood,hydrolysable flavonoids, total anthocyanins, Monomeric btl4mtypeo)t rvn xdto,uigaSoxhlet a using oxidation, prevent to -butyl-4-methylphenol) β ° crtn qiaet10gdw. g equivalent/100 -carotene Energy ;M udooe150 Quadropole MS C; + ° opez-Garc ,rsetvl.Tecleto n rcsigo h data the of processing and collection The respectively. C, ´ ( 9  × 0 fw g 100 lipids o.8,Is ,2019 3, Iss. 84, Vol. srpre ntentiinlcmoiinsec- composition nutritional the in reported As ° kcal uiga es r ni civn white achieving until hr, 2 least at during C aoeodcnet eedtrie accord- determined were contents Carotenoid ´ a n Basurto-Pe and ıa, ) +  ( ° 2 = ;ahcnetwsmaue ycal- by measured was content ash C; ×  × (4 itr fiber dietary µ ° /L.Rslswr xrse in expressed were Results g/mL). ;axlaytmeaue280 temperature auxiliary C; × .5mm 0.25 (protein r ora fFo Science Food of Journal a(07 n o tal. et Rop and (2007) na ˜ )  × +  h xrcinper- extraction The m carbohydrates)) 0.25 0 fw g 100 / z g f3 o800), to 30 of µ )(Agilent, m) β  -carotene ° Cfor 491 ° (1) C;

Food Chemistry × (2) 0.1% Sample values 100 3 A × 50 Sample A − DPPH A DPPH A = ) % ( C for 20 min after which 2.5 mL of ] 1% (m/v). After shaking, the mix- ° 6 versus concentration, the EC with three colours (white, yellow, and red). Completely open Completely 700 nm [Fe(CN) 3 The reducing power of each extract was deter- wittrockiana × is the absorbance of the DPPH solution and ) was calculated from the graph of the DPPH radical mol/L). After 1 hr in the dark, at room temperature, ab- Viola 50 5 − DPPH DPPH radical scavenging effect were added to 2.7 mL10 of the DPPH methanolic solution (6.09 To 1.0 mLtions, of were pansies added extract 2.5and solutions, 2.5 mL at mL of different of phosphate K concentra- buffer 0.2 M (pH 6.6) scavenging effect percentage versus extract concentration. Reducing power. mined by the procedure described by Fernandes et al. (2018). sorbance was read atby 517 the nm. percentage Antioxidant of activity scavenging effect, was according expressed to Eq. (2): tures were incubated at10% 50 trichloroacetic acid (m/v) wasvolume added of with 2.5 further mL stirring.tube, of A to which the 2.5 mixture mL was of distilled transferred water to and 0.5 another mL test of FeCl is the absorbanceconcentration in providing 50% the of(EC DPPH presence radical of scavenging the effect sample. The extract (m/v) were added. TheFrom absorbance the values graph were Abs read at 700 nm. A Half open Flowering stages Flowering L; the extract µ Vol. 84, Iss. 3, 2019 r Bud g cyanidin-3-glucoside/g dry weight µ Journal of Food Science C for 30 min, under agitation (IKA, RCT Model B, Staufen, ° Antioxidant activity. The total monomeric anthocyanins, total flavonoids and hy- g Cy 3-glu/g dw), flavonoids in mg of quercetin equivalent/g µ concentration range varied between 0.09 andsolution; 1.75 the mg extract/mL dilutions were performed with extraction solution) 492 determined by(2018). the Pansies procedure extract described diluted by solutions Fernandes (300 et al. DPPH (2,2-diphenyl-1-picrylhydrazyl)The radical DPPH scavenging radical activity. scavenging activities of the extracts were ( dry weight (mg QE/gacid dw), equivalent/g hydrolysable dry tannins in weightgallic mg (mg acid of TAE/g equivalent/g dw), tannic dry and weight (mg TRC GAE/g in dw). mg methodologies used by Fernandes etwere al. performed (2018). in All measurements triplicate.cyanins The were expressed results in for monomeric antho- foil under freezing until further analysis. drolysable tannins contents, as(TRC) well of as the edible the flowers total extracts were reducing quantified following capacity the freeze drier (Coolsafe, Lynge,obtained Denmark) were redissolved for with 2 the same days. solventa (water:acetone) The concentration to extracts of 50 mg extract/mL and covered with aluminum 37 Germany) at 1000 rpm.and The placed water:acetone in extracts a were rotaryto filtered evaporator remove (Stuart, acetone. Then, RE300DB, Stone, all UK) extracts were frozen and stored in a Figure 1–Flowering stages (bud, half open flower and completely open flower) of sample were extracted with 50 mL of water:acetone (6:4, v/v) at Nutritional composition of pansies . . .

Food Chemistry tg fflwrn bd hwdtelws ausadreached and first values lowest the at the showed length in and (bud) presented width, flowering weight, are of the flowering stage expected, during As colors 1. three Table with pansies for oos u eut eesmlrt hs ecie yWeryszko- by described (2012). those Sulborska to and similar Chmielewska were three results the Our between observed colors. were differences (C), were (E) structural walls stomata No folded found. with mesophyll cells epidermis in and (D), air bundles Furthermore, was vascular intercellular mesophyll them. covered large The with between thin, striate. cells, were spaces branched parallel cells irregular forming by papillae cuticle composed the of was layer of it a mesophyll structure, walls by and this the detail (A) more that petals in detected all observing Conical By of found. 2). epidermis were (Figure (B) adaxial structure the their in to papillae relation in examined were test. Levene’s by tested was variances the of homogeneity were The they the not). (if in test Games–Howell variances or namely, identical) (if were groups performed, test different was difference significant analysis honestly Tukey’s hoc the post a detected variances, treatments, were of between differences homogeneity of significant not if Additionally, or respectively. existence if the ( on determine differences depending to significant out were carried variance normality there were of The Welch Analysis ANOVA data. test. Shapiro–Wilk or the by (ANOVA) verified of was treatment data statistical the of the for used was irsoi nlsso ase petals pansies of analysis Microscopic Discussion and Results analysis Statistical 0.5. that of concentration absorbance extract an gave the to walls. folded corresponding with ( determined cells mesophyll epidermis were and (E) (A) mesophyll; epidermis petal adaxial the the in in bundles vascular papillae branched conical (D) the stomata; with a petal with the epidermis of the section of longitudinal fragment petals: (C) pansies yellow of 2–Structure Figure . . . pansies of composition Nutritional hsclanalysis Physical egt,lnts its H n ae ciiy(a activity water and pH, widths, lengths, Weights, yellow of Petals IL), Chicago, Inc., (SPSS 18.0 v. software, Statistical SPSS The Viola × wittrockiana P < opeeyoe flowers open completely .5 ewe samples, between 0.05) w values ) ifrn ooswr eetd u ihu n pattern. any without of but pansies detected, between were as colors well different as stages flowering between differences nte pce fvoa eadn Hada and pH Regarding viola. Damalas, of study. species for another present mm) (15.2 mm) the (18.8 width length detected lower and in were reported (2014) analyzed stages Fotiadis and flower cultivars Koutroubas, in distinct differences the significant at among No weight stage. from and flower prominent half-open dimensions less to and bud stage half-open flower the the open from completely notable open. most to was flower completely dimensions were in flowers increase the The when values maximum their urtoa composition Nutritional otns n oe br(. /0 w ausfrpansies. for values dw) dw) g g g/100 g/100 (64.5 (9.3 higher de- carbohydrates fiber reported who lower and (2013) and (2012), dw) contents, Vieira al. g protein. et g/100 of g Rop (16.8 g/100 dw by protein 8.11 g as ash g/100 well dw; 6.7 as g tected flowers, g/100 pansy 3.22 for fat dw; dw) dw; g g in g/100 g/100 is 15.4 47.7 dw) protein (carbohydrates g macronutrients of g/100 values 10.5 similar to Navarro-Gonz 6.3 Gonz and ash Javier, by Luna-Recio, described dw; dw; ones g g the g/100 g/100 with 23.17 accordance to 5.21 9.15 to protein dw; 4.48 dw; g g fat g/100 g/100 43.2 55.6 to to 17.2 42.0 fiber (carbohydrates results weight bud of dry and range to flower Our respectively. converted flower, white red open 31 open completely completely between for and fw, yel- varied g open kcal/100 energy completely fw). 52 the g caloric and and g/100 The bud 0.37 respectively. 1.16 white flower, between in to fw, low ranging (0.92 g macronutrient, ash g/100 proteins abundant 1.31 and fol- fw), and less fw), g fw), the g g/100 was g /100 4.66 Fat g g/100 to 2.06 abundant (1.50 8.78 to fiber most to (0.95 dietary second (3.94 total (Rop by the samples flowers lowed were all edible in Carbohydrates for authors macronutrients 2012). other al., by fw, described et g been g/100 91.3 main has and the 85.0 as was between Water ranging 2. pansies, Table in in constituent presented is stages flowering tinct h urtoa opsto fpniso ifrn oosa dis- at colors different of pansies of composition nutritional The o.8,Is ,2019 3, Iss. 84, Vol. il arvensis Viola r ora fFo Science Food of Journal lz(08,woreported who (2018), alez ´ oes oee,ti is this however, flowers; lzBri,Periago, alez-Barrio, ´ w e significant few , 493 B);

Food Chemistry c,A a,B a,A a,b,B b,A b,A b,A c,B c,A,B b,B b,A b,A,B a,A a,B a,A a,A a,A a,A a,A a,A a,A a,A a,A a,A 1 1 8 2 6 4 8 9 1 5.64 11.42 6.30 6.18 5.45 5.48 0.001 0.001 0.001 0.12 0.15 0.18 0.09 0.07 0.08 ± ± ± ± ± ± ± ± ± fw) Energy ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 47 31 31 46 51 48 52 48 52 . (kcal/100 g ∗ 0.62 0.63 0.68 6.03 6.21 6.01 53.12 45.98 49.84 58.38 48.24 53.69 0.982 0.978 0.984 a,A a,A a,A a,A a,A a,A a,A a,A a,A 0.05). Uppercase letters 0.07 0.03 0.08 0.09 0.08 0.19 0.14 0.12 0.13 < Ash ± ± ± ± ± ± ± ± ± -value (g/100 g fw) P 1.03 1.00 0.92 1.16 1.16 1.10 1.08 1.02 0.94 b,A b,A a,A b,A a,A b,C b,A b,A a,B a,A a,b,A b,A 0.1 0.7 0.1 0.1 0.1 0.1 0.1 0.1 0.2 a,A a,A a,A b,B b,A,B b,A a,A a,A a,A a,b,A a,A b,B ± ± ± ± ± ± ± ± ± Total 5.01 3.30 3.08 3.79 4.43 3.22 0.001 0.001 0.001 0.05 0.07 0.04 0.08 0.05 0.13 4.5 4.5 1.5 4.6 4.0 4.3 4.4 4.7 3.8 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± dietary fiber (g/100 g fw) ∗ 0.20 0.20 0.20 6.10 6.15 6.32 The fatty acid composition of pansies at different Flowering stage 18.33 14.97 13.84 19.28 20.46 22.00 0.985 0.983 0.984 a,A a,A a,A a,A a,A a,A a,A a,A a,B ˜ aes, Barros, Carvalho, & Ferreira, 2010; Pires, Dias, 0.1 1.3 1.9 0.1 1.6 0.7 1.9 2.3 0.4 ± ± ± ± ± ± ± ± ± 7.7 4.2 3.9 7.1 8.8 4.8 7.7 6.6 8.0 0.05). Uppercase letters present the differences among different colors at each (g/100 g fw) Carbohydrates < Fatty acids. enzymes required for their production. Moreover, bothtioned were men- to reduce someblood pressure heart and cholesterol disease profile risk (Miyoshiet et factors, al., al., 2014; as Miura 2008; triglycerides 2010; Ramel, Shidfar, Martinez, Keshavarz, Hosseyni, Kiely,Singer Ameri, Bandarra, et & & al., Yarahmadi, 1990). Thorsdottir, 2008; detected Furthermore, in high myristic contents (C14:0) in acid redtadecanoic and was acid yellow pansies, also (C17:0) as in welloverall as white fatty hep- pansies. acid When profile considering for the pansies, it was found that this flower flowering stages and colors isacids presented were identified in in Table 3. pansies.linoleic Eighteen The predominant fatty acid fatty acids (C18:2n6), were linolenic followed acid (C18:3n3). by Similar results palmitic were reportedthors by acid other (Guimar au- (C16:0) and Barros, & Ferreira, 2017). Theacid highest relative (C18:2n6) amounts of were linoleic detectedacid in (C18:3n3) red in budsof completely palmitic (51.0%), open acid of yellow (C16:0) linolenic flowersThe in first (23.0%) completely two open and fatty red acidssynthesized flowers are by (17.4%). essential the fatty human acids, organism as due they to cannot the be lack of desaturase Nutraceutical composition of three different colors at distinct flowering stages a,A a,A a,A a,B a,A b,B a,C a,B a,A -value P , water activity. w a,B b,A,B a,A 0.02 0.09 0.01 0.04 0.04 0.09 0.09 0.04 0.03 a,A,B a,B a,A a,A a,A a,A a,A a,B a,A,B b,A,B a,B a,A Fat ± ± ± ± ± ± ± ± ± 1.57 2.17 0.04 0.10 0.001 3.61 0.001 0.05 2.01 0.07 0.03 1.37 1.52 0.06 0.001 0.05). a of three different colors at distinct flowering stages. (g/100 g fw) ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± < 0.75 0.80 0.67 0.37 0.36 0.45 0.54 0.47 1.31 wittrockiana × b,B a,A b,A a,A a,A b,B a,A a,A b,B -value P Viola 0.05 0.06 0.03 0.01 0.13 0.06 0.14 0.01 0.03 wittrockiana ± ± ± ± ± ± ± ± ± Protein × (g/100 g fw) 1.46 1.19 1.36 1.05 1.10 2.03 0.95 0.96 2.06 Viola flower. In fact, they stated that a,A a,A a,A a,A a,A a,B a,A a,A a,A,B Vol. 84, Iss. 3, 2019 r 0.3 1.8 7.0 1.5 1.7 0.3 0.3 2.1 0.7 RedRedRed 0.12 Red 7.79 Red 18.39 6.08 0.982 ± ± ± ± ± ± ± ± ± WhiteWhiteWhite 0.15 White 7.94 White 15.86 6.15 0.985 Yellow 0.984 Moisture standard deviation. Lowercase letters present the differences among different flower stages of a single color ( (g/100 g fw) standard deviation. fw, fresh weight. 85.1 91.3 86.5 ± ± 0.05). Crataegus monogyna < stage -value open open open Flowering P Half openCompletely 85.3 Half openCompletely 85.3 Half openCompletely 89.5 Journal of Food Science Concerning flowering stages, no consistent patterns were ob- w Dietary fiber is not included. Values are expressed as mean flowers at different flowering stageswithout showed significant low energy differences contents, betweenlow calorie them, diets. being suitable for nutritional composition may be observed duringof the different development flowers, includinggenerally pansies. the completely For open nutritionaltein flowers presented purposes, content, the but highest pro- the lowest total dietary fiber. Nevertheless, all tion of stagestage 1 6 (small (fully bud with opena flower petals decrease at still in anthesis)dw) covered protein by that (Dafny-Yelin et content sepals) was al., of to accompanied 2005). petals So, by considerable (203.2 variations to in 88.6 the g/100 g moisture and protein contents decreased, whileand ash, carbohydrates, fat increased whenpattern comparing for buds protein with was found flowers. A in similar roses, namely, from the transi- fiber decreased, while incontent red was pansies detected. an Barros, increasefailed in Carvalho, to and carbohydrates Ferreira detect (2011)development a of also pattern of macronutrient turnover during the served between pansies with differenttein and colors. fat An contents increase wasthe in observed last pro- in stage white of and flowering yellow pansies (completely on open flower) while dietary 494 flower stage ( ∗ Values are expressed as: mean Lowercase letters present the differences among different flower stages of a single color ( Red Bud 85.0 Yellow Bud 85.6 Color White Bud 85.6 Table 2–Nutritional composition of ∗ pHa Yellow 6.18 Length (mm) Yellow 21.02 Width (mm) Yellow 7.86 ParametersWeight (g) Color Yellow Bud 0.18 Half open Completely open Table 1–Physico-chemical characterization of Nutritional composition of pansies . . . present the differences among different colors at each flower stage (

Food Chemistry urtoa opsto fpnis... . . pansies of composition Nutritional

Table 3–Fatty acids composition (g fatty acids/100 g fatty acids) of the oils extracted from Viola × wittrockiana of different colors at distinct flowering stages.

White Yellow Red Fatty acid Bud Half open Completely open Bud Half open Completely open Bud Half open Completely open SFA C12:0 2.0 ± 2.5a,A 1.1 ± 0.3a,A 1.1 ± 0.3a,A 3.2 ± 0.7a,A 4.2 ± 0.9a,B 16.1 ± 5.4b,B 2.2 ± 0.3a,A 4.9 ± 3.2a,B 23.4 ± 0.9b,C C14:0 2.2 ± 1.4a,A 6.9 ± 4.0b,A 5.0 ± 0.9a,b,A 13.8 ± 6.4a,B 14.8 ± 5.8a,B 11.8 ± 5.6a,B 5.0 ± 0.7a,A 8.6 ± 1.5b,A,B 17.8 ± 0.4c,C C15:0 0.4 ± 0.4a,A 0.2 ± 0.1a,A 0.2 ± 0.3a,A 0.3 ± 0.2a,A 0.5 ± 0.4a,A 0.2 ± 0.2a,A nd 0.4 ± 0.4a,A 0.3 ± 0.1a,A C16:0 15.0 ± 2.1a,A 15.4 ± 1.9a,A 16.6 ± 3.7a,A 16.6 ± 0.3a,A 17.0 ± 2.3a,A 16.5 ± 1.0a,A 14.8 ± 1.1a,A 14.7 ± 2.1a,A 17.4 ± 0.6b,A C17:0 12.3 ± 9.4a,B 8.9 ± 2.9a,A,B 14.9 ± 3.1a,B 2.6 ± 0.7c,A 1.4 ± 0.2b,A 0.5 ± 0.2a,A 1.2 ± 0.3a,A 12.0 ± 9.8b,B 1.0 ± 0.6a,A C18:0 3.0 ± 0.4a,b,A 4.5 ± 1.7b,A 2.4 ± 1.2a,A 3.2 ± 1.6a,A 4.2 ± 0.8a,A 2.9 ± 0.5a,A 3.0 ± 0.2a,A 3.8 ± 0.5b,A 3.6 ± 0.2b,B C20:0 0.6 ± 0.6a,A 1.2 ± 0.4b,A nd 1.0 ± 0.5a,b,A 1.3 ± 0.2b,A 0.5 ± 0.4a,A 1.0 ± 0.2a,A 0.6 ± 0.7a,A 0.4 ± 0.2a,A C22:0 1.6 ± 0.4b,A 2.1 ± 0.8b,A 0.6 ± 0.5a,A 1.8 ± 0.5b,A 2.0 ± 0.5b,A 1.0 ± 0.2a,A 1.3 ± 0.2b,A 1.2 ± 0.7a,b,A 0.7 ± 0.1a,A C24:0 2.7 ± 3.6a,A 2.3 ± 1.2a,A 1.5 ± 0.6a,A 2.1 ± 1.2a,A 2.0 ± 1.1a,A 1.4 ± 1.2a,A 1.4 ± 0.4a,b,A 2.4 ± 1.2b,A 1.0 ± 0.1a,A MUFA C16:1n7 nd 0.6 ± 0.3A nd 0.3 ± 0.2a,A 0.2 ± 0.2a,A 0.3 ± 0.2a 0.3 ± 0.1A nd nd C18:1n9 6.8 ± 1.6a,b,A 7.8 ± 1.5b,B 5.3 ± 1.4a,B 10.8 ± 3.4b,B 11.5 ± 2.5b,C 3.6 ± 0.7a,A 6.2 ± 2.0b,A 3.6 ± 0.6a,A 4.4 ± 0.5a,b,A,B PUFA C18:2n6 37.4 ± 5.8a,B 33.6 ± 1.5a,A 33.2 ± 3.8a,B 29.1 ± 1.6a,A 25.6 ± 11.3a,A 21.9 ± 1.9a,A,B 51.0 ± 3.9c,C 32.8 ± 5.6b,A 18.7 ± 0.7a,A C18:3n3 14.6 ± 1.7a,A 14.3 ± 0.5a,A 19.3 ± 3.6b,B 13.8 ± 3.0a,A 15.4 ± 2.2a,A 23.0 ± 1.5b,C 12.7 ± 3.1a,A 14.1 ± 3.0a,A 11.0 ± 0.3a,A A a,A a,A a a,A o.8,Is ,2019 3, Iss. 84, Vol. C18:4n3 0.2 ± 0.4 nd nd 1.1 ± 1.2 nd 0.4 ± 0.3 nd 0.8 ± 1.0 0.4 ± 0.3 SFA 39.9 42.7 42.2 44.5 47.3 50.8 29.9 48.6 65.5 MUFA 7.4 9.1 5.3 11.5 11.7 3.9 6.4 3.6 4.4 PUFA 52.7 48.2 52.5 44.0 41.0 45.3 63.7 47.7 30.0 PUFA/SFA 1.32 1.13 1.24 0.99 0.87 0.89 2.13 0.98 0.46 n6/n3 2.6 2.3 1.7 2.1 1.7 0.9 4.0 2.3 1.7

nd, not detected; mean ± standard deviation (n = 3). Lowercase letters present the differences among different flower stages of a single color (P-value < 0.05). Uppercase letters present the differences among different colors at each flower stage (P-value < 0.05). r ora fFo Science Food of Journal 495

Food Chemistry - t. c,B b,B a,C c,C b,C a.A a,A c,A b,B 50 β g 0.02 0.01 0.01 0.02 0.01 0.01 0.01 0.01 0.01 Hyper- µ ± ± ± ± ± ± ± ± ± (mg/ml) Reducing Power EC 0.92 0.84 0.67 1.71 0.99 0.58 0.48 0.73 0.64 0.05). Uppercase < b,B a,B a,C c,C b,A a,A a,A c,A b,B 50 -value 0.01 0.02 0.01 0.01 0.01 0.01 0.01 0.02 0.01 P r( ± ± ± ± ± ± ± ± ± (mg/ml) DPPH EC 0.43 0.38 0.38 0.48 0.30 0.20 0.17 0.34 0.26 a,A b,A c,A a,A b,A c,A a,B a,b,B b,B 0.7 2.9 0.8 0.3 0.1 0.6 24 44 42 g Cy-3 glu/g dw, respectively) were ± ± ± ± ± ± ± ± ± gCy-3 µ µ ( glu/g dw) 6.3 1.5 3.1 Monomeric 303 353 402 anthocyanins 11.1 35.9 22.9 a,A b,C c,C b,B a,A c,B b,C a,B a,A flowers revealed significant differences between 1.6 1.0 1.0 0.6 0.7 5.0 1.1 4.0 4.0 ± ± ± ± ± ± ± ± ± Flavonoids 40.7 77.7 47.3 42.2 82.5 68.4 51.5 47.9 124.5 (mgQE/gdw) In white, yellow, and red pansies, other bioactive compounds Concerning flowering stages, significant differences in pansies of lowed by anRed increase pansies in showed the the highestdw), values contents hydrolysable of of TRC tannins (13.8 (30.1 bioactive(68.4 mg mg compounds. GAE/g mg TAE/g dw), QE/gthe and dw) completely flavonoids in openmonomeric the flower anthocyanins, bud stage. stage,carotene/g the Regarding dw slight carotenoids highest and decreasing and obtained contents 402 until in (1300 completely open flowers. These different patterns Regarding anthocyanins, Skowyra et al. (2014) reportedsults similar to re- ours, mentioning thatin violanin the was the three major anthocyanin differenthighlighting the coloured higher pansies content (red,weight) in in yellow, comparison red and to (11.40 violet), yellow mg/gpetals. (4.69 mg/g freeze-dried freeze-dried weight) showed similar values,21.8 namely mg the GAE/g TRC,flower, dw ranging respectively); (yellow from hydrolysable bud55.7 5.3 and mg tannins to completely TAE/g betweenrespectively); dw open and 19.2 (completely white flavonoids between opendw and 40.7 (white red and bud and and 124.5 completely white mg open white QE/g flowers, flower, respectively). different colors were observed. Whitemuch and similar yellow pansies behavior showed TRC, than hydrolysable red tannins, ones,pletely and open increasing flavonoids flower the from stage.patterns values On bud were observed. of contrary, to The in increasebe com- stated red related in to pansies, TRC the opposite values accumulationfull-flowering of might phenolic stage compounds during that the maytions, also such be as relatedattraction the to of intensification ecological pollinators of func- line (Langenheim, antifungal with 1994). defenses Our those and(2012), results of the are who Bagdonaite, in Martonfi, reportedicum Repcak, that perforatum and the Labokas bioactivetwo developmental compounds stages (budding of anding full-flowering stages), the be- growth and development of the reproductive parts fol- whereas anthocyanins are responsible for thepurple-colored most petals. red-, blue-, However, these and authorscultivar of found chrysanthemum a with “Kastelli” redcentrations colored-petals and of high carotenoids, con- accumulate probably reddish because carotenoids this that cultivar are might absent in yellow petals. of different colors at distinct flowering stages. a,A b,B c,C a,A,B a,A b,B b,B a,A a,A 0.05). 1.4 2.5 5.0 1.2 2.7 4.0 4.7 0.5 1.6 < ± ± ± ± ± ± ± ± ± Opun- tannins -value Hydrolyzable 21.5 35.5 55.7 26.5 23.7 42.8 30.1 20.4 19.2 ; 48.9% to P (mg TAE/g dw) wittrockiana × , extract concentration providing 50% of DPPH radical scavenging effect and the extract concentration that gave an c,A c,B b,B a,B b,C a,C c,B b,A a,A 50 - Viola 28 36 26 30 8 6 30 36 12 β ± ± ± ± ± ± ± ± ± g µ Total ( -carotene/g dw and 303 to 404 278 132 804 576 873 carotenoids 1073 1133 1300 β carotene/g dw) Opuntia ficus-indica g µ Vol. 84, Iss. 3, 2019 a,B b,B c,C a,A b,A c,B b,C a,b,B a,A r 0.8 0.6 0.5 0.5 0.6 1.2 1.1 1.1 1.1 , an increase in linolenic acid (26.8% ± ± ± ± ± ± ± ± ± ) and linolenic acids (from 3.7% to 6.2% standard deviation. TRC, total reducing capacity. DPPH, 2,2-diphenyl-1-picrylhydrazyl. GAE, gallic acid equivalent. TAE, tannic acid equivalen TRC (mg ) were reported (Ammar, Ennouri, Bali, & ± GAE/g dw) 21.8 13.3 11.6 stage Opuntia stricta open open open Crataegus monogyna Flowering Completely Half openCompletely 7.6 Half openCompletely 12.5 Half open 13.0 Journal of Food Science g Cy-3 glu/g dw, respectively). Regarding total carotenoids, µ Opuntia ficus-indica flowers, from vegetative to full flowering stages, an increase in Total reducing capacity, total carotenoids, hydrolysable Concerning flowering stages, significant differences were found absorbance of 0.5 inletters the present reducing the power differences assay. dw, among dry different weight. colors Lowercase letters at present each the flower stage differences among ( different flower stages of a single colo 496 the yellow pansies showedsies. always According higher to Park values et al. thanresult (2015), most white from yellow accents the pan- in presence flowers of carotenoids (especially, xanthophylls), anthocyanins, with red pansies showing thetypes highest of values compounds for both (873 to402 1300 sented in Table 4.during Quantitative flowering differences and inbeen between these observed. Regarding pansies compounds the of colorferences of in different the values colors flower, the were have major detected dif- in carotenoids and monomeric Total reducing capacity, totaltotal flavonoids, carotenoids, and monomeric hydrolysable anthocyanins tannins, of contents of three pansies different colors and at distinct flowering stages are pre- ever, in red65.5% pansies and SFA PUFA decreased increased from 63.7% significantly to from 30.0%. 29.9%tannins, total to flavonoids, and monomeric anthocyanins. for Attia, 2014). In general terms,bud to MUFA completely percentage open decreased flower from and in all 31.2% color in pansies (28.4%, white, 66.1%, yellow, and red pansies, respectively); how- tia palmitic (from 38.2% to59.5% 43.0% for for thors in other flower species,stages particularly, between of bud and flower to 29.5%), and aobserved (Barros decrease et in al., 2011). linoleic Moreover, acid in two (15.6 species to of 14.2%) were pansies, respectively. Contrary, in red pansies palmiticincreased acid from (C16:0) 14.8% to51.0% 17.4% to and linoleic 18.7%. acid Similar decreased from patterns were observed by other au- (Table 3). Insies general, showed different the(C18:3n3) patterns main increased during for fatty flowering. 32.4% Linolenic acids and acid 66.4% detected in in white pan- and yellow ratios lower than 4.0recommended for (varying the between human diet 0.9helping (Department to and of ensure 4.0), Health, that 1994), which flowers are are considered a healthy food. showed higher values of PUFAall and SFA than pansies MUFA. Inratios at general, higher the than 0.45 three (ranging from flowering 0.46 to stages 2.13) showed and n-6/n-3 PUFA/SFA Values are expressed as: mean QE, quercetin equivalent. Cy-3 glu, cyanidin-3-glucoside. EC Yellow BudRed 5.3 Bud 13.8 White Bud 7.3 Color Table 4–Nutraceutical composition of dried Nutritional composition of pansies . . .

Food Chemistry nixdn ciiyptenfo h u ocmltl pnstage open completely the EC & to (lower in bud of Zhang, the increase from (Zhao, content an pattern showed activity materials high pansies antioxidant white plant a and Yellow of had 2014). activity Yang, pansies antioxidant responsible red compounds the of major for assay), buds TRC the by (predicted that phenols fact power), related be reducing the might for This to activity. mg/mL antioxidant highest 0.48 the and of indicative DPPH for mg/mL (0.17 nomto nohreil oesarayconsumed. flower- already the flowers improve pansies edible to in other necessary on are knowledge information studies the more So, increase although behavior, pansies. to yellow ing and helped white study in this stage flower bud tannins, open the hydrolysable completely from TRC, increased to anthocyanins The monomeric pansies. and red flavonoids, contents in anthocyanins The observed monomeric acids. were and palmitic carotenoids and linolenic, total linoleic, highest of val- contribution due the mainly the pansies predominated, to SFA red and PUFA in white constant. whereas remained in observed, ues contents calorie was protein low pansies in yellow to increase and an appropriate flowering, being car- During by ash, diets. followed and macronutrient, main proteins, and yellow, the bohydrates, white, was In water flowering. pansies, during red pan- colors of different compositions with nutraceutical sies and nutritional the in changes products. new production as flowers the edible enhance of chef’s to commercialization consumers, industry and for food useful and be producers, can different cuisine, nu- This with about flowering. pansies during information of colors more compositions provide nutraceutical as and well tritional as flowers edible as EC lowest the 4). by (Table characterized were assays pansies power red reducing of Buds and activity DPPH the scavenging using stages, flowering radical three at and colors different with Conclusion activity Antioxidant 2006). al., et Sood pattern 1976; consistent Uemoto, flowers. a & follow opened not (Sakata fully do and anthocyanins half of contents the the in So, decrease a by followed ering, flowers in content of anthocyanins total in increase an reported to (2006) 303 and 22.9, to com- 1.5 35.9, to to- to bud of (6.3 2011). the 402 content stage from flower al., the open increased et flowering, pletely anthocyanins (Salem during monomeric decreased pansies tal in they all while flowers, in flowering, However, red of during and also increased flowers yellow were compounds orange colors phenolic in the different example, with For as specie 2011). species flower development same other the in the found during of compounds flowers bioactive of of contents and . . . pansies of composition Nutritional ns neednl fteslto sdi h extraction. yellow the than in pansies used solution red the for of activity independently ones, detected antioxidant who of (2014), al. values et higher Skowyra Different by white. reported and been have red results by the followed showed activity, pansies antioxidant were yellow highest flowers, patterns open no completely colors, In different observed. of pansies Comparing content. iey.Smlrrslswr on nptl fegtcliasof cultivars eight of petals in found Rosa were results Similar tively). h rsn okdmntae htteewr significant were there that demonstrated work present The pansies of value the indicate obtained results the conclusion, In flowers in determined was pansies of activity antioxidant The oedamascene Rose µ × y3gugd nwie elw n e ase,respec- pansies, red and yellow, white, in dw glu/g Cy-3 g hybrida 50 aus,acmaidb nices nttlphenols total in increase an by accompanied values), Shizre l,21) ncnrr,So tal. et Sood contrary, On 2010). al., et (Schmitzer and oebourboniana Rose atau tinctorius Carthamus ttefis tgso flow- of stages first the at atau tinctorius Carthamus Slme al., et (Salem 50 values Gonz asagr . aasa . cuz . eslaee . us,M 21) Discrimination (2011). M. Musso, & P., Heiselmayere, H., Schulz, M., Baranska, S., Gamsjaeger, ozo .R,Pgis,A,Bns,M,Tnt,M . eihn,F,Xa,J,&Tni,R. Tundis, & J., Xiao, F., Menichini, C., M. Tenuta, M., Bonesi, A., Pugliese, R., M. Loizzo, contents phenolic Total (2014). F. (2014). Chen, K. & T. X.-R., Lim, Xu, D.-P., Xu, H.-B., Li, S., Li, A.-N., Li, enne,L,Csl . eer,J . eer,E . aav,J . aahs,E (2018). E. Ramalhosa, & A., J. Saraiva, L., E. Pereira, A., J. Pereira, S., Casal, L., Fernandes, Vainstein, . . . E., Pichersky, M., Shalit, M., Ovadis, N., Menda, I., Guterman, M., Dafny-Yelin, L N., Carazo, bioac- and composition the Comparing (2011). R. F. C. I. Ferreira & M., A. Carvalho, of L., Barros, concentrations in Variation (2012). J. Labokas, & M., Repcak, P., Martonfi, E., Bagdonaite, agnem .H 19) ihrpattreod:Apyoeti vriwo hi ecological their of overview phytocentric A terpenoids: plant Higher (1994). H. J. Langenheim, J J., M. Kotze, (2011). Standardization for Organization International (1994). Health, Department aaa,C . otobs .D,&Ftai .(04.Ocrec fErpa edpansy field European of Occurrence (2014). S. Fotiadis & D., S. Koutroubas, A., C. Damalas, Aquino-Bola ( AOAC Guimar ma,I,Enui . ai . ti,H 21) hrceiaino w rcl pear prickly two of Characterization (2014). H. Attia, & O., Bali, M., Ennouri, I., Ammar, References Contributions Author Acknowledgments OC( AOAC n ..rvee n dtdteatce ...... n J.P. with and associated were J.P. J.S., administration. and E.R., project J.S., S.C., E.R., P.B., S.C., J.S., article; study; E.R., the the supervised S.C., edited draft; and reviewed original J.P. the and curation; prepared data and performed wrote L.F. E.R. the and P.B., performed L.F. S.C., L.F., methodology; investigation; the provided P.B. and J.S., J.P., (UID/ CIMO under to FEDER support and Sci- financial for Portugal) for AGR/00690/2013). Foundation (FCT, PT2020 the Technology to Programme Fur- grateful and Agreement. are ence Partnership by authors PT2020 the co-financed the thermore, and within funds FEDER, the national through support and FEDER/007265) QUI/00062/2013) financial (UID/QUI/50006/2013-POCI/01/0145/ UID/ Unit the (FCT research for LAQV Unit SFRH/BD/95853/2013 FCT/MEC research sup- grant QOPNA and financial to research Fernandes the the Luana for to by Portugal) provided (FCT, port Technology and ence 21) hmclcmoiino h dbeflwr,pny( pansy flowers, edible the of composition Chemical (2018). fcrtni n aoodcneti easo as utvr ( cultivars pansy of petals in spectroscopy. content Raman flavonoid and carotenoid of 21) dbeflwr:Arc oreo htceiaswt nixdn n hypoglycemic and antioxidant with phytochemicals of source rich properties. A flowers: Edible (2016). plants flowers. wild and edible 51 of capacities antioxidant and feto liaecaigo h hsc-hmcladmcoilqaiyo ase ( pansies of quality microbial and physico-chemical the on wittrockiana coating alginate of Effect rose of stages advanced the during spectrum protein development. in petal Changes proteome: Flower (2005). A. de%20los%20p%C3%A9talos%20de%20flores%20comestibles%20204-232.pdf) p https://upcommons.upc.edu/bitstream/handle/2117/18886/Actividad%20antioxidante%20 los de antioxidante 22 of tivity Products in compounds bioactive major roles. dielsi Dasineura of leaves and stages flower Standardization. different for (12966-2) Organization acids fatty International of Switzerland: esters Geneva, methyl 1–15). of pp. Preparation ed., 2: (1st esters—Part methyl acid fatty of chromatography Office. Stationary M. H. England: 6277–6284. ( Quality ( squash edible in od hraetcl rcsei applications. cosmetic or pharmaceutical, food, .L,&Verdalet-Guzm & L., J. International AOAC of analysis of methods International. AOAC clcnttet n iatvt of bioactivity and constituents ical pce oesgoigi uii tfu oeigstages. flowering four at Tunisia in growing flowers species 59 ( il arvensis Viola niriu majus Antirrhinum .. .. n ..cnetaie h ril;LF,SC,E.R., S.C., L.F., article; the conceptualized E.R. and S.C., L.F., Sci- for Foundation Portuguese the acknowledge authors The 181–188. , 448–454. , lzBri,R,Prao .J,Ln-ei,C,Jve,G-.F,&Navarro-Gonz & F., G.-A. Javier, C., Luna-Recio, J., M. Periago, R., alez-Barrio, ´ e,R,Bro,L,Crah,A . eria .C .R 21) tde nchem- on Studies (2010). R. F. C. I. Ferreira, & M., A. Carvalho, L., Barros, R., aes, ˜ ora fCeia Ecology Chemical of Journal p.8881.TeNtelns Springer. Netherlands: The 818–821). (pp. 1990). 03.Ttldeayfie nfos nyai–rvmti ehd(985.29). method enzymatic–gravimetric foods, in fiber dietary Total 2003). , , 36 35 rteu monogyna Crataegus o,E . Urrutia-Hern N., E. nos, ˜ uigstorage. during ) 302–308. , ora fArclua n odChemistry Food and Agricultural of Journal 302–308. , pz . laao .P,Rdio .C,&Hge,A 20) Actividad (2005). A. Huguet, & C., M. Rodrigo, P., M. Almajano, D., opez, ´ res . ono,S . ofan .H 21) oaie soitdwith associated Volatiles (2010). H. J. Hoffmann, & D., S. Johnson, A., urgens, ¨ nOetaa Greece. Orestiada, in ) fca ehd faayi fAA International AOAC of analysis of methods Official Dpea Cecidomyiidae). (Diptera: Viola uubt pepo Cucurbita snwsucso iatv compounds. bioactive of sources new as ) Planta o.8,Is ,2019 3, Iss. 84, Vol. ora fRmnSpectroscopy Raman of Journal tlsd oe oetbe.EIA-SB – Rtivdfrom (Retrieved 1–5 EUITAB-ESAB. comestibles. flores de etalos ´ n .(03.Pyiohmclprmtr n nixdn compounds antioxidant and parameters Physicochemical (2013). 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