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Agrociencia

ISSN: 1405-3195

[email protected]

Colegio de Postgraduados

Mxico

Reyes-Luengas, Alberto; Salinas-Moreno, Yolanda; Ovando-Cruz, Manuel E.; Arteaga-Garibay

Ramn I.; Martnez-Pea, Marcos D.

ANLISIS DE CIDOS FENLICOS Y ACTIVIDAD ANTIOXIDANTE DE EXTRACTOS ACUOSDE VARIEDADES DE JAMAICA (Hibiscus sabdariffa L.) CON CLICES DE COLORES DIVERS

Agrociencia, vol. 49, nm. 3, abril-mayo, 2015, pp. 277-290

Colegio de Postgraduados

Texcoco, Mxico

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RESUMEN

El creciente mercado de bebidas funcionales requiere la ca-

racterizacin especfica de las fuentes de bioactivos, para

seleccionar aquellas con la mayor cantidad del compuesto

necesario en la formulacin. La composicin de cidos fe-

nlicos y contenido de antocianinas y proantocianidinas,

de extractos acuosos de tres variedades de jamaica (Hibiscus

sabdariffa L.), con rojo oscuro (Sudn), rojo claro (Crio-

lla Nayarit) y blanco (Alma Blanca) en sus clices, fueronevaluados mediante HPLC-DAD. Adems, se determin su

actividad antioxidante. El diseo experimental fue comple-

tamente al azar y con tres repeticiones. El contenido mayor

de cidos fenlicos totales se detect en Sudn y Alma Blan-

ca (.. y .. mg EAG gMS). La fraccin

de cidos fenlicos libres (AFL) fue abundante y con com-

plejidad alta en los clices de los tres colores. El cido clo-

rognico y sus dos ismeros predominaron en las fracciones

AFL y los cidos fenlicos glucosilados (AFG) representaron

entre . y . % del total de cidos fenlicos en las tres

variedades. Los clices blancos tenan la cantidad mayor de

cido cafeico y derivados de protocatquico. La actividad

antioxidante mayor (p.) se present en el extracto de

Sudn (IC. g mL). El color de los clices puede

ser un indicador del tipo predominante de fenoles en H.

sabdariffa L. y ayudar en la seleccin de variedades para

ABSTRACT

The growing market of functional beverages requires

characterize specifically the sources of bio-actives to select

those with the highest quantity of the compound needed

in the formulation. The composition of phenolic acids and

the content of anthocyanins and proanthocyanidins, from

aqueous extracts of the calyxes of three varieties of jamaica

(Hibiscus sabdariffa L.) were assessed using HPLC-DAD:

dark red (Sudan), light red (Criolla Nayarit) and white (AlmaBlanca). Antioxidant activity was also determined. The

experimental design was completely randomized with three

replications. The highest contents of total phenolic acids

were detected in Sudan and Alma Blanca (.. and

.. mg GAE gDM). The fraction of free phenolic

acids (FPA) was abundant and was highly complex in the

calyxes of the three colors. Chlorogenic acid and its two

isomers were predominant in the FPA, and the glycosylated

phenolic acids (GPA) accounted for . to . % of the

total phenolic acids in the three varieties. The white Alma

Blanca calyxes contained more caffeic and protocatechuic

acid derivatives than the red calyxes of the other varieties. The

highest antioxidant activity (p.) was found in the Sudan

extract (CI. g mL). The color of the calyxes can

indicate the predominant type of phenols in H. sabdariffa

L. and aid in the selection of varieties for formulations with

particular nutraceutical properties.

Key words: Hibiscus sabdariffa L., phenols, calyx, color,

antioxidant activity.

ANLISIS DE CIDOS FENLICOS Y ACTIVIDAD ANTIOXIDANTE DEEXTRACTOS ACUOSOS DE VARIEDADES DE JAMAICA (Hibiscus sabdariffaL.)

CON CLICES DE COLORES DIVERSOS

ANALYSIS OF PHENOLIC ACIDS AND ANTIOXIDANT ACTIVITY OF AQUEOUS

EXTRACTS OF JAMAICA (Hibiscus sabdariffaL.) VARIETIES WITH CALYXESOF DIFFERENT COLORS

Alberto Reyes-Luengas1, Yolanda Salinas-Moreno2*, Manuel E. Ovando-Cruz3,Ramn I.Arteaga-Garibay4, Marcos D. Martnez-Pea4

1Departamento de Ingeniera Agroindustrial. Universidad Autnoma Chapingo. 56200. Km13.5 Carretera Los Reyes-Texcoco. Chapingo, Mxico. 2Laboratorio de Calidad de Cultivospara uso Humano y Pecuario. Campo Experimental Centro Altos de Jalisco. Instituto Na-cional de Investigaciones Forestales Agrcolas y Pecuarias (INIFAP). 47600. Km 8, CarreteraTepatitln-Lagos de Moreno. Tepatitln de Morelos, Jalisco. ([email protected]). 3CampoExperimental Valles Centrales. INIFAP. Melchor Ocampo No. 7, Colonia Agencia Municipal,Santo Domingo Barrio Bajo 7, Etla, Oaxaca. 4Laboratorio de Recursos Genticos Microbianos.Centro Nacional de Recursos Genticos. INIFAP. 47600. Boulevard de la biodiversidad No

400 Tepatitln de Morelos, Jalisco.

*Autor responsable vAuthor for correspondence.Recibido: mayo, 2014. Aprobado: marzo, 2015.Publicado como ARTCULO en Agrociencia 49: 277-290. 2015.

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AGROCIENCIA, 1 de abril - 15 de mayo, 2015

VOLUMEN 49, NMERO 3

obtener formulaciones con propiedades nutracuticas par-

ticulares.

Palabras clave: Hibiscus sabdariffaL., fenoles, antocianinas, cli-

ces, color, actividad antioxidante.

INTRODUCCIN

La jamaica (Hibiscus sabdariffaL.), o flor de

jamaica como se le conoce en Mxico, es unaplanta anual cultivada en los climas tropica-

les y subtropicales del mundo. Sus clices se aprove-chan para elaborar bebidas fras o calientes, merme-ladas, salsas, y aderezos. En la medicina tradicional,los extractos acuosos de la decoccin de los clicesdeshidratados de jamaica se utilizan para el controlde la presin arterial, alteraciones hepticas o fiebre

(Ali et al., ). Estos beneficios teraputicos seatribuyen a la diversidad de los fitoqumicos de losclices (Rodrguez-Medina et al., ; Ramirez-Rodriguez et al., ; Hopkins et al., ), enlos que predominan los fenoles. Entre los fenolesms abundantes estn las antocianinas (Tsai et al.,), que son responsables del color rojo brillan-te de los extractos de los clices y son estudiadasampliamente (Galicia-Flores et al., ; Ramrez-Rodrguez et al., ). Los cidos fenlicos son otro grupo importan-te de compuestos en los clices de la jamaica y se

encuentran en forma libre o conjugada. En frutasy hortalizas estn principalmente en forma libre(Shahidi y Naczk, ), y en los cereales predomi-nan en forma conjugada o ligada (Liu, ).

En extractos acuosos de jamaica se identific elcido clorognico y sus ismeros I y II como loscidos fenlicos principales (Fernndez-Arroyo etal., ; Ramirez-Rodriguez et al., ), junto conlos derivados de los cidos protocatquico y glico(Ramirez-Rodriguez et al., ) y el cido cafeico(Huang et al., ). Otros fenoles del tipo flavo-

noide, como quercetina y sus glucsidos, y los glu-csidos de miricetina (Fernndez-Arroyo et al. ;Ramirez-Rodriguez et al. ) y proantocianidinas(Syago-Ayerdi et al., ), del tipo catequina,epigalocatequina y epigalocatequin-galato (Huanget al., ) tambin estn identificados en la florde jamaica. El color de los clices de jamaica est asociadoal contenido y tipo de fenoles. Los clices oscuros

INTRODUCTION

Jamaica (Hibiscus sabdariffa L.), or flor de

jamaica, as it is known in Mexico, is an annualplant cultivated in tropical and subtropical

climate worldwide. Its calyxes are used to make

cold or hot beverages, jams, sauces and dressings.In traditional medicine, aqueous extracts obtainedby decoction of the dehydrated calyxes are used tocontrol blood pressure, hepatic disorders and fevers(Ali et al., ). These therapeutic benefits areattributed to diverse phytochemicals present in thecalyx (Rodrguez-Medina et al., ; Ramirez-Rodriguez et al., ; Hopkins et al., ), of

which phenols are predominant. Anthocyaninsare among the most abundant phenols (Tsai et al.,), they are responsible for the bright red color

of the extracts and are extensively studied (Galicia-Flores et al., ; Ramrez-Rodrguez et al., ). Phenolic acids are another important groupof compounds in the jamaica calyxes. Thesecompounds can be found in free or conjugatedform. In fruits and vegetables they are mainly infree form (Shahidi and Naczk, ), whereas incereals the conjugated or linked form predominates(Liu, ). In aqueous extracts of jamaica, chlorogenic acidand its isomers I and II as the principal phenolicacids were identified (Fernndez-Arroyo et al.,

; Ramirez-Rodriguez et al., ), togetherwith derivatives of protocatechuic and gallicacids (Ramirez-Rodriguez et al., ) and caffeicacid (Huang et al., ). Other flavonoid-typephenols, such as quercetin and its glycosides, andthe glycosides of myricetin (Fernndez-Arroyoet al., ; Ramirez-Rodriguez et al., ) andproanthocyanidins (Syago-Ayerdi et al., ) ofthe catechin, epigallocatechin and epigallocatechin-galate types (Huang et al., ) are also identifiedin jamaica flower calyxes.

The color of the jamaica calyxes is associated withthe content and type of phenols. Dark calyxes havea higher content of anthocyanins than light-coloredcalyxes, whereas the white or green calyxes lack theseflavonoids (Christian and Jackson, ) or haveminimal quantities (Salinas-Moreno et al., ).Characterization of the phenolic acids from jamaicaaqueous extracts is limited (Rodrquez-Medina etal., ; Ramrez-Rodrguez et al., ) and have

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ANLISIS DE CIDOS FENLICOS Y ACTIVIDAD ANTIOXIDANTE DE EXTRACTOS ACUOSOS DE VARIEDADES DE JAMAICA (Hibiscus sabdariffaL.)

279REYES-LUENGAS et al.

tienen ms antocianinas que los clices claros, perolos blancos o verdes carecen de estos flavonoides(Christian y Jackson, ) o estn en cantida-des mnimas (Salinas-Moreno et al., ). La ca-racterizacin de los cidos fenlicos de extractosacuosos de jamaica es limitada (Rodrquez-Medina

et al., ; Ramrez-Rodrguez et al., ) y seha realizado en variedades con clices coloridos,por lo que se desconoce la composicin de losblancos.

La actividad antioxidante asociada a las bebidaselaboradas a partir de la decoccin acuosa de los c-lices de jamaica (Prenesti et al., ) se atribuye alos fenoles que contienen, y destacan las antocianinas(Syago-Ayerdi et al., ) y los cidos fenlicos.De estos ltimos, el cido clorognico y sus deriva-dos son los ms importantes (Fernndez-Arroyo et

al., ). El mercado creciente de las bebidas funcionalesrequiere caracterizar los fenoles de cada variante, paraelegir la ms adecuada para el diseo de bebidas concaractersticas nutraceticas particulares.

Por lo tanto, los objetivos del presente estudiofueron: ) analizar los cidos fenlicos solubles (libresy conjugados) en los extractos acuosos de variedadesde jamaica con color diferente de clices medianteHPLC-DAD; ) cuantificar flavonoides del tipo an-tocianinas y proantocianidinas; y, ) determinar laactividad antioxidante de los extractos.

MATERIALESYMTODOS

Material biolgico

Las variedades de jamaica Sudn, con clices rojo oscu-

ro, Criolla Nayarit con clices rojo claro y Alma Blanca con

clices blancos fueron estudiadas. Sudn y Alma Blanca fue-

ron cultivadas en Santa Mara Cortijo, Jamiltepec, estado de

Oaxaca; la Criolla Nayarit, fue cultivada en Huajicori, estado

de Nayarit.

Preparacin de los extractos acuosos

Los extractos se prepararon con . g de clices secos y

mL de agua destilada. La mezcla hirvi min, se separ el l-

quido de los clices por decantacin y la extraccin se repiti en

las mismas condiciones. Los extractos se juntaron, la solucin se

filtr con papel Whatman No. y se afor a mL con agua

destilada. Las siguientes variables se evaluaron en este extracto.

focused on colored calyxes, and thus the compositionof the white varieties is unknown. The antioxidant activity associated with thebeverages made from aqueous decoction of the

jamaica calyxes (Prenesti et al., ) is attributedto the phenols they contain. Of these, anthocyanins

(Syago-Ayerdi et al., ) and phenolic acids areoutstanding. Of the latter, chlorogenic acid andits derivatives are the most important (Fernndez-

Arroyo et al., ). The growing market of functional beveragesrequires characterization of the phenols of each variantto select the most suitable for the design of beverages

with particular nutraceutical characteristics. Therefore, the objectives of this study were: ) toanalyze the soluble (free and conjugated) phenolicacids in the aqueous extracts of jamaica calyxes of

different colors using HPLC-DAD, ) to quantifyanthocyanin-type flavonoids and proanthocyanidins;and ) to determine the antioxidant activity of theextracts.

MATERIALSANDMETHODS

Biological material

The jamaica varieties Sudan with dark red calyxes, CriollaNayarit with pink calyxes and Alma Blanca with white calyxes

were studied. Sudan and Alma Blanca were cultivated in Santa

Mara Cortijo, Jamiltepec, state of Oaxaca; Criolla Nayarit was

cultivated in Huajicori, state of Nayarit.

Preparation of aqueous extracts

Extracts were prepared with . g dry calyxes and mL

distilled water. The mixture was boiled for min and the liquid

separated from the calyxes by decantation. The extraction was

repeated in the same conditions. The extracts were filtered

together through Whatman No. filter paper and gauged to

mL with distilled water. The following variables were determined

with this extract.

Phenolic acids

Free, glycosylated and esterified phenolic acids were

determined by the method of Bakan et al. (). To determine

free phenolic acids, two liquid-liquid extractions were carried

out with mL ethyl acetateto mL aqueous extract after

adjusting pH to with N HCl. The two organic phases were

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VOLUMEN 49, NMERO 3

cidos fenlicos

Los cidos fenlicos libres, glucosilados y esterificados se

determinaron con el mtodo de Bakan et al. (). Para de-

terminar los cidos fenlicos libres se realizaron dos extraccio-

nes lquido-lquido con mL de acetato de etilo a mL del

extracto acuoso, despus de ajustar su pH a , con HCl N.Las dos fases orgnicas se juntaron y concentraron a sequedad

en un rota-evaporador (Heidolph instruments, mod. Laborata

digital, Germany). El residuo se resuspendi en mL de

metanol grado HPLC. Las muestras se mantuvieron en frascos

mbar a C hasta su anlisis. La fase acuosa restante se

afor a mL con agua destilada y se dividi en dos volme-

nes iguales.

Para la fraccin de cidos fenlicos glucosilados (AFG), a

uno de los volmenes se adicionaron mL de HCl N y se

coloc h a C; el otro volumen se emple para la fraccin de

cidos fenlicos esterificados (AFE), al cual se aadieron mL

de NaOH N, se dej reaccionar h en oscuridad. Despus se

ajust a el pH de los extractos y se procesaron en la misma

forma que los cidos fenlicos libres. Los cidos fenlicos to-

tales en cada fraccin se cuantificaron mediante la prueba Fo-

lin Ciocalteu (Singleton y Rossi, ). A L del extracto

acuoso se adicionaron L de reactivo de Folin-Ciocalteu

N, se agit y se dej reaccionar min en oscuridad. La reac-

cin se neutraliz con L de NaCOal % y se afor a

mL con agua destilada. La muestra se agit y qued en oscu-

ridad min. La absorbancia se midi en un espectrofotmetro

(PerkinElmer, Lamda UV/Vis, USA) a nm. Una curva

estndar de cido glico ( a g L) fue realizada, y losresultados se expresaron en mg equivalentes de cido glico por

g de MS (mg EAG gMS).

Anlisis de cidos fenlicos por cromatografa

lquida de alta resolucin acoplada a detector

con arreglo de diodos (HPLC-DAD)

Los cidos fenlicos se analizaron con un equipo

PerkinElmer serie (USA), integrado por bomba cuaterna-

ria, degasificador, detector de UV/Vis con arreglo de diodos

(DAD), con auto-muestreador y columna analtica Hypersil

ODS. ( mm), con tamao de partcula de m. El

mtodo de Glowniak et al. () fue usado, y como fase mvil

se emple una mezcla de metanol: cido actico: agua (::

v/v/v). La velocidad de flujo fue mL min, el tiempo de

corrida min y el volumen de inyeccin mL. La deteccin

se realiz a nm (derivados dep-hidroxibenzoico) y nm

(derivados de hidroxicinmico). La temperatura de la columna

se mantuvo en C.

put together and concentrated to dryness in a rotary evaporator

(Heidolph Instruments, mod. Laborata digital, Germany).

The residue was re-suspended in mL HPLC grade methanol.

The samples were kept in amber-colored recipients at C

until analysis. The remaining aqueous phase was gauged to

mL with distilled water and divided into two equal volumes.

For the fraction of glycosylated phenolic acids (GPA), mLN HCl was added to one of the volumes and left h at C. The

other volume was used for the esterified phenolic acid fraction

(EPA); mL N NaOH were added to this volume and left to

react for h in darkness. Afterward, the pH of the extract was

adjusted to and processed in the same way as the free phenolic

acids. Total phenolic acids were quantified in each fraction using

the Folin Ciocalteu assay (Singleton and Rossi, ). To L

of the aqueous extract, N Folin-Ciocalteu reagent was added,

shaken and left to react for min in darkness. The reaction was

neutralized with L % NaCOand gauged to mL

with distilled water. The sample was shaken and left in darkness

for min. Absorbance was measured in a spectrophotometer

(PerkinElmer, Lamda UV/Vis, USA) at nm. A gallic

acid standard curve ( to g L) was constructed and the

results were expressed in mg gallic acid equivalents per g of DM

(mg GAE gDM).

Phenolic acid analysis by high performance

liquid chromatography coupled to a diode

array detector (HPLC-DAD)

Phenolic acids were analyzed with PerkinElmer series

(USA) equipment comprising a quaternary pump, degasifier,UV/Vis diode array detector (DAD), auto-sampler and a

Hypersil ODS. analytical column ( mm) with

particle size of m. The method of Glowniak et al. () was

utilized, and a mixture of methanol: acetic acid: water (::

v/v/v) was used as the mobile phase. Flow rate was mL min,

running time was min, and injection volume was L.

Detection was done at nm (derivatives ofp-hydroxybenzoic

acid) and nm (derivatives of hydroxycinnamic acid).

Column temperature was kept at C.

A diode array detector was used to obtain the UV spectrum

of each of the peaks of the phenolic acid chromatogram.

Identification was achieved by comparing retention times with

commercial standards and their respective UV spectra. To

determine which of the chromatogram peaks corresponded to

chlorogenic acid, a pinch of commercial standard was added to the

sample. Once the chlorogenic acid peak was located, the literature

and a library of UV spectra of different phenolic acids reported

for Hibiscus sabdariffaL. were consulted, and the derivatives of

the acids were identified. For quantification, standard curves of

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El detector con arreglo de diodos se us para obtener el

espectro de UV de cada pico del cromatograma de cidos fe-

nlicos. La identificacin se realiz por comparacin de los

tiempos de retencin con los estndares comerciales y sus res-

pectivos espectros en UV. Para identificar cul de los picos en

el cromatograma corresponda al cido clorognico, se agreg a

la muestra una pizca del estndar comercial. Una vez ubicadoel pico correspondiente al cido clorognico, se us la literatura

y una librera de espectros de UV de los diferentes cidos fen-

licos reportados en Hibiscus sabdariffa L., y se identificaron los

derivados de cidos. Para la cuantificacin se prepararon curvas

patrn de los cidos cafico, clorognico, ferlico, glico, pro-

tocatquico,p-cumrico,p-hidroxibenzoico y sirngico (Sigma,

MN) en las mismas condiciones que las muestras.

Fenoles extractables totales y antocianinas totales

Para cuantificar los fenoles extractables totales (FET) se us

el mtodo de Folin-Ciocalteu (Singleton y Rossi, ). Las an-

tocianinas totales se determinaron al medir la absorbancia de los

extractos acuosos en un espectrofotmetro (Perkin Elmer Lamb-

da UV/Vis, USA) de acuerdo con lo descrito por Abdel-Aal y

Hucl (), y las adaptaciones de Salinas-Moreno et al. ().

Una curva patrn de cianidina -glucsido (Polyphenols, Nw)

fue realizada y los resultados se expresaron en mg equivalentes de

cianidina--glucsido por g de MS (mg CGE gMS).

Proantocianidinas

Las proantocianidinas se evaluaron con el mtodo de Wallacey Giusti (). Una alcuota de L de la muestra se hizo reac-

cionar con L de metanol y L de solucin de DMAC

(-dimetilamino cinamaldehido al % en una mezcla : (v/v)

de metanol y cido sulfrico N). La reaccin dur min a

temperatura ambiente, en oscuridad, y se ley la absorbancia a

nm; la concentracin fue calculada con una curva patrn

de catequina (Sigma, MN). Los resultados se expresaron en g

equivalentes de catequina por mg de MS (g EC mgde MS).

Actividad antioxidante

Para evaluar la actividad antioxidante se us el mtodo del

radical libre ,-difenil--picrilhidrazil (DPPH) (Sigma Aldrich

Co) descrito por Soler-Rivas et al. () con una solucin

M de DPPH en metanol al %. En una cubeta de cuar-

zo se colocaron mL de extracto y . mL de DPPH. La ab-

sorbancia se monitore cada min por min a una longitud

de onda de nm. La absorbancia de referencia (A) fue ob-

tenida al sustituir el volumen de extracto por metanol al %.

caffeic, chlorogenic, ferulic, gallic, protocatechuic, p-coumaric,

p-hydroxybenzoic and syringic acids (Sigma, MN) were prepared

under the same conditions as those of the samples.

Total extractable phenols and total anthocyanins

To quantify total extractable phenols (TEP), the methodof Folin-Ciocalteu (Singleton and Rossi, ) was used. Total

anthocyanins were determined by measuring absorbance of the

aqueous extracts in a spectrophotometer (Perkin Elmer Lambda

UV/Vis, USA) following the method described by Abdel-Aal and

Hucl (), with adaptations by Salinas-Moreno et al. ().

A standard curve for cyanidin -glucoside (Polyphenols, Nw)

was constructed, and the results were expressed in mg cyanidin-

-glucoside equivalents per g of DM (mg CGE g DM).

Proanthocyanidins

Protoanthocyanidins were assessed with the method of

Wallace and Giusti (). A L aliquot of the sample was

made to react with L methanol and L DMAC

solution ( % -dimethylaminocinnamaldehyde in a mixture (v/v)

of methanol and N sulfuric acid). The reaction lasted min at

room temperature in the dark. Absorbance was read at nm.

The concentration was calculated with a reference catechin curve

(Sigma, MN). Results were expressed in g catechin equivalents

per mg of DM (g CE mgDM).

Antioxidant activity

To evaluate antioxidant activity, the method used was the

free radical ,-diphenyl--picrylhydrazyl (DPPH) (Sigma

Aldrich Co) described by Soler-Rivas et al. () with a

M solution of DPPH in % methanol. In a quartz cell

were placed mL of extract and . mL DPPH. Absorbance

was monitored every min for min at a wavelength of

nm. The reference absorbance (A) was obtained by

substituting the extract volume for % methanol. The

percentage of DPPH reduction was obtained with the equation

DPPH (%)(AAn) / A, where A and An were the

reference and sample absorbances, respectively. The data were

used to determine the IC parameter, which represents the

concentration (g mL) of phenolic compounds required to

reduce the DPPH free radicals by % (Einbond et al., ).

Experimental design and statistical analyses

The experimental design was completely randomized.

With the data about phenolic composition, an ANOVA and

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El porcentaje de reduccin de DPPH se obtuvo de la expre-

sin: DPPH(%)(AAn) /A, dondeAyAnfueron las

absorbancias de referencia y de la muestra, respectivamente.

Los datos se usaron para determinar el parmetro IC, que

representa la concentracin (g mL) de compuestos fenlicos

requerida para reducir % del radical libre DPPH (Einbond

et al., ).

Diseo experimental y anlisis estadsticos

El diseo experimental fue completamente al azar. Con los

datos de composicin fenlica se realiz un ANDEVA y pruebas

de comparacin de medias (Tukey; p.). Todas las determi-

naciones se realizaron por triplicado. Los anlisis se efectuaron

con SAS System versin ..

RESULTADOSYDISCUSIN

cidos fenlicos

El contenido de cidos fenlicos totales (AFT)fue diferente entre las variedades analizadas. Sudntuvo el contenido mayor (p.) y Criolla Na-yarit el menor (.. y .. mg EAGgMS). Los clices blancos de Alma Blanca tuvie-ron contenido mayor (p.) de AFT que los deCriolla Nayarit, coloreados y con antocianinas (Fi-gura ). La fraccin ms abundante de cidos fen-licos en las variedades analizadas fue la de los cidos

fenlicos libres (AFL). Un patrn similar fue repor-tado para frutas y verduras (Shahidi y Naczk, ).Los AFL en las variedades Sudn y Alma Blanca fue-ron similares (p.), pero mayores que en Crio-lla Nayarit. El contenido mayor (p.) de cidosfenlicos glucosilados (AFG) correspondi a Sudny el menor lo tuvieron Criolla Nayarit y Alma Blan-ca. La fraccin AFE fue la menor de las tres, con elcontenido mayor (p.) en Sudn y el menor en

Alma Blanca.La fraccin AFL mostr entre y picos de ci-

dos fenlicos o de sus derivados. La identidad de losderivados se obtuvo al comparar sus respectivos es-pectros en UV con los estndares comerciales y hubocoincidencia del espectro, pero no as del tiempo deretencin (Figura ).

En la fraccin de AFL el cido clorognico y susismeros I (cido -cafeoil qunico) y II (cido -ca-feoil qunico) fueron los ms abundantes, los siguiel cafeico y los derivados del cido protocatquico

comparison of means were performed (Tukey; p.). All of

the determinations were done in triplicate. The analyses were run

in SAS system version . software.

RESULTSANDDISCUSSION

Phenolic acids

Total phenolic acids (TPA) content was differentamong the analyzed varieties. Sudan had the highestcontent (p.) and Criolla Nayarit had the lowest(.. and .. mg GAE g DM).

White calyxes of Alma Blanca had higher content(p.) of TPA than those of Criolla Nayarit,

which are colored and have anthocyanins (Figure). The most abundant fraction of phenolic acidsin the analyzed varieties was that of free phenolic

acids (FPA). A similar pattern was reported for fruitsand vegetables (Shahidi and Nacsk, ). FPA inthe varieties Sudan and Alma Blanca were similar(p.), but higher than in Criolla Nayarit. Thehighest content (p.) of glycosylated phenolicacids (GPA) was found in Sudan and the lowest inCriolla Nayarit and Alma Blanca. The EPA fraction

SudnCriolla Nayarit

Alma Blanca

a

c

b

a

b

a

a

b b a b c

DSH(AFT)

DSH(AFL)

DSH(AFG)

DSH(AFE)

AFT AFL AFG AFE

12

11

10

9

87

6

5

4

3

2

1

0

-1

mgEAGg

MS

Fracciones de cidos fenlicos

Figura . Contenido de cidos fenlicos totales (AFT) y susfracciones libres (AFL), glucosilados (AFG) y es-

terificados (AFE) en extractos acuosos de tres va-riedases de jamaica. DSH: diferencia significativahonesta. Barras con letras diferentes son estadsti-camente diferentes (Tukey, p.).

Figure . Content of total phenolic acids (AFT) and theirfree (AFL), glycosylated (AFG) and esterified (AFE)fractions in aqueous extract of three varieties ofjamaica. DSH: honest significant difference. Barswith different letters are statistically different(Tukey; p.).

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(Cuadro ). Alma Blanca destac por sus contenidoselevados de cido cafico y derivados de protocat-quico. Los contenidos de cido clorognico y sus is-meros fueron menores que , y g gde clorognico, ismero I e ismero II en extractosacuosos de clices molidos de jamaica reportados por

Fernndez-Arroyo et al. (). Las variedades de clices coloridos tuvieron deri-vados de cido glico, ausentes en Alma Blanca, encantidades ms altas que los derivados del protoca-tquico. Tambin hubo derivados de los cidosp-cu-mrico y ferlico en cantidades pequeas. El conte-nido de cidos fenlicos totales fue parecido en AlmaBlanca y Sudan y ms alto que en Criolla Nayarit.Los valores obtenidos por HPLC fueron similares alos cuantificados con el mtodo de Folin-Ciocalteau(Figura ). Alma Blanca tuvo un contenido alto de

derivados del cidop-hidroxibenzoico, respecto a lasvariedades con clices color rojo. El cido clorognico y sus ismeros (I y II) tam-bin predominaron en la fraccin de AFG, similara la fraccin de AFL en la proporcin mayor delclorognico sobre sus ismeros. Adems el conte-nido de estos cidos tuvo el orden SudnAlmaBlancaCriolla Nayarit. El cido cafeico fue el se-gundo ms importante en la fraccin AFG y las varie-dades tuvieron el mismo orden por su contenido enlos AFL. La variedad Sudn mantuvo la presencia dederivados del cido sirngico en esta fraccin, como

en la de AFL; las otras variedades no tuvieron deriva-dos de este cido.

Los AFE representaron todos aquellos cidos fe-nlicos no ligados a componentes de pared celular,pero poseen alguna molcula unida a su estructuramediante enlace ster (Liu, ). En las variedadesanalizadas destac la cantidad de derivados de sirn-gico y p-hidroxibenzoico. Las variedades coloridastuvieron cantidad alta de cido cafeico, que podraprovenir de la hidrlisis del cido clorognico, quees inestable al pH alcalino al que el extracto se some-

te para su anlisis. La variedad Criolla Nayarit tuvoun contenido alto de derivados de protocatquico,Sudn tuvo un valor varias veces menor y en AlmaBlanca no se detect en la fraccin AFE. La variedadSudan tuvo derivados de cido sirngico en las tresfracciones de cidos fenlicos analizadas. En la fraccin AFL de las variedades analiza-das el cido clorognico represent ., . y. % del total en Alma Blanca, Sudn y Criolla

was the lowest of the three, with the highest content(p.) in Sudan and the lowest in Alma Blanca.

The FPA fraction showed to peaks ofphenolic acids or their derivatives. The identity ofthe derivatives was obtained by comparing theirrespective UV spectra with commercial standards.

There was coincidence in the spectrum but not inretention time. (Figure ). In the FPA fraction, chlorogenic acid and itsisomers I (-caffeoyl quinic acid) and II (-caffeoylquinic acid) were the most abundant, followed bycaffeic acid and the derivatives of protocatechuicacid (Table ). Alma Blanca was outstanding forits high content of caffeic acid and derivatives ofprotocatechuic acid. The contents of chlorogenicacid and its isomers were less than the , and g gchlorogenic, isomer I and isomer

II in aqueous extracts of ground jamaica calyxesreported by Fernndez-Arroyo et al. (). The jamaica varieties with colored calyxes hadlarger quantities gallic acid derivatives, which wereabsent in Alma Blanca, than of protocatechuicderivatives. There were also small quantities ofp-coumaric and ferulic acid derivatives. The totalcontent of phenolic acids was similar in Alma Blancaand Sudan and higher in Criolla Nayarit. The valuesobtained by HPLC were similar to those quantified

with the Folin-Ciocalteau method (Figure ). AlmaBlanca had a high content of p-hydroxybenzoic acid

derivatives, relative to the varieties with red calyxes.Chlorogenic acid and its isomers (I and II) were

also predominant in the GPA fraction, similar to theFPA fraction in that chlorogenic acid was present ina larger proportion than its isomers. Moreover, thecontent of these acids were in the order SudanAlmaBlancaCriolla Nayarit. Caffeic acid was the secondin importance in the GFP fraction and had thesame order as with FPA contents. The Sudan varietymaintained the presence of syringic acid derivativesin the fraction, as in the FPA fraction. The other

varieties did not have derivatives of this acid. The EPA represented all of those phenolic acidsnot linked to components of the cell wall, but havesome molecule joined to its structure by an esterbond (Liu, ). In the varieties analyzed, thequantities of syringic and p-hydroxybenzoic acidderivatives were outstanding. The colored varietieshad a high quantity of caffeic acid. This acid couldhave originated from the hydrolysis of chlorogenic

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VOLUMEN 49, NMERO 3

Figura . Perfil cromatogrfico a nm de los cidos fenlicos libres del extracto acuoso de los clices de las variedades de

jamaica Sudn (A), Criolla Nayarit (B) y la mezcla de estndares (C). La identidad de los picos es: ) derivado del cidoglico; ) derivado dep-hidroxibenzoico; ) derivado de sirngico; ) cido -cafeoil qunico (ismero I); ) derivadode protocatequico; ) derivado de p-cumrico; ) cido -cafeoil-qunico (ismero II); ) cido clorognico; ) noidentificado; ) cido cafeico; ) cido sirngico; ) derivado del cido vanlico; ) derivado del cido ferlico;) cido glico; ) cido protocatquico; ) cido p-hidroxibenzoico; ) cido p-cumrico; ) cido ferlico;) cido sinpico; ) cido salislico. Los picos con las letras a y b se identificaron como derivados del cido cafeico.

Figure . Chromatographic profile, at nm, of free phenolic acids from aqueous extract of the calyxes of the jamaica varietiesSudan (A), Criolla Nayarit (B) and the mixture of standards (C). Identity of the peaks: ) derivative of gallic acid;) derivative ofp-hydroxybenzoic; ) derivative of syringic acid; ) -caffeoyl quinic acid (isomer I); ) derivative ofprotocatechuic; ) derivative of p-coumaric; ) -caffeoyl-quinic (isomer II); ) chlorogenic acid; ) not identified;) caffeic acid; ) syringic acid; ) derivative of vanillic acid; ) derivative of ferulic acid; ) gallic acid;) protocatechuic acid; )p-hydroxybenzoic acid; )p-coumaric acid; ) ferulic acid; ) synaptic acid; ) salicylicacid. The peaks with letters a and b were identified as derivatives of caffeic acid.

80

70

60

50

40

30

20

10

1

2

3

4

5

6

7

8

9

10

11 1213

0 2 4 6 8 10 12 14 16 18 20 22

350

300

250

200

150

100

50

0.0

0 2 4 6 8 10 12 14 16 18 20 22

80

70

60

50

40

3020

10

1

2

3

4

5 6

78

9

10 11 1213

0 2 4 6 8 10 12 14 16 18 20 22

14

15

8

16

10

11

17 18

19 20

Absorbancia(mA

U)

Tiempo de retencin (min)

A

B

C

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Cuadro.cidosfenlicosenlasfraccioneslibres,glucosiladasyesterific

adasdeextractosacuososdeclicesdevariedadesdejamaica(gg MS).

Table.Phenolicacidsinfree,g

lycosylated,andesterifiedfractionsofaqueousextractsfromcalyxesofdifferentvarietiesofjamaica(gg DM).

cidos

Fenlicos

DGal

Dp-H

D-Ptc

Srg

Clg

Clg-I

Clg-II

Caf

p-C

Dp-C

DF

Total

(gg1)

LibresAB

-

301.7

259.5

-

1879.324

.3

746.4

9.1

1096.844.1

4

93.929.1

-

21.3

12

4810.9

Sudn

321.2

-

159.1

310.2

2147.8278

.2

1117.8

9.1

719.5

4.2

47.5

1.7

-

43.5

22

4888.6

CN

226.9

117.9

142.6

-

1171.314

.7

735.457.2

659.4

9.1

1

06.914.2

-

-

-

3160.4

Glicosilados

AB

-

-

58.7

-

685.30

.6

437.7

2.5

507.1

4.6

91.74.9

-

4

-

1784.5

Sudn

-

58

82.2

1149.377

.6

836.9

9.0

748.634.1

47.31.5

-

38

-

2960.3

CN

-

28.8

-

446.21

.0

527.618.2

386.916.4

75.94.1

-

-

-

1465.4

Esterificados

AB

-

294.5

433

47.30.9

59.31.0

-

-

869.1

Sudn

-

271.3

40

556.1

408.10.3

-

144.4

-

1419.9

CN

-

130.3

178.8

329.1

329.10.7

386.7

16

-

1021.3

AB:Almablanca;CN:CriollaN

ayarit:DGal:derivadosdeglico;Dp-H:derivadosdelcidop-hidroxibenzoico;D-Ptc:derivadosdeprotocatquico;Srg:cidosirngico;Clg:cidoclo-

rognico;Clg-I:ismerodecloro

gnicoI;Clg-II:ismerodeclorognicoII;

Caf:cafeico;p-C:p-cumrico;Dp-C:deriv

adodep-cumrico;DF:derivadodeferlicovA

B:Almablanca;

CN:CriollaNayarit:DGal:deriv

ativesofgallicacid;Dp-H:derivativesofp-hydroxybenzoicacid;D-Ptc:derivativesofprotocatechuicacid;Srg:syringicacid;Clg:chlorogenicacid;Clg-I:

chlorogenicisomerI;Clg-II:chlo

rogenicisomerII;Caf:caffeicacid;p-C:p-coumaricacid;Dp-C:derivativeofp-coumaricacid;DF:derivativeofferulicacid.

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VOLUMEN 49, NMERO 3

Nayarit. Al considerar los ismeros, la proporcincreci hasta ., . y . % en ellas. En AlmaBlanca el cido cafeico fue el cuarto en importancia(. %), pero en las variedades coloridas los de-rivados del cido glico ocuparon esa posicin. Lapredominancia del cido clorognico y sus ismeros

en los extractos acuosos analizados coincidi con loinformado por Rodrguez-Medina et al. () yRamrez-Rodrguez et al. (), quienes obtuvie-ron el mismo resultado con tcnicas de HPLC-MS.Pero este resultado contrast con lo observado porHuang et al. (), ya que identificaron una frac-cin de Hibiscusspp rica en polifenoles, con cidosprotocatquico (. %) y cafeico (. %) comolos ms abundantes. Las divergencias podran de-berse al disolvente usado para obtener los extractos,pues el etanol, en contraste con el agua, favorece la

extraccin del protocatquico (Chao y Yin, ).Adems, la gentica de los materiales usados encada caso puede ser diferente.

Entre los cidos fenlicos en los clices de ja-maica, el protocatquico es el ms estudiado por susefectos como antioxidante que puede proteger de laoxidacin a las clulas hepticas de ratas (Tseng et al.,), e inducir apoptosis en clulas afectadas porleucemia en humanos (Tseng et al., ). Adems,el cido clorognico se ha relacionado con una reduc-cin significativa de las presiones sistlica y diastlica(Watanabe et al., ).

Fenoles extractables totales, antocianinas yproantocianidinas en los clices de jamaica

Los FET de una matriz vegetal o alimento son to-dos aquellos factibles de solubilizarse y ser extradoscon el disolvente usado (Saura-Calixto et al., ).En el estudio presente el contenido de FET fue es-tadsticamente diferente (p.) entre las varie-dades analizadas. El contenido mayor correspondia Sudn y el menor a Alma Blanca (Cuadro ). El

contenido de FET en los tres genotipos fue similar alreportado en variedades con colores de clices seme-jantes (Christian y Jackson, ).

El contenido de antocianinas totales fue mayoren Sudn y menor en Alma Blanca (p.). Lasvariedades con color diferente al blanco presenta-ron valores similares ( a mg ECG gMS) alos obtenidos por Juliani et al. () en variedadescultivadas en Senegal. En Alma Blanca el contenido

acid, which is unstable in the alkaline pH to whichthe extract was subjected for analysis. The varietyCriolla Nayarit had a high content of protocatechuicacid derivatives, Sudan had a value several timeslower, and it was not detected in the EPA fractionof Alma Blanca. The variety Sudan had derivatives

of syringic acid in the three phenolic acid fractionsanalyzed. In the FPA fraction of the analyzed varieties,chlorogenic acid accounted for ., . and. % of the total, in Alma Blanca, Sudan andCriolla Nayarit. If the isomers are considered, theproportion increases to ., . and . %,respectively. In Alma Blanca, caffeic acid was thefourth most important (. %), but in the coloredvarieties, the derivatives of gallic acid occupied thisposition. The predominance of chlorogenic acid

and its isomers in the aqueous extracts analyzedcoincided with reports by Rodrguez-Medina etal. () and Ramrez-Rodrguez et al. (),

who found the same result with the HPLC-MStechniques. This result, however, contrasted withobservations of Huang et al. (), who quantifieda fraction of Hibiscus spp. rich in polyphenols, in

which protocatechuic (. %) and caffeic (. %)acids were the most abundant. The divergences maybe due to the solvent used to obtain the extractssince ethanol, unlike water, favors extractionof protocatechuic acid (Chao and Yin, ).

Moreover, the genetics of the materials used mayhave been different in each case. Among the phenolic acids in the calyxes of

jamaica, protocatechuic acid is the most studiedbecause of its effects as antioxidant, capable ofprotecting oxidation of the rat liver cells (Tseng etal., ) and to induce apoptosis in cells affected byleukemia in humans (Tseng et al., ). Moreover,chlorogenic acid has been associated with significantreduction of systolic and diastolic blood pressure(Watanabe et al., ).

Total extractable phenols, anthocyanins andproanthocyanidins in jamaica calyxes

Total extractable phenols (TEP) of a plant orfood matrix are all of those that are feasibly solubleand extractable with a given solvent (Saura-Calixtoet al., ). In our study the TEP was statisticallydifferent (p.) among the analyzed varieties.

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Cuadro . Contenido de polifenoles extractables en clices secos de tres variedades de Hibiscus sabdariffa L.Table . Content of extractable polyphenols in dry calyxes of three Hibiscus sabdariffaL. varieties.

Sudn Criolla Nayarit Alma Blanca DHS

Fenoles extractables totales 36.51.25 a 15.50.10 b 13.50.32 c 1.123Antocianinas 10.990.34 a 4.520.33 b 0.200.03 c 0.641Proantocianidinas 1.040.03 a 0.430.04 b 0.240.05 c 0.077

Valores por fila, seguidos de la misma letra, no son estadsticamente diferentes (Tukey, .). DHS: diferencia honestasignificativa. mg EAG gMS; mg ECG gMS; mg EC gMSvValues in a row followed by the same letter arenot statistically different (Tukey,.). DHS: honest significant difference. mg GAE gDM; mg ECG gDM;mg EC gDM.

fue mayor que lo informado por Christian y Jack-son, (). En las variedades con clices rojos, laintensidad del color est relacionada con su conte-nido de antocianinas (Christian y Jackson, ;Salinas et al., ). La variedad Sudn, con clices

oscuros, present . veces ms antocianinas queCriolla Nayarit, con clices claros. Las antociani-nas en los FET en las variedades con clices rojosrepresentaron %, y en la de clices blancos %.Segn Syago-Ayerdi et al. (), cerca de y % de los FET en jamaica de clices rojos, sonantocianinas y cidos fenlicos.

Las proantocianidinas (PAs) fueron los fenolesmenos abundantes en los extractos analizados. Elmayor contenido se observ en la variedad Sudn(p.) y el menor en Alma Blanca. Su presencia

en clices de jamaica fue reportada por Obouaye-ba et al. () al monitorear los fitoqumicos enextractos acuosos de jamaica. El mtodo analticoutilizado (DMAC) detecta con mayor precisinmonmeros de flavan--ols, y reacciona slo conunidades terminales, sin discriminar monmeros,dmeros o trmeros (Hummer y Schreier, ).

As, las PAs en las jamaicas analizadas pueden serde tipo flavanoles, porque Huang et al. () re-portaron presencia de catequina, epigalocatequina yepigalocatequin-galato en un extracto concentradode jamaica. Las proantocianidinas se consideran fe-

noles con actividad antioxidante que ayudan en laprevencin de problemas cardiovasculares, porqueprotegen el msculo cardaco (Natella et al., ).

Actividad antioxidante

El ICes la concentracin del extracto que redu-ce % del radical libre. Los valores bajos de ICseasocian con el poder antioxidante mayor (Einbond

The highest content was found in Sudan and thelowest in Alma Blanca (Table ). The TEP contentin the three genotypes was alike to that reported forvarieties with colored calyxes similar (Christian and

Jackson, ).

The content of total anthocyanins was highestin Sudan and lowest in Alma Blanca (p.). Thevarieties that were not white had values ( to mgECG gDM) similar to those obtained by Julianiet al. () in varieties cultivated in Senegal. In

Alma Blanca, the content was higher than thatreported by Christian and Jackson (). Invarieties with red calyxes, the intensity of the coloris related with their anthocyanin content (Christianand Jackson, ; Salinas et al., ). The Sudanvariety, with dark red calyxes, had . times more

anthocyanins than Criolla Nayarit, with light redcalyxes. The anthocyanins in red calyxes accountedfor % of the TEP, and in white calyxes %.

According to Syago-Ayerdi et al. (), nearly and % of the TEP in red jamaica calyxes areanthocyanins and phenolic acids. The proanthocyanidins (PAs) were the leastabundant phenols in the analyzed extracts. Sudan

was the variety with the highest content (p.),and Alma Blanca had the lowest. Their presence in

jamaica calyxes was reported by Obouayeba et al.() when keeping track of phytochemicals in

aqueous extracs of jamaica. The analytical methodused (DMAC), detect flavan--ol monomers withgreater precision and, reacting only with terminalunits, it does not discriminate monomers, dimersor trimers (Hummer and Schreier, ). It isthus possible that the PAs in the jamaicas analyzedare flavanol types, as Huang et al. () reportedthe presence of catechin, epigallocatechin andepigallocatechin-galate in a concentrated extract of

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VOLUMEN 49, NMERO 3

et al., ). El patrn de la reduccin del radicalDPPH mostrado por Alma Blanca y Criolla Nayaritfue distinto al de Sudn (Figura ).

La reduccin por los extractos de Alma Blancay Criolla Nayarit fue casi lineal, en tanto que la deSudan fue exponencial. La variedad Sudn present

ICmenor que Alma Blanca y Criolla Nayarit, esdecir, tiene poder antioxidante mayor, atribuido asu contenido alto de antocianinas, ms que a suscidos fenlicos, que fueron similares a los de AlmaBlanca. El ICde los extractos de Criolla Nayarity Alma Blanca fue similar (p.). La actividadantioxidante de Alma Blanca se atribuye a los ci-dos fenlicos, pues prcticamente carece de anto-cianinas. La variabilidad entre mtodos para medirla actividad antioxidante en extractos acuosos de

jamaica y la forma de expresar los datos, dificulta

la comparacin de resultados. Segn Prez-Ramrezet al. () el ICpara una bebida de jamaica fue. mg de clices secos mL y los clculos conla concentracin de FET de la variedad Sudn danun ICde . ug mL

; por lo tanto, con po-der antioxidante mayor. Los resultados mostraron elpotencial antioxidante (cidos fenlicos) de las va-riedades de Hibiscus spp. de clices blancos similaral de las variedades con clices de otro color.

CONCLUSIONES

Independientemente de su color, los clices sonfuente abundante de cidos fenlicos y otros flavo-noides con actividad antioxidante. El color de los c-lices puede ser un indicador del tipo predominantede fenoles y ayudar a la seleccin de las variedadespara obtener formulaciones con propiedades nutra-cuticas particulares.

LITERATURACITADA

Abdel-Aal, E.-S. M., and P. Hucl. . A rapid method forquantifying total anthocyanins in blue aleurone and purple

pericarp wheat. Cereal Chem. : -.Ali, B. H., N. A. Wabel, and G. Blunden. . Phytochemical,

pharmacological and toxicological aspects of Hibiscussabdariffa L.: A review. Phytother. Res. : -.

Bakan, B. A., C. Bily, D. Melcion, B. Cahagnier, B. C.Regnault-Roger, B. J. R. Philogene, and D. Richard-Molard. . Possible role of plant phenolics in theproduction of trichothecenes by Fusarium graminearumstrains on different fractions of maize kernels. J. Agric.Food Chem. : -.

jamaica. Proanthocyanidins are considered phenolswith antioxidant activity that aids in the preventionof cardiovascular problems by protecting the cardiacmuscle (Natella et al., ).

Antioxidant activity

IC is the concentration of an extract whichreduces free radicals by %. Low IC values areassociated with greater antioxidant power (Einbondet al., ). The pattern of reduction of the DPPHradical exhibited by Alma Blanca and Criolla Nayarit

was different from that of Sudan (Figure ).Reduction by the Alma Blanca and Criolla Nayarit

extracts was almost linear, whereas that of Sudan wasexponential. The Sudan variety had a lower ICthan Alma Blanca and Criolla Nayarit, meaning

that it has greater antioxidant power, attributedto its high content of anthocyanins rather than itsphenolic acids, which were similar to those found in

Alma Blanca. The ICof the Criolla Nayarit andAlma Blanca varieties was similar (p.). Theantioxidant activity of Alma Blanca, which is similarto that of varieties of other colors, is attributed tothe phenolic acids since it almost completely lacksanthocyanins. Differences in methods for measuringantioxidant activity in aqueous extracts of jamaicaand how the data are expressed make it difficult to

90

80

70

60

50

40

30

20

10

00 50 100 150 200 250

-1Concentracin de fenoles (mg mL )

-1Sudn (IC =113.3 mg mL ; b)50

ReduccindeDPPH(

%)

-1Criolla N. (IC =142.0 mg mL ; a)50-1Alma B. (IC =139.7 mg mL ; a)50

Figura . Actividad antioxidante (IC) de extractos acuososde variedades de H. sabdariffaL. con color diferen-te de clices.

Figure . Antioxidant activity (IC) of aqueous extractsfrom varieties of H. sabdariffaL. with calyxes ofdifferent colors.

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Chao, Y-C., and M.-C. Yin. . Antibacterial effects of rosellecalyx extracts and protocatechuic acid in ground beef andapple juice. Foodborne Pathog. Dis. : -.

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compare results. For example, Prez-Ramrez et al.() reported that the ICof a jamaica beverage

was . mg dry calyxes mLwater. For comparison,respective calculations performed from the TEPconcentration in the variety Sudan, give an ICvalueof . ug mL; therefore, its antioxidant power

is lower. The results showed that the antioxidantpotential (phenolic acids) of the varieties of whitecalyx Hibiscusspp. is similar to that of the varieties

with calyxes of other colors.

CONCLUSIONS

Regardless of their color, jamaica calyxes area source of abundant phenolic acids and otherflavonoids with antioxidant activity. The color of thecalyxes may be an indicator of the predominating type

of phenols and could aid in the selection of varietiesfor formulations with particular nutraceuticalproperties.

End of the English version

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