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A NOVEL POTENTIAL INGREDIENT FOR FUNCTIONAL FOODS: PUNICALAGINS of POMEGRANATES ITU Istanbul Technical University Department of Food Engineering Artemis KARAALİ Neşe ŞAHİN, Mine GÜLTEKİN 2 nd International Congress on Functional Foods and Nutraceuticals, 4-6 May 2006, İstanbul
Transcript

A N

OV

EL P

OTE

NTI

AL

ING

RED

IEN

T FO

R

FUN

CTI

ON

AL

FOO

DS:

P

UN

ICA

LAG

INS

of P

OM

EGR

AN

ATE

S

ITU

Ista

nbu

lTec

hn

ical

Un

iver

sity

Dep

artm

ent

of F

ood

Eng

inee

rin

g

Art

emis

KA

RA

ALİ

Neş

e ŞA

HİN

, M

ine

LTEKİN

2n

dIn

tern

atio

nal

Con

gres

son

Fu

nct

ion

alFo

ods

and

Nu

trac

euti

cals

, 4-6

May

20

06

, İst

anbu

l

2

Out

line

of th

ePr

esen

tatio

n

Pom

egra

nate

Oth

erP

heno

licC

ompo

unds

in P

omeg

rana

tes

Phe

nolic

san

dH

ealth

Pun

ical

agin

sof

Pom

egra

nate

Exp

erim

enta

lStu

dyM

etho

dsR

esul

tsan

dD

iscu

ssio

nC

oncl

usio

n

3

Pom

egra

nate

(Punicagranatum

)

•Pom

egra

nate

is o

neof

the

olde

stkn

own

fruits

.•C

onsi

dere

da

holy

fruit

in th

eQ

uran

, and

grow

nin

Tur

key,

Iran

, Med

iterr

anea

n, M

iddl

eE

ast a

ndA

rabi

cco

untri

esan

dU

SA.

•Mai

nly

cons

umed

as fr

esh

fruit

(see

ds) o

rjui

ce.

•Che

mic

alco

mpo

sitio

ndi

ffers

depe

ndin

gon

cu

ltiva

r, gr

owin

gre

gion

(soi

land

clim

ate)

, ha

rves

tmat

urity

, cul

tura

lpra

ctic

es,

stor

age

perio

dan

dco

nditi

ons(

Tabl

es1

and

2).

0.61

gA

sh

16.5

7 g

Sug

ars

Valu

epe

r100

g

Prox

imat

es

0.6

gFi

ber

17.1

7 g

Car

bohy

drat

es0.

30g

Fat

0.95

gP

rote

in80

.97

gM

oist

ure

68 k

cal

Cal

orie

s

Tabl

e1.

Foo

dV

alue

Per 1

00 g

of E

dibl

ePo

rtion

(USD

A N

utrit

iona

lNut

rient

Dat

abas

efo

rSta

ndar

d R

efer

ence

, 200

5).

3 m

gM

agne

sium

259

mg

Pot

assi

um3

mg

Sod

ium

0.3

mg

Iron

8 m

gP

hosp

horu

s

3 m

gC

alci

um

Valu

epe

r100

g

Min

eral

s

0.59

6 m

gP

anto

then

icac

id

0.10

5 m

gV

itB

6

Vita

min

s

4.6

mcg

Vit

K0.

6 m

gV

itE

108

IUP

roV

itA

6.1

mg

Asc

orbi

cA

cid

0.3

mg

Nia

cin

0.03

mg

Rib

ofla

vin

0.03

mg

Thia

min

eLi

pids

50 m

cgα-

Car

oten

e40

mcg

Β-C

arot

ene

Oth

ers

0.06

3 g

Fatty

acid

s, to

tal

poly

unsa

tura

ted

0.03

8 g

Fatty

acid

s, to

tal

satu

rate

d

Tabl

e2.

Foo

dV

alue

Per 1

00 g

of E

dibl

ePo

rtion

(USD

A N

utrit

iona

lNut

rient

Dat

abas

efo

rSta

ndar

d R

efer

ence

, 200

5).

6

Phen

olic

sof

Pom

egra

nate

s

Pom

egra

nate

sar

equ

iteric

hin

hyd

roly

sabl

eta

nnin

san

dan

thoc

yani

ns.

Maj

orhy

drol

ysab

leta

nnin

sin

pom

egra

nate

sar

ega

llota

nnin

s, e

llagi

cac

idta

nnin

san

dga

llagy

ltan

nins

(thes

ear

ete

rmed

“pun

ical

agin

”s).

7

Phe

nolic

com

poun

dsha

vebe

ensh

own

toac

tas

free

radi

cals

cave

nger

san

dto

have

antio

xida

ntac

tivity

.H

owev

er, t

heir

bioa

vaila

bilit

ies

and

the

exac

tm

echa

nism

of th

eira

ctio

nin

the

hum

anbo

dy is

not

yet

to

tally

eluc

idat

ed.

Ita

subj

ects

tillu

nder

wid

esc

ient

ific

inve

stig

atio

n.

Phen

olic

sof

Pom

egra

nate

s

8

Pol

yphe

nols

wer

esh

own

toha

veth

eab

ility

toin

hibi

tlo

w-d

ensi

tylip

opro

tein

(LD

L) o

xida

tion

and

mac

roph

age

foam

cell

form

atio

n. T

here

fore

they

are

hypo

thes

ized

toha

ve:

Ant

i-ath

erog

enic

effe

cts,

redu

cing

the

inci

denc

eof

ca

rdio

-vas

cula

rand

cere

bro-

vasc

ular

dise

ases

(Kul

karn

iet

al.,

200

4, L

iet a

l., 2

006)

.A

ntip

rolif

erat

ive

activ

ityag

anis

tora

l, co

lon

and

pros

tate

canc

ers

(See

ram

, et a

l., 2

005)

, re

duci

ngca

ncer

mor

talit

y(K

ulka

rnie

t al.,

200

4, L

iet a

l, 20

06).

Hea

lthC

laim

sfo

rPol

yphe

nols

9

•Apo

ptop

icef

fect

sag

anis

thum

anco

lon

canc

erce

lls(S

eera

m, e

t al.,

200

5).

•The

rape

utic

effe

cton

col

ic, c

oliti

s, d

iarr

hea,

dy

sent

ery,

leuc

orrh

ea, m

enor

rhag

ia, o

xyur

iasi

s,

para

lysi

s, re

ctoc

ele

and

head

ache

(Sch

uber

tet a

l.,

1999

).•T

hera

peut

icef

fect

on in

fect

ion

of m

ale

orfe

mal

ese

xual

orga

ns, m

astit

is, a

cne

(Sin

ghet

al.,

200

2).

•Ant

ivira

lact

ivity

agan

istg

enita

lher

pes

viru

s(S

ingh

et

al.,

2002

).

Hea

lthC

laim

sfo

rPol

yphe

nols

10

•Fur

ther

ther

apeu

ticac

tions

such

as v

erm

ifuga

l, ta

enic

idal

, ast

ringe

nt, a

ntis

pasm

odic

, ant

iyhs

teric

, di

uret

ic, c

arm

inat

ive,

sud

ofiri

c, g

alac

togo

gue,

em

men

agog

ue(S

chub

erte

t al.,

199

9).

•Fol

licul

itis,

pie

, alle

rgic

derm

atiti

s, ty

mpa

nitis

, di

arrh

ea, d

ysen

tery

(Sin

ghet

al.,

200

2).

•Neu

ropr

otec

tive

effe

cts

thro

ugh

impr

ovin

glip

idpr

ofile

sin

dia

betic

patie

nts

(Sch

uber

tet a

l., 1

999)

.

Hea

lthC

laim

sfo

rPol

yphe

nols

11Fi

gure

1.C

hem

ical

stru

ctur

eof

pun

ical

agin

s(G

ilet

al.,

200

0).

Puni

cala

gins

of P

omeg

rana

tes

•The

new

term

inve

nted

for

pom

egra

nate

tann

ins

is

“pun

ical

agin

s”.

•Itis

foun

dto

be q

uite

abun

dant

in

the

pom

egra

nate

juic

e(<

2g/L

). •It

is a

sup

erio

rant

ioxi

dant

resp

onsi

ble

fort

hefre

e-ra

dica

lsc

aven

ging

abilit

yof

the

pom

egra

nate

juic

e.

12

Oth

erPh

enol

icC

ompo

unds

in

Pom

egra

nate

s

•Ella

gic

acid

deriv

ates

•Ant

hocy

anin

s(D

elph

inid

in, c

yani

din,

pel

argo

nidi

n3-

gluc

osid

e, 3

,5-d

iglu

cosi

des)

Figu

re2

.Che

mic

alSt

ruct

ure

of E

llagi

cAc

id(G

ilet

al.,

200

0).

13Expe

rimen

talS

tudy

MAT

ERIA

LS:

Pom

egra

nate

s: p

eela

ndse

edex

tract

s

Juic

es: F

resh

juic

epr

epar

edin

the

labo

rato

ryC

omm

erci

alju

ice

sam

ple

Pee

lpow

der

See

dp

owd

eraf

ter

rem

oval

of f

atty

mat

ter

Peel

ing

pom

egra

nate

sPe

elin

gpo

meg

rana

tes

Extr

actio

nof

ant

ioxi

dant

frac

tions

in m

etha

nol

Extr

actio

nof

ant

ioxi

dant

frac

tions

in m

etha

nol

Filtr

atio

nan

dco

ncen

trat

ion

unde

rvac

uum

(40C

)Fi

ltrat

ion

and

conc

entr

atio

nun

derv

acuu

m(4

0C)

1. F

orPo

meg

rana

teSe

edan

dPe

elSa

mpl

es(S

ingh

et a

l., 2

002)

.Met

hods

Pres

sing

and

extr

actin

gth

eju

ice

loca

lised

in s

acs/

com

mer

cial

juic

ePr

essi

ngan

dex

trac

ting

the

juic

elo

calis

edin

sac

s/co

mm

erci

alju

ice

Cen

trifu

ging

for2

min

at 1

400

rpm

Cen

trifu

ging

for2

min

at 1

400

rpm

Peel

ing

Peel

ing

2. F

orPo

meg

rana

teJu

ice

Sam

ple

(Gil

et a

l., 2

000)

3. “

Phen

olic

prof

iling

”: A

naly

sis

of In

divi

dual

Phen

olic

Com

poun

dsby

HPL

C

(Sin

ghet

al.,

200

2 an

dG

ilet

al.,

200

0)

20 µ

l sam

ple

inje

cted

into

HPL

C e

quip

ped

with

PAD

ded

ecto

r, C

18 s

upel

cosi

lcol

umn

(5 µ

m p

artic

lesi

ze)

Solv

entA

:3%

ace

ticac

id+9

7% w

ater

Solv

entB

: 3%

ace

ticac

id+2

5% a

ceto

nitr

ile+7

2% w

ater

20 µ

l sam

ple

inje

cted

into

HPL

C e

quip

ped

with

PAD

ded

ecto

r, C

18 s

upel

cosi

lcol

umn

(5 µ

m p

artic

lesi

ze)

Solv

entA

:3%

ace

ticac

id+9

7% w

ater

Solv

entB

: 3%

ace

ticac

id+2

5% a

ceto

nitr

ile+7

2% w

ater

Extr

acts

diss

olve

din

M

eOH

:Wat

er(6

:4 v

/v)

Extr

acts

diss

olve

din

M

eOH

:Wat

er(6

:4 v

/v)

Cen

trifu

ged

Juic

ein

MeO

H:W

ater

(6:4

v/v

) C

entr

ifuge

dJu

ice

inM

eOH

:Wat

er(6

:4 v

/v)

Peel

and

seed

extr

acts

Peel

and

seed

extr

acts

Dis

solv

ing

in M

eOH

:Wat

er(6

:4 v

/v)

Dis

solv

ing

in M

eOH

:Wat

er(6

:4 v

/v)

Sam

ple

(300

µl)+

10 fo

ld-d

ilutio

nw

ithFC

reag

ent(

1.5

ml)+

7.5%

Na2

C03

(1.2

ml)

Sam

ple

(300

µl)+

10 fo

ld-d

ilutio

nw

ithFC

reag

ent(

1.5

ml)+

7.5%

Na2

C03

(1.2

ml)

Stan

ding

for1

.5 h

at r

oom

tem

pera

ture

Stan

ding

for1

.5 h

at r

oom

tem

pera

ture

Abs

orba

nce

mea

sure

men

tat 7

65 n

mby

UV-

visi

ble

spec

trop

hoto

met

erA

bsor

banc

em

easu

rem

enta

t 765

nm

byU

V-vi

sibl

esp

ectr

opho

tom

eter

4. D

eter

min

atio

nof

Tot

al P

heno

ls(F

olin

-Cio

calte

uM

etho

d, S

ingh

et a

l., 2

002)

.

Juic

eJu

ice

5. D

eter

min

atio

nof

Ant

ioxi

dant

Act

ivity

(DPP

H M

etho

d, M

oren

oet

al.,

199

8)

100 µl

ext

ract

s/ju

ice

+ 3.

9 m

l DPP

H s

olut

ion

100 µl

ext

ract

s/ju

ice

+ 3.

9 m

l DPP

H s

olut

ion

Vigo

urou

sM

ixin

gVi

gour

ous

Mix

ing

Stan

ding

for3

0 m

inat

room

tem

pera

ture

Stan

ding

for3

0 m

inat

room

tem

pera

ture

Abs

orba

nce

mea

sure

men

tat 5

15 n

mby

UV-

visi

ble

spec

trop

hoto

met

er(%

of r

adic

alsc

aven

ged

byte

st s

ampl

e)

Abs

orba

nce

mea

sure

men

tat 5

15 n

mby

UV-

visi

ble

spec

trop

hoto

met

er(%

of r

adic

alsc

aven

ged

byte

st s

ampl

e)

19

Tota

l Ph

enol

icC

onte

nts

Tabl

e2.

Phen

olic

cont

ento

f pom

egra

nate

peel

san

dse

edex

tract

s

galli

cac

ideq

uiva

lent

3.1

46.5

Phe

nolic

s(m

g/g)

2.88

See

d48

.66

Pee

l

Phe

nolic

s(m

g/g)

(Sin

ghet

al.,

200

2)P

omeg

rana

te

Res

ult

san

dD

iscu

ssio

n

20

Tota

l Ph

enol

icC

onte

nts

Figu

re5.

Tota

l phe

nolc

onte

ntof

pom

egra

nate

peel

and

seed

extra

cts.

tota

l phe

nolic

con

tent

01020304050

mg GA/g

peel

seed

21

galli

cac

ideq

uiva

lent

2480

1860

Phen

olic

s(m

g/L)

2161

.88

2426

.39

Fres

hju

ice

2941

.01

C

omm

erci

alju

ice

Phen

olic

s(m

g/L)

(Alp

er e

t. A

l., 2

005,

G

ilet

al.,

200

0).

Pom

egra

nate

Tabl

e3

.Phe

nolic

cont

ent

of p

omeg

rana

teju

ices

.

Tota

l Ph

enol

icC

onte

nts

22

Tota

l Ph

enol

icC

onte

nts

Figu

re6.

Tota

l phe

nolc

onte

ntof

pom

egra

nate

juic

es*h

ydro

stat

icpr

essu

reus

edin

the

com

mer

cial

prod

ucts

crus

hes

the

who

lefru

it,ex

tract

ing

also

the

ella

gita

nnin

sin

the

peel

s

tota

l phe

nolic

con

tent

0

500

1000

1500

2000

2500

3000

mg GA/L

fresh

juic

e

com

mer

cial

juic

e

23

Rad

ical

Scav

engi

ng

Act

ivit

y

12.2

at 2

5 pp

m54

.0 a

t 25

ppm

Ant

ioxi

dant

Act

ivity

(%)

23.2

at 1

00 p

pmS

eed

81.0

at 5

0 pp

mP

eel

Ant

ioxi

dant

Act

ivity

(%)

(Sin

ghet

al.,

200

2)P

omeg

rana

te

Tabl

e4

.Ant

ioxi

dant

activ

ityof

pom

egra

nate

seed

and

peel

exra

cts

24

Antio

xidan

t Acti

vity

0102030405060 % scavenging activity

peel

seed

Rad

ical

Scav

engi

ng

Act

ivit

y

Figu

re7.

Antio

xida

ntac

tivity

of p

omeg

rana

tepe

elan

dse

edex

tract

s.

25

Rad

ical

Scav

engi

ng

Act

ivit

y

Tabl

e5

.Ant

ioxi

dant

activ

ityof

pom

egra

nate

juic

es.

54.8

at 5

mg/

mL

32.5

at 5

mg/

mL

Ant

ioxi

dant

Act

ivity

(%)

-Fr

esh

juic

e26

.4

Com

mer

cial

juic

e

Ant

ioxi

dant

Act

ivity

(%)

(Gil

et a

l., 2

000)

Pom

egra

nate

26

Rad

ical

Scav

engi

ng

Act

ivit

y:

Antio

xida

nt A

ctiv

ity

0102030405060

% scavenging activity

Figu

re8.

Antio

xida

ntac

tivity

of p

omeg

rana

teju

ices

*hyd

rost

atic

pres

sure

used

in th

eco

mm

erci

alpr

oduc

tscr

ushe

sth

ew

hole

fruit,

extra

ctin

gal

soth

eel

lagi

tann

ins

in th

epe

els

Fres

hju

ice

Com

mer

cial

juic

e

27

Ph

enol

icP

rofi

ling

Figu

re3.

Phen

olic

com

posi

tion

of fr

esh

pom

egra

nate

juic

e.

28Fi

gure

4.Ph

enol

icco

mpo

sitio

nof

com

mer

cial

pom

egra

nate

juic

e.

Ph

enol

icP

rofi

ling

29

Con

clu

sion

s

Pom

egra

nate

, ju

ice

and

othe

rpom

egra

nate

prod

ucts

prom

ise

toha

vehe

alth

-pro

mot

ing

effe

cts.

The

fact

orre

spon

sibl

efo

rthe

high

antio

xida

ntac

tivity

in th

epo

meg

rena

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unic

alag

in”,

whi

chis

an

ella

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nnin

.

Pom

egra

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peel

sm

aybe

a ri

chso

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and

raw

mat

eria

lfor

natu

rala

ntio

xida

nts.

30

Ref

eren

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nym

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2005

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Stan

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D.M

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Ant

ioxi

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Activ

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Juic

ean

dIt

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with

Phen

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nan

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ngAn

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Carp

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Food

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551-

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aney

uki,

T., M

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A., P

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002.

Ant

ioxi

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Activ

ities

of

Pom

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nate

Frui

tEx

trac

tan

dIt

sAn

thoc

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din,

Cya

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n, a

ndPe

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50, 1

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02.O

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Prop

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Seed

Oil

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Ferm

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J. E

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6, 1

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Antip

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Apo

ptop

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dAn

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Activ

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Puni

cala

gin,

Ella

gic

Acid

and

A To

tal P

omeg

rana

teTa

nnin

Extr

act

are

Enha

nced

InCo

mbi

natio

nw

ithO

ther

Poly

phen

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as F

ound

InPo

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teJu

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