Transcript שקופית 1 - תשלובת ד"ר רימון

‫‪Migal, Kiryat Shmona‬‬
‫בירור הרכב החומרים מונעי החמצון בחלקים שונים של‬
‫פרי הרימון ע"י סריקה של ‪ 29‬זני רימונים שונים‬
‫יום עיון למגדלי רימונים‪ ,‬מכון וולקני‪ ,‬פברואר ‪2008‬‬
‫מוקדש לזכרו של משה זמירי ז"ל‬
Atherosclerosis lesion
development
Thrombus formation
Carotid Intima-Media Thickness
Percent of mortality
In western countries
OTHERS 25%
CHD 54%
CANCER 21%
Recent biological studies have been proven that certain compounds in the
pomegranate juice has been shown to:
Reduce blood pressure; Reduce LDL oxidation; Reduce the atherosclerosis formation
Moreover: consumption of
pomegranate juice
can lead to reduction of the
atherosclerosis lesion
Prof. Miki Aviram
Pomegranate Juice Consumption by Healthy
Copper Ion-Induced LDL Oxidation
(nmol TBARS/ mg LDL protein)
Volunteers Reduces LDL Oxidation
A.
30
*
20
*
10
0
0
1
Time (weeks)
2
Pomegranate Juice Consumption by Patients with Carotid Artery
Stenosis Reduces Carotid Intima-Media Thickness (IMT)
A. Control
Carotid IMT (mm)
3
2
1
0
4
B. Pomegranate Juice
2
0
Baseline
After 1 year
These activities are attributed to the Pomegranate high levels of antioxidant
content of several Juices
Total
activity and
its high Polyphenols
total polyphenol content
Total polyphenols content of several fruit juices
7
5
4
3
2
1
ra
Ju nbe
ic rr
e y
G
re
en
Te
a
O
ra
ng
e
Ju
ic
e
C
lu
Ju ebe
ic rr
e y
B
e
in
W
ed
R
e
Ju gra
ic na
e
te
0
Po
m
Polyphenols (mM)
6
Most of the data derived from one pomegranate cultivar called ‘wonderful’
Our main objectives:
1. To compare the levels of antioxidants,
total polyphenols, anthocyanins and
four major hydrolysable tannins in 29
pomegranate cultivars.
2. To verify where the bioactive compounds
are localize in the fruits (arils, lamellas,
seeds, peels), and examine their
chemical nature and their antioxidant
activity.
P.G. 112-13
P.S. 21
C13
PG128-29
PG 100-1
PG 103-4
PG 104-5
PG 108-9
PG 109-10
PG 112-13
PG 118-19
PG 119-20
PG 120-21
PG 102-3
PG 101-2
PG 105-6
PG 106-7
PG 114-15
PG 116-17
PG 121-22
PG 123-24
PG 127-28
PG 128-29
PG 130-31
A17
C13
C14
C133
ERS
-
2006
PS21
PS23
XBS
Camel
Two different juices and two different homogenates were pretreated from
each cultivar:
1. Arils juice
2. Juice prepare by Juice extractor
3. Homogenate prepared from the whole fruit
4. Homogenate prepared from the peels
Antioxidant
capacity
*10^3 (mg/L)
Total
Polyphenols
*10^3 (mg/L)
Total
anthocyanins
*10^2 (mg/L)
Punicalagin
m/z 1083
*10^2 (mg/L)
2
Antioxidant activity
1
0
2
Total polyphenols
R2 = 0.76**
1
0
3
2
Total anthocyanins
R2 = 0.46**
1
Punicalagin
1.4
R2 = 0.03
0.7
0
Punicalin
m/z 781
(mg/L)
Antioxidant activity,
total polyphenols,
total anthocyanins,
and the level of
hydrolysable tannins
in aril juices
8
Punicalin
6
R2 = 0.00
Punicalagin
4
Gallagic acid
m/z 601
*10 (mg/L)
2
3
2
Gallagic acid
R2 = 0.01
1
Ellagic acid
0
Gallagic acid
** significant by Pirson test ,α=0.01
No positive correlation was found between the acid level and the content of
organic acids in the aril juices
acid level
(%)
Acid level
2
1
pH level
0
Citric acid
level (%)
Ascorbic acid
level (mg/L)
pH level
R2 = 0.07
4
3
2
Acetic acid
level (%)
R2 = 0.08
Citric acid level
2
R2 = 0.05
1
0
20
16
12
8
4
0
0.8
0.6
Obtained by HPLC
Ascorbic acid level
R2 = 0.00
Acetic acid level
R2 = 0.48**
0.4
0.2
0
** significant by Pirson test ,α=0.01
Positive correlation was found between the TSS and the content of
Glucose and fructose in the aril juices
TSS (%)
Antioxidant
capacity
(mM)
10
8
Antioxidant contents
6
4
2
0
18
17
16
15
14
13
%TSS
R2 = 0.68**
Glucose
level (%)
Glucose level
6
R2 = 0.58**
5
Obtained by HPLC
4
Fructose
level (%)
Fructose level
6
R2 = 0.58**
5
4
** significant by Pirson test ,α=0.01
Punicalin Gallagic acid Punicalagin LDL Oxidation Anthocyanin
Ellagic Acid
Antioxidant
Total
m/z 781
m/z 301
m/z 1083
m/z 601
Pigment
Inhibition
Polyphenols capacity
*10^2 (mg/L)
(mg/L)
*10^2 (mg/L)
(mg/L)
*10^2 (mg/L) *10^3 (mg/L) *10^4 (mg/L)
*10 (%)
1
Antioxidant activity
0.5
0
6
4
Total polyphenol
R2 = 0.61**
2
0
4
3
2
1
0
8
6
4
2
0
20
18
6
4
2
0
40
36
~
~
8
Total anthocyanin
R2 = 0.00
Antioxidant activity,
total polyphenols,
total anthocyanins,
and the inhibitory effect on
LDL oxidation in juices
prepared by juice extractor
LDL oxidation inhibition
Lipid peroxides, R2 = 0.71**
TBARS,
R2 = 0.69**
Punicalagin
R2 =
0.63**
Gallagic acid
R2 =
0.64**
4
0
6
4
Punicalin
R2 =
0.58**
2
0
15
Ellagic Acid
10
R2 = 0.68**
5
The antioxidant was increased
by 5 fold compare to juice
prepared only from arils
0
** significant by Pirson test ,α=0.01
Antioxidant
capacity
*10^4 (mg/L)
Total
Polyphenols
*10^4 (mg/L)
Punicalagin Anthocyanin
Pigment
m/z 1083
*10^4 (mg/L) *10^2 (mg/L)
6
5
4
3
2
1
0
Total polyphenol
1
Ellagic Acid
Punicalin
m/z 301
m/z 781
*10^4 (mg/L) *10^2 (mg/L)
R2 = 0.90**
0.5
0
4
3
Total Anthocyanin
Antioxidant activity,
total polyphenols,
total anthocyanins,
and the inhibitory effect on
LDL oxidation in homogenates
prepared from the whole fruit
R2 = 0.05
2
1
0
3
Punicalagin
R2 = 0.79**
2
1
0
Gallagic acid
m/z 601
*10^3 (mg/L)
Antioxidant activity
4
3
Ellagic Acid
R2 = 0.44**
2
1
0
1.2
Punicalin
R2 = 0.54**
0.6
0
8
6
Gallagic acid
R2 = 0.55**
The antioxidant was increased
by 20 fold compare to juice
prepared only from arils
4
2
0
** significant by Pirson test ,α=0.01
Antioxidant activity
0.5
0
1.5
1
Total Polyphenol
R2 = 0.91**
0.5
0
8
6
Total anthocyanin
R2 = 0.01
4
Antioxidant activity,
total polyphenols,
total anthocyanins,
and the inhibitory effect on
LDL oxidation in homogenates
prepared from the peels
2
0
4
3
Punicalagin
R2 = 0.39**
2
Punicalin
Ellagic Acid
m/z 781
m/z 301
*10^4 (mg/L) *10^2 (mg/L)
1
0
8
Gallagic acid
m/z 601
*10^3 (mg/L)
Punicalagin Anthocyanin
Antioxidant
Total
m/z 1083
capacity
Polyphenols
Pigment
*10^5 (mg/L) *10^2 (mg/L) *10^4 (mg/L) *10^5 (mg/L)
1
0
6
The antioxidant was increased
by 40 fold compare to juice
prepared only from arils
Ellagic Acid
R2 = 0.49**
4
2
0
Punicalin
R2 = 0.72**
1
8
6
4
Gallagic acid
R2 =
0.89**
2
0
** significant by Pirson test ,α=0.01
The antioxidant level is not correlated to the % of the peels from the fruits
Antioxidant
capacity
*10^2 (mM)
4
3
2
1
0
Fruit weight
*10^2 (gr)
Antioxidant contents
6
Fruit weight
R2 = 0.01
4
2
Peel weight
*10^2 (gr)
0
3
Peel weight
R2 = 0.01
2
1
Aril weight
*10^2 (gr)
0
4
3
2
1
0
Aril weight
R2 = 0.06
Summary
1. In arils juice- the antioxidant activity appear to be related to polyphenols
and anthocyanins. This is accordance with other data demonstrated that in
colored figs, apple, peaches and grapes the antioxidant level is significantly
higher then in green fruits.
2. In homogenate prepared from the whole fruit, the level of the antioxidant
increased by 20-fold suggested that the peels contributed significantly to
antioxidant activity.
3. The predominant type of polyphenolic compounds extracted from the peels
are the hydrosalbe tannins, composed mainly from punicalagin isomers, punicalin,
gallagic acid and ellagic acid.
The study enlarge our knowledge about the natural variation of the antioxidant
content and the compounds that give this activity.
It also identify the location and concentration of some phytochemical that have
antioxidant activity in pomegranate fruits.
In the future, such information will enable breeders to select and breed
genotypes having higher levels of health beneficial compounds and also
provide useful information for addressing consumer choices for healthier products
Students
Migal, Kiryat Shmona
Revital Zulker
Ira Glazer
Elinor Shwarzt
Collaborators:
Dr. Doron Holland, Newe Ya’ar Research Center
Prof. Miki Aviram, Technion, Faculty of Medicine, Haifa
Dr. Hamutal Borochov, Mop Arava, Eilat
Dr. Igal Bar-Ilan, Migal, Kiryat Shmona
Prof. Ruth Ben-Arie, The fruit storage laboratory, Kiryat Shmona
‫תודה על ההקשבה‬
‫‪H‬‬
‫‪80‬‬
‫‪70‬‬
‫‪60‬‬
‫‪50‬‬
‫‪40‬‬
‫‪30‬‬
‫‪20‬‬
‫‪10‬‬
‫‪0‬‬
‫)‪Antioxidant capacity (mM‬‬
‫רמת האנטיאוקסידנטיות במיצים מסחריים‬
‫‪M‬‬
‫רמת הפוליפנולים במיצים מסחריים‬
‫‪10‬‬
‫‪6‬‬
‫‪4‬‬
‫‪2‬‬
‫‪H‬‬
‫‪0‬‬
‫‪M‬‬
‫)‪Total Polyphenols (mM‬‬
‫‪8‬‬
‫‪ – NF‬נטוראפוד‬
‫מיץ אורגני מסחרי‬
‫‪ D2‬מיץ רימונים נסחט במסחטה‬
‫‪ PJ‬מיץ מיקי מהול לשתייה‬
‫‪ -M‬תרכיז רימונים ממולי‬
‫‪ -71‬מיץ טרי מגרגירים בלבד‬
‫‪ -H‬כנ"ל‬
‫יין רימונים‪ -‬יקב דלתון‬
‫מיץ טבעי‪ -‬מעמוס‬
The identity of the hydrolysable-tannins
obtained by using
Multiple Reaction Monitoring (MRM)
method of LC-MS-MS, according to their
mother and daughter ions.
Multiple Reaction
Monitoring (MRM)
The Mother ion was fragmented by
argon using different collision energies
to daughter ions. The level of the typical
daughter ion was then measured.
Q1
Q2
Q3
Single
Ion m/z
Collision
Single daughter
Ion m/z