בירור הרכב החומרים מונעי החמצון בחלקים שונים של פרי הרימון ע"י סריקה של 29 זני רימונים שונים

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1 Migal, Kiryat Shmona בירור הרכב החומרים מונעי החמצון בחלקים שונים של פרי הרימון ע"י סריקה של 9 זני רימונים שונים יום עיו ן ל מגדל י רי מונ ים, מכון ו ול קני, פברואר 8 מוקד ש לזכר ו של משה ז מי רי ז"ל

2 Atherosclerosis lesion development Thrombus formation Carotid Intima-Media Thickness Percent of mortality In western countries OTHERS 5% CANCER % CHD 5%

3 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

4 Pomegranate Juice Consumption by Healthy Volunteers Reduces LDL Oxidation A. * Time (weeks) * (nmol TBARS/ mg LDL protein) Copper Ion-Induced LDL Oxidation

5 Pomegranate Juice Consumption by Patients with Carotid Artery Stenosis Reduces Carotid Intima-Media Thickness (IMT) A. Control Carotid IMT (mm) B. Pomegranate Juice Baseline After year

6 These activities are attributed to the Pomegranate high levels of antioxidant activity and its high total polyphenol content Total polyphenols content of several fruit juices 7 Polyphenols (mm) 5 Pomegranate Juice Red Wine Blueberry Juice Cranberry Juice Green Tea Orange Juice

7 Most of the data derived from one pomegranate cultivar called wonderful

8 Our main objectives:. To compare the levels of antioxidants, total polyphenols, anthocyanins and four major hydrolysable tannins in 9 pomegranate cultivars.. 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. - C P.S. PG8-9

9 PG - PG - PG - PG - PG -5 PG 5- PG -7 PG 8-9 PG 9- PG - PG -5 PG -7 PG 8-9 PG 9- - PG - PG - PG - PG 7-8 PG 8-9 PG - A7 C C C ERS PS PS XBS Camel

10 Two different juices and two different homogenates were pretreated from each cultivar:. Arils juice. Juice prepare by Juice extractor. Homogenate prepared from the whole fruit. Homogenate prepared from the peels

11 Antioxidant activity Total polyphenols R =.7** Total anthocyanins R =.**. Punicalagin R =..7 8 Punicalin R =. Gallagic acid R =. Antioxidant activity, total polyphenols, total anthocyanins, and the level of hydrolysable tannins in aril juices Punicalagin Gallagic acid ** significant by Pirson test,α=. Ellagic acid PG - PG 7-8 PG -7 PG 9- PG 8-9 PG -5 PG - PG 5- PG - PG -7 PG - PG - PG -7 PG 8-9 PG -5 PG - PG - PG - PG - PG 8-9 PG 9- PG - Eve Orange PG -5 PG - PG 5- PG - PG - Gallagic acid m/z * (mg/l) Punicalin m/z 78 (mg/l) Punicalagin m/z 8 *^ (mg/l) Total anthocyanins *^ (mg/l) Total Polyphenols *^ (mg/l) Antioxidant capacity *^ (mg/l)

12 No positive correlation was found between the acid level and the content of organic acids in the aril juices Acid level R =.8 acid level (%) ph level R =.7 ph level Citric acid level R =.5 Citric acid level (%) 8 Ascorbic acid level Ascorbic acid level (mg/l).8.. PG - PG 7-8 PG -7 PG 9- PG 8-9 PG -5 PG - PG 5- PG - PG -7 PG - ERS Camel PG 8-9. Acetic acid level R =. Acetic acid level (%) PG -5 PG - PG - PG - PG - PG 8-9 PG 9- PG - C A7 C PS PS C XBS R =.8** Obtained by HPLC ** significant by Pirson test,α=.

13 Positive correlation was found between the TSS and the content of Glucose and fructose in the aril juices Antioxidant contents %TSS Glucose level Fructose level R =.8** R =.58** R =.58** Obtained by HPLC PG - PG 7-8 PG -7 PG 9- PG 8-9 PG -5 PG - PG 5- PG - PG -7 PG - ERS Camel PG 8-9 PG -5 PG - PG - PG - PG - PG 8-9 PG 9- PG - C A7 C PS PS C XBS ** significant by Pirson test,α=. Fructose level (%) Glucose level (%) TSS (%) Antioxidant capacity (mm)

14 Antioxidant activity.5 Total polyphenol R =.** 8 8 ~ ~ Total anthocyanin R =. 8 LDL oxidation inhibition Lipid peroxides, R =.7** TBARS, R =.9** Punicalagin R =.** Gallagic acid R =.** Punicalin R =.58** 5 Ellagic Acid R =.8** 5 ** significant by Pirson test,α=. PG - PG 9- PG - PG -5 PG -7 PG 8-9 PG - PG - PG 7-8 PG - PG - PG - PG 8-9 PG -5 Orange PG 8-9 PG - PG - PG - Eve PG - Antioxidant activity, total polyphenols, total anthocyanins, and the inhibitory effect on LDL oxidation in juices prepared by juice extractor The antioxidant was increased by 5 fold compare to juice prepared only from arils PG - PG 5- Ellagic Acid m/z (mg/l) Punicalin m/z 78 *^ (mg/l) Gallagic acid m/z (mg/l) Punicalagin m/z 8 *^ (mg/l) LDL Oxidation Inhibition * (%) Anthocyanin Pigment *^ (mg/l) Total Polyphenols *^ (mg/l) Antioxidant capacity *^ (mg/l)

15 PG -7 PG 9- PG - PG 8-9 PG 7-8 PG - PG -7 PG 8-9 PG -5 PG - PG 5- PG - PG 5- PG -7 Eve PG 8-9 PJ PG - Orange PG - PG - PG Punicalagin Ellagic Acid Punicalin Gallagic acid PG - PG -5 PG 9- PG -5 PG - PG - Antioxidant activity Total polyphenol R =.9** Total Anthocyanin R =.5 R =.79** R =.** R =.5** R =.55** Antioxidant activity, total polyphenols, total anthocyanins, and the inhibitory effect on LDL oxidation in homogenates prepared from the whole fruit The antioxidant was increased by fold compare to juice prepared only from arils ** significant by Pirson test,α=. PG - PG - Gallagic acid m/z *^ (mg/l) Punicalin m/z 78 *^ (mg/l) Ellagic Acid m/z *^ (mg/l) Punicalagin m/z 8 *^ (mg/l) Anthocyanin Pigment *^ (mg/l) Total Polyphenols *^ (mg/l) Antioxidant capacity *^ (mg/l)

16 Antioxidant activity.5.5 Total Polyphenol R =.9**.5 8 Total anthocyanin R =. Punicalagin R =.9** 8 Ellagic Acid R =.9** Punicalin R =.7** PG -7 PG -5 PG - PG - PG - PG - PG - PG 8-9 PG - PG 7-8 PG 9- PG -7 PG - PG 9- PG 8-9 PG - PG 5- Orange PG - PG - PG - 8 PG -5 PG - PG 5- PG -5 PG - Eve PG 8-9 PG -7 Gallagic acid R =.89** Antioxidant activity, total polyphenols, total anthocyanins, and the inhibitory effect on LDL oxidation in homogenates prepared from the peels The antioxidant was increased by fold compare to juice prepared only from arils ** significant by Pirson test,α=. Gallagic acid m/z *^ (mg/l) Punicalin m/z 78 *^ (mg/l) Ellagic Acid m/z *^ (mg/l) Punicalagin m/z 8 *^5 (mg/l) Anthocyanin Pigment *^ (mg/l) Total Polyphenols *^ (mg/l) Antioxidant capacity *^5 (mg/l)

17 The antioxidant level is not correlated to the % of the peels from the fruits Antioxidant contents Antioxidant capacity *^ (mm) Fruit weight R =. Fruit weight *^ (gr) Peel weight R =. Peel weight *^ (gr) Aril weight R =. Aril weight *^ (gr) PG -7 PG -5 PG - PG - PS PG - PG - PG 8-9 PG - PG 7-8 PG 9- PG -7 PG - PG 9- PG 8-9 XBS PG 5- A7 ERS PG - PG - PG -5 PG - PS C PG - C CEve PG 8-9 PG -7

18 Summary. 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.. In homogenate prepared from the whole fruit, the level of the antioxidant increased by -fold suggested that the peels contributed significantly to antioxidant activity.. 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.

19 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

20 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

21 תודה על ההקשבה

22 NF נטוראפוד מיץ אורגני מסחרי D מיץ רימונים נסחט במסחטה PJ מיץ מיקי מהול לשתייה M- תרכיז רימונים ממולי 7- מיץ טרי מגרגירים בלבד H- כנ"ל יין רימונים- יקב דלתון מיץ טבעי- מעמוס Antioxidant capacity (mm) Total Polyphenols (mm) D NF NF D PJ 7 רמת האנטיאוקסידנטיות במיצ י ם מסחריים U PJ U M 7 H spring D רמת הפוליפנולי ם במיצים מסח ריים M spring H D

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24 The identity of the hydrolysable-tannins obtained by using Multiple Reaction Monitoring (MRM) method of LC-MS MS-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. Q Q Q Single Ion m/z Collision Single daughter Ion m/z