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Migal, Kiryat Shmona

Migal, Kiryat Shmona. בירור הרכב החומרים מונעי החמצון בחלקים שונים של פרי הרימון ע"י סריקה של 29 זני 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.

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Migal, Kiryat Shmona

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  1. Migal, Kiryat Shmona בירור הרכב החומרים מונעי החמצון בחלקים שונים של פרי הרימון ע"י סריקה של 29 זני רימונים שונים יום עיון למגדלי רימונים, מכון וולקני, פברואר 2008 מוקדש לזכרו של משה זמירי ז"ל

  2. Atherosclerosis lesion development Thrombus formation Carotid Intima-Media Thickness Percent of mortality In western countries OTHERS 25% CHD 54% CANCER 21%

  3. Moreover: consumption of pomegranate juice can lead to reduction of the atherosclerosis lesion 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 Prof. Miki Aviram

  4. Pomegranate Juice Consumption by Healthy Volunteers Reduces LDL Oxidation A. 30 * * 20 Copper Ion-Induced LDL Oxidation (nmol TBARS/ mg LDL protein) 10 0 0 1 2 Time (weeks)

  5. A. Control 3 2 1 Carotid IMT (mm) 0 B. Pomegranate Juice 4 2 0 Baseline After 1 year Pomegranate Juice Consumption by Patients with Carotid Artery Stenosis Reduces Carotid Intima-Media Thickness (IMT)

  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. 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 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 C13 PG128-29 P.S. 21

  9. PG 104-5 PG 102-3 PG 100-1 PG 101-2 PG 103-4 PG 105-6 PG 109-10 PG 112-13 PG 106-7 PG 108-9 PG 118-19 PG 114-15 PG 116-17 PG 120-21 PG 119-20 - - PG 123-24 PG 127-28 PG 121-22 PG 128-29 PG 130-31 ERS C13 C14 C133 A17 XBS PS21 Camel PS23 2006

  10. 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

  11. Antioxidant activity Antioxidant activity, total polyphenols, total anthocyanins, and the level of hydrolysable tannins in aril juices Antioxidant capacity *10^3 (mg/L) 2 1 0 Total polyphenols 2 Total Polyphenols *10^3 (mg/L) R2 = 0.76** 1 0 3 Total anthocyanins *10^2 (mg/L) 2 Total anthocyanins R2 = 0.46** 1 Punicalagin Punicalagin m/z 1083 *10^2 (mg/L) R2 = 0.03 1.4 0.7 0 Punicalagin Punicalagin Punicalin 8 Punicalin m/z 781 (mg/L) R2 = 0.00 6 4 2 Gallagic acid 3 Gallagic acid m/z 601 *10 (mg/L) R2 = 0.01 2 1 Ellagic Ellagic acid acid Eve PG 105-6 PG 102-3 PG 103-4 Orange Gallagic acid Gallagic acid PG 101-2 PG 100-1 PG 108-9 PG 106-7 PG 104-5 PG 127-28 PG 121-22 PG 123-24 PG 130-31 PG 114-15 PG 120-21 PG 128-29 PG 118-19 PG 112-13 PG 116-17 PG 109-10 PG 201-212 PG 119-20 PG 200-211 PG 205-216 PG 204-215 PG 206-217 PG 203-214 PG 202-213 0 ** significant byPirson test, α=0.01

  12. pH level R2 = 0.07 4 Acid level pH level 3 R2 = 0.08 2 acid level (%) 2 1 0 Ascorbic acid level 20 16 R2 = 0.00 12 Ascorbic acid level (mg/L) 8 Citric acid level 4 0 2 Acetic acid level R2 = 0.05 0.8 Citric acid level (%) R2 = 0.48** 1 0.6 Acetic acid level (%) 0.4 0 0.2 5 6 1 9 4 2 7 3 31 28 17 10 29 22 15 13 24 21 20 13 21 14 19 17 23 0 - - - - - - - - 133 XBS ERS - - - - - - - - - - - - A C C Camel 104 101 105 100 106 102 108 103 PS PS C 130 127 116 109 128 121 118 114 112 123 120 119 PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG No positive correlation was found between the acid level and the content of organic acids in the aril juices Obtained by HPLC ** significant byPirson test, α=0.01

  13. 10 Antioxidant contents 8 Antioxidant capacity (mM) 6 4 2 0 %TSS 18 17 R2 = 0.68** TSS (%) 16 15 14 13 Glucose level 6 R2 = 0.58** Glucose level (%) 5 4 Fructose level 6 R2 = 0.58** Fructose level (%) 5 4 5 6 1 9 4 2 7 3 31 28 17 10 29 22 15 13 24 21 20 13 21 14 19 17 23 - - - - - - - - 133 XBS ERS - - - - - - - - - - - - A C C Camel 104 101 105 100 106 102 108 103 PS PS C 130 127 116 109 128 121 118 114 112 123 120 119 PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG Positive correlation was found between the TSS and the content of Glucose and fructose in the aril juices Obtained by HPLC ** significant byPirson test, α=0.01

  14. Antioxidant activity 1 Antioxidant capacity *10^4 (mg/L) Antioxidant activity, total polyphenols, total anthocyanins, and the inhibitory effect on LDL oxidation in juices prepared by juice extractor 0.5 0 Total polyphenol 6 Total Polyphenols *10^3 (mg/L) R2 = 0.61** 4 2 0 Total anthocyanin 4 Anthocyanin Pigment *10^2 (mg/L) R2 = 0.00 3 2 1 0 LDL oxidation inhibition 8 LDL Oxidation Inhibition *10 (%) Lipid peroxides, R2 = 0.71** 6 TBARS, R2 = 0.69** 4 2 0 Punicalagin 20 Punicalagin m/z 1083 *10^2 (mg/L) 18 R2 = 0.63** 6 4 2 0 Gallagic acid 40 Gallagic acid m/z 601 (mg/L) 36 R2 = 0.64** 8 4 0 Punicalin Punicalin m/z 781 *10^2 (mg/L) 6 R2 = 0.58** 4 2 0 Ellagic Acid Ellagic Acid m/z 301 (mg/L) 15 R2 = 0.68** 10 5 0 Eve Orange PG 100-1 PG 103-4 PG 102-3 PG 205-216 PG 106-7 PG 202-213 PG 108-9 PG 201-212 PG 204-215 PG 200-211 PG 123-24 PG 109-10 PG 128-29 PG 127-28 PG 118-19 PG 114-15 PG 130-31 PG 112-13 PG 120-21 PG 121-22 PG 203-214 ~ ~ ~ ~ The antioxidant was increased by 5 fold compare to juice prepared only from arils ** significant byPirson test, α=0.01

  15. 6 Antioxidant activity 5 Antioxidant capacity *10^4 (mg/L) 4 Antioxidant activity, total polyphenols, total anthocyanins, and the inhibitory effect on LDL oxidation in homogenates prepared from the whole fruit 3 2 1 0 Total polyphenol 1 Total Polyphenols *10^4 (mg/L) R2 = 0.90** 0.5 0 Total Anthocyanin 4 3 R2 = 0.05 Anthocyanin Pigment *10^2 (mg/L) 2 1 0 Punicalagin 3 R2 = 0.79** Punicalagin m/z 1083 *10^4 (mg/L) 2 1 0 Ellagic Acid 4 R2 = 0.44** Ellagic Acid m/z 301 *10^2 (mg/L) 3 2 1 0 Punicalin 1.2 R2 = 0.54** Punicalin m/z 781 *10^4 (mg/L) 0.6 0 Gallagic acid 8 Gallagic acid m/z 601 *10^3 (mg/L) 6 R2 = 0.55** 4 2 0 PJ Eve PG 105-6 PG 101-2 PG 103-4 PG 108-9 Orange PG 106-7 PG 118-19 PG 121-22 PG 127-28 PG 130-31 PG 109-10 PG 128-29 PG 112-13 PG 102-3 PG 116-17 PG 104-5 PG 205-216 PG 206-217 PG 100-1 PG 203-214 PG 120-21 PG 123-24 PG 204-215 PG 200-211 PG 119-20 PG 114-15 PG 201-212 PG 202-213 The antioxidant was increased by 20 fold compare to juice prepared only from arils ** significant byPirson test, α=0.01

  16. Antioxidant activity 1 Antioxidant capacity *10^5 (mg/L) 0.5 Antioxidant activity, total polyphenols, total anthocyanins, and the inhibitory effect on LDL oxidation in homogenates prepared from the peels 0 Total Polyphenol Total Polyphenols *10^4 (mg/L) 1.5 R2 = 0.91** 1 0.5 0 Total anthocyanin 8 Anthocyanin Pigment *10^2 (mg/L) 6 R2 = 0.01 4 2 Eve Orange PG 108-9 0 PG 101-2 PG 105-6 PG 100-1 PG 106-7 PG 102-3 PG 103-4 PG 127-28 PG 104-5 PG 109-10 PG 123-24 PG 121-22 PG 119-20 PG 128-29 PG 112-13 PG 206-217 PG 120-21 PG 130-31 PG 201-212 PG 114-15 PG 116-17 PG 205-216 PG 204-215 PG 203-214 PG 200-211 PG 118-19 PG 202-213 Punicalagin The antioxidant was increased by 40 fold compare to juice prepared only from arils 4 Punicalagin m/z 1083 *10^5 (mg/L) R2 = 0.39** 3 2 1 0 Ellagic Acid 8 Ellagic Acid m/z 301 *10^2 (mg/L) R2 = 0.49** 6 4 2 0 Punicalin Punicalin m/z 781 *10^4 (mg/L) R2 = 0.72** 1 0 Gallagic acid Gallagic acid m/z 601 *10^3 (mg/L) 8 R2 = 0.89** 6 4 2 0 ** significant byPirson test, α=0.01

  17. Antioxidant contents 4 3 Antioxidant capacity *10^2 (mM) 2 1 0 Fruit weight 6 R2 = 0.01 Fruit weight *10^2 (gr) 4 2 0 Peel weight 3 R2 = 0.01 Peel weight *10^2 (gr) 2 1 0 Aril weight 4 R2 = 0.06 Aril weight *10^2 (gr) 3 7 3 2 1 9 6 5 4 24 21 22 29 28 10 17 20 31 13 13 19 15 21 17 23 - - - - - - - - 2 Eve XBS ERS - - - - - - - - - - - - A C C 106 102 101 100 108 105 104 103 PS PS C 128 114 123 121 127 109 116 120 119 130 112 118 PG 203-214 PG 206-207 1 PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG PG 0 The antioxidant level is not correlated to the % of the peels from the fruits

  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. 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.

  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. רמת האנטיאוקסידנטיות במיצים מסחריים 80 70 60 50 Antioxidant capacity (mM) 40 30 20 10 0 H M U 2 D 71 PJ NF D 4 spring רמת הפוליפנולים במיצים מסחריים 10 8 6 Total Polyphenols (mM) 4 2 0 H M U 2 D 71 PJ NF D 4 spring NF – נטוראפוד מיץ אורגני מסחרי D2 מיץ רימונים נסחט במסחטה PJ מיץ מיקי מהול לשתייה M- תרכיז רימונים ממולי 71- מיץ טרי מגרגירים בלבד H- כנ"ל יין רימונים- יקב דלתון מיץ טבעי- מעמוס

  23. 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 Single Ion m/z Q3 Single daughter Ion m/z Q2 Collision

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