K. Kaderides, L. Papaoikonomou, I. Patsopoulou, L. Sorovakou, A.M. Goula

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1 K. Kaderides, L. Papaoikonomou, I. Patsopoulou, L. Sorovakou, A.M. Goula Department of Food Science and Technology, School of Agriculture, Forestry and Natural Environment, Aristotle University, Thessaloniki, Greece

2 Pomegranate is an ancient fruit originating from the Middle East and nowadays the global pomegranate production is around 2 million tons The most important growing regions are: China Iran Egypt Turkey Spain U.S.A. 24% Peel 14% Seeds 62% Juice

3 Component Content (%) Total solids 96. Moisture 4. Total sugars Proteins 8.72 Crude Fiber 21.6 Fat 9.4 Ash 5. Total phenolics 8.1 Phenolic compound Aguilar et al., 28; Ullah et al., 212 Antioxidant activity Anti-mutagenic activity Anti-hypertension activity Anti-inflammatory activity Anti-atherosclerotic activity Content (mg/1 g dry matter) Ellagic acid Catechin Punicalagin Gallic acid Protocatechol 4.17 Vanilline 3.91 Caffeic acid 6.46 Ferulic acid 5.89 p-coumaric acid Others 8.2 Rowayshed et al., 213; Gullon et al., 216

4 Pomegranate Peels Fodder Food Industry Ice cream Tea 24 kg Cosmetics 1 kg Seeds 14 kg

Pomegranate Peels Drying Solvent Grinding

5 Pomegranate Peels Drying Solvent Grinding Ultrasound-assisted extraction Fodder Filtration Recycled solvent Evaporation Wall material Drying Phenolics Emulsification Encapsulation by spray drying Food additives Microcapsules of phenolics Kaderides et al., 215

6 Increase of their stability during storage and passage through the gastrointestinal tract Improvement of color Masking of astringency Suitability for use as an additive in functional foods Fang & Bhandari, 21

Method of encapsulation Extrusion Encapsulation efficiency 89.

emulsion using a rotating disk reactor Freeze drying 79.

, 214 75.5% 97.22% 63.19% da Rosa et al., 214 Saikia et al.

7 Method of encapsulation Extrusion Encapsulation efficiency 89.39% Rapid extraction of supercritical solution Formation of multiple emulsion using a rotating disk reactor Freeze drying 79.78% Belščak-Cvitanovic et al., 211 Santos et al., % Akhtar et al., % 97.22% 63.19% da Rosa et al., 214 Saikia et al., 215 Marin et al., % 99.8% Bustamante et al., 217 Kaderides et al., 215 Freeze drying liposomes Spray drying Reference

1 Good rheological properties at high concentration 2 Disperse or emulsify the active material and stabilize the emulsion produced 3 Chemical non reactivity with the active core materials 4 Seal and

8 1 Good rheological properties at high concentration 2 Disperse or emulsify the active material and stabilize the emulsion produced 3 Chemical non reactivity with the active core materials 4 Seal and hold the active material within its structure during processing/storage 5 Provide maximum protection to the active material against environmental conditions 6 Acceptable solubility of the solvent to the food industry Desai & Park, 25

Phenolic extract Wall material References Spray drying Pomegranate peel extract Maltodextrin; Whey protein; Skim milk powder Maltodextrin Modified starch Kaderides et al., 215 Cam et al.

9 Phenolic extract Wall material References Spray drying Pomegranate peel extract Maltodextrin; Whey protein; Skim milk powder Maltodextrin Modified starch Kaderides et al., 215 Cam et al., 214 Bustamante et al., 217 Carrot extract Maltodextrin Ersus &Yurdagel, 28 Grape juice Maltodextrin; Soy protein; Whey protein Moser et al., 217 Olive leaf extract Chitosan Kosaraju et al., 26 Barberry extract Maltodextrin; Gum Arabic; Gelatin Mahdavi et al., 216 Mutle plant extract Maltodextrin; Gum Arabic; Mesquite gum Pavón-García et al., 211 Soybean extract Maltodextrin; Starch Georgetti et al., 28 Βlack currant extract Maltodextrin; Inulin Bakowska & Kolodziejczyk, 211 Bilberry extract Whey protein concentrate Betz et al., 212 Apple extract; Olive leaf extract Sodium caseinate; Lecithin Kosaraju et al., 28 Co-crystallization Yerba Mate extract Calcium alginate Deladino et al., 28 Green tea EGCG extract Gelatin Shutava et al., 29 Blackcurrant extract Glucan Xiong et al., 26 Freeze drying Cloudberry extract Maltodextrin DE 5-8 & DE18,5 Laine et al., 28 Grape pomace extract Maltodextrin; Gum Arabic Stoll et al., 216 Blueberries extract Maltodextrin Celli et al., 216 Hibiscus tea extract Pullulan Gradinaru et al., 23

Carbohydrates Wall material with appropriate encapsulation

Replacement with alternative products Insoluble dietary

Soluble dietary fibres Assist regular bowel movements and

Insoluble dietary fibres Slow down the metabolism rates of

10 Carbohydrates Wall material with appropriate encapsulation properties Undesired taste Unnatural additives Proteins Replacement with alternative products Insoluble dietary fibres Complex carbohydrates Strong absorption properties Soluble dietary fibres Assist regular bowel movements and flushing the intestinal system of undesirable materials Insoluble dietary fibres Slow down the metabolism rates of sugars and form a lining gel within the intestines Natural dietary fibres

11 Production of oranges in E.U. Country Tons (213) Spain Italy Greece 914. Portugal 26.3 Highly polluted wastewater 5% by-product (peel, seed, pulp) 5% juice Wastes Food industry (source of dietary fiber) Animal feed Fertilizer No economic value

12 Product Activity Reference Encapsulated pomegranate peel extract Hazelnut paste Antioxidant Kaderides et al., 215 Ice cream Antioxidant Cam et al., 214 Pomegranate peel extract Shrimps Antimicrobial Basiri et al., 215 Meat pate Antimicrobial Hayrapetyan et al., 212 Curd Antioxidant Sandhya et al., 218 Pork meat Antioxidant Qin et al., 213 Beef meatballs Antioxidant Turgut et al., 217 Bread Antioxidant Paari et al., 212; Altunkaya et al., 213 Sunflower oil Antioxidant & Antimicrobial Cooked chicken Antioxidant & patties Antimicrobial Iqbal et al., 28; Kanatt et al., 21 Naveena et al., 28

13 Product Activity Reference Encapsulated pomegranate peel extract Hazelnut paste Antioxidant Kaderides et al., 215 Ice cream Antioxidant Cam et al., 214 Pomegranate peel extract Shrimps Antimicrobial Basiri et al., 215 Meat pate Antimicrobial Hayrapetyan et al., 212 Curd Antioxidant Sandhya et al., 218 Pork meat Antioxidant Qin et al., 213 Beef meatballs Antioxidant Turgut et al., 217 Bread Antioxidant Paari et al., 212; Altunkaya et al., 213 Sunflower oil Antioxidant & Antimicrobial Cooked chicken Antioxidant & patties Antimicrobial Iqbal et al., 28; Kanatt et al., 21 Naveena et al., 28

The exploitation of pomegranate and orange wastes based on: Ultrasound-assisted extraction of phenolic compounds from pomegranate peel Encapsulation of extract by

14 The exploitation of pomegranate and orange wastes based on: Ultrasound-assisted extraction of phenolic compounds from pomegranate peel Encapsulation of extract by spray drying using orange juice industry by-product as wall material Study of: Incorporation of crude and encapsulated extract in foods: a. Cake b. Smoothie c. Sunflower oil

15 Orange Juice production Juice Orange wastes Hot water 1 C Boiling 2 min Washing Drying 6 C, 48 h Grinding Sieving Orange wastes powder

16 1. Extraction temperature: 35 C 2. Solvent type: Water 3. Solvent/Solid ratio: 32/1 4. Amplitude level: 4% (5 W) 5. Pulse duration/pulse interval ratio: 7/6 6. Extraction time: 1 min Kaderides et al., W, 2 khz VCX 13 Sonics and Materials (Danbury, CT, USA) with Ti Al V probe (13 mm)

17 Pomegranate Peel Washing Drying 4 oc, 48 h Solvent Filtration Ultrasoundassisted extraction Grinding Wall material (Orange wastes powder) Evaporation Emulsification Encapsulation by spray drying

18 1. Wall material: Orange wastes 2. Inlet air temperature: 162 C 3. Feed solids concentration: 5 % w/w 4. Ratio of core to wall material: 1/9 5. Drying air flow rate: 17.5 m3/h 6. Flow rate of compressed air for atomization:.8 m3/h Concurrent Two fluid nozzle atomization Peristaltic pump for feed Buchi, B-191, Buchi Laboratoriums-Technik, Flawil, Switzerland

19 PRODUCT: a. Cake Shelf-life test at 25 C for 18 days b. Smoothie Shelf-life test at 4 C for 12 days c. Sunflower oil Shelf-life test at 6 C for 2 days ADDITIVE: Phenolic extract (Crude and encapsulated) 3 samples were tested: Measurement of: Sample with encapsulated extract Antioxidant activity Sample with crude extract Total phenolics content Control sample Oxidation stability

20 Shelf-life test at 25 C for 18 days Shelf-life test at 4 C for 12 days Cake Smoothie Phenolics concentration: 5 ppm Phenolics concentration: 5 ppm Eggs 26. % Pineapple 42.8 % Sugar 22.9 % Banana 4.8 % Margarine 19.1 % Water 14.1 % Flour 19.1 % Spearmint 1.4 % Milk 13. % Cilantro.8 % Phenolics concentration: 5 ppm Sunflower oil Shelf-life test at 6 C for 2 days Phenolics concentration: 1 ppm

21 Results

85 mg GAE/g DM Optimum conditions for maximum encapsulation yield Wall

22 Optimum conditions for maximum extraction yield Solvent T (oc) A (%) t (min) Pulse (sec/sec) L/P Water /6 32/1 Yield: mg GAE/g DM Optimum conditions for maximum encapsulation yield Wall material T (oc) Orange wastes powder 162 Qa (%) Qs (m3/h) 5.8 w/c 9/1 Yield: 12.96%

23 Crude extract mg Punicalagin/g dry peel Standard Peak: 1. Punicalagin B 2. Punicalagin A

24 Crude extract Standard mg punicalagin/g dry peel Peak: 1. Punicalagin B 2. Punicalagin A Kharchoufi et al., 218 Ultrasound Ass. Extraction Lu et al., 28 Zivkovic et al., 218 Conventional Extraction Gullon et al., 216 Cam & Hisil, 21 Pressurised Water Extraction

25 Day Day 7 Day 14 Time (day) Punicalagin Scavenging Total (mg/g dry capacity Phenolics peel) (%) (mg GAE/g dry peel) Day 21

26 Day Day 7 Day 14 Time (day) Punicalagin Scavenging Total (mg/g dry capacity Phenolics peel) (%) (mg GAE/g dry peel) Day 21 Gil et al., 2

27 Shelf-life test at 6 C for 2 days Folin Ciocalteu Method Oil (1 ppm),5,45 Total Phenolic content (mg GAE/g oil) Total Phenolic content (mg GAE/g oil) Folin Ciocalteu Method Oil (5 ppm) Encapsulated TPC,4 Blank,35 Crude TPC,3 p-value =.912,25,2,15,1, p-value =.86,9,8,7,6,5 Encapsulated TPC,4 Blank,3 Crude TPC,2,1 2 5 Time (days) p-value =.14 1 Encapsulated TPC 6 Crude TPC 5 Blank 9 Blank DPPH Method Oil (1 ppm) Scavenging capacity (%) Scavenging capacity (%) 9 15 Time (days) DPPH Method Oil (5 ppm) Encapsulated TPC 8 p-value =.1 7 Crude TPC Time (days) Time (days) 15 2

2 18 Blank 16 14 Encapsulated TPC 12 Crude TPC 1 Shelf-life test at 6 C for 2 days p-value

PV (meq O2/kg oil) Peroxide value, PV (meq O2/kg oil) Peroxide Value Oil (5 ppm) 2 18 Blank

28 2 18 Blank Encapsulated TPC 12 Crude TPC 1 Shelf-life test at 6 C for 2 days p-value = Time (days) Peroxide Value Oil (1 ppm) Peroxide value, PV (meq O2/kg oil) Peroxide value, PV (meq O2/kg oil) Peroxide Value Oil (5 ppm) 2 18 Blank 16 Encapsulated TPC 14 Crude TPC 12 p-value = Time (days)

Cake (5ppm) 3 Blank Encapsulated TPC 2,5 Crude TPC 2 p-value =.

29 Folin Ciocalteu Method Cake (5ppm) 1 9 Scavenging capacity (%) 8 Blank p-value =.1 7 Encapsulated TPC Crude TPC Total phenolic content (mg GAE/g Cake) DPPH Method Cake (5ppm) 3 Blank Encapsulated TPC 2,5 Crude TPC 2 p-value =.4 1,5 1, Time (days) ANTIOXIDANT ACTIVITY Time (days) TOTAL PHENOLICS CONTENT Shelf-life test at 25 C for 18 days 2

Peroxide value, PV (meq O2/kg Cake extract) Peroxide Value Cake (5ppm) 25 2 15

30 Peroxide value, PV (meq O2/kg Cake extract) Peroxide Value Cake (5ppm) p-value = Time (days) Shelf-life test at 25 C for 18 days

DPPH Method Smoothie (5ppm) 1 8 Blank p-value =.

(5ppm) 4 2 2 4 6 8 1 12 Time (days) ANTIOXIDANT ACTIVITY

GAE/g Smoothie) Scavenging capacity (%) 12 1,8 1,6 1,4 1,2 Blank

31 DPPH Method Smoothie (5ppm) 1 8 Blank p-value =.22 6 Encapsulated TPC Crude TPC Folin Ciocalteu Method Smoothie (5ppm) Time (days) ANTIOXIDANT ACTIVITY Shelf-life test at 4 C for 12 days 14 Total phenlic content (mg GAE/g Smoothie) Scavenging capacity (%) 12 1,8 1,6 1,4 1,2 Blank p-value =.42 1 Encapsulated TPC,8 Crude TPC,6,4, Time (days) TOTAL PHENOLICS CONTENT 14

32 An integrated approach for utilization of pomegranate peels is suggested based on the spray drying encapsulation of their phenolics compounds using an alternative wall material (orange wastes) Pomegranate peel extract was found to be rich in punicalagin The extract (crude and encapsulated) was found efficient in improving the shelf-life of sunflower oil, cake and smoothie The extract changed the texture and the color of cake significantly

33 Thank you! This work is supported by the General Secretariat for Research and Technology (GSRT) and the Hellenic Foundation for Research and Innovation (HFRI)

K. Kaderides, E.M. Kapantai, A.M. Goula

K. Kaderides, E.M. Kapantai, A.M. Goula Department of Food Science and Technology, School of Agriculture, Forestry and Natural Environment, Aristotle University, Thessaloniki, Greece Pomegranate is an