Title Implementation of valuable compounds from olive mill wastewater as additives in functional foods & cosmetics

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BioOlea Conference, Corfu 21 February 2014 Phenoliv AB Spin-off Company Lund, Sweden Title Implementation of valuable compounds from olive mill wastewater as additives in functional foods & cosmetics Dr. Charis M. Galanakis Chemist

Food Wastes - Byproducts For more than 40 years were considered as substrates for treatment, minimization & prevention In European legislations (442/1975/EEC, 689/1991/EEC) are defined as "wastes" due to the fact that they are removed from production process as undesirable materials Today the term food by-products is used within the food industry Prospect for the recovery & recycling of high added-value compounds within food chain via new products 2

Olive mill wastewater 3 Phase-olive oil production Olive oil [Ι] Olive kernel oil [ΙI] High pollution index: Wastewater [ΙII] COD 220 g/l BOD 100 g/l ph 3.0-5.9 Solids 80 g/l Phenols 20 g/l 3

Components of Olive Mill Wastewater 3-phase mixture (a) Water (b) Oil Typical content (c) Solids (a) 83-94% water (b) 4-16% organic (c) 0.4-2.5% inorganic Salts Phosphoric compounds Sugars Volatile acids Nitrogenous compounds Fats Phenols (hydroxytyrosol) Dietary fibers (pectin) 4

5-Stage Universal Recovery Processing Goals Yield maximization Adaption Clarification of valuable compounds Ensuring limited diminution of functional properties Ensuring edibility of the final product Ensuring sustainability of the process I. Macroscopic Pre-treatment Wet milling, thermal &/or vacuum concentration mechanical pressing, electro-osmotic dewatering II. Macro- & Micro- molecules Separation Alcohol precipitation, ultrafiltration, isoelectric solubilization - precipitation, III. Extraction Solvent, acid, alkali, microwave- assisted, ultrasonics, pervaporation IV. Isolation & Purification Nanofiltration, adsorption chromatography V. Product Formation Spray drying, emulsions Galanakis C. M. (2012). Recovery of high added-value compounds from food wastes: Conventional, emerging technologies and commercialized applications. Trends in Food Science & Technology, 26(2), 68-87. Galanakis, C.M. (Eds). Food Waste Recovery: Processing Technologies & Techniques. Elsevier, ISBN: 9780128003510. Expected publication: 2015 5

Patented Methodology Olive mill wastewater Macroscopic pretreatment 1. Centrifugation 2. Condensation (2-3 h, 60-80 o C) Macro- & Micro-molecules separation Extraction 4. Alcoholic precipitation (85% EtOH + 10 min boiling) Ethanolic extract Polyphenols Purification - Isolation 5. Membrane Technology Phenoliv 3. Extraction (EtOH +/- acids) Encapsulation Insoluble residue Pectin Pectinoliv 6. Spray drying Condensed wastewater Water & fats removal Pectin solubilization Oleuropein conversion to hydroxytyrosol Dietary fiber precipitation (pectin) Clarification pectin from ions & phenols phenols from higher MW compounds Tornberg & Galanakis (2008). Olive waste recovery. WO2008/082343 6

Process Originality A simple & physical methodology for the recovery of dietary fibers & polyphenols in 2 products from OMW Fiber extraction prior precipitation A part of the ethanol is used in both stages Citric acid extraction converts oleuropein to hydroxytyrosol Lower quantity of extract is required for the health claims Application of the phenol rich ethanolic extract directly as additive to drinks Preservation of polyphenols 7

Competitive Advantages Zero or negative cost of the source EtOH recycling via condensation of the extract a) Green b) Sustainable process c) Cheap Full conversion of wastewater to high added-value products Generation of a relative crude product containing a mixture of polyphenols Development of tailor-made applications as food additives 8

Phenoliv Pilot Plan Condensation Electric must condenser Pneumatikakis winery, Chania, Greece 2 tn olive mill wastewater were condensed up to 280 Kg (26-30 o C) for 15 h Extraction Condensed waste were mixed for 1 h with 96% EtOH Insoluble residue removed with a decanter, EtOH removed with a vacuum condenser SwePharm, Lund Spray Drying The extract was mixed with maltodextrin solution (coating material) Dryer Anhydro Lab S3 (35 Kg water/h) Karlshamn, Sweden 30 Kg powder containing 4% phenols 9

Application Studies Lundolive Meat products preservation Pectinolive Fat replacement in meatballs 100 mg polyphenols/l improved the red colour of meat within 72 h of preservation Pectinolive restricted oil uptake during deep fat frying of the product bl T1 T2 OL 50 OL 75 OL 100 Galanakis, C.M., Barbier, C., & Tornberg, E. (2011). 11 th Int. Congress on Engineering & Food, 22-26 May, Athens, Greece. Galanakis, C. M., Tornberg, E., & Gekas, V. (2010). LWT- Food Science and Technology, 43(7), 1018-1025. 10

Applications under investigation Chips Sparkling water & drinks Lundolive Meat products Oils Bakery products Cosmetics ΜΕΠΑΕ(2013-2015): A study for the implementation of olive mill waste-polyphenols in foods & cosmetics 11

g ABTS/g active compound g DPPH/g active compound Antioxidant Activity of Olive Polyphenols (PP) (a) 160 140 120 100 80 60 40 20 0 Polyphenols 1 Polyphenols 2 Tocopherols α-tocopherol Ascorbic acid (b) 18 16 14 12 10 8 6 4 2 0 Polyphenols 1 Polyphenols 2 Tocopherols α-tocopherol Ascorbic acid ABTS & DPPH assays PP possess higher antioxidant activity per g active compound compared to tocopherols mixture, α- tocopherol & ascorbic acid (unpublished results) 12

Addition of Antioxidants in Olive Oils Peroxide values of olive oil samples prior & after their thermal oxidation Polyphenols reduced oil oxidation even at 500 mg/l compared to ascorbic acid (effective at 1000 mg/l) & tocopherols (not effective) (unpublished results) 13

Polyphenols as active compounds in cosmetics Sample Peaks in nm [Absorption] Benzophenone-3TXT (0.001%) 241.0 [0.5159] 286.5 [0.7377] 323.0 [0.4736] Lundolive2 (0.001%) 279.0 [0.0196] TiO 2 (0.01%) 278.0 [1.6793] 309.0 [1.7326] 328.5 [1.9191] 350.0 [1.3392] TiO 2 (0.01%) Lundolive2 (0.01%) 279.5 [2.6747] 309.0 [1.7439] 326.0 [1.8510] 350.0 [1.2202] TiO 2 (0.01%) Lundolive1 (0.01%) 275.0 [1.7089] 309.0 [1.4843] 325.0 [1.4956] 350.0 [1.2327] UVA (315-400 nm), UVB (280-315 nm) Polyphenols could be effective as SPF-boosters with TiO 2 (unpublished results) 14

Products in the market Natural preservative in Swedish chocolates One bar of chocolate contains the daily recommended quantity of polyphenols (5 mg) Corresponds to 20 g extra virgin olive oil Health claims approval from the EFSA Polyphenols from olive oil have an antioxidant activity that may help maintain healthy LDL cholesterol level and lipid peroxidation 15

Forthcoming products in the market Addition of Lundolive in vegetarian smoothies rich in ω-3-fatty acids (α-linolenic acid) health claims for both polyphenols & ω-3-fatty acids Protection against ω-3-fatty acids oxidation 16

Polyphenols market 1 billion euro annual turnover in Europe (2010), corresponding to 2,200 tn of polyphenols The market of olive polyphenols is growing (2% per total, meaning 30 tn and revenue > 20 million dollars) Polyphenols cost (100%) comes up to ~4000 /Kg Lundolive (10%) price is today 200 /Kg & the company's goal is to gradually reduce it to 100 & 40 / Kg after developing a full scale production line 17

Contribution to the Sustainable Development & Extroversion The production of functional components in combination with a process that decontaminate environment from hazardous waste Get rid of OMW with no charge (1,500,000 tn OMW can generate 30,000 tn Lundolive/year) Promote extroversion through partnerships between Greek & Scandinavian food companies (i.e. mills, meat, chocolate-industries) Promoting of the beneficial effects of olives, olive oil & Greek diet relying on the competitive advantages of innovative products 18

www.phenoliv.com 19

Thank You for Your Attention! Information: chemlab.gr phenoliv.com charisgalanakis.com https://www.iseki-food.net/sigs/sig5 Linkedin: Galanakis Laboratories Phenoliv AB Charis Galanakis Food waste recovery 20