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Factorial design of phenolic extraction process from two phase olive mill waste K. Tzathas, A. Vlysidis, G. Lyberatos, A. Vlyssides School of Chemical Engineering National Technical University of Athens 6 th International Conference on

  1. Factorial design of phenolic extraction process from two phase olive mill waste K. Tzathas, A. Vlysidis, G. Lyberatos, A. Vlyssides School of Chemical Engineering National Technical University of Athens 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  2. Scope of the study Scope of the study  Valorisation of the OMW from II-phase olive mills  Recovery of high added value compounds  Residual Oil  Phenolic compounds  Decrease the environmental impact of this primary industrial field in Greece and in Mediterranean countries  Necessary pretreatment steps to diminish inhibition phenomena to the biological processes that follow  Anaerobic digestion and/or composting  Increase their sustainability  Developing novel processes leading to a range of added value products  Under the concept of zero waste biorefineries 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  3. INTRODUCTION 0 χλμ 500 More than 95% of the global production derives NATIONAL GEOGRAPHIC MAPS FRANCE from the Mediterranean area (600 BC) SPAIN (200 BC) ITALY TURKEY (600 BC.) GREECE (4000 BC) (1500 BC.) SYRIA Sicilia (4000 BC.) (600 BC) ALGERIA CYPRUS (600 BC) Crete MAROCO (2000 BC.) (2500 BC) (600 BC) ISRAEL (600 π . Χ .) (4000 BC.) JORDAN TUNISIA LYBIA (600 π . Χ .) EGYPT (1500 π . Χ ..) SPAIN ITALY CREECE TURKEY TUNISIA SYRIA MAIN OLIVE OIL PRODUCERS (in thousands tons per year) 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  4. SYSTEMS FOR OLIVE OIL EXTRACTION The image part with relationship ID rId3 was not found in the file. Pressing system Three-phase system Two-phase system Olives Olives Olives (1000 kg) (1000 kg) (1000 kg) Cold Cold Washing Cold Washing Washing Washing Washing Washing water water water wastewater wastewater wastewater Hot Hot Crushing and malaxing Crushing and malaxing Crushing and malaxing water water solid phase solid phase POMACE Centrifugation Centrifugation Pressing Wooden POMACE (550 kg) (Three-phase (Two-phase Screening (550 kg) Stones 55% moisture liquid decander) decander) 55% moisture phase liquid oily phase phase Centrifugation Washing Washing Oil Oil oily water water washing washing phase Washing Oil sludge phase water washing POMACE Olive oil Olive oil OMWW OMWW (800 kg) (200 kg) (0.2 tn) (210 kg) (1-1.6 tn) 75% moisture Olive oil OMWW (0.9 tn) (200 kg) 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  5. The Greek situation regarding Greek situation Greek situation the olive oil and OMW Greek Oliv Greek Olive Mills per e Mills percentage entage distribution distribution • In Greece there are 150 M olive trees cultivated East Macedonia and Thrace Central Macedonia in ~765.000 hectares West Macedonia • The annual production of each tree rises up to Thessaly 300 kg of olives North Aegean Central Greece • The 1/3 of Greek farmers are working on Ionian West Greece cultivation of olives Attica • Olives and olive oil production in Greece rise up Peloponnese 1.750.000 t and 400.000 t, respectively -There are around 2500 olive oil mills in Greece South Aegean - 2100 centrifugal systems (most of them III-phases) Crete - Replaced from II-phases -There are 20 pomace processing plants 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  6. Com Composition of II-phase OMW osition of II-phase OMW  Moisture: 67.5%  COD: 30 kg/m 3  Oil content: 10% (db)  TPC: 30.5 mg/g db 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  7. Experimental Me Experimental Methodology f thodology for e r extracting the tracting the phenolic com phenolic compounds ounds  Acid hydrolysis process  Lab scale experiments with initial amount of OMW 100 g (67.5% of moisture)  Design of a 2 4 factorial experiment in order to measure the effect of four important process parameters  Dilution of OMW (X1)  Quantity of a strong acid (X2)  Hydrolysis Time (X3)  Temperature (X4) At the end of each run the aqueous phase was Dilution H 2 SO 4 % Time (min) Temperature Level ( o C) separated and analyzed (v/w) (v/w) -TPC (Folin method) 1 3 3 60 70 - Phenolic compounds (HPLC) 0 2 2 45 60 Hydroxytyrosol - Tyrosol - Oleuropein -1 1 1 30 50 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  8. Experimental R Experimental Results sults HYDROXY- Χ 1 Χ 2 Χ 3 Χ 4 TYROSOL OLEUROPEIN TYROSOL Run TPC (mg/g) Strong Acid Dilution (mL) Time (min) Temperature (oC) mg/g Dry mg/g Dry mg/g Dry (mL) 1 100 (-1) 1 (-1) 30 (-1) 50 (-1) 5.12 3.61 1.24 19.33 2 100 (-1) 1 (-1) 30 (-1) 70 (+1) 5.93 3.94 1.27 20.61 3 100 (-1) 1 (-1) 60 (+1) 50 (-1) 5.82 3.76 1.36 19.97 4 100 (-1) 1 (-1) 60 (+1) 70 (+1) 6.01 3.56 1.15 20.59 5 100 (-1) 3 (+1) 30 (-1) 50 (-1) 6.34 3.29 1.16 19.31 6.26 3.74 1.33 20.29 6 100 (-1) 3 (+1) 30 (-1) 70 (+1) 7 100 (-1) 3 (+1) 60 (+1) 50 (-1) 6.74 3.81 1.42 20.10 8 100 (-1) 3 (+1) 60 (+1) 70 (+1) 6.83 3.99 1.33 21.90 9 300 (+1) 1 (-1) 30 (-1) 50 (-1) 5.40 3.92 1.38 21.02 10 300 (+1) 1 (-1) 30 (-1) 70 (+1) 5.14 3.86 1.37 20.46 11 300 (+1) 1 (-1) 60 (+1) 50 (-1) 5.24 4.30 1.27 22.99 12 300 (+1) 1 (-1) 60 (+1) 70 (+1) 6.01 3.95 1.22 21.17 13 300 (+1) 3 (+1) 30 (-1) 50 (-1) 6.27 3.61 1.41 22.52 6.25 3.47 1.34 21.25 14 300 (+1) 3 (+1) 30 (-1) 70 (+1) 15 300 (+1) 3 (+1) 60 (+1) 50 (-1) 6.64 4.85 1.64 25.05 16 300 (+1) 3 (+1) 60 (+1) 70 (+1) 6.41 5.14 1.73 27.73 17 200 (0) 2 (0) 45 (0) 60 (0) 6.04 3.89 1.36 21.28 6.08 3.76 1.41 22.27 18 200 (0) 2 (0) 45 (0) 60 (0) 6.00 3.92 1.37 20.90 19 200 (0) 2 (0) 45 (0) 60 (0) 20 200 (0) 2 (0) 45 (0) 60 (0) 6.07 3.93 1.30 21.62 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  9. Modelling Results from the Factorial Design  Regarding the TPC, the significance of the model was adequate  R 2 between the experimental and model predictions is 0.91  Important parameters are the dilution, the hydrolysis time and the acid addition Υ TPC = β 0 + β 1 Χ 1 + β 2 Χ 2 + β 3 Χ 3 + β 4 Χ 4 + β 5 Χ 1 Χ 2 + β 6 Χ 1 Χ 3 + β 7 Χ 2 Parameter Estimate t Ratio β 0 21.52 Χ 3 + β 8 Χ 1 Χ 4 + β 9 Χ 2 Χ 4 + β 10 Χ 3 Χ 4 β 1 1 . 255 5.74 β 2 0 . 75 3.43 β 3 0 . 91875 4.2 β 4 0 . 23125 1.06 Experimental values β 5 0 . 6125 2.8 and model prediction β 6 0 . 54125 2.48 for TPC (mg/g) β 7 0 . 50625 2.32 β 8 ‐0 . 35375 -1.62 β 9 0, . 9125 1.33 β 10 0 . 1775 0.81 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  10. Central composite design for the most important parameters •Finally, a central composite design was implemented 3 2 examining the most important parameters of the factorial design • Dilution of OMW (X1) • Addition of strong acid (X2) • Hydrolysis time (X3) was decided to be examined separately as a kinetic study on the optimum conditions. • Both hydrolysis time (X3) and temperature (X4) was set at level +1 of the FD experiment Level Η 2 Ο (mL) H2SO4 (mL) 1.414 350 3.5 1 335.36 3.35 0 300 3 -1 264.64 2.65 - 1.414 250 2.5 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  11. Experimental and modelling results of the CCD Η 2 Ο (mL) H2SO4 (mL) TPC OLEUROPEIN HYDROXYTY TYROSOL Χ 1 Χ 2 (mg/g) (mg/g) ROSOL (mg/g) (mg/g) 1 1 15.8 1.12 5.55 3.67 1 -1 21.2 1.46 4.68 4.53 -1 1 22.3 1.27 5.8 4.52 -1 -1 17.81 1.04 6.1 2.99 1.4142 0 19 1.25 5.15 3.58 -1.4142 0 21.1 1.37 6.1 3.94 0 1.4142 24.3 1.64 6.7 4.71 0 -1.4142 23.33 1.46 5.4 4.19 0 0 21.22 1.22 5.98 3.51 0 0 20.74 1.24 5.86 3.58 Best model predictions are acquired for hydroxytyrosol output Experimental and Model predictions for the hydroxytyrosol (mg/g) (R 2 =0.86) Parameter Estimate t Ratio Υ hydroxytyrosol = β 0 + β 1 Χ 1 + β 2 Χ 2 + β 3 Χ 1 Χ 2 + β 4 Χ 1 Χ 1 + β 5 Χ 2 Χ 2 β 0 5 .895 β 1 ‐0.3767 -4.03 β 2 0 . 3011 3.22 β 3 0 . 2925 2.21 β 4 - 0 .2113 -2.02 β 5 0 .00125 0.01 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  12. Optimum extraction conditions and validation on the optimum value Optimum conditions For X1 was 0.104 (304 mL) and For X2 was 1.414 (3.5 mL) Y hydroxytyrosol = 6.4 mg/g HYDROXYTYROSOL TYROSOL OLEUROPEIN TPC (mg/g) mg/g Dry mg/g Dry mg/g Dry 6.51 4.93 1.40 24.82 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

  13. Reco covery of phenolic com of phenolic compounds fr ounds from om treat treated OMW d OMW Liquid aqueous phase Micro & ultrafiltration Ion-Exchange from centrifugation Evaporation Resins 4-6 g/L of phenolic High Concentrated compounds ~ 80 g/L of Extraction rich in 3-4% of sugars phenolic Phenolic Compounds compounds 6 th International Conference on Sustainable Solid Waste Management, Naxos Island, Greece, 13–16 June 2018

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