assessment and dissemination of strategies for the extraction of area/booklets/booklet olive... ·...

Assessment and dissemination of strategies for the extraction ofbioactive compounds from tomato, olive and grape processing residues

(BIOACTIVE-NET)

“HANDBOOK on BIOACTIVE COMPOUNDSfrom OLIVE PROCESSING RESIDUES”

BIOACTIVE-NET MANUALHANDBOOK on BIOACTIVE COMPOUNDS from OLIVE PROCESSING RESIDUES

Visit our Website: www.bioactive-net.com

© BIOACTIVE-NET

HANDBOOK on BIOACTIVE COMPOUNDSfrom OLIVE PROCESSING RESIDUES

3

The aim of this handbook is to provide an overview about the bioactive compoundsin olive processing residues, the extraction techniques and the applicationpossibilities in the food and cosmetic industry.

The Bioactive-net manual is a collection of three publications as part of theproject BIOACTIVE-NET. The aim of this project is to collect most relevantknowledge and technologies related to bioactive compounds in tomato, oliveand grape processing residues, from techniques of extraction, to applicationfields and economic feasibility of the extraction, and make it accessible to thepublic.

This publication has been carried out with the support of the European Commission,priority 5 (food quality and safety): contract number FOOD-CT-2006-43035,Specific Support Action (SSA) “Assessment and dissemination of strategies forthe extraction of bioactive compounds from tomato, olive and grape processingresidues”. It does not necessarily reflect the views of the Commission and in noway anticipates the Commission’s future policy on this area.

Additionally, BIOACTIVE-NET foresees dissemination workshops addressed totomato processors, olive oil mills and wine producers in the South Europeancountries. The Bioactive-net manual constitutes a key part of that disseminationaction and will be available on the project website and upon request.

www.bioactive-net.com

HANDBOOK on BIOACTIVE COMPOUNDSfrom OLIVE PROCESSING RESIDUES

The BIOACTIVE-NET project is a Specific Support Action (SSA) funded by theEuropean Commission under the 6th Framework Program.

The primary objective of bioactive-net is to assess and disseminate to the SMEprocessors, strategies for the extraction of bioactive compounds from tomato,olive, and grape processing residues and therefore allow the:

Creation of a broad information platform regarding the extraction of bioactivecompounds from tomato, olive, and grape processing residues as well as theirapplication facilities in the food and cosmetic industry.

Implementation of dissemination workshops in the south European countries(Spain, Italy, Greece and France) aimed at transferring know-how and evaluatingeconomic feasibilities of the extraction of bioactive compounds to the residuegenerating companies (SMEs), to the technology providers, to the industrialresidue extractors and to the end-users of the respective natural ingredients.

Strengthening of the European market on natural ingredients, which has anenormous economic potential due to high availability of the raw materials.

Increase of the competitiveness of the European food industry by pre-emptingthe competition in the use of bio-active compounds derived from natural,renewable and economic source processing residue.

Increase of the use of bio-active compounds in the European diet.

Project details:Type of instrument: Specific Support Action (SSA)Priority 5: Food Quality and SafetyProject number: 043035Project duration: 2 years (01.11.2006 – 31.10.2008)

Membres de BIOACTIVE-NET:

This handbook has been developed by Elvira Casas (ainia), Marianna Faraldi (Tecnoalimenti)and Marie Bildstein (ttz Bremerhaven) for inclusion in the Bioactive-net manual.

[email protected]

Tecnoalimenti S.C.p.A. (Italy)

ANFOVI - L’organisme de formation desVignerons Indépendants (France)

Union of Agricultural Cooperatives in Peza (Greece)

VIGNAIOLI PIEMONTESI S.C.A (Italy)

AMITOM - Mediterranean InternationalAssociation of the Processing Tomato (France)

CCAE - Confederación de CooperativasAgrarias de España (Spain)

ainia centro tecnológico (Spain)

Project coordinator: ttz Bremerhaven (Germany)

© BIOACTIVE-NET

BIOACTIVE-NET

4 5

TABLE of content

4 Application fields for bioactive compounds

in food and cosmetic products ................................................................................ 24

4.1. Legislation ............................................................................ 24

4.2. Olive polyphenols ................................................................. 25

4.3. Hydroxytyrosol .................................................................... 26

4.4. Oleuropein ........................................................................... 26

4.5. Existing and potential markets

for natural ingredients from olive processing residues ............ 26

5 Assessment of the economic feasibility for the extraction

of olive bioactive compounds from processing residues ................ 27

5.1. Extraction of bioactive compounds from

olive processing residues ...................................................... 28

5.1.1. Minimum Hypothesis ................................................... 28

5.1.2. Intermediate Hypothesis .............................................. 30

5.1.3. Maximum Hypothesis................................................... 30

5.2. Economic analysis of the extraction of olive bioactive

compounds from processing residues .................................... 31

5.2.1. Extraction of Polyphenol containing

oil and olive fibres ....................................................... 31

5.2.2. Extraction of Polyphenol powder and olive fibres ......... 35

5.3. Yearly profit and break-even point ........................................ 37

6 Acknowledgements..................................................................... 37

7 References ................................................................................. 38

8 Other related projects and links .................................................. 40

1 Introduction ......................................................................................................................... 8

2 Bioactive compounds in olive processing residues ........................ 9

2.1. Olive Polyphenols ................................................................. 9

2.2. Hydroxytyrosol .................................................................... 10

2.3. Oleuropein........................................................................... 10

3 Best Available Techniques for the extraction and purification

of bioactive compounds from olive processing residues ................ 11

3.1. Pre-treatment of the olive processing residues ...................... 11

3.1.1. Tray dryer .................................................................. 12

3.1.2. Drum dryer ................................................................ 12

3.1.3. Fluid bed dryer............................................................ 12

3.1.4. Milling and Homogenisation

the olive processing residues ................................................ 13

3.2. Extraction of the dried and homogeneous

olive processing residues ...................................................... 14

3.2.1. Solvent extraction ....................................................... 14

3.2.2. Supercritical Fluid Extraction (SFE)

SC-CO2 extraction ..................................................... 16

3.3 Purification of the extracts ................................................... 17

3.3.1. Chromatographic techniques........................................ 17

3.3.2. Membranes filtration techniques ................................... 20

3.3.3. Crystallization.............................................................. 22

3.4. Drying of the purified bioactive extracts................................ 22

3.4.1. Freeze Drying.............................................................. 23

3.4.2. Spray Drying................................................................ 23

3.4.3. Rotary Vacuum Drying................................................. 24

6 7

1. INTRODUCTION

At European scale, 3 million tons olives are processed in olive oil per year (witha yield in oil of about 60.000 tons). Olive oil production is divided into threeactivity fields:

Oil mills, which process the olives into oil, waste water and solid waste.Refineries, where the non-consumable oil is refined.Plants where the oil cake is processed and residual oil is extracted from thewaste resulting from olive oil extraction.

The wastes obtained after the oil milling process, as showed in the followinggraphic, are mainly Olive Mill Waste Water (OMWW), alpeorujo, pomace andmargine.

8 9

The European olive oil milling leads to 4,5 millions m3 of waste water per year.The 12.000 oil mills, mostly of small dimensions, some isolated, others insertedin urban contexts have evident difficulties to unify to recover the wastes.

Following the General EU Legislation on wastes (Directive 2006/12/EC), Member Statesshall take the necessary measures to ensure that waste is recovered or disposed ofwithout endangering human health and without using processes or methods whichcould harm the environment. Currently the olive processing residues are disposedagainst payment from the olive oil mills or sold to oil extractors at low price.

Is it possible to gain more added value from the olive processing residues forthe companies? Can the health benefits from olive be obtained from oliveprocessing residues? Various research studies demonstrated the high level oforganic substances contained in olive pomace and Olive Mill Waste Water.

2. BIOACTIVE COMPOUNDS in OLIVE processing residues

The main bioactive compounds extractable from olive processing residues arePolyphenols and more specifically Hydroxytyrosol and Oleuropein.

2.1. Olive Polyphenols

Ingredient descriptionPolyphenols are a group of chemical substances found in plants, characterizedby the presence of more than one phenol group per molecule. Polyphenols aregenerally further subdivided into tannins, and phenylpropanoids such as ligninsand flavonoids. Tannin chemistry originated in the importance of the eponymouslynamed tannic acid to the tanning industry; lignins to the chemistry of soil andplant structure; and flavonoids to the chemistry of plant secondary metabolitesfor plant defense, and flower color (e.g. from anthocyanins).

Notable sources of polyphenols include berries, tea, beer, wine, olive oil, chocolate/cocoa,walnuts, peanuts, yerba mate, and other fruits and vegetables. High levels of polyphenolscan generally be found in the fruit skins.

Metabolic known effectsResearch indicates that polyphenols may have antioxidant characteristics with potentialhealth benefits. They may reduce the risk of cardiovascular disease and cancer.

OLIVE

Crushing & grinding Crushing & grindingMilling

Washing

Malaxation Malaxation Malaxation

Pressing Paste centrifugation Paste centrifugation

Oil & Water Pomace Oil &aqueousphase

Aqueousphase +

oil

Pomace Olive oil Pomace &vegetable

water(alperujo)

Centrifugation

Olive oil Vegetablewater

Decantation

Olive oil Vegetablewater

TRADITIONALPROCESS

THREE PHASEPROCESS

TWO PHASEPROCESS

Water

Polyphenols have also been investigated as a source of additional health benefit inorganic products, but no conclusion was made. It is believed that polyphenols can bindwith nonheme iron (from plant sources) and decrease its absorption by the body.

The suggested daily dosage of polyphenols is 100mg. Therefore it would benecessary to consume 200ml olive oil a day (~40 teaspoons or one glass).

Amount of extractable moleculeThe Polyphenols concentrations in oil oscillate from 100 to 1000 mg totalpolyphenols per kg olive oil. In alpeorujo the polyphenols concentration is around8500mg polyphenols /kg alpeorujo.

2.2. Hydroxytyrosol

Ingredient descriptionHydroxytyrosol is a phytochemical. It is responsible for the bitter taste of extra virginolive oil. The wastewaters generated during olive processing contain high levels ofhydroxytyrosol, most of which can be recovered to produce hydroxytyrosol extracts.

Metabolic known effectsHydroxytyrosol has antioxidant properties. It is believed to be the antioxidantwith the highest free radical scavenging capacity: double than that of quercetin(wine antioxidant) and more than 3 times that of epicatechin (tea antioxidant).

Studies by Visioli et al (2000) showed that a low dose of hydroxytyrosol reducesthe consequences of sidestream smoke-induced oxidative stress in rats.

Amount of extractable moleculeThe hydroxytyrosol concentration is around 400 µ g/kg in OMWW (Olive MillWaste Water).

2.3. Oleuropein

Ingredient descriptionOleuropein is a polyphenolic fraction derived from the fruit, leaves, bark androots of the olive tree, which help make it strongly resistant to damage frominsects and other factors.

Oleuropein is known as an iridoid, a type of plant chemical found throughoutthe olive tree and in olive oil.

10 11

Metabolic known effectsWithin Oleuropein is a chemical agent called elenolic acid, which has been shownto assist the body's immune defense. Research studies have found that elenolic acidhelps the body to balance levels of friendly bacteria and support the immune system.

Like resveratrol, the polyphenolic component of red wine, oleuropein imparts someimportant antioxidant benefits that may help prevent the oxidation of LDL cholesteroland support healthy heart function. The oxidation of LDL cholesterol—the so-called "bad" cholesterol—can severely damage the walls of arteries. Along withresveratrol, oleuropein is high on the list of beneficial components of the acclaimedMediterranean Diet (which includes olive oil), believed to be responsible for thereduced incidence of heart disease in those who habitually partake in the Diet.

3. BEST AVAILABLE TECHNIQUES for the extractionand purification of BIOACTIVE COMPOUNDS from oliveprocessing residues

To extract bioactive compounds out of olive processing residues different stepshave to be followed:

Pre-treatment of the olive processing residues.Extraction of the dried and homogeneous olive processing residues.Purification of the extracts.Drying of the purified extracts.

3.1. Pre-treatment of the olive processing residues

Olive processing residues contain a lot of water. Depending on the extraction processit is necessary or not to dewater the samples before the extraction process. Forsupercritical fluid extraction, for example dewatering of the olive residues is necessary.Moreover the drying of the olive residues facilitates the transport and storage of thebioactive rich residues. To enable an efficient extraction, it is also necessary to millthe dried residues in order to guaranty the homogeneity of the extractor feed.

Several techniques and equipments may be applied in order to prepare oliveprocessing residues. Some of the most used drying and milling techniques arepresented here.

3.1.1. Tray dryer

Olive residues are spread out thinly on trays, inside a cabin which is connected to asource of air heated by gas diesel or biomass to remove the moist vapours. Dependingon the cabinet design, there are batch tray dryers, semi-continuous tray dryers andcross flow chambers.

3.1.2. Drum dryer

Olive residues are spread over the surface of a heated drum. The drum rotates andthe residues remain on the drum surface during the major part of the rotation, whilethe drying process takes place, and then they are scraped off.

The following table compares tray drying and drum drying processes used witholive processing residues:

3.1.3. Fluid bed dryer

The feed of wet material is dried by intimate contact with hot air when thematerial is in a fluidized state.

The dryer comprises:

A top fluidising chamber.A bottom air distribution chamber.A specially designed perforated plate.

12 13

Door No. 1 Door No. 2Trays

Trays

Figure 1: Tray dryer.

Drum drying ++ ++ long

Dryingtime

Tray drying + + long

Operatingeasiness

Energyconsumption

Initialinvestment

Technique

easy

easy

Figure 2: Fluid-bed drying process.

3.1.4. Milling and Homogenisation of the olive processing residues

Sometimes it may be necessary to reduce the size of the particles (by milling)and to mix the residues by a homogenisation process in order to improve theextraction process.

Milling of the dried olive processing residuesMilling is used to convert the shred material into fine particles. One possible millingtechnique for the pre-treatment of the olive processing residues is the hammermill.

A hammermill is essentially a steel drum containing a vertical or horizontal cross-shaped rotor on which pivoting hammers are mounted (see figure 3). The hammersare free to swing on the ends of the cross. The rotor is spun at a high speedinside the drum while material is fed into a feed hopper. The material is impactedby the hammers on the ends of the rotating cross and thereby is shredded andexpelled through screens in the drum.

Figure 3: Hammermill.

14 15

Homogenisation of the dried olive processing residuesHomogenisation is a process that makes a mixture the same throughout theentire substance. A mixing phase is required and enough in order to homogenisethe dried and milled residues.

3.2. Extraction of the dried and homogeneous olive processing residues

The operation of extraction consists of the separation of one or more speciesfrom a solid or liquid matrix based on the different relative solubility that suchsubstance or substances present in a certain solvent respect to the rest of thecomponents of the matrix. In other words, extraction works according to theprinciple that soluble components can be separated from insoluble or less solublecomponents by dissolving them in a suitable solvent. Raw materials that are suitablefor extraction may contain solids only, solids and a solution, or solids and a liquid.

3.2.1. Solvent extraction

Conventional solid-liquid extractionThis technique implies the contact of the plant solid matrix with a liquid solvent.The selection of the solvent will be determined by the chemical and physicalproperties of the target substances. In particular, the thermal stability and the polarcharacter of the substance have special relevance. The solvent temperature mustbe chosen accurately depending on the raw material and on the thermal resistanceof the solutes we want to recover. In order to facilitate the transfer of the targetsubstances to the liquid, the plant feedstock is normally treated mechanically.

This process is used to extract oils. It is not suitable for thermolabile substances.Some organic solvents that may be used as extraction agent are toxic and canleave traces in the end product. Ethanol can be used to replace some toxic ordangerous organic solvents. In addition, all the solvent extraction requires apurification stage after extraction, such as filtration or centrifugation.

Ultrasound and microwave assisted extraction are similar to conventionalextraction, with the addition of ultrasound or microwave in order to increasethe yields, reduce the volume of solvent and reduce the working time.

Sonicated-assisted extraction (ultrasound)Sound waves with frequencies higher than 20 kHz can improve the extractionyield of plant material because they involve alternative expansions and compressionsof matter inducing the creation of bubbles in liquids.

The most relevant parameter to control in sonicated assisted extraction isfrequency, because small changes of this parameter can affect dramatically theyield of extraction. Ultrasounds cause a greater penetration of solvent into cellularmatrices improving mass transfer.

Ultrasound-assisted extraction has been used to extract nutraceutical such asessential oils, lipids, antioxidants, steroids and terpenoids. It allows processconditions to be milder compared to traditional solvent extraction, so it isrecommended for thermolabile substances.

Microwave-assisted extraction (MAE)Microwaves are electromagnetic waves that interact with matter, in particular withpolar molecules to generate heat. They can therefore, penetrate water and biologicalmatrices heating up the whole at homogeneous rate. Radiation produces superheatingof water within plant cells and causes the rupture of the cellular wall facilitating thetransfer of interesting substances to the bulk phase inside the extraction vessel andthe penetration of the solvent into the plant matrix. Microwaves can thereforeimprove the extraction yields of nutraceuticals. The volume of solvent needed, andthe extraction time are reduced.

The effectiveness of MAE depends strongly on the polarity of the solvent and on theparticle size and the distribution of the vegetable material. It can be applied to extractpolar components, but it is not suitable for dry materials or too wet matrices using non-polar solvents. (Water, methanol and ethanol are polar enough to be employed). In addition,MAE requires a purification stage after extraction, such as filtration or centrifugation.

Solventextraction

Filtration

Drying

Spent biomass

EvaporationExtract

Fresh or dry biomass Solvent

Solvent Extract

Ultrasounds

Figure 4: Sonicated-assisted extraction process diagram.

16 17

Accelerated solvent extraction (ASE)Accelerated solvent extraction is a solid-liquid extraction carried out at hightemperatures (that improve the diffusivity of the solvent accelerating the extraction)and pressures (to maintain the solvent in liquid phase), below the critical point ofthe solvent. Most of the solvents used in conventional solid-liquid extraction aresuited for ASE (including water) to recover polar compounds from plant material.

3.2.2. Supercritical Fluid Extraction (SFE) SC-CO2 extraction

The supercritical state is reached by bringing the fluid to a temperature andpressure beyond its critical point. Supercritical fluids present characteristics ofboth gases and liquids and properties that make them especially suitable forextraction processes.

Supercritical fluids have higher diffusion coefficients and lower viscosity andsurface tension than conventional solvents. The dissolving capacity of supercriticalfluids depends on its density, so the selectivity of extraction can be changed byadjusting the temperature and/or the pressure of extraction. After the extractiontime, the pressure is reduced, or the temperature is increased, so the solubilityof the extract decreases and it can be separated.

The most used solvent is CO2, which is cheap, safe, non-toxic and its supercriticalconditions may be fairly easily reached. It can be used to extract polyphenolssuch as resveratrol and other natural antioxidants from grape peels and stalks.It is suitable for thermolabile substances and it can be also used for polarsubstances if some modifiers are added to it (methanol, ethanol, water, acetone…).

The following table compares the different extraction techniques exposed.

Ultrasound High

Conventional Polar & Non-polar High

Extractionmethod

+

+

Compoundsextracted

Humiditytolerance

Purificationrequirements

Polar & Non-polar

Microvawe High+Polar

CO2 supercritical Low–Non-polar

Subcritical Medium-high++Polar

3.3 Purification of the extracts

After the extraction processes, recovering a biological product from the interferencesand the impurities requires some purification steps, to obtain the product accordingto the final specifications. The purification simply aims at obtaining the targetmolecule pure enough in the shortest possible time.

3.3.1. Chromatographic techniques

Chromatography is a very special purification process because, it can separatecomplex mixtures with great precision (even very similar components can beseparated). In fact, chromatography can purify basically any soluble or volatilesubstance. It can be used to separate delicate products because the conditions arenot typically hard. For these reasons, it can be used to separate mixtures of olivebioactive compounds.

Another advantage of these techniques is that the separated compounds areimmediately available for identification or quantification. On the other hand, someinstrumentation is expensive and not easily portable and some work is needed toavoid the contamination of the column.

Separation by chromatography depends on the differential partition ofcompounds between a stationary phase (the adsorbent) and a mobile phase(the buffer solution). Normally, the stationary phase is packed into a verticalcolumn of plastic, glass or stainless steel, and the buffer is pumped throughthis column. The sample to be fractionated is pumped on the top of thecolumn and the various sample components travel with different velocitiesthrough the column and are subsequently detected and collected at thebottom of the column. In general, bio-molecules are purified using purificationtechniques that separate them according to differences in their specificproperties, as shown in following table.

Size

Molecular property exploited

Gel filtration (sometimes called size-exclusion)

Ion exchange chromatography

Type of chromatography

Charge

Adsorption chromatographyLigand specificity

18 19

Partition chromatographyThe stationary phase is usually a liquid, which can be mechanically coated orchemically bonded on a comparatively inert solid support. The molecules toseparate are held in this stationary phase (shown in figure 5). Reverse phase (RP)chromatography, when the stationary phase is less polar than the mobile phase,is a good example of liquid-liquid chromatography.

The advantages of this technique are the high recovery, the large volumes andthe easiness to scale-up.

Size-exclusion or gel filtrationThe separation in gel filtration depends on the different abilities of thesample molecules to enter inside the pores which contain the stationaryphase. Very large molecules do not enter and move through thechromatographic bed faster. Smaller molecules, which can enter inside thegel pores, move slower through the column, because they spend a part oftheir time in the stationary phase. Molecules are eluted according to theirdecreasing molecular size.

Figure 5: partition chromatography.

Sample molecules carriedby mobile phase

Sample molecules heldin sorbed solvent

Figure 6: Size-Exclusion chromatography mechanism.

The disadvantages of this simple and effective method are its low capacity andthe fact that it does not work very well for crude mixtures, so this process canbe applied in the final “polishing” step.

Ion exchange chromatographyThe basis for ion exchange chromatography is the competitive binding ofcompounds with differences in charge, to an oppositely charged chromatographicmedium, the ion exchanger.

By using this technology, large volumes can be processed.

Adsorption chromatographyA bio specific adsorbent is prepared by coupling a specific ligand, on a solidsurface, that will only interact with the molecules that can selectively bind to it,(the ones that are needed to be separated). Molecules that are not bound eluteunretained. The retained compound can later be released in a purified state. Thiskind of technique is used in Fine Chemistry.

The following table compares the main characteristics of the different chromatographytechnologies that have been explained before.

Figure 7: Ion-exchange chromatography mechanism.

Adsorptionchromatography

Laboratoryscale X

Ion exchangechromatography

Size-exclusionchromatography

Partitionchromatography

Technique

X X X

Large scale X X X X

Selectivity High Low High High

Resolution High Low High High

Capacity High Low High High

Recoveryyields

High(close to 100%) High Low (50-60%) High

Operatingeasiness

Easy Simple, fast Lengthy procedure Easy

Costs ++ + ++ ++

Finechemistryapplications

X NO X X

Industrialchemistryapplications X X X X

20 21

Chromatographic methods such as reverse phase, ion-exchange and adsorptionchromatography compete reasonably well with affinity-based purifications, especiallywhen high purity is not required.

3.3.2. Membranes filtration techniques

Membranes selectively filter gases or liquids in solutions or mixtures into theirdifferent components. The membrane micropores are sized to allow somemolecules and particles to go through and block others. Thus membranes arevery specific, with their molecular structure tailored according to the particularspecies to be separated.

Membrane filtration is regarded as BAT in the BREF (Best Available TechniquesReference Document) for the Food, Drink and Milk Industries due to the reducedwater consumption and waste water pollution that its use entails. There are threemain technological processes depending on the size of the components to retain:microfiltration, ultrafiltration and reverse osmosis.

MicrofiltrationMicrofiltration is a low-pressure cross-flow membrane process for separatingcolloidal and suspended particles in the diameter range 0.1-10 µ m.Microfiltration is a purely physical process in which particles are capturedon the surface on the membrane. Any particle larger than the pore size ofthe membrane cannot squeeze through. Membrane filters are widely usedin biotechnology and food and beverage applications where sterile productis required.

UltrafiltrationUltrafiltration membranes retain particles in the range 0.01-0.1 µ m and operatein the pressure range 0.5-10 bar. It has become the best method to concentrate,largely replacing size-exclusion chromatography in these applications. UF membranesare highly used in biopharmaceutical applications.

The major advantages of ultrafiltration over competing purification techniquessuch as chromatography are:

High throughput of product.Relative easiness of scale-up.The equipments are easy to clean and sanitise.

Reverse osmosisOsmosis explains the phenomenon whereby if a semi-permeable membraneseparates two salt solutions of different concentration, water will migratefrom the weaker solution, through the membrane, to the more concentratedsolution, until the solutions have the same salt concentration. Reverse osmosisinvolves applying pressure to reverse the natural flow of water, forcing thewater to move from the more concentrated solution to the weaker. Thesemi-permeable membrane is porous, allowing water to pass through, butblocking the passage of the bulkier salt molecules. The result is water withoutsalt on one side of the membrane.

Reverse osmosis mainly remove water and molecular compounds smaller in sizethan water molecules. Reverse osmosis is a high-efficient technique forconcentrating/separating low-molecular-weight substances in solution. For thispurposes, it requires an energy source and is fairly expensive.

3.4.1. Freeze Drying

It is also known as lyophilization. It is used to preserve a perishable material orto make it easier to transport. Freeze-dryed product can be rehydrated quicklyand easily. This process includes the following steps:

Freezing the material.Reducing the surrounding pressure.Adding enough heat to allow the frozen water in the material to sublimedirectly from the solid phase to gas.

3.4.2. Spray Drying

Spray drying is the most widely used industrial process, involving particle formationand drying. It is highly suited for the continuous production of dry solids in powder,granulated or agglomerated form from a liquid feedstock. It is an ideal process whenthe final product must comply with precise quality standards regarding particle sizedistribution, residual moisture content, bulk density, and particle shape.

Spray drying involves the atomization of a liquid feedstock into a spray of droplets andcontacting the droplets with hot air in a drying chamber. Evaporation of moisture fromthe droplets and formation of dry particles proceed under controlled temperature andairflow conditions (it is shown in the following drawing).

Drying occurs very fast, so this process is very useful for materials that can bedamaged by long time exposures to heat. Spray drying has been identified in theBest Available Techniques Reference Document for the Food, Drink and MilkSectors as best available technique for drying because of its reduced energy andwater consumption and its reduced emissions of dust.

22 23

Figure 8: Spray drying process

3.3.3. Crystallization

Crystallization is a technique used to purify solid compounds. It is based on theprinciples of solubility. As a general rule, compounds (solutes) tend to be moresoluble in hot liquids (solvents) than they are in cold liquids. If a saturated hotsolution is allowed to cool, the solute is no longer soluble in the solvent and itforms crystals of pure compound. Impurities are excluded from the growingcrystals and the pure solid crystals can be separated from the dissolved impuritiesby filtration. High purity products are obtained so, this process is particularlyused at pharmaceutical level.

The following table compares the purification techniques explained in the present chapter.

3.4. Drying of the purified bioactive extracts

The bioactive compounds need to be dried enough to be safely stored until they arerequired for further processing. The whole drying process needs to be gentle to reducethe risk of bioactive compounds degradation. Different drying methods can be used.

Scale-up

Techni-que

Partitionchromato

Size-exclusionchromato

Ionexchangechromato

Adsorp-tion

chromato

Micro-filtration

Ultra-filtration

Reverseosmosis

Crysta-llization

Relativelyeasy

Moredifficult

Moredifficult

Moredifficult

Relativelyeasy

Relativelyeasy

Relativelyeasy

Used

Equipment FlexibleRelatively

inexpensiveExpensive

Complexand

expensive

Simple,reliable, easy to maintain

Suitable Suitable SuitableNot

suitableSuitable

Pressuresensitivecompounds

Suitable

Suitable Suitable Suitable Suitable Suitable Suitable SuitableTemperaturesensitivecompounds

Suitable

Selectivity High Low High Low LowHigh Low High

ParticleIndependent Dependent Independent Independent

Dependent(0,1-10 µ m)

Dependent(0,01-0,1µ m)

Dependent Independent

Purificationtime

Fast Fast Low Long Long Long Long

Costs ++ + ++ ++ + +Fairlycostly

++

Energyrequirements

Low Low Low LowQuitelow

High LowQuitelow

Efficiency High Low High High High Medium



Notsuitable

Notsuitable

3.4.3. Rotary Vacuum Drying

Wet feed is loaded as a batch and is heated indirectly while being agited by a paddle.The operation is normally carried out under vacuum. Solvent recovery is possible bycondensing the vapours generated during the drying operation.

Other advantages of this process are:

Granular / pasty wet materials can be handled.Low temperature operation is possible: it is ideal for materials that would bedamaged or changed if they were exposed to high temperatures. The vacuumremoves moisture while preventing the oxidation or explosions that can happenwhen some materials are mixed with air.Mode of heating is indirect.High energy efficiency.Closed operation: solvents can be recovered, it is safer and minimizes theproduct loss caused by atmospheric contaminants, dusting, oxidation, discoloration,and chemical change.

24 25

4. Application fields for BIOACTIVE COMPOUNDSin food and COSMETIC PRODUCTS

4.1. Legislation

From a legislative point of view, natural ingredients are regulated as Food additivesand/or Cosmetic products.

Food additivesFood additives used as ingredients during the manufacture or preparation of foodand which are part of the finished product are covered by the scope of theDirective 89/107/EEC. Prior to their authorisation, food additives are evaluatedfor their safety by the Scientific Committee on Food, an expert panel that advisesthe European Commission in questions relating to food.

All authorised food additives have to fulfil purity criteria which are set out indetail in three Commission Directives:

Commission Directive 95/45/EC laying down specific purity criteria concerningcolours for use in foodstuffs.Commission Directive 95/31/EC laying down specific criteria of purity concerningsweeteners for use in foodstuffs.Commission Directive 96/77/EC laying down specific purity criteria on foodadditives other than colours and sweeteners.

Cosmetic productsCouncil Directive 76/768 of 27 July 1976 on the approximation of the laws ofthe Member States relating to cosmetic products.

Restrictions and prohibitions on ingredients that can be used in cosmetics areincluded in various lists under the EU Cosmetics Directive.

Decision 96/335/EC updated by Commission Decision 2006/257/EC, establishan inventory and a common nomenclature of ingredients employed in cosmeticproducts. The inventory is purely indicative and shall not constitute a list ofsubstances authorized for use in cosmetic products.

4.2. Olive polyphenols

State of the competitionThe following companies have been identified as bulk olive polyphenols suppliers:

Possible applicationItalian botanicals manufacturer Indena and Japanese cosmetics giant Kanebo havejoined forces to develop an olive fruit extract designed to provide moisturizingand anti-ageing properties for the skin care category. This is one example of theapplication of olive polyphenols in the cosmetic industry.

Price per kg

OLIVEPURE®35% polyphenols Naturex

PurityOrigin ofpolyphenols

CompanyProduct

Olive fruits >35% 312.50$/kg

PROLIVOLSTMSEPPIC

Margines (watersoluble part ofolives, obtainedfrom the pulp ofolives during theolive oil process)

>35% 380.41$/kg

Olive polyphenols can also be incorporated in health food products (biscuits,bakery products, dietetic products) and nutraceuticals.

4.3. Hydroxytyrosol

State of the competitionTwo companies offering bulk supply of hydroxytyrosol have been found: Lachifarmain Italy and Genosa in Malaga (Spain).

Possible applicationHydroxytyrosol can be used in food (as natural additives alternatively to synthesiscompounds), chemical, pharmaceutical and cosmetic industries.

4.4. Oleuropein

State of the competitionThe supply of bulk oleuropein is not exploited by a lot of companies so far, asshown in the following Table where only one company is represented.

Possible applicationOleuropein can be used in the food, chemical, pharmaceutical and cosmeticindustries for its antibacterial properties.

4.5. Existing and potential markets for naturalingredients from olive processing residues

During the market observation only one enterprise was located which concerns itselfwith the subject of the natural production of olive polyphenols from olive processingresidues and not from olive fruits. This method of approach is a world novelty.

Different companies are currently extracting and marketing oleuropein fromolive leaves. It would be interesting to estimate the part of leaves gained duringthe washing stage before the milling of the olives to evaluate the economicinterest of the extraction of oleuropein out of the washing residues.

26 27

Price per kg

OLIVEPURE®leaf 15% Naturex

PurityOrigin ofoleuropein

CompanyProduct

Olive leaves >15% 42.12$/kg

5. Assessment of the ECONOMIC FEASIBILITY for theextraction of OLIVE BIOACTIVE COMPOUNDS fromprocessing residues

The objective of this chapter is to demonstrate the net benefit of the extractionof bioactive compounds from olive processing residues in light of the costs forthe industrial extraction of bioactive compounds and the revenues derivingfrom their selling.

Two extraction processes have been used as calculation example to determine theeconomic feasibility of the extraction of bioactive compounds from olive processingresidues:

Example 1: Supercritical Fluid Extraction (SFE)/SC-CO2 extraction.Example 2: Solvent extraction.

Considering the SFE and Solvent extraction, the following flow-charts show theprocess steps required to extract compounds and the necessary equipments.

Belt Dryer

Hammermill

Supercriticalfluid extractor

Spray Dryer

Drying

Milling

Extraction

Drying

Olive processing residues (Pomace)

Example 1:

SUPERCRITICAL FLUID EXTRACTION

Belt Dryer

Hammermill

Homogenisator

Stirred Tank

Drying

Milling

Homogenisation

Extraction

Olive processing residues (Pomace)

Example 2:

SOLVENT EXTRACTION

Decanter

Ultrafiltration

Evaporator

Spray Dryer

Decantation

Filtration

Evaporation

Drying

7.3t Pomace / day(moisture ~55%)

It is interesting to underline that the pre-treatment (drying and milling of thepomace) must be necessarily done during harvesting season (from Novemberto February, 4 months) to preserve the product. The extraction and the dryingare two operations that can be done all year over (either 110 days in the minimumhypothesis; 200 days in the intermediate hypothesis or 330 days in the maximumhypothesis; 24 hours a day), reducing in this way the effort during the short andhard period of the olive harvesting and partially overlapping the problem of theseasonality of the extracts obtainable from olive.

It is also very important to note that the calculations developed in this chapterinclude only labour, energy, maintenance, quality control and consumable costs.Costs such as costs of sales and marketing, shipping, handling and storing, recoverycosts of solvent have not been considered. However, even if not exhaustive, thefollowing evaluations give a good idea of the economic feasibility of two differentextraction methods.

5.1. Extraction of bioactive compounds from olive processing residues

Starting from the production data of olive processing residues provided bythe European industrial Associations of the sector participating to theproject, three hypotheses have been chosen (minimum, intermediate andmaximum) to provide more adapted information to companies with differentproduction sizes.

5.1.1. Minimum Hypothesis

The minimum hypothesis considers a quantity of 800 tons/year of olive processingresidues to treat in single olive processing companies. Small olive mills have a yearlywork of 110 days corresponding to the harvesting season lasting from November toFebruary (4 months).

28 29

Example 1:

SUPERCRITICAL FLUID EXTRACTION

Example 2:

SOLVENT EXTRACTION

With SFE it is possible to extract,during the 110 days of yearlyproduction, around 145kg of oilcontaining polyphenols and 2 500kgof fibre per day. The oil can be soldas such or further processed intopolyphenol powder leading to101kg/day.

With solvent extraction it ispossible to extract, during the 110days of yearly production, around145kg of oil containing polyphenolsand 2 500kg of fibre per day. Theoil can be sold as such or furtherprocessed into polyphenol powderleading to 101kg/day.

Beltdryer

7.3t Pomace / day(moisture ~55%)

PRE-TREATMENT (during harvesting season):

3.6t dry Pomace / day

(moisture ~10%)

Spraydryer

101 kg polyphenolspowder / day

DRYING(all year over):

Beltdryer

PRE-TREATMENT (during harvesting season):

3.6t dry Pomace / day

(moisture ~10%)

Hammermill

Supercritical fluidextractor

2.5t fibre/day

EXTRACTION(all year=110days):

EXTRACTION(all year=110days):

47.3t dry milledPomace / day

Addition watersolvent 1:12

(all yearover=110 days)

Homegenisator

SolventExtractor

Decanter

145.5 kg polyphenolscontaining oil / day

2.5t fibre / day

3.6t dry milledPomace / day

(all yearover=110 days)

145 kg polyphenolscontaining oil / day

DRYING (all year over):

Spray dryer

101 kg polyphenolspowder / day

Hammermill

5.1.2. Intermediate Hypothesis

The intermediate hypothesis considers a quantity of 50.000 tons/year of olive processingresidues to treat in single cooperatives. Cooperatives are working 200 days a year. Theolive processing residues have to be pre-treated during the harvesting season lastingfrom November to February (4months) before the extraction that can be performedduring the 200 days of yearly production.

30 31

With SFE it is possible to extract, during

the 200 days of yearly production, around

5t of oil containing polyphenols and 87.5t of

fibre per day. The oil can be sold as such or

further processed into polyphenol powder

leading to 3.5t/day.

With solvent extraction it is possible to

extract, during the 200 days of yearly

production, around 5t of oil containing

polyphenols and 87.5t of fibre per day. The oil

can be sold as such or further processed into

polyphenol powder leading to 3.5t/day.

5.1.3. Maximum Hypothesis

The maximum hypothesis considers an amount of 100.000 tons/year of oliveprocessing residues to pre-treat during the harvesting season lasting from Novemberto February (4months). The maximum hypothesis for the cost calculation considersthat the extraction of bioactive compounds is performed by a big and centralizedextractor working 330 days a year for the extraction of the processing residuesof the whole region.

Example1:SUPERCRITICAL FLUID EXTRACTION

Example 2:SOLVENT EXTRACTION

With SFE it is possible to extract, during

the 330 days of yearly production, around

6t of oil containing polyphenols and 106t

of fibre per day. The oil can be sold as such

or further processed into polyphenol powder

leading to 4.2t/day.

With solvent extraction it is possible to

extract, during the 330 days of yearly

production, around 6t of oil containing

polyphenols and 106t of fibre per day. The oil

can be sold as such or further processed into

polyphenol powder leading to 4.2t/day.

5.2 Economic analysis of the extraction of olive bioactivecompounds from processing residues

5.2.1. Extraction of Polyphenol containing oil and olive fibres

5.2.1.1. Cost analysis

The costs of the different treatment steps have been evaluated.

Drying costs

The drying phase is the same in both processes (SFE and solvent). Theproduction costs have been calculated for different capacities of belt dryerdepending on the hypothesis considered. The labour costs, the energy costsand the quality control analysis costs are proportional to the olive residuesamount to treat yearly.

Milling costs

The milling phase is the same in both processes (SFE and solvent) and requiresa Hammermill.

Yearly DRYING costs (�)

MinimumHypothesis(800t/year)

IntermediateHypothesis

(50 000t/year)

MaximumHypothesis

(100 000t/year)

24 399 82 730 165 460

Minimum Hypothesis

Capacity of the Dryer: 20 000 kg/day

Initial Investment: 80 000 �

Intermediate Hypothesis



Maximum Hypothesis

Capacity of the Dryer: 2x 500 000 kg/day

Initial Investment: 1 200 000 �



Yearly DRYING costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

24 399 82 730 165 460






Maximum Hypothesis

Capacity of the Dryer: 2x 500 000 kg/day




Minimum Hypothesis

32 33

Yearly MILLING costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

38 819 243 432 483 116

Yearly MILLING costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

38 819 243 432 483 116

Minimum Hypothesis

Capacity of the Mill: 5 000 kg/day


Minimum Hypothesis

Capacity of the Mill: 5 000 kg/day



Capacity of the Mill: 3x 50 000 kg/day





Maximum Hypothesis



Maximum Hypothesis



Homogenisation costs

The homogenization is a phase necessary only in the solvent extraction process.It requires the investment in a Homogenisator.

No homogenisation necessary!!

Yearly HOMOGENIZING costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

59 520 1 944 420 2 415 186

Minimum Hypothesis

Capacity of the Homogenisator: 720 000 kg/day


Intermediate and Maximum Hypothesis

Capacity of the Homogenisator: 3 600 000 kg/day


Extraction costs

The initial investment of the two extraction processes is extremely different:160 000 ¤ to 240 000 ¤ for the solvent extraction and 6 000 000 ¤ to230 000 000 ¤ for the SFE extraction. Besides, the solvent consumption ishigher for the non-supercritical extraction. A CO2 consumed toward sampleextracted ratio of 30 and a CO2 recycling of 80% have been considered.For the solvent extraction 90% solvent recovery has been considered.

Maximum Hypothesis

Capacity of the SFE Extractor: 115x 1 320 kg/day



Yearly EXTRACTION costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

717 949 36 645 662 66 522 267

Yearly EXTRACTION costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

227 409 8 556 760 17 101 764

Minimum Hypothesis



Minimum Hypothesis







Capacity of the SFE Extractor: 2x 1 000 000 kg/day


Decantation costs

The decantation is a phase necessary only in the solvent extraction process.

No decantation necessary!!

Yearly DECANTATION costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

107 123 158 113 521 833

Minimum Hypothesis

Capacity of the Decanter: 2x 25 000 kg/day



Capacity of the Decanter: 2x 1 200 000 kg/day


Other costs

The cost of supervision and an average transport of 15km have been consideredfor the maximum hypothesis.

Yearly OTHER costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

6 747 16 728 413 136

Yearly OTHER costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

8 891 496 093 1 367 186











34 35

Total investment and production costs

Summing the pre-treatment, the extraction and the purification costs, the followinginvestment in equipments and operation costs are required.

TOTAL yearly cost (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

790 058 37 467 917 68 538 029




(50 000t/year)

MaximumHypothesis

(100 000t/year)

464 017 11 002 183 21 100 495

TOTAL initial investment (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

6 213 455 194 650 000 236 127 273

TOTAL initial investment (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

1 128 727 18 560 000 22 773 636

5.2.1.2. Benefit analysis

The process provides different kinds of natural extracts which can be sold in rawstate as food integrator or for cosmetic formulations. Market price of polyphenoloil has been estimated around 30¤/kg while olive fibres are estimated at 4.50¤/kg.

Produits de l’extraction (kg)


Polyphenols Fibres

16 000 280 000 Produits de l’extraction (kg)


Polyphenols Fibres

16 000 280 000


(50 000t/year) 1 000 000 17 500 000


(50 000t/year) 1 000 000 17 500 000

MaximumHypothesis

(100 000t/year) 2 000 000 35 000 000

MaximumHypothesis

(100 000t/year) 2 000 000 35 000 000

Yearly revenue (�)



(50 000t/year)

HypothèseMaximale

(2 500t/an)

1 740 000 108 750 000 217 500 000




(50 000t/year)

MaximumHypothesis

(100 000t/year)

1 740 000 108 750 000 217 500 000

Products of the extraction (kg) Products of the extraction (kg)

5.2.2. Extraction of Polyphenol powder and olive fibresTo obtain polyphenol powder, the polyphenol containing oil have to be further processed.The olive fibres obtained from SFE or solvent extraction can be sold as such.

5.2.2.1. Cost analysis

The costs related to a further drying phase were estimated.

Drying costs

A carrier is needed to spray dry an oil.

Yearly SPRAY DRYING costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

53 433 689 200 1 365 730

Yearly SPRAY DRYING costs (�)



(50 000t/year)

MaximumHypothesis

(100 000t/year)

53 433 690 080 1 367 490

Minimum Hypothesis

Capacity of the Spray Dryer: 500 kg/day


Minimum Hypothesis

Capacity of the Spray Dryer: 500 kg/day



Capacity of the Spray Dryer: 2x 3 000 kg/day





Maximum Hypothesis



Maximum Hypothesis









Total investment and production costs

By adding the pre-treatment, the extraction, the purification costs and the furtherdrying costs, the following investment in equipments and operation costs are required.




(50 000t/year)

MaximumHypothesis

(100 000t/year)

843 491 38 157 117 69 903 759




(50 000t/year)

MaximumHypothesis

(100 000t/year)

517 450 11 692 263 22 467 985

TOTAL initial investment



(50 000t/year)

MaximumHypothesis

(100 000t/year)

6 313 455 195 350 000 237 177 273

TOTAL initial investment



(50 000t/year)

MaximumHypothesis

(100 000t/year)

1 228 727 19 260 000 23 823 636

36 37

5.3. Yearly profit and break-even point

MaximumHypothesis

(100 000t/year)


(50 000t/year)


Example 1:SUPERCRITICALFLUIDEXTRACTION

Polyphenolcontaining oil;Olive fibres

Yearly profit (¤)without amortization

949 942 71 282 083 148 961 971

Yearly profit (¤)including a 5 yearsamortization

-270 058 33 132 083 102 681 971

Polyphenolpowder;Olive fibres

805 562 64 908 673 136 227 820

-434 438 26 618 673 89 737 820

Example 2:SOLVENTEXTRACTION

Polyphenolcontaining oil;Olive fibres

1 275 983 97 747 818 196 399 505

1 147 983 97 395 818 195 917 505

Polyphenolpowder;Olive fibres

1 131 603 91 373 527 183 663 594

983 603 90 881 527 182 971 5945.2.2.2. Benefits analysis

Considering an average price of about 35.00 ¤/kg for polyphenol powder and thedescribed incomes related to selling olive fibres, the following annual revenues canbe estimated.

Produits de l’extraction (kg)


Polyphenols Fibres

11 116 280 000 Produits de l’extraction (kg)


Polyphenols Fibres

11 116 280 000


(50 000t/year) 694 737 17 500 000


(50 000t/year) 694 737 17 500 000

MaximumHypothesis

(100 000t/year) 1 389 474 35 000 000

MaximumHypothesis

(100 000t/year) 1 389 474 35 000 000

Products of the extraction (�) Products of the extraction (�)




(50 000t/year)

MaximumHypothesis

(100 000t/year)

1 649 053 103 065 789 206 131 579




(50 000t/year)

MaximumHypothesis

(100 000t/year)

1 649 053 103 065 789 206 131 579

The financial loss for the minimum hypotheses for the supercritical fluid extractiondemonstrate the non-feasibility of this process. The other cases studied provedto be economically feasible. Nevertheless, the advantage of SFE over solventextraction in term of residual solvent in the end product and also the cost ofbuilding the treatment center, the transport for conveyance of the by-products,the packaging, and the environment have to be considered.

6. ACKNOWLEDGEMENTS

The BIOACTIVE-NET members gratefully acknowledge the financial support ofthe European Commission for the completion of the Bioactive-net manual.

The publication of this handbook would have not been possible without theinputs of all experts contributing from the eight BIOACTIVE-NET memberorganizations and from external collaborators who kindly supported the consortiumwith their valuable contributions.











Andreadakis I. M., Largo Consumo: “Un esame liscio come l’olio”, Giugno 2005,pg.141.

Pellucci E., Largo Consumo: “Fra le macchine enologiche e oleicole”, Ottobre2005, pg.51.

Pellucci E., Largo Consumo: “extravergine in sofferenza”, Novembre 2005,pg 37.

Collie A., Agrofood Industry hi-tech: “Polyphenols and cognition”, February2006, pg.17.

Galli C., Visioli F., Tecnologie Alimentari: “Nuovi sviluppi nella ricerca suicomponenti minori di rilevanza nutrizionale nell’olio di oliva extravergine”,Maggio 2006, pg.37.

Macciola V., De Leonardis A., Ingredienti Alimentari: “natura e proprietàantiossidanti di fenoli estratti da un olio extra-vergine di oliva e suoisottoprodotti”, Maggio 2006 pg.6.

Louise Prance, Cosmetics design-europe.com: “Natural Ingredients driveself-tan market”, 07/02/2007

Simon Pitman, Cosmetics design.com: ”Italian and Japanese companies joinforces on olive extract”; 9/02/2006

38 39

7. REFERENCES

Porcelli G., Folliero G., “Additivi e coloranti negli Alimenti”, Bulzani Editore,Roma 1977: Antiossidanti, p 15.

C. Di Pinto, “Lo smaltimento delle acque reflue nei frantoi” Accademia Nazionaledell’olivo, Spoleto 1986: difficoltà che si presentano nel raggiungimento deilimiti di legge e tecnologie attualmente a disposizione per la depurazione delleacque di vegetazione, p69

Baldi A., Romani A., Vincieri F. F., “Olive oil Quality, Regione Toscana assessoratoall’agricoltura, foreste, caccia, pesca, 1992”: studio di alcuni composti polifenolicipresenti nelle drupe di Olea Europea L.

Galli C., Blasevich M., Petroni A., “Olive oil Quality, Regione Toscana assessoratoall’agricoltura, foreste, caccia, pesca, 1992”: il 3,4 di-idrossi-fenil-etanolo, estrattodall’olio di oliva, inibisce l’aggregazione delle piastrine umane e la sintesi dimetaboliti della ciclo e lipossigenasi, in vitro.

Cerruti G., “Il Rischio Alimentare” 1993: antiossidanti, sinergisti, sequestranti,cap. 8.5.

Visioli F., Galli C., Life Sciences: “Oleuropein protects low density lipoproteinfrom oxidation”, October 1994.

Demicheli M. C., Bontoux L., Joint research center: “Olive oil industry: marketprospects and technological breakthroughs”, October 1995.

Saija A., Trombetta D., Tomaino A., Lo Cascio R., Princi P., Uccella N., Bonina F.,Castelli F., International Journal of Pharmaceutics: “In vitro evaluation of theantioxidant activity and biomembrane interaction of the plant phenols oleuropeinand biomembrane”, 1998.

Visioli F., Romani A., Malinacci N., Zarini S., Conte D., Vincieri F., Galli C., J.Agric.Food Chem: “Antioxidant and other biological activities of olive millwaste waters”, July 1999.

Agrofood Industry hi-tech: “Overview of poliphenols”, March 2004, pg.10.

Danzing L., International Food Ingredients: “Colour”, March 2004, pg. 42.

Redaelli R., Pisacane V., Berardo N., Agrofood Industry hi-tech: “Antioxidantsin Italian oat cultivars”, November 2004, pg.38.

G. Sivakumarl, C. Briccoli Bati, N. Uccella, Kimiya Prirodnikh Soedinenii: “HPLC-MS Screening of the Antioxidant Profile of Italian Olive Cultivars”, February2005.

40

8. OTHER RELATED PROJECTS AND LINKS

TDC Olive “Setting-up a network of Technology Dissemination Centres tooptimise SMEs in the olive and olive oil sector” was funded under the 6thFP. The main objective of the project was the setting up of a European networkof Technology Dissemination Centres (TDCs) for table olive processing andolive oil production SMEs in order to improve through a training programof olive mills´ and table olives producers´ technical competence andcompetitiveness.

(Project number: FOOD-CP-2004-505524)

Gateway to the European Union. www.europa.eu

Community Research & Development Information Service. www.cordis.europa.eu

ttz Bremerhaven. www.ttz-bremerhaven.de

ainia centro tecnológico. www.ainia.es

Confederación de Cooperativas Agrarias de España. www.ccae.es

AMITOM - Mediterranean International Association of the Processing Tomato.www.amitom.com

VIGNAIOLI PIEMONTESI S.C.A (Italy). www.vignaioli.it

Union of Agricultural Cooperatives in Peza (Greece). www.pezaunion.gr

ANFOVI - L’organisme de formation des Vignerons Indépendants (France).www.anfovi.com

Tecnoalimenti S.C.p.A. (Italy). www.tecnoalimenti.com

assessment and dissemination of strategies for the extraction of area/booklets/booklet olive... ·...

Documents