Athens May 23th,

2015

"Technological aspects to
produce high quality EVOO"

Servili Maurizio
Full Professor of Food Science and Technology
DSA3 Section of Food Technology and Biotechnology,
university of Perugia, Italy

QUALITY

NEW APPROACHES
TO
QUALITY
THE OLIVE OIL PRODUCTION

NEW
EXTRACTION TECHNOLOGY

OLIVE BY-PRODUCTS
OLIVE POMACES AND
VEGETATION WATERS (OVW)

New approaches to the olive

industry
Traditional
products

Traditional olives and extra-virgin olive oils with a quality

only guaranteed by merchandise limits
Innovative products

HIGH QUALITY PRODUCTS

HIGH QUALITY OLIVES AND EXTRA-VIRGIN OLIVE OILS.
HIGH QUALTIY SECONDARY PRODUCTS FOR THE ANIMAL FEEDING.
FUNCTIONAL FOODS CONTAINIG OLIVE BIOACTIVE PHENOLS.

Just two questions

1. What is an extra virgin olive oil

(EVOO)
from
a
marketable
point
of
view
?
It s a superior category virgin olive
oil
2. Nowadays, how does the merchandise
quality really protect the consumers of the
EVOO ?...
by
ensuring:

a low state of alteration;

the purity.

Origin area

Hedonistic quality

Quality
is the grade with which
a pool of intrinsic
characteristics satisfies
the human requirements
(UNI EN ISO 9000:2000)

Health

Safety

 MERCHANDISE QUALITY

EVOO

Virgin Olive Oil

0,8
20
2,50
0,22
0,01

2,0
20
2,60
0,25
0,01

>2
_
_
_
_

Md = 0
Mf > 0

Md 3,5
Mf > 0

Md > 3,5
Mf = 0

250
1,5
0,15
0,2

250
1,5
0.15
0,2

300
1,5
0,50
0,3

Acidic composition (%)

Miristic (%)

0,05

0,05

0,05

Linolenic (%)

1,0

1,0

1,0

Arachidic (%)
Eicosanoic (%)
Behenic (%)
Lignoceric (%)
Sum of the (E)-linolenics isomers (%)

0,6
0,4
0,2
0,2
0,05

0,6
0,4
0,2
0,2
0,05

0,6
0,4
0,2
0,2
0,05

0,5
0,1

0,5
0,1

Acidity(%)
Peroxide value(meq O
2/Kg)
K232
K270
K

EVOO ALTERATION STATE

Sensory Analysis : defects median value (Md)

Sensory analysis: "fruity" median value (Mf)

Alkyl esters:

Lampante Olive Oil

FAEE 35

Ethyl esters FAEE (mg/Kg)

Waxes(mg/Kg)
Saturated Fatty Acids in position 2 of the triglyceride (%)
Stigmastadyene
mg/kg
Difference ECN42 HPLC and ECN42 teoric calculation

Sterols composition (%)

colesterol (%)
Brassicasterol (%)

VOO GENUINENESS
0,5
0,1

Campesterol (%)

4,0

4,0

4,0

Stigma-sterol (%)
sitosterol (%)

< camp
93,0

< camp
93,0

_
93,0

-7-stigmas
- tenol(%)

0,5

0,5

0,5

Total sterols (%)

Eritrodiol and uvaol (%)

>1000
4,5

>1000
4,5

>1000
4,5

bioactive compounds

hydrophilic
tocopher
terpene
acids
fatty
squalene
acids
sterols
polyphenols
ols

CHEMICAL COMPOSITION OF EXTRA VIRGIN OLIVE OIL (EVOO)

PHOSPHOLIPIDS

GLYCERIDS

ALIPHATIC ALCOHOLS
AND TERPENIC

98,5-99,5 %
MAG 0.1-0.2
DAG%2-3 %

TRIGLYCERIDES
(TAG) 97-98%

OOO 40,7 %
POO 21 %
OLO 7,2 %
PLO

6,4 %

SOO 3,7 %
POP

2,8 %

MINOUR
COMPOUND
S

0,5-1,5%

VOLATILE
COMPOUNDS
PIGMENTS
STEROLS
HYDROCARBONS
TERPENIC ACIDS
TOCOPHEROLS
HYDROPHILICH
PHENOLS

OLEIC acid
It protects the gastric mucous, reduces the secretion of hydrochloric acid
and the risk of gastro-duodenal ulcers;
It inhibits the secretory activity of the pancreas (pancreatitis) and bile;
improves the emptying of bile from the gall bladder, prevents the formation
of gall stone, facilitates the absorption of fat-soluble vitamins and calcium,
exerts a laxative, in particular fasting; helps to correct the chronic
constipation;
It inhibits the synthesis and metabolism of cholesterol and total related to
lipoprotein LDL ("bad cholesterol"), triglycerides, the PA and the arachidonic
acid (n-6) that has pro-inflammatory activity;
It does not decrease the "good cholesterol" HDL, the "scavenger" that
prevents the accumulation of fat in the walls of arteries.

oleic acid

Claim of EFSA regarding the nutrition and health (UE Reg. 432/2012).
Replacing dietary saturated fats with unsaturated fats, contributes to the maintenance of normal
levels of blood cholesterol. Oleic acid is an unsaturated fat. This claim may be used only for food
with a high content of unsaturated fatty acids as specified in the indication "HIGH UNSATURATED
FAT" in the Annex to Regulation (CE) n. 1924/2006.
Claim of the Food and Drug Administration (FDA) for the use in the labels of olive oil and
products made from olive oil (USA 2004).
Scientific evidence suggests that: eating 2 tablespoons (23 grams) of olive oil per day, it is
possible to reduce the risk of coronary heart disease because of its content in
monounsaturated fatty acids (oleic ac. ). To achieve this beneficial effect, the olive oil has to
replace a similar amount of polyunsaturated fatty acids without increasing the amount of total daily
calories. Label: a portion of this product [name of the food] contains [x] grams of olive oil..

(%)
Myristic (C14:0)

0.0 - 0.1

Palmitic (C16:0)

7.0 20.0

Palmitoleic
(C16:1)

0.3 - 3.5

Eptadecanoic
(C17:0)

0.0 - 0.4

Eptadecenoico
(C17:1)

0.0 - 0.4

Stearic (C18:0)

1.0 - 4.0

Oleic (C18:1 -9)

47.0 84.0

Linoleic (C18:2 6)

3.0 21.0

Linolenic (C18:3
-3)

0.2 - 1.5

Arachidic (C20:0)

0.1 - 0.7

11-Eicosenoic
(C20:1)

0.1 - 0.1

fatty acids

High content of monounsaturated fatty acids.

Behenic (C22:0)

0.0 - 0.3
Optimal
ratio oleic ac. / linoleic ac.
Lignoceric (C24:0) 0.0 0.4
(7-11).

Optimal ratio 6: 3 (10:1).

VARIABILITY OF THE FATTY ACIDS (%) EVALUATED ON 243 EVOO

INDUSTRIAL PLANTS SAMPLES. Unpublished data.

Fatty acids

80.5 %

79,7 %

70,4 %
54.4 %

51.2 %

22.8 %

22.4 %

12.6 %
5.6 %

MUFA

OLEIC ACID

SFA

PUFA

VARIABILITY OF THE FATTY ACIDS (%) EVALUATED ON 20

EVOOS FROM GREECE.
Fatty acids

74.6
69.5
62.8

73.5
67.5
61.1

21.6
18.3

17.9
12.2

15.6
8.0
MUFA

Oleic acid

SFA

PUFA

16000

(mg/kg)

14000
12000

STEROLS

10000
8000
6000
4000

PA
LM

d
se
e
e

G
ra
p

RA
PE

O
LI
SU
VE
N
FL
O
SU
W
CO
N
ER
FL
RN
H
O
II G
W
ER
H
O
H
LE
II G
IC
H
LI
N
O
LE
IC
PE
AN
U
T
SO
N Y
BE
A

PA
LM
IS
TI
CO
CO
N
CO
U
T
CO
A
BU
TT
ER

2000

Claim of 'EFSA regarding the nutrition and health (EU Reg. 432/2012).
The sterols / stanols ratio contributes to the maintenance of normal levels of blood cholesterol. A
claim must be accompanied by information to the consumer that the beneficial effect is obtained
with
the daily intake of at least 0.8 g of plant sterols / stanols.
In vivo studies have shown that phytosterols (-sitosterol, in particular) reduce the concentration
of total cholesterol and LDL cholesterol, reduce the growth and induce apoptosis, tumor cells of the
prostate; are also effective in the natural treatment of benign prostatic hyperplasia (Klippel et al,
1997; Carbin et al, 1990; Wilt et al., 1999; Von Holtz et al., 1998; Jones et al., 1997; Law 2000; Plat

7000

(mg/kg
)

6000
Squalene

5000

4000

3000

2000

1000

Extravirgin
olive oils

Refined
olive oils

Pumpkin
seed oils

Rise
seed oils

Seed oils

Squalene is a hydrocarbon intermediate of cholesterol biosynthesis. In vivo and in vitro studies have
shown that stimulating the acyl-coenzyme A, it regulates the process of absorption, synthesis, esterificate
and eliminate cholesterol;
It shows antioxidant activity similar to that of trans retinol; increases efficiency of statins in reducing
cholesterol; it reduces the risk of cancer. (Chan et al. 1996; Newmark 1999; Martin-Moreno et al. 1999;
Trichopoulou et al. 1995; Landa et al. 1994; Khono et al. 1995; Kelly et al., 1999; Owen et al., 2000; Cornelli
et al., 2003; Reddy et al., 2009; Naziri et al, 2013).

10000 (mg/kg)

Triterpenic acids

oleanolic acid

9000
TERPENIC ACIDS

8000
7000
6000
5000
4000
3000
2000
1000
0

Extraavirgin olive
oil

Virgin olive oil

Olive pomace oil

Triterpenic acids. In vitro and in vivo studies have demonstrated that acids pentacyclic triterpenes
(oleanolic, ursolic, maslinic and betulinic) show different biological activities: anti-inflammatory,
hepatoprotective, anticancer, antiviral, anti-HIV, anti-microbial, antifungal, anti-diabetic, gastroprotective
and anti-hyperlipemia (Liu, 1995; Xu, Zeng, Wan, e Sim, 1996; Dedoussis et al., 2004, Kalogeropoulus

et al., 2010).

TOCOPHEROLS

Claim of EFSA regarding the nutrition and health (EU Reg. 432/2012).
Vitamin E helps to protect cells from oxidative stress.

Tocopherols. The -tocopherol, in particular, in addition of having vitamin activity (vitamin E or

anti-sterility), are important antioxidants: they protect cells and tissues from oxidative and
inflammatory processes that are at the basis of aging, degenerative diseases and cancer (Baldioli
et al., 1996; Servili et al., 2009; Tsimidou et al., 2010; Tsimidou et al., 2012)

VARIABILITY OF THE -TOCOPHEROL (mg/kg) EVALUATED ON 882 EVOO

INDUSTRIAL PLANTS SAMPLES. Unpublished data.
-TOCOPHEROL

EVOO
538.0

266.7

90.5

VARIABILITY OF THE -TOCOPHEROL (mg/kg) EVALUATED ON

20 EVOOS FROM GREECE.
-TOCOPHEROL

545.0

306.
2

177.0

EVOO POLYPHENOLS
Flavonoids

Lignans

Phenolic Acids
R1
COOH

HO
R2
R1

COOH

HO
R2
7'

2'

1'

3'

HO 4'

8'

5'

Secoiridoids

COOCH3

5
9

10

6'

1'

HO

5'
2'

CH2OH
4' OH
OH
3'

R = H: ligstroside
R = OH: oleuropein

EVOO PHENOLIC COMPOSITION

Phenolic acids and derivatives: p-hidroxybenzoic, ferulic, cinnamic, benzoic,
(acetoxyethyl)-1,2-dihydroxybenzenic.

Flavones: apigenin, luteolin

Phenolic alcohols:

3,4 DHPEA ((3,4-Dihdroxyphenyl)

ethanol)
p-HPEA
((p-Hydroxyphenyl) ethanol)
(3,4-Dihdroxyphenyl) ethanol-glucoside

Secoiridoids:

DHPEA-EDA (Dialdehydic form of decarboxymethyl elenolic acid linked to 3,4-DHPEA)

p-HPEA-EDA (Dialdehydic form of decarboxymethyl elenolic acid linked to

p-HPEA)
3,4
DHPEA-EA (Oleuropein aglycon)
Ligstroside aglycon
Oleuropein
p-HPEA-derivative

Dialdehydic form of oleuropein aglycon

ialdehydic form of ligstroside aglycon

Lignans:

(+)-1-Acetoxypinoresinol
(+)-Pinoresinol

Hydroxy-isocromans: Verbascoside

4-

3'
HO 4

1'
6'
O

7'

5'

7
6
9

10

3'

8'

HO 4'

COOCH3
4

1'

8'

6'
O
5'

HO 4' 5'
3

OH

7'

3'
4

10

1 O

2'

HO

HO

6'

OH
HO 4' '
(p-HYDROXYPHENYL)
ETHANOL
5

6'
O

O
7

Hydrophilic Polyphenol
COOCH3

4
5

1 O

2'
3'

HO 4'

8'

1 O

DIALDIALDEHYDIC FORM OF
DECARBOXYMETHYL
ELENOLIC ACID LINKED TO p-HPEA
(p-HPEA-EDA) = OLEOCHANTAL

8'

7'

1'

(p-HPEA-EA)
2'

7'

1'

LIGSTROSIDE AGLYCON

3'

2'

1'
6'
O

7'

(3,4-DHPEA-EA)

8'

HO

5'

(p-HPEA = tyrosol)

OLEUROPEIN AGLYCON

6
9

10

1 O

2'
3'

1'

8'

6'
OH
HO 4'
5'
(3,4-DIHYDROXYPHENYL)
ETHANOL
(3,4-DHPEA = hydroxityrosol)

8
O
DIALDIALDEHYDIC FORM OF DECARBOXYMETHYL
ENOLIC ACID LINKED TO 3,4-HPEA (3,4 DHPEA-EDA)

(+)-1- ACETOXYPINORESINOL

7'

(+)-1-

OLIVE BIOPHENOLS (OBP) PHARMACOLOGICAL PROPERTIES.

Obied et al., 2012

1. Antioxidant :

OBP have RONS scavenging, reducing power, and metal chelating activities, induce
endogenous antioxidant enzymes such as catalase, superoxide dismutase, quinone reductase, glutathione
peroxidase, glutathione reductase, glutathione S-transferase, and g-glutamylcysteine synthetase.

2. Anti-inflammatory :

OBP act against cardiovascular diseases (CVD) and some types of cancer by
inhibition of proinflammatory enzymes, phosphoinositide 3 kinase, tyrosine kinases, and downregulation of various
proinflammatory cytokines, tumor necrosis factor alpha, interleukins including and monocyte chemotactic protein-1.

OBP have: 3.1.

many
reported pharmacological
3.Cardiovascular
Blood pressure-antihypertensive
activities; 3.2. Platelet and endothelial function;
activities based on preclinical (in vitro, ex
4.Immunomodulatory
: vivo)
OBP have studies
been shown to in
modulate
immune function,
particularly
vivo,
and
in
addition
to
inflammatory processes associated with the immune system.
few clinical: 5.1.
studies.
activities
5. Gastrointestinal
GastroprotectiveThese
effects; 5.2. Modulation
of digestive enzymes.
6. suggest
Endocrine : high
6.1. Antidiabetic
effects6.2. Osteoprotective
effects6.3.
Other endocrine effects.
potential
for the
prevention
7. Respiratory
: OBP antioxidant and
properties against lung diseases.
and treatment
ofanti-inflammatory
diseases
and the
8.Autonomic : 8.1. Cholinergic effects; 8.2. Adrenergic effects.
promotion of human health.
3.3. Atherosclerosis; 3.4. Other cardioprotective properties.

9. Central nervous system :

9.1. Neuroprotective effects.; 9.2. Analgesic and antinociceptive

effects;
9.3. Behavioral effects.

10. Antimicrobial and chemotherapeutic :

10.1. Antibacterial properties. 10.2.

Antifungal properties.
10.3. Antiviral properties. 10.4. Antiprotozoal and antiparasitic activities.
11. Anticancer and chemopreventive : Biophenols can directly control cell growth at
different stages of carcinogenesis via inducing apoptosis or inhibiting proliferation by diverse mechanisms.

HEALTH EFFECTS OF EVOO PHENOLIC COMPOUNDS

Recently , the NDA Panel of the European Food Safety Authority (EFSA), has allowed
the health claim to the VOO phenolic compounds.

there is evidence of a cause and effect between the consumption of

olive oil polyphenols (standardised by the content of hydroxytyrosol
and its derivatives) and protection of LDL particles from oxidative
relationship damage

Inhibition of oxidation of LDL

cholesterol.

According to the Panel, 5 mg of hydroxytyrosol and its derivatives should be taken

every day, provided by a moderate consumption of olive oil, nothing that some
olive oils have a concentration too low to make this amount of polyphenols
remaining in the context of a balanced diet.
(EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA). Polyphenols in olive
related health claims. EFSA Journal 2011; 9 (4): 2033).

VARIABILITY OF THE POLYPHENOLS (mg/kg) EVALUATED ON 737 EVOO

INDUSTRIAL PLANTS SAMPLES. Unpublished data.

Hydrophilic Polyphenol

EVOO
995.
5

544.
5

168.
0
total
phenols

788.7

385.
0

196.
4
93.1

102.
5
derivatives
of
hydroxytyro
sol

46.2
derivative
s of
tyrosol

sum of
lignans

VARIABILITY OF THE POLYPHENOLS (mg/kg) EVALUATED ON 20

EVOOS FROM GREECE.

Hydrophilic Polypheno

707
.1
525.
0
289.0
Total
phenols

621
.3
439.
2
246.
3 53.5
derivatives of
hydroxytyrosol

derivatives of
tyrosol

42.7
lignans

"sensory" molecules

carotenoids
chlorophylls
and
oleic compounds
acid
hydrophilic
volatile
phenols
phaeophytins

CHEMICAL AND ENZYMATIC PATHWAY INVOLVED IN

EVOO VOLATILE COMPOUNDS GENESIS
volatile compounds

LOX pathway

Homolytic cleavage
of
13hydroperoxides
Conversion of
aminoacids

EVOO
volatile
compoun
ds

Fatty acids
metabolism

Sugar
Autoxidati
fermentati on
on

LIPOXIGENASE (LOX) PATHWAY INVOLVED IN C6 AND C5 EVOO VOLATILES

PRODUCTION. Angerosa et al., 2004
hexanal ADH hexan-1-ol

linoleic acid

LOX

13-hydroperoxides

HPL

isomerase

AAT

hexyl acetate

ADH
2-hexen-1-ol ( E )
2-hexenal( E )

3-hexenal (Z)

linolenic acid
13-alkoxy radical

ADH

AAT

3-hexenyl acetate ( Z )
3-hexen-1-ol ( Z )
pentene radical

pentene dimers

2-penten-1-ol
1-penten-3-ol

2-pentenal
1-penten-3-one

volatile compounds

RELATIONSHIPS BETWEEN EVOO VOLATILE

SENSORY PERCEPTIONS. Angerosa et al., 2004.
Compound

Odour Quality

COMPOUNDS

AND

HUMAN

References

Aldehydes
volatile compounds
sweet, pungent, floral
Servili et al., (2001); Reiners e Grosch (1998).
propanal
green, apple, cut grass
Servili et al., (2001); Morales et al., (1997).
hexanal
green, apple, floral
Servili et al., (2001); Morales et al., (1997).
(E)-2-pentenal
green, pleasant
Morales et al., (1997).
(Z)-2-pentenal
bitter, almonds, green, green apple-like, fatty, bitter almond
Servili et al., (2001); Guth e Grosch, (1991); Morales
(E)-2-hexenal
et al., 1997;Reiners e Grosch (1998).
like, cut grass
green, fruity, sweet
Morales et al., (1997).
(Z)-2-hexenal
artichoke, green, floral
Morales et al., (1997).
(E)-3-hexenal
Servili et al., (2001); Morales et al., (1997) Ullrich e
green leaves, grassy, green, apple-like, leaf-like, cut grass
(Z)-3-hexenal
Grosch, (1988); Reiners e Grosch (1998).
cut
grass
Servili et al., (2001);
2,4-hexadienal
Alcohols
ethanol
hexan-1-ol
(E)
-3-hexen-1-ol
(Z)-3-hexen-1-ol
(E)
-2-hexen-1-ol

alcoholic, ripe apple, floral

fruity, aromatic, soft, cut grass
fruity, fatty, pungent, cut grass

Servili et al., (2001); Reiners e Grosch (1998)

Ramstad e Nestrick, 1980; Servili et al., (2001);
Morales et al., (1997)
Servili et al., (2001); Bedoukian, (1971).
Morales et al., (1997); Bedoukian, (1971) Reiners e

banana, leaf-like, green-fruity, pungent

green, grassy, fruity, fatty, pungent

Grosch (1998).
Morales et al., (1997); Bedoukian, (1971).

Esters
ester
Servili et al., (2001).
methyl acetate
sweet, strawberry, apple
Morales et al., (1997).
ethyl propanoate
fruity
Reiners e Grosch (1998).
ethyl isobutyrate
fruity
Reiners e Grosch (1998).
ethyl 2-methylbutyrate
fruity
Reiners e Grosch (1998).
ethyl 3-methylbutyrate
Ramstad e Nestrick, (1980); Servili et al., (2001);
green-banana, fruity, green, green leaves, floral, ester
(Z)-3-hexenyl acetate

hexyl acetate
3-methylbutyl acetate

Guth e Grosch, (1991); Morales et al., (1997).

sweet, fruity, floral

banana

Servili et al., (2001); Morales et al., (1997).

Morales et al., (1997).

volatile compounds

139
.7
76.
8
27.
3
18.
2

aldehydes

5.5
1.2

alcohols
mg/kg

mg/kg

VARIABILITY OF THE VOLATILE COMPOUNDS (mg/kg) EVALUATED ON 227 EVOO

INDUSTRIAL PLANTS SAMPLES. Unpublished data.

3.
5

0.8
0.02

esters

SENSORY PROPERTIES OF EVOO PHENOLIC COMPOUNDS

Andrewes et al., 2003.

- Tyrosol (p-HPEA):
sticking astringency, not bitter (e.t.t*.: 4.4-18)
- Hydroxityrosol (3,4-DHPEA-EDA):
astringent, bitter, burning/stinging/numbing mostly on tongue (e.t.t*.: 0.41.6)
- Oleuropein aglycon (3,4-DHPEAEA):
very bitter, very astringent (e.t.t*.: 0.050.2)
- Ligstroside aglycon (p-HPEAEA):
astringent, bit burning (pungent), bitter (e.t.t*.: 0.050.2)

hydrophilic phenols

- Oleochantal (p-HPEA-EDA):
strong burning (pungent), mostly at the back of throat, slightly bitter, astringent
(e.t.t*.: 0.4-1.6)
* e.t.t.= estimated taste threshold (mM)

QUALITY

NEW
NEW APPROACHES
TO
THE
OLIVE OILTECHNOLOGY
PRODUCTION
EXTRACTION

NEW VOO
EXTRACTION TECHNOLOGY

OLIVE BY-PRODUCTS
OLIVE POMACES AND
VEGETATION WATERS (OVW)

HOW CAN WE PRODUCE A

ENVIRONMENT, AGRONOMICIC
PRACTICES
GEOGRAPHICAL LOCATION, CULTIVAR,
IMPLANT SYSTEM OF THE ORCHARD,
IRRIGATION

HIGH QUALITY
EVOO

EXTRACTION TECHNOLOGY
CRUSHING, MALAXATION, OIL EXTRACTION,
VALORIZATION OF EVOO BY-PRODUCTS,
PRODUCTION OF FUNCTIONAL FOODS

FATTY ACIDS COMPOSITION (%) OF EVOOs OF SEVERAL ITALIAN OLIVE CVs.

GROWN IN ITALY. Unpublished data.

Biancolilla Cerasuola Coratina Frantoio Leccino Peranzana

Palmitic
11,61
9,86
12,36
12,34
13,23
12,27
Palmitoleic
0,52
0,22
0,51
1,01
1,25
0,8
Margaric
0,12
0,02
0,08
0,01
0,01
0,07
Heptadecenoic
0,2
0,03
0,05
0,02
0,09
0,11
Stearic
2,23
2,54
2,1
1,65
1,53
1,86
Oleic
74,10
76,83
75,43
75,77
77,96
76,45
Linoleic
9,81
9,34
7,94
8,04
4,54
7,21
Linolenic
0,69
0,51
0,72
0,55
0,68
0,58
Arachidic
0,39
0,36
0,31
0,29
0,28
0,33
The fatty acids composition was evaluated according to the current official method (UE Reg., 1989, 2003)
Values are the mean of five different VOO samples (n = 5) standard deviations.

FATTY ACIDS COMPOSITION (%) OF EVOOs OF SEVERAL OLIVE CVs. GROWN

IN TWO DIFFERENT GEOGRAPHICAL AREAS. Inglese et al., 2010.

Arbequina

Coratina

Leccino

Argentina

Italy

Argentina

Palmitic

20,66

13,60

16,29

12,36

17,39

13,23

Palmitoleic

3,69

1,10

0,67

0,51

1,16

1,25

Margaric

0,04

0,09

0,05

0,08

0,05

0,01

0,20

0,20

0,08

0,05

0,09

0,09

Stearic

1,53

2,30

1,77

2,1

1,71

1,53

Oleic

53,4

69,5

71,5

75,4

68,5

78,0

Linoleic

18,72

11,60

7,99

7,94

9,19

4,54

Linolenic

1,16

0,70

1,27

0,72

1,43

0,68

Arachidic

0,29

0,40

0,37

0,31

0,33

0,28

Heptadecenoic

Italy

Argentina

Italy

The fatty acids composition was evaluated according to the current official method (UE Reg., 1989, 2003)
Values are the mean of five different VOO samples (n = 5) standard deviations.

PHENOLIC COMPOSITION (mg/kg) OF EVOOs OF SEVERAL ITALIAN OLIVE Cvs.

GROWN IN ITALY. Servili et al., 2004.

Coratina

Leccino

1,80,8
7,91,8
3,20,1 12,31,6
390,611,0 67,6 15,5
168,45,2 12,5 1,2
(+)-1-Acetoxypinoresinol27,51,1
4,9 0,5
(+)-Pinoresinol
11,70,8 28,41,7
3,4-DHPEA-EA
287,19,2 47,2 15,0
Total phenols
890,371,2180,8 21,
8
3,4-DHPEA
p-HPEA
3,4-DHPEA-EDA
p-HPEA-EDA

Moraiolo Carolea
1,20,0
2,60,0
291,39,1
83,21,0
16,51,0
24,80,3
174,36,8

593,842,1

Canino

Frantoio

5,71,23
2,10,2
1,20,2
4,11,02 1,70,2
1,60,8
273,56,2 264,15,9 133,03,0
163,38,4 104,55,2
48,60,6
9,41,0
28,01,2
12,11,0
13,21,2
11,7 0,9
8,10,9
124,93,1 109,26,3 97,71,0
605,350,1 510,238,2 302,220,3

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al., 1992.
Values are the mean of five different VOO samples (n = 5) standard deviations.

PHENOLIC COMPOSITION (mg/kg) OF EVOOs OF SEVERAL SPANISH OLIVE Cvs.

GROWN IN SPAIN. Unpublished data.

Arbequina

Cornicabra

Hojiblanca

3,4-DHPEA-EA

1,60,4

6,01,0

2,41,3

p-HPEA

2,00,6

12,61,0

7,7 0,5

3,4-DHPEA-EDA

95,17,0

142,7 5,4

120,63,2

p-HPEA-EDA

49,81,8

50,72,4

27,31,2

(+)-1-Acetoxypinoresinol

16,91,1

5,5 0,5

13,7 0,9

6,20,4

9,0 0,9

6,5 0,3

44,51,2

138,6 6,0

95,32,1

216,115,2

365,025,3

273,5 18,2

(+)-Pinoresinol
3,4-DHPEA-EA
Total phenols

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992.
Values are the mean of five different VOO samples (n = 5) standard deviations.

PHENOLIC CONCENTRATION (mg/kg of total phenols) OF OLIVES OF FOUR

TYPICAL ITALIAN Cvs. GROWN IN ITALY. Servili et al., 2007.
300,0

Frantoio

Coratina

Leccino

Moraiolo

250,0

200,0

150,0

100,0

50,0

0,0
07/20 08/08 08/24 09/11

09/26 10/10

10/24 11/07 11/13

11/27

PHENOLIC COMPOSITION (mg/kg) OF EVOOs OF SEVERAL OLIVE Cvs. GROWN

IN THE SAME AREA (SICILY, ITALY) WITH DIFFERENT ORCHARDS IMPLANT
SYSTEMS. Unpublished data.

super
intensive

traditional
Nocellara
1,51,8
p-HPEA
8,81,6
3,4-DHPEA-EDA
204,915,5
p-HPEA-EDA
52,41,2
(+)-1-acetoxypinoresinol 31,7 0,5
(+)-pinoresinol
119,2 1,7
3,4-DHPEA-EA
76,715,0
Total Phenols
495,221,8
3,4-DHPEA

Biancolilla
0,70,1
1,9 0,3
103,03,7
41,7 3,2
46,6 3,3
103,1 6,5
64,4 4,2
361,3 9,8

Cerasuola
3,0 0,3
8,6 0,7
202,9 5,8
83,8 6,5
29,9 2,2
165,210,8
106,0 8,8
599,516,6

Arbequina
1,60,1
15,0 0,4
19,41,2
28,11,0
15,61,7
5,3 0,3
44,4 0,8
129,4 2,6

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al., 1992.
Values are the mean of five different VOO samples (n = 5) standard deviations.

FATTY ACIDS COMPOSITION (%) OF EVOOs FROM OLIVE TREES (LECCINO Cv. )
GROWN UNDER FULL IRRIGATION (FI), DEFICIT IRRIGATION (DI) OR SEVERE DEFICIT
IRRIGATION (SI). Servili et al., 2007.

FI

DI

SI

Myristic

0.01 0.001a

0.01 0.001b

0.01 0.001a

Palmitic

14.0 0.8a

13.7 1.5 a

14.6 1.8 a

Palmitoleic

1.4 0.2a

0.8 0.1b

1.2 0.3a

Margaric

0.04 0.01a

0.03 0.008a

0.03 0.005a

Eptadecenoic

0.1 0.02a

0.04 0.01b

0.1 0.01ab

Stearic

1.0 0.6a

2.1 1.1 a

1.0 0.7a

Oleic

77.3 1.2 a

76.1 2.7a

76.1 2.1 a

Linoleic

4.9 0.3a

5.9 0.5b

5.9 0.5b

Linolenic

0.6 0.07a

0.7 0.06a

0.6 0.1a

Arachic

0.3 0.02a

0.3 0.04a

0.3 0.04a

Eicosenoic

0.2 0.03a

0.2 0.02a

0.2 0.04a

Behenic

0.1 0.04a

0.1 0.05a

0.02 0.02b

Lignoceric

0.1 0.01a

0.05 0.009a

0.1 0.01a

The fatty acids composition was evaluated according to the current official method (UE Reg., 1989, 2003)
Values are the mean of five different VOO samples (n = 5) standard deviations. Different letters indicate
significant differences (P < 0.05) between irrigation treatments within each year.

PHENOLIC COMPOSITION (mg/kg) OF EVOOs FROM OLIVE TREES (LECCINO Cv. )

GROWN UNDER FULL IRRIGATION (FI), DEFICIT IRRIGATION (DI) OR SEVERE DEFICIT
IRRIGATION (SI) IN 2003 AND 2004. Servili et al., 2007.

FI

DI

SI

3,4 -DHPEA

2.3 0.3 a

2.4 0.7 a

3.5 0.9 a

p-HPEA

7.7 0.8 a

7.4 1.4 a

3.1 0.4 b

3,4 -DHPEA-EDA

130.1 25.9 a

291.7 40.3 b

318.5 39.1 b

p-HPEA-EDA

80.2 11.8

128.2 25.6 b

129.9 24.1 b

4.4 0.8 a

3.8 0.9 a

6.0 2.6 a

(+)-1-pinoresinol

44.8 6.7 a

49.7 5.6 a

47.2 7.9 a

3.4 DHPEA-EA

114.1 19.2 a

139.6 20.1 a

185.5 23.0 b

Total phenols

383.6

35.0 a

622.8 52.1 a

693.7 51.9 b

(+)-1-acetoxypinoresinol

e phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992.

Values are the mean of five different VOO samples (n = 5) standard deviations. Different letters indicate significant differences (P
<0.05) between irrigation treatments within each year.

VOLATILE COMPOSITION (g/kg) OF EVOOs FROM OLIVE TREES (LECCINO Cv. )

GROWN UNDER FULL IRRIGATION (FI), DEFICIT IRRIGATION (DI) OR SEVERE DEFICIT
IRRIGATION (SI).
Servili et al., 2007. The volatile compounds were determined in duplicate by HS-SPME-GC-MS as reported by Servili et al., 2001.

FI

DI

SI

1-penten-3-one

855.8 55.0a

869.3 43.0a

786.0 117.0 a

pentanal

70.1 32.0a

117.2 23.0a

137.9 49.2a

2-pentenal

88.3 12.1 a

72.0 5.4a

57.5 8.4b

hexanal

492.1 94.0a

500.1 75a

372.1 52.0a

trans-2-hexenal

33091 3644.0a

27676.3 2286b

22632.62364.4c

nonanal

65.3 13.0 a

77.0 23.3 a

71.1

1-butanol

51.8 13.0 a

21.0 3.6a

20.1 4.6a

2-methyl-1-butanol 341.4 43.0a

308.9 23.1 a

307.5 21.6 a

18.4 a

1-pentanol

33.9 0.7a

23.5 2.4b

27.8 5.0b

1-penten-3-ol

615.4 136.1 a

769.0 50.0a

706.0 124.8 a

1-hexanol

128.4 18.3 a

126.7 26.0a

177.9 38.3 b

(E)-2-hexen-1-ol

7915.7 102.1 a

2519.2 42.5 b

1834.0 202.1 c

(Z)-2-hexen-1-ol

562.8 65.2 a

648.2 29.0a

786.3 114.0 b

(E)-3-hexen-1-ol

13.4

0.4a

8.7 0.4b

9.4

1.2 b

(Z)-3-hexen-1-ol

164.9 41.2 ab

202.4 25.0a

137.8 21.8 b

1-hexen-3-ol

11.1 2.0ab

7.7 1.2 a

13.9 3.5b

9.7 0.5a

43.4 1.6 b

hexyl acetate

9.8

0.6a

PROCESS CONTROL ACCORDING TO THE NATIONAL QUALITY SYSTEM

HARVESTED AT A SUITABLE DEGREE OF
MATURATION ACCORDING TO THE CULTIVAR.

OLIVES

Crushing

WORKED AT AN OPTIMAL SANITARY STATE.

DIFFERENTIATED EFFECT ON THE
CONSTITUTIVE PARTS OF THE DRUPES BY:

Malaxation

TEETH CRUSHER

BLADE
Control
of CRUSHER
TIME, O2
and
CRUSHERLEVEL
WITH A LOW
TURNS NUMBER
TEMPERATURE.

Extraction

STONING

DOUBLE STOKER

THREE-PHASES
CENTRIFUGATION WITH
REDUCED WATER ADDITION
concentrates
EVOO dried
anddestoned
purified extracts
pomaces:
for:

HIGH QUALITY
EVOO
EVOO
OVWs

bio-energy
feed industries
and compost
as integrators;
production;
cosmetic
zootechnic
industries
industries
as as
natural
integrative
antioxidants;
feeding:
EVOOofindustries
supplement
high biological
for increasing;
value compounds
polyphenols
acids and
concentration;
polyphenols).
POMACES the(fatty
food industries for producing functional products

HOW CAN WE PRODUCE A

ENVIRONMENT, AGRONOMICIC
PRACTICES
GEOGRAPHICAL LOCATION, CULTIVAR,
IMPLANT SYSTEM OF THE ORCHARD,
IRRIGATION

HIGH QUALITY
EVOO

EXTRACTION TECHNOLOGY
CRUSHING, MALAXATION, OIL
EXTRACTION, VALORIZATION OF EVOO
BY-PRODUCTS, PRODUCTION OF
FUNCTIONAL FOODS

OLIVE STORAGE

Evolution of polyphenols content (resorcin ppm), bitter and fruity

sensations in VOO extracted from olives at time 0 and stored for
different periods (days)

OLIVE STORAGE

NOT STORED FOUR DAYS STORAGETEN DAYS STORAGE

MEAG + EEAG

6,5

8,0

71,6

MEAG

5,2

5,7

47,0

EEAG

1,3

2,3

24,6

0,2

0,4

0,5

EEAG/MEAG

ALCHIL ESTERS COMPOSITION ACCORDING

TO THE TIME (DAYS) OF OLIVES STORAGE.

How could these process innovations be oriented

?
Target

improvement of EVOO
health and sensory properties by

controlling of the endogenous oxidoreductases

activity
during the EVOO mechanical extraction process.
crushing with a differentiated
effect on the constitutive parts
of the drupes

Controlling of O2 level
and temperature during
the malaxation

THE
OIL
DISTRIBUTION
COSTITUTIVE
IN THE
OIL OLIVE
CONTENT
IN THE
FRUIT
FRUIT
PARTS

Cryo-SEM in Coratina Cv. olive.Servili et al.,

2006

Fruit outer
surface, in
prespective

Seed
Seed
2,0-4,0%
35-40
1-2 %
%
Stone
11,0-24,5
%
Mesocarp
70,0-81,5 %
Epicarp
Epicarp +
+ Mesocarp
Mesocarp
98-99
30 % %
Epicarp
1,5-3,5
%

surface Cryo-fracture
Internal tissues
perfectly preserved with
parenchyma cells oil
droplets containing

Detail: On the cross cryo-fracture plane,

under the external surface there is a
compact cuticle layer and very close, a
single layer of epidermic and the first
three layers of parenchyma cells.

DISTRIBUTION OF THE MOST IMPORTANT ENZYMATIC ACTIVITY IN THE FRUIT

Seed

PG: 66.5 83.0

%
PME:
98.0 99,5 42.5
% - 82.0
LPO:
% - 99,5
PPO: 98.0
% POD: 34,5
56,5 %

LPO: lipoxygenses

Stone
Mesocarp
Epicarp

PG: 8.0 12.0

% 0.5 2.0
PME:
% LPO: 12.5 PPO:50,5
0.5 %
2
% POD: 34,5
76,5 %
LPO: 3,5 PG:15,5
9.0 %24.5
%

PPO: polyphenoloxidases POD: peroxydases

PME: pectin-methyl-esterases PG: polygalacturonases

EA
p
-H
P
N
EA

Li
zh
g
Ve
en
st
ro
rb
id
si
as
e
d
co
e
si
g
lu
d
e
co
si
d
D
(+
e
O
em
l
)e
et
u
1
ro
-A
h
yl
p
ce
ei
ol
to
n
e
u
xy
r
op
p
in
ei
or
n
(+
es
)i
no
P
in
l
or
es
in
ol

3,
4D
H
P

EA
p
-H
P
N
EA

Li
z
h
g
V
en
st
er
ro
id
b
si
as
e
d
co
e
si
gl
d
u
e
co
si
d
D
(+
e
O
em
l
)eu
et
1
ro
-A
h
yl
p
ce
ei
ol
to
n
e
xy
u
ro
p
p
in
ei
or
n
(+
es
)in
P
ol
in
or
es
in
ol

3,
4D
H
P

Phenols
distribution
(mg/100g)
in the
Phenols
distribution
(%) in the different
constitutive
parts of
the fruit. Servili parts
et al., 2007 of the fruit
different constitutive
CORATINA Cv.
FRANTOIO Cv.

SEE
seed
D

STONE
stone

PUL
pulp
P

Total activities (U/mg d.w.) and corresponding contribution (%) of

the constitutive parts of the olives. Servili et al., 2007.

enzymatic activity (U/mg d.w.)

FRANTOIO
Cv.

% of contribution to the
total enzymatic activity
CORATINA
Cv.

0.25

10
20
30
0.75

0.00
40
50
60
70 minutes
0.00

0.50

0.25

10
20

MCounts
1.00

30
40

3-Esen-1-olo,
(Z)2-Esen-1-olo,
(Z)-

3-Esenil acetato, (Z)-

SEED
2-Esenale, (E)-

1-Penten-3-olo

Hexanale

OF CRUSHED OLIVE SEED AND PULP.

1-Pentanolo
Esil acetato
2-Penten-1-olo, (E)-Esanolo
3-Esen-1-olo,1(E)-

0.75

Pentanale

MCounts
1.00

2-Pentenale, (E)-

2-Esenale, (E)1-Pentanolo
Esil acetato
3-Esenil acetato, (Z)1-Hexanol
3-Esen-1-olo, (E)2-Esen-1-olo, (Z)-

Esanale

0.50

Acido acetico estere butilico

1-propanolo
2-Pentenale, (E)1-Penten-3-olo

Pentanale

HS/GC-MS CHROMATOGRAM OF THE VOLATILE COMPOSITION

Servili et al., 2007

PULP

50
60
70 minutes

Volatile compounds produced by LOX pathwhay from crushed

olive seed and pulp in Frantoio and Coratina Cvs (g/g). Servili et
al., 2007.

The volatile compounds were determined in duplicate by HS-SPME-GC-MS as reported by Servili et al., 2001. Results are mean value
of three indipendent determinations standard deviation

centrifugazon
centrifugation
es

Reduced pastes
water washing

malaxation

Control of time,
temperature, and
O2 concentration

crushing

Differentiated
effect on the
several costitutive
parts of the fruit

ION
T
A
OV
ING
INN RUSH
IN C

DIFFERENTIATED EFFECT ON THE

CONSTITUTIVE PARTS OF THE DRUPES

STONING
TEETH CRUSHER
BLADE CRUSHER
CRUSHER WITH A LOW TURNS NUMBER
DOUBLE STOKER

HAMMER CRUSHER

BLADE CRUSHER

DISC CRUSHER

DOUBLE STOKER

PHENOLIC COMPOSITION (mg/kg) OF EVOOs (FRANTOIO Cv.) OBTAINED BY

DIFFERENT CRUSHING METHODS. Servili et al., 2007.

200,0

60,0

150,0

40,0

100,0

20,0

50,0

3-4 DHPEA-EA
p-HPEA-EDA
3-4 DHPEA-EDA
Sum of phenolic fractions

First ripening stage

0,0

Stoned

p-HPEA

Blade +
Pre-crusher

3,4 DHPEA

Hammer

0,0

Low turn number

80,0

Blade +
Pre-crusher

250,0

Hammer

100,0

Stoned

300,0

Low turn number

120,0

Second ripening stage

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992. Results are mean value of three indipendent

LIGNANS COMPOSITION (mg/kg) OF EVOOS (FRANTOIO Cv.) OBTAINED BY

DIFFERENT CRUSHUNG METHODS. Servili et al., 2007.
50,0
45,0
40,0
35,0
30,0
25,0
20,0
15,0
10,0

Second ripening stage

Results are mean value of three indipendent determinations standard deviation

Stoned

Blade
Pre-crusher

Hammer

Low turn number

First ripening stage

Stoned

Low turn number

Pinoresinol

Pre-crusher

Acetoxypinoresinol

Blade +

0,0

Hammer

5,0

VOLATILE COMPOSITION (mg/kg) OF EVOOs (FRANTOIO Cv.) OBTAINED BY

DIFFERENT CRUSHING METHODS. Servili et al., 2007.

Hammer

Blade
Low turn
Stoned
+ pre-crusher number crusher

aldheydes
Pentanal

236,5 4,0

273,4 2,1

Hexanal

280,0 2,9

511,4 35,7

17,9 1,0

66,5 6,7

553,7 0,3

579,6 5,3

2-Pentenal (E )

10,7 0,3

13,2 0,9

94,8 1,8

16,6 1,0

2-Hexenal (E )

43600,6 327,0

44718,9 207,8

39811,6587,4

52228,1521,0

19,4 0,1

42,0 3,5

341,6 14,4

88,9 5,4

0,0 0,0

0,0 0,0

158,2 10,0

72,0 3,7

167 5,2

94,5 4,7

23,3 0,7

62,6 1,4

166 11,3

91,4 5,1

52,4 3,5

104 7,4

2,4-Esadyenal (E,E )
2-Heptenal (E )

alcohols
1-Pentanol
2-Penten-1-ol (E )
1-Penten-3-ol

960,3 53,2

1-Hexanol

1788 57

899 43,3

522 49,2

300 28,2

2152 74

512 41

1501 56,0

49,2 2,3

77,0 5,1

9,9 0,2

20,4 0,5

3-Hexen-1-ol (Z )

88,4 22,2

103,6 10,1

3-Hexen-1-ol (E )

22,2 0,2

20,2 0,1

The volatile compounds were determined in duplicate by HS-SPME-GC-MS as reported by Servili et al., 2001. Results are mean value of three
indipendent determinations standard deviation.

NEW HAMMER
CRUSHER

NEW CRUSHER WITH

SLATES

undergo the phenomenon of

coalescence

Small drops of oil

Cell wall

Olive oil

LPO
PPO

Phenols
evolution

DURING THE
MALAXATION

POD

Volatile
compounds
generation

How could these reactions be controlled ?

NEW APPROACH TO THE MALAXATION

PROCESS
Control of the gaseous
exchange

Technological parameters:
TIME

TEMPERATURE

O2 CONTROL

18

120

100

O2 evolution

16

CO2 evolution

PHENOLIC
COMPOSITION OF
PASTES
(CORATINA Cv.)
MALAXED AT
DIFFERENT O2
CONCENTRATIONS.

14
80

12
10

60

8
40

6
4

20

2
0

Servili et al., 2008.

0
0'

5'

10'

15'

20'

25'

30'

35'

40'

0'

5'

10'

15'

20'

25'

30'

35'

40'

1400

3800
3600

1300

3400
1200
3200

0 kPa

1100
3000
1000

2800

O2 30 kPa
O2 50 kPa
O2 100
kPa

2600

TOTAL PHENOLS
0'

10'

(mg/100g d.w.)
20'

30'

900

0'

40'

110

70

105

68

100

65

95

63

90

60

85

58

80

55

75

p-HPEA-EDA

53

(mg/100g d.w.)

70
0'

10'

20'

30'

40'

3,4-DHPEA-EDA

50

LIGNANS
0'

10'

(mg/100g d.w.)
20'

30'

40'

30'

40'

(mg/100g d.w.)
10'

20'

120

O2 evolution

100

80

VOLATILE
COMPOSITION
OF
PASTES
(CORATINA CV.) MALAXED AT DIFFERENT O2
CONCENTRATIONS. Servili et al., 2008.

60

40

20

0
0'

5'

10'

15'

0 kPa

20'

25'

30'

35'

40'

O2 30 kPa

C6 and C5 ALDEHYDES

(g/kg f.w.)

O2 50 kPa

O2 100
kPa

C6 and C5 ALCOHOLS

(g/kg f.w.)

120

O2 evolution

0 kPa

100

O2 30 kPa

80

60

O2 50 kPa
40

PHENOLIC
COMPOSITION
OF
EVOOs FROM PASTES (CORATINA
Cv.) MALAXED AT DIFFERENT O2
CONCENTRATIONS. Servili et al., 2008.

O2 100 kPa

20

0
0'

5'

10'

15'

20'

25'

30'

35'

40'

PHENOLIC COMPOUNDS(mg/kg)

O2 = 0 kPa

O2= 30 kPa

O2= 50 kPa

3,4-DHPEA

6,8 (0.7)a

3,2 (0.8)b

4,4 (0.7)b

1,4 (0.2)c

5,9 (0.5)bc

7,8 (0.9)b

4,3 (0.4)c

478,9 (16.2)a

437,7 (14.3)b

343,1(11.5)c

229,9 (9.2)d

144,2 (1.8)a

135,3 (1.59)b

126,2 (1.4)c

125,1 (3.1)c

30,8 (0.94)a

25,8 (2.8)b

p-HPEA
3,4-DHPEA-EDA
p-HPEA-EDA
(+)-1-acetoxypinoresinol

10,0(1.1)a

O2= 100 kPa

29,2 (0.4)ab

27,1 (0.5)ab

(+)-pinoresinol

8,1 (0.03)ab

8,0 (0.04)a

8,6 (0.4)b

7,9 (0.1)a

3,4-DHPEA-EA

475,6(13.9)a

361,9(14.1)b

339,2 (6.9)b

170,6 (2.3)c

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992. The phenolic content is the mean
value of three independent experiments standard deviation. Values in each row bearing the same superscripts are not significantly
(P < 0.05) different from one another.

120

O2 evolution

100

0 kPa
O2 30 kPa

80

60

O2 50 kPa

40

O2 100 kPa

20

VOLATILE COMPOSITION OF EVOOS

FROM
PASTES
(CORATINA
Cv.)
MALAXED
AT
DIFFERENT
O2
CONCENTRATION.
Servili et al., 2008.

0
0'

5'

10'

15'

20'

25'

30'

35'

40'

O2 = 0 kPa
ALDEHYDES(g/Kg)
2-Pentenal E
()
Hexanal
2-Hexenal E
()
ALCOHOLS(g/Kg)
1-Pentanol
2-Penten-1-ol E( )
1-Penten-3-ol
1-Hexanol
3-Hexen-1-ol E
()
3-Hexen-1-ol Z)
(
2-Hexen-1-ol E( )

548,5(16.3)ab
1187,0 (9.9)a
51565,0
(827.3)a
40,0 (5.7)a
87,5 (0.7)a
890,0 (2.8)a
2326,0(49.5)a
25,5 (0.7)ab
561,0 (4.2)a
3654,5(30.4)a

O 2= 30 kPa

O2= 50 kPa

O 2= 100 kPa

509,7 (5.8)b
1624,3 (30)bc
52900,0
(565.7)ab

636,7 (17.9)c
1532,1 (27.3)b
54340,5
(355.7)b

613,0 (51.2)ac
1744,0(121.2)c
53920,0
(332.1)b

54,3
67,0
82,0
3694,2
31,6
513,6
5905,0

39,4
(5)a
105,8 (5.7)c
1093,5(33.7)c
1788,0 (57.2)c
20,0 (1.9)b
486,3 (11.1)b
3350,1 (80.5)c

48,0 (3.2)ab
105,0 (8.3)c
1185,0 (91.2)c
2170,0(123.1)a
21,0 (1.9)b
498,0 (31.2)b
4185,0 (35.6)d

(5)b
(0.2)b
(1.2)b
(2)b
(3.8)a
(9.6)b
(321)b

The volatile compounds were determined in duplicate by HS-SPME-GC-MS as reported by Servili et al., 2001. The volatile content is the
mean value of three independent experiments standard deviation. Values in each row bearing the same superscripts are not
significantly (P < 0.05) different from one another.

NEW APPROACH TO THE MALAXATION

PROCESS
Control of the gaseous
exchange

Technological parameters:
TIME

02 CONTROL

TEMPERATURE

PHENOLIC COMPOSITION (mg/kg) OF VOOs OBTAINED MALAXING AT DIFFRERENT TEMPERATURES

Taticchi et al., 2012

Coratina

Ogliarola

AND O 2 LEVELS.

Peranzana

Moraiolo

ACTIVITY OF THE OLIVE POLYPHENOLOXIDASE

TEMPERATURES. Taticchi et al., 2012

100

100

80

80

60
40
20
0

60
40
20
0

20

30

35

40

45

50

60

20

70

30

35

40

45

50

60

70

50

60

70

TEMPERATURE (C)

TEMPERATURE (C)

Ogliarola

Moraiolo

100

100

80

80

PPO ACTIVITY (%)

PPO ACTIVITY (%)

DIFFERENT

Peranzana

PPO ACTIVITY (%)

PPO ACTIVITY (%)

Coratina

(PPO) AT

60
40
20
0

60
40
20
0

20

30

35

40

45

TEMPERATURE (C)

50

60

70

20

30

35

40

45

TEMPERATURE (C)

THERMAL
STABILITY
OF THE OLIVE POLYPHENOLOXIDASE (PPO) AT
DIFFERENT TIMES AND TEMPERATURES. Taticchi et al., 2012.
Coratina

Peranzana

90

90

PPO ACTIVITY (%)

100

PPO ACTIVITY (%)

100
80
70
60
50
40

80
70
60
50
40

30
0

20

40

30

60

TIME (min)

20

40

60

TIME (min)

Moraiolo

100

100

90

90

PPO ACTIVITY (%)

PPO ACTIVITY (%)

Ogliarola

80
70
60
50
40

80
70
60
50
40
30

30
0

20

TIME (min)

40

60

20C

30C

20

TIME (min)

40C

40

60

Aldheydes
VOLATILE COMPOSITION
(g/kg) OF VOOs OBTAINED
MALAXING AT DIFFERENT
TEMPERATURES.
Taticchi et al., 2012.

Alcohols

Esters

20 C

25 C

35 C

Evolution of lipoxigenase () and hydroperoxide-lyase actitvity ().

according to the temperature. Salas e Sanchez, 1999 .

OPTIMIZATION OF TEMPERATURE AND PARTIAL PRESSURE OF THE O2 DURING

THE MALAXATION ACCORDING TO THE OLIVE VARIETIES. Taticchi et al., 2012

TRIALS

TEMPERATURES

O2 PARTIAL PRESSURE (KPa) IN

THE HEAD SPACE OF THE
MALAXER

20 C

61.3

24 C

37.2

24 C

85.4

30 C

21.3

30 C

101.3

30 C

61.3

30 C

61.3

30 C

61.3

30 C

61.3

10

36 C

85.4

11

36 C

37.2

12

40 C

61.3

AVERAGE VALUES OF PHENOLIC COMPOUNDS (mg/kg) EVALUATED ON EVOO ACCORDING

TO THE DIFFERENT MALAXING CONDITIONS. Selvaggini et al., 2014

cv.
Coratina

total derivatives of
derivatives of sum of
lignans
phenols oleuropein ligstroside

cv.
Itrana

total derivatives of
derivatives of sum of
lignans
phenols oleuropein ligstroside

cv.
Peranzana

total derivatives of
derivatives of
phenols oleuropein ligstroside

sum of
lignans

cv. Ogliarola

derivatives of sum of
total derivatives of
lignans
phenols oleuropein ligstroside

EVOO PHENOLIC COMPOSITION (g/kg) ACCORDING TO DIFFERENT MALAXING

CONDITIONS. Selvaggini et al., 2014.
MIN

MEAN

MAX

MIN

CORATIN
A

MEAN

MAX

OGLIAROL
A

SATURATED ALDHEYDES

271.0

327.2

427.0

630.0

804.8

1007.
5

UNSATURATED ALDEHYDES
(C6)

3577
4.1

45706.1

57351.
0

39565.
0

49014.9

55772
.4

SATURATED ALCOHOLS

619.5

1015.1

1944.7

958.5

1160.9

1434.
5

UNSATURATED ALCOHOLS
(C 6)

1561.
0

2198.6

2753.5

2157.5

2726.3

3641.
5

UNSATURATED
ALCOHOLS(C5)

413.3

614.1

806.2

220.6

373.7

477.2

ESTERS

22.0

93.5

65.5

ITRANA

65.1

PERANZANA

126.7

187.5

SATURATED ALDHEYDES

295.5

803.0

1245.4

433.0

623.0

986.5

UNSATURATED ALDEHYDES
(C6)

31618
.9

43719
.5

56206.
0

25239
.5

31047
.3

37918.2

SATURATED ALCOHOLS

1577.
5

2945.
6

4367.5

788.0

1199.
5

2213.5

UNSATURATED ALCOHOLS (C
6)

295.4

524.0

748.8

1614.
0

2468.
1

4825.0

UNSATURATED
ALCOHOLS(C5)

4007.
1

5485.
4

7790.5

358.1

498.5

630.1

1092.

1407.

MODELLING BY RESPONSE SURFACES

OBTAINED BY OPTIMIZING OF THE
MALAXING CONDITIONS. Selvaggini et al., 2014.

Peranzan
33C
a; 54 KPa

Coratina
25C ; 21 KPa

Oxygen

Temperatur
e (C)

(kPa)

Desiderabilit
y

Oxygen

Temperatur
e (C)

(kPa)

Desiderabilit
y

MODELLING BY RESPONSE SURFACES

OBTAINED BY OPTIMIZING OF THE
MALAXING CONDITIONS. Selvaggini et al., 2014.

Itrana

32C ; 21
KPa

33C ; 21 KPa

Ogliarola

Oxygen
(kPa)

Oxygen
(kPa)

Temperatur
e (C)

Temperatur
e (C)

Itrana
Desiderabilit
y

Moraiolo
Desiderabilit
y

MALAXERS WITH A
CONTROLLED
GASEUS EXCHANGE

Comparison by traditional malaxation and flash thermal conditioning (FTC).

Traditional
Malaxation

Decanter

Separator

Pump load
Drain pump

S
T1 =
T2

T3

Crusher
P1

P2

FTC

Pump load

Decanter

Pause

EVOO-Line

S
T1

Drain pump

T3

Crusher

P1

T2

P2

Separator

malaxation

crushing

Optimization of the crushed paste

temperature

Pomace oil content from Cv Ottobratica olive pastes processed at 25 C and 30 C by a

traditional malaxation (control) and by a FTC followed by 15 and 20 min of malaxation.
Servili et al., 2014.
Pomace Moisture content (%)

25 C

Unpublished data.

Pomace Oil content (%)

25 C

30 C

30 C

Marketable parameters of VOOs from Cv. Ottobratica olive pastes processed at 25 C

and 30 C by a traditional malaxation (control) and by a FTC followed by 15 and 20
min of malaxation. Unpublished data.
Control 30 min

Free Acidity
(g of oleic acid/100g of
oil)

Peroxide value
(meq of O2/kg of oil)

(g of oleic acid/100g of
oil)

FTC 20 min

0,32 0,02

0,34 0,01

0,38 0,02

6,1 0,2

5,8 0,3

5,5 0,1

Control 30 min

Free Acidity

FTC 15 min

0,27 0,01

FTC 15 min
0,25 0,01

FTC 20 min
0,26 0,01

25 C

30 C

Phenolic composition (mg/kg) of VOO from olive pastes processed at 25 C and 30 C by

a traditional malaxation (control) and by a FTC followed by 15 and 20 min of
malaxation. Servili et al., 2014. Unpublished data.

Cv. Moraiolo

25 C

30 C

25 C

Cv. Coratina

25 C
25C

30 C

total phenols
derivatives of oleuropein
lignanns
derivatives of ligstroside

30 C
30C

Phenolic composition of VOO (mg/kg) from olive pastes processed at 25 C and 30 C by

a traditional malaxation (control) by a FTC followed by 15 and 20 min of malaxation.
Unpublished data.

Cv. Cellina di Nard

25 C

30 C

Cv. Ottobratica
total phenols
derivatives of oleuropein
lignanans
derivatives of ligstroside

25 C
25

30 C
30

Volatile compounds (g/Kg) of VOO from olive pastes processed at 25 C and 30 C by a

traditional malaxation (control) and by a FTC followed by 15 and 20 min of malaxation.
Unpublished data.

Cv. Peranzana

25 C

30 C

25 C

30 C

Cv. Coratina
aldehydes
alcohols

25 C

25

30 C

30C

esters

Volatile compounds (g/Kg) of VOO from olive pastes processed at 25 C and 30 C by a

traditional malaxation (control) and by a FTC followed by 15 and 20 min of malaxation.
Unpublished data.

Cv. Cellina di Nard

25 C

30 C

Cv. Ottobratica

aldehydes
alcohols
esters

25 C

30 C

CV.
Cortina

Cv. Moraiolo

Cv. Cellina di
Nard

Cv.
Ottobratica

Cv.
Peranzana

The model with 5 principal significant

components explains the 92% of the
total variance of the data (each
component explains 34%, 31%, 17%,
7%, and 3 % respectively).
CO: Cv. Coratina; MO: Cv. Moraiolo; PE: Cv.
Peranzana; CE: Cv. Cellina di Nard; OT: Cv.
Ottobratica.
1: Control malaxed at 25C for 30 min; 2: FTC
malaxetd at 25C for 15 min; 3: FTC malaxed at
25C for 20 min; 4: Control malaxed at 30C for
30 min; 5: FTC malaxed at 30C for 15 min ; 6:

Score plot and loading plot of

the first two principal
components of the PCA model
built with all samples and all
variables. Unpublished data.

Pastes cooling effect on EVOO phenolic composition

(mg/kg) (Peranzana Cv.). Unpublished data.
total phenols
derivatives of oleuropein
derivatives of ligstroside
lignans
A: pastes (control)
malaxed at
25 C for 30 min.
B: Pastes cooled at 15 C
by FTC and malaxed at 25
C for 30 min.
C: pastes (control)
malaxed at
30 C for 30 min.
D: pastes cooled at 15 C

Pastes cooling effect on EVOO volatile composition (g/kg)

(Peranzana Cv.).

Unpublished data.

aldehydes
alcohols
esters

A: pastes (control)
malaxed at
25 C for 30 min.
B: Pastes cooled at 15
C by FTC and malaxed
at 25 C for 30 min.
C: pastes (control)
malaxed at
30 C for 30 min.
D: pastes cooled at 15
C by FTC and malaxed
at 25 C for 30 min.

malaxation

crushing

Optimization of the crushed paste

temperature and simultaneous

TECHNOLOGICAL INNOVATION IN VOO MECHANICAL EXTRACTION

PROCESS

CENTRIFUGATION

REDUCTION OF PHENOLIC

INCREASE OF SOLID-LIQUID

ANTIOXIDANTS LOSS

SEPARATION EFFICIENCY

DOUBLE OIL EXTRACTION;

TWO PHASES DECANTER
DIFFERENTIAL VELOCITY
THREE PHASES DECANTER

COCHLEA VARIATION;

AT LOW WATER CONSUME

COCHLEA STRUCTURE;

DECANTER
A:
cochlea
B: drum
C: carter

Decanter section
pomace
s
water
oil

oil
Malaxe
d paste
entranc
e

Oil
escape

1th
room

water

2nd
room

Pomace
sdischar
ge
Oil tranfer form
the 1th to the 2nd

water
pomace
s

INNOVATIONS IN THE
VOO EXTRACTION BY
CENTRIFUGATION

TRADITIONAL THREE PHASE

Traditional three phases
SYSTEM

TWO PHASE SYSTEM

Two
phases
OIL
oil

MALAXED PASTES
POMACE
Pomace
+ VEGETATION
vegetationWATER
waters

OIL
oil

Water
addition
WATER
ADDITION
10.5:1 v:w
v:w

feed

MALAXED PASTED

VEGETATION
WATER
Vegetation
waters
POMAC
pomace
E

THREE PAHASES SYSTEM WITH

Traditional
three
phases
A REDUCED WATER
FEEDING
OIL
oil
WATER
Water
addition
0.1-02
:1
10.5:1 v:w
v:w

feed
MALAXED
PASTES

VEGETATION
Vegetation
waters
WATER

POMACE
pomace

TECHNOLOGICAL INNOVATION IN VOO MECHANICAL EXTRACTION

PROCESS

trditional
3 phases *
Free fatty acids

innovative
2 phases
3 phases s
**

0.15

0.20

0.28

5.80

5.20

2.90

Oleic acid %

Peroxides value
(meq O2/Kg oil)

Total phenols

303.0

448.
0

69.60

108.5

(p.p.m)
Oleuropein
derivatives
(p.p.m)

495.0

116.4

*WATER ADDITION DURING PASTES FEEDING INTO THE DECANTER

**
PASTES FEEDING INTO THE DECANTER WITHOUT WATER ADDED
VOO quality according to the centrifugation system

TECHNOLOGICAL INNOVATION IN VOO MECHANICAL EXTRACTION

PROCESS
Phenolic composition and induction period of virgin olive oils obtained from the cultivars

Centrifugation system

Coratina and Oliarola with two phases and three phases centrifugation system (1).

Coratina cultivar

Oliarola cultivar

two phases

three phases

two phases

three phases

3,4 DHPEA(2)

0,87 0,02a

0,58 0,08b

0,66 0,11c

0,50 0,11c

p-HPEA

3,74 0,07a

2,34 0,08b

3,30 0,10c

4,22 0,10d

Vanillic acid

0,41 0,01a

0,19 0,01b

0,26 0,01c

0,14 0,05d

Caffeic acid

0,16 0,01

0,12 0,02

427,2 13,8

30,09 1,03

0,09 0,01

0,21 0,03

18,53 0,68

3,4 DHPEA-EDA

522,2 13,5

p-HPEA-EDA

78,16 0,52a

67,26 2,55b

20,99 0,82c

22,40 0,33c

38,41 0,10a

35,62 1,11b

48.00 3,40c

46,72 5,78c

351,7 11,0a

244,9 13,6b

68,01 6,00c

52,04 3,11d

Total polyphenols(3)

673 4a

585 7b

304 5c

263 4d

Induction period [h]

17,8 0,1

p-HPEA-ester
(+)-1-Acetoxypinoresinol
3,4 DHPEA-EA

15,5 0,2

5,2 0,1

4,6 0,1

(1)
Mean values of three indipendent determination. Values, in each row, bearing the same superscrits are not significantly (P<0.05)
different from one another.
(2)
Evalueted by HPLC and expressed as mg Kg -1
(3)
Evaluated colorimetrically and expressed in mg Kg -1 as 3,4-DHPEA equivalent.

De Stefano et al.(1999).

LIQUID-LIQUID SEPARATION
THE ROTATIONAL SPEED IS ABOUT 6500 RPM

BELLS
CONTAINING THE
FILTRANT
VESSELS

FILTRATION
DIATOMEE
FLOUR
CONTAINER

FLTERED
OIL
ESCAPE

SPECULS

REASSEMBLINGFEEDING ELECTROPUMP

CLOUDY
OIL
ENTRANC
E

FILTRATION

DIATOMEE DETAIL: THEY ARE USUALLY USED AS FILTRANT ORGANIC

MATERIALS FOR THE CLOUDY OILS FILTRATION DETTAGLIO DI

FILTRATION

BRILLANTANT FILTRATION

COMPENSATION DRIVING
FORCE PLATES
FILTRATING
SET

HIDROSTATIC
CLOSURE
HAND
CLOSURE

FLTERED
OIL
FEEDING ELECTROPUMP
BRILLANT
OIL

FILTRATION BY PRESSURE: IT IS OFTEN USED AS

LAST CLEAN FILTRATION BEFORE THE OIL BOTTLING.

BRILLANTANT FILTRATION

VOO SHELF-LIFE
CHEMICAL COMPOSITION (FATTY ACIDS, ANTIOXIDANTS)
PACKAGING
STORAGE CONDITIONS (TIME, TEMPERATURE, OXYGEN, LIGHT)

The influence of the exposure to the light on the quality of the extravirgin olive oil was
studied as a function of the storage time.

SAMPLE C

SAMPLE A

low polyphenols content

and high oleic acid
percentage

POLYPHENOLS
DEPLETION OF EVOOs A AND B

medium polyphenols content

and low oleic acid percentage

SAMPLE D

SAMPLE B

high polyphenols
content
and oleic acid
percentage

very low polyphenols

content
and low oleic acid
percentage

SCHEME OF THE STATISTICAL CENTRAL DESIGN

B+
A

DB+BA

CA+BA

C+
A

C+
B

DC+DB

DC+CA

D+
B

A+
D

D+
C

The ONION DOE was built applying the chemometric package MODDE. 9.1 (Umetrics AB, Ume, Sweden).

COMPOSITION OF THE FATTY ACIDs (TIME 0, %)

DB+B
A
DC+C
A
CA+B
A
DC+D
B
C+B
A+D
D+C
B+A
D+B
C+A
D
C
B
A

64,
8

77,
7

The evaluations were carried out according to the Official Analysis Method (EU Reg. 1989/03). Data are the mean of
three independent analytical determinations S.D.

TOCOPHEROLS AND HYDROPHILIC PHENOLS (TIME ZERO, mg/kg)

1476,7

682,5

128,3
18,1

C+A

D+B

B+A

D+C

A+D

C+B

DC+DB

CA+BA

DC+CA

DB+BA

The evaluations were carried out according to method reported by Selvaggini et al., (2006) and Psomiadu and
Tsimidu, (1998) for polyphenols and tocopherols, respectively. Data are the mean of three independent analytical

PLS MODEL
X:
ALL
THE
DETERMINATIONS.

ANALYTICAL

Y: EXPOSURE TIME
LIGHT SOURCE.

TO

THE

TOTAL EXPLAINED VARIANCE

(WITH THREE LATENT SIGNIFICANT
VARIABLES): 92%

The PCA and the PLS

models were built applying
the
SIMCA
13.0.0.0
chemometric
package
(Umetrics
AB,
Ume,
Sweden).

TRANSPARENT

UVAG

SCREENED BLACK

DIRECT LIGHT EXPOSITION (6 WEEKS)

DIFFUSE LIGHT EXPOSISITON (6 MONTHS)

MERCHANDISE
PARAMETERS

MERCHANDISE
PARAMETERS

PHENOLIC PROFILE

QUALITY

OLIVE BY-PRODUCTS
NEW
APPROACHES
TO
OLIVE
POMACES AND
THE OLIVE OIL PRODUCTION
VEGETATION WATERS (OVW)

NEW
EXTRACTION TECHNOLOGY

OLIVE BY-PRODUCTS
OLIVE POMACES AND
VEGETATION WATERS (OVW)

VOO 2.7%

16853,2

55180
873,0
Pomac
e

olive vegetation
(OVW)
water

virgin olive oil

47.8%

47.8%

(VOO) 2.7%

49.5%

(OVW) 49.5%

26283

408,6

Total
phenols
(mg/K g)

2439
mean

olive vegetation water

olive pomace

max

min

Total
phenols
(mg/kg)

6788,3

105,5
mean

max

min

Total
phenols
(mg/L)

1751,6
mean

max

min

POTENTIAL APPLICATIONS OF EVOO BY-PRODUCTS

VIRGIN OLIVE POMACES for :

bio-energy production;
compost production;

zootechnic industries as integrative feeding: supplement of

high biological value compounds (fatty acids and
polyphenols).
OLIVE VEGETATION WATERS (OVWs) crude phenolic concentrates (OVW CPC)
and purified extracts (OVW PEs) for:

feed industries as integrators.

Production of functional foods enriched with phenolic compounds.

In food industries, as an additive of natural origin with antioxidant
and antimicrobial activities.

Mourvaky et al.,
2007

Pauselli et al., 2007

Servili et al et al., 2007.

Dal Bosco et al.,

2012

Servili et al.,
Servili et al., 2011,
2012.

unpublished data.

Merchandise value of the oil extracted from dried virgin olive pomaces.
et al., 2007

3,7

Acidity (g oleic ac./100 g oil)

Peroxide value (meq O2 /Kg oil)

Acidic composition
(%) of the oil extracted
from dried virgin olive
pomaces.
Servili et al., 2007

Miristic

10,1

0,1

Palmitic

12,3

Palmitoleic

0,9

Margaric

0,1

Heptadecenoic

0,1

Stearic

1,9

Oleic

75,5

Linoleic

8,3

Linolenic

0,6

Arachic

0,2

Gadoleic

0,3

Behenic

0,0

Servili

PHENOLIC COMPOSITION (mg/100 g d.m.) OF DRIED POMACES.

Servili et al., 2007.

3,4-DHPEA

35,1 0,2

p-DHPEA

13,1 1,1

Verbascoside

526,2 15,4

3,4-DHPEA-EDA

758,2 17,8

p-HPEA-EDA

181,4 0,8

(+)-1-Acetoxypinoresinol
(+)-1-Pinoresinol
Sum of the phenolic fractions

89,5 0,2
19,5 1,3
1623,0 23,6

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992.
The phenolic content is the mean value of three independent experiments standard deviation.
Values in each row bearing the same superscripts are not significantly (P < 0.05) different from
one another.

FATTY ACIDS COMPOSITION OF THE COW BUFFALO MILK LIPIDIC FRACTION.

Unpublished data.

Feeding with
Rmse
stoned olive
pomace integration
1,0
5,2
4,7

Control

Caproic
Caprilic
Caprinic
Lauric
Miristic
Palmitic
Stearic
Oleic
Palmitoleic
Linoleic
- Linolenic
- Linolenic
Stearidonic
Arachidonic
Eicosapentaenoic
Docosaesaenoic

3,2
4,1
4,3
14,4
34,1
7,3
16,8
1,0
2,2
0,43
0,06
0,5
0,2
0,07
0,04

2,7
3,7
4,0
14,3
33,5
8,6
17,8
1,3
2,3
0,46
0,09
0,5
0,2
0,06
0,03

0,8
1,0
0,6
1,0
2,9
0,9
2,6
0,3
0,4
0,08
0,02
0,11
0,08
0,03
0,03

TOCOPHEROLS, TOCOTRIENOLS AND RETINOL COMPOSITION (mg/g), AND

OXIDATIVE STABILITY (mg MDA/g) OF THE COW BUFFALO MILK LIPIDIC
FRACTION. Unpublished data.
Feeding
with stoned
Control
olive pomace Rmse
integration

-tocopherol

6,8

8,2

1,0

1,4

1,7

0,2

0,2

0,2

0,0

-tocotrienol

0,2

0,4

0,1

Total tocopherols

8,6

10,4

1,1

Retinol

2,5

3,2

0,3

TBARS

15,1

12,1

1,8

-tocopherol
-tocopherol

HYDROXYTYROSOL (3,4-DHPEA) CONCENTRATION (EVALUATED BY LC-ESIMS/MS ANALYSIS) OF THE MILK OF COW BUFFALOS FEEDED WITH DESTONED
OLIVE POMACE INTEGRATION. Servili et al., Unpublished data.

max

mean

min

FATTY ACIDS COMPOSITION OF THE COW BUFFALO MOZZARELLA LIPIDIC

FRACTION. Unpublished data.
Feeding with stoned olive
pomace integration
3.89A
2.44B
3.01A
2.24B
4.12a
3.60b
0.09
0.08
18.66a
16.92b
39
36.07
6.82
8.05
14.06B
20.90A
1.52
1.69
0.52
0.47
0.07
0.08
0.06
0.08
0.0009
0.0005
11.00A
8.39B
60.97a
55.29b
27.92B
36.31A
77.69A
71.63B
22.31B
28.37A
18.77B
24.65A
3.53
3.71
3.33
2.76
0.74
0.81
3.76a
2.73b
4.95A
3.68B
3.68a
3.14b

Control
C6:0
C8:0
C10:0
C12:0
C14:0
C16:0
C18:0
C18:1n9
C18:2n6
C18:3n3
C20:4n6
C20:5n3
C20:6n3
SCFA
MCFA
LCFA
SAT
INS
MUFA
PUFA
n6
n3
n6/n3

atherogenic index
thrombogenic index

RMSE
0.38
0.21
0.23
0.02
1
2
0.92
1.57
0.32
0.53
0.01
0.03
0
0.59
2.31
2.31
1.79
1.79
1.61
0.75
0.3
0.1
0.54
0.44
0.3

Animals and dietary treatments

Feeding
40 days ration supplemented with
stoned pomace in post-extraction

Post-feeding
30 days standard ration

Control
30 days standard ration

Unifeed base: hay, ryegrass, straw, corn mash, starch and protein
commercial core.

PHENOLIC COMPOSITION (mg/100 g d.m.) OF DRIED STONED POMACES .

Unpublished data.

3,4-DHPEA

523,7

4,5

3,4-DHPEA-EDA

568,1

11,1

p-HPEA

113,0

0,1

p-HPEA-EDA

33,7

1,0

Verbascoside

1082,
8

91,6

139,1

1,9

(+)-Pinoresinol

113,1

0,1

Total phenol

2573,
6

92,4

(+)-1Acetoxypinoresinol

Merchandise value of the oil

extracted from dried virgin olive
pomaces. Unpublished data.
Free Acidity
(g of oleic acid/100g
of oil)

1,25

Peroxide value

7,0

(meq. of O2/kg of oil)

Effect of the dietary treatment on the concentration of -tocopherol

and hydroxytyrosol bovine milk. Unpublished data.

Effect of the dietary treatment on the fatty acid composition of

bovine milk.
Unpublished data.

SFA

MUFA Oleic acid PUFA CLA (C:18:2,C9,T11)

control

feeding

postfeeding
The results are the means of five samples standard deviation (vertical lines).

Animals and dietary treatments

Feeding II

Feeding I
90 days ration supplemented with
stoned pomace

60

days ration supplemented with stoned

pomace + 30 days standard ration

Control
90 days standard ration

Unifeed base: hay, ryegrass, straw, corn mash, starch and protein
commercial core.

Effect of the dietary treatment on the concentration (g/kg) of hydroxytyrosol

in the Longissimus dorsi meat. Unpublished data.

Control

pomace feeding

pomace feeding
(90 days) 800

700

Effect of the dietary

treatment on the
concentration (g/kg)
of
of -tocopherol on
meat from
Longissimus dorsi
Unpublished
* The muscle.
results are the means
of ninedata.
samples (3
for the contol; 3 forthe feeding 90 days ; 3 for
the feeding 60 days.

(60 days)

693,4
624,2

588,0

600
500
400
300
200
100
0

Control

pomace
feeding

pomace feeding
(60 days)

Effect of the dietary

treatment on the fatty
acid composition (%) on
meat from Longissimus
dorsi muscle. Unpublished data.

Control

pomace
feeding

pomace
feeding

(90 days)

(60 days)

Effect of the dietary

treatment on the
concentration of
conjugated dienes
(mol/mg) on meat from
Longissimus dorsi muscle.
* The results are Unpublished
the means of
nine samples (3 for
data.
the contol; 3 forthe feeding 90 days ; 3 for the
feeding 60 days.

Control

pomace
feeding

pomace
feeding

Effect of the dietary

treatment on the TBARs of
meat from Longissimus
dorsi muscle. Unpublished data.

pomace
feeding

pomace
feeding

(90 days)

(60 days)

Effect of the dietary

treatment on the DPPH in
the Longissimus dorsi
meat. Unpublished data.

* The results are the means of nine samples (3 for the

contol; 3 forthe feeding 90 days ; 3 for the feeding 60

(mg of fresh / ml MR)

Control

Control

pomace
feeding

pomace
feeding

POTENTIAL APPLICATIONS OF EVOO BY-PRODUCTS

VIRGIN OLIVE POMACES for :

bio-energy production;
compost production;

zootechnic industries as integrative feeding: supplement of

high biological value compounds (fatty acids and
polyphenols).
OLIVE VEGETATION WATERS (OVWs) crude phenolic concentrates (OVW CPCs)
and purified extracts (OVW PEs) for:

feed industries as integrators.

Production of functional foods enriched with phenolic compounds

In food industries, as an additive of natural origin with antioxidant
and antimicrobial activities.

OVW COMPOSITION.

Servili et al., 2011

PARAMETERS

VALUES

Total sugars (g/l)

13.6

Reducing sugars (g/l)

10

Ash (g/l)

6.3

Phenolic compounds (g/l)

Solids (%)

pH

5.5

COD (mg/l)

129000

DIRECT MEMBRANES FILTRATION OF FRESH OVWS IN AN INDUSTRIAL SCALE.

Servili et al., 2011.

OVW

All the process

was carried out
with controlled
temperature
and under N2
atmosphere

Enzymatic treatment

Microfiltration cut-off 0.1-0.3 m

permeate
concentra
te
Added to
the
pomaces
for
compost
production

Ultrafiltration cut-off 7 kDa

permeate
concentra
te

inverse osmosis (IO)

OVW CPC

Low
environment
al impact

permeate

PHENOLIC COMPOSITION (g/L) OF OVW AND OVW PHENOLIC CONCENTRATE.

Servili et al., 2011.

OVW

OVW CPC

3,4-DHPEA

0,01

(0,01)

0,03 (0,003)

p-HPEA

0,02

(0,04)

0,01 (0,001)

3,4-DHPEA-EDA

4,10

(0,1)

16,90 (1,7)

Verbascoside

0,70

(0,1)

2,40 (0,2)

Total phenols

4,90

(0,2)

19,30 (1,7)

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992. The phenolic
content is the mean value of three independent experiments standard deviation. Values in each row bearing the
same superscripts are not significantly (P < 0.05) different from one another.

OLIVES
(Moraiolo, Peranzana, Ogliarola and Coratina Cvs.)

Crushing
OVW CPC
(5%) ADDITION

Malaxation
(25C , 40 min)

Extraction

POMACES

OVWs

EVOO

NOLIC COMPOSITIONS (mg/kg) OF CONTROL EVOO AND FUNCTIONAL EVOO. Servili et al., 20
CONTROL
EVOO

OVW CPC
EVOO

CONTROL
EVOO

Moraiolo
3.4-DHPEA
p-HPEA

3.4-DHPEA-EDA
p-HPEA-EDA

6,5
10,3

114,4

OVW CPC
EVOO

Coratina

(0.32)a

11,0

(0.6)b

1,9

(0.1)a

2,9

(0.2)b

(0.5)a

11,7

(0.9)a

6,3

(0.4)a

5,3

(0.5)a

(5.4)a

251,7

(12)b

281,7

(13.4)
a

480,8

103,0

(7.2)a

119,0

(8.9)a

216,0

(+)-1-acetoxypinoresinol

13,2

(0.9)a

15,4

(1.1)a

13,2

(0.7)a

14,4

(1.2)a

(+)-1-pinoresinol

15,0

(1.1)a

17,4

(1.2)a

18,4

(1.2)a

18,8

(1.3)a

3.4-DHPEA-EA

Total phenols

136,3

392,7

(6.8)a

(11.4)
a

140,6

566,8

(7.1)a

(16.7)
b

278,3

815,8

Peranzana

(10.8)a

(23.9)a

(22.2)
a

220,1

(39.1)
b

297,0

1039,3

(19.9)a

(24.1)a

(50.1)
b

Ogliarola

3.4-DHPEA

2,6

(0.1)a

5,2

(0.3)b

1,7

(0.1)a

5,5

(0.3)b

p-HPEA

4,5

(0.2)a

5,1

(0.2)b

9,1

(0.4)a

7,5

(0.4)a

3.4-DHPEA-EDA

69,6

(3.3)a

173,2

(8.2)b

56,9

(2.7)a

137,9

(6.6)b

p-HPEA-EDA

48,4

(2.4)a

52,1

(2.6)a

72,3

(3.6)a

80,2

(4.01)a

(+)-1-acetoxypinoresinol

17,7

(0.9)a

17,1

(0.9)a

12,5

(0.6)a

15,0

(0.8)b

(+)-1-pinoresinol

19,5

(0.9)a

19,9

(0.9)a

22,1

(1.1)a

25,8

(2.6)a

3.4-DHPEA-EA

Total phenols

148,4

310,6

(7.4)a

(8.6)a

151,9

424,5

(7.6)a

(11,6)B

182,9

357,4

(12.2)a

(10.3)
a

213,3

485,3

(15.2)a

(17.3)
b

The phenols concentration was evaluated by HPLC previously reported by Montedoro et al.,1992. The phenolic content is the mean value of three independent experiments
standard deviation. Values in each row bearing the same superscripts are not significantly (P < 0.05) different from one another.

VOLATILE COMPOSITIONS (g/kg) OF CONTROL EVOO AND FUNCTIONAL EVOO. Servili et al., 201
CONTROL
EVOO

OVW CPC
EVOO

CONTROL
EVOO

Moraiolo
Hexanal

671

(E)-2-Pentenal

28.0

(E)-2-Hexenal

8678.0

(61,6)a
(1,4)a

(632,2)a

OVW CPC
EVOO

Coratina

512.0
13.0

8509.0

(49,4)b
(2,8)b

(207,9)a

1815.0
209.5

116300.
0

(33,9)a
(4,9)a

(1414,2)a

1574.0

(12,7)a

188.5

(10,6)a

116950.
0

(2757,7)a

1-Penten-3-ol

613.5

(0,7)a

628.0

(1,4)a

689.5

(6,4)a

754.0

1-Pentanol

148.5

(2,1)a

147.5

(0,7)a

37.5

(0,7)a

28.5

(3,5)b

(E)-2-Penten-1-ol

474.5

(1106,5)a

506.5

(2,1)a

478.0

(1,4)a

524.5

(2,1)a

1-Hexanol

14385.
0

(959,2)a

(E)-3-Hexen-1-ol

72.0

(Z)-3-Hexen-1-ol

1125.5

(318,2)a

15235.
0

(4,2)a

(E)-2-Hexen-1-ol

Acetic acid, hexyl ester

Hexanal
(Z)-3-Hexen-1-ol, acetate
(E)-2-Pentenal

(E)-2-Hexenal

43.0
1129
310.0
301

59320

(0,7)a

12785.
0

(1470,8)a

3208.5

(217,6)a

3062.0

(7,1)a

23.0

(1,4)a

23.0

(1,2)a

1606.5

(2,1)b

245.5

(6,4)a

272.0

(4,2)a

45.0
1237
432.0
306

58150

(106,1)a
(2,1)a
(115,9)a
(37,1)b
(9,2)a

(1046,5)a

5232.5
1141
11.0
101

30650

(21,9)a

6052.0

(103,9)a
(0,9)a
(6,2)a

(1983,8)a

32595

(56,9)a

711

(45,7)a

148

1-Pentanol

214

(19,4)a

276

(25,7)b

12

(2,1)a

13

(E)-2-Penten-1-ol

577

(49,9)a

647

(56,9)a

150

(9,7)a

191

(E)-3-Hexen-1-ol

98

(Z)-3-Hexen-1-ol

2093

(460)a
(8,1)a
(201,1)a

6095
86
2562

(443,1)a
(7,1)a
(245,2)a

(E)-2-Hexen-1-ol

10720

Acetic acid, hexyl ester

1764

(114,2)a

1522

(98,5)b

(Z)-3-Hexen-1-ol, acetate

3640

(235,6)a

3328

(215,4)a

(724,7)a

10739

(732,2)a

824
89

1460
10
-

(17,2)a

919
9.0
117

741

6314

(280)a

Ogliarola

1-Penten-3-ol

1-Hexanol

(205,4)a

85.0

15275.
Peranzana0

(4,2)a
(106,4)a
(20,5)a
(4,2)a

(829,6)a

(36,8)a

(79,3)a
(5,8)a

(127,5)a
(2,1)a
-

184

672
113

1297
15
-

(89,4)b
(1,4)a
(11,3)a

(2109,7)a
(14,2)b
(2,1)a
(12,3)b

(63,5)a
(10,3)b

(113,2)a
(1,3)b
-

OLIVE VEGETATION WATER PURIFIED

COMPOSITION (mg/g). Servili et al., 2011

EXTRACT

(OVW

3,4 DHPEA

47. 2.2
9

p -HPEA

4.7 1.1

PE)

3-4 DHPEA-EDA

595.234.8

Verbascoside

14.51.9

Total polyphenols

662.334.9

The results are the mean value of three indipendent determinations standard deviation.) The phenolic
fractions have been determined by HPLC according to Montedoro et al., 1992 and expressed as mg 100g1.
(1)

PHENOLIC

PHENOLIC COMPOSITION (mg/Kg) OF THE YOGURT CONTAINING OVW PE.

et al., 2011

YOGURT
+
100 mg/L OVW PE

INITIAL
STATE

YOGURT
+
200 mg/L OVW PE

3,4 DHPEA (mg/kg)

20.5 1.0 d

26.6 1.2 c

p-HPEA

0.8 0.1 c

3.1 0.2 a

53.9 3.7 c

138.78.4 a

1.9 0.1 a

4.1 0.3 b

77.23.9 c

172.53.9 a

3,4 DHPEA

30.41.3 b

43.9 2.3 a

p-HPEA

4.1 1.4 d

1.8 0.1 b

3-4 DHPEA-EDA
Verbascoside

Total phenols
AFTER 1 MONTH OF
STORAGE

3-4 DHPEA-EDA

11.70.1
d

68.2 4.1 b

Verbascoside

n.d.

n.d.

Total phenols

47.0
1.9 d

113.94.7 b

ab, Values in the same row with different superscript letters differ significantly (P<0.01). nd: not found.

Servili

VOLATILE COMPOUNDS FOUND BY HS-SPME-GC/MS IN THE HEAD SPACE OF

THE YOUGURT WITH OVW PE. Servili et al., 2011.

Aldheydes
Acetaldehyde
Hexanal
Nonanal

Fatty acids
Acetic acid
Butanoic Acid
Hesanoic acid

Chetons
2,3-Butanedione
3-Hidroxy-2-butanone Alcohols
2,3-Pentanedione
4-Penten-1-ol
1-Pentanol
3-Penten-2-ol
1-Hexanol
1-Heptanol
2-Chetons
2-Ethyl-1-hexanol
Acetone
1-Octanol
2-Butanone
2-Heptanone
2-Nonanone
2-Undecanone

Monoterpens
Limonene
6-Methyl-5-hepten-2-one

Lattons
d-Decalactone

Aromatic compounds
Ethyl-benzene
Acetophenone
Phenol
Phenyl-ethyl-alcohol

VOLATILE COPMOSITION (g/kg) OF THE YOGURT HEAD SPACE WITH AND WITHOUT AN
OVW PE ADDITION. Servili et al., 2011
YOGURT CONTROL

YOGURT
100 mg/L OVW PE

YOGURT
200 mg/L OVW PE

Acetaldehyde

19485.4 413.3a

28168.3398.4c

15731.6333.7e

Diacetyl

4520.0 80.0a

4040.0 409.0b

4906.0 69.0c

Acetoin

11400.0 241.8a

13350.5188.8b

13300.0 282.1b

74.02.3c

127.84.1d

Initial State

2-Pentanone

103.81.9

2-Heptanone

27.91.0a

33.01.2b

38.61.7d

2-Nonanone

16.10.7ac

18.21.5bc

20.31.1b

1-Pentanol

17.90.8a

8.50.4c

14.80.4d

1-Hexanol

18.00.8a

14.70.6b

20.90.9c

1-Octanol

4.70.2a

6.00.6b

5.20.2ad

AFTER 1 MONTH OF STORAGE

Acetaldehyde

18000.0381.8b

24000.0339.4d

12000.0254.6f

Diacetyl

4551.5 81.9a

4068.1 73.2b

4872.0 74.0c

Acetoin

11356.9160.6a

12164.4172.0c

13700.0193.7b

2-Pentanone

108.13.8a

80.62.9c

124.83.1d

2-Heptanone

32.51.2b

32.21.1b

38.11.3d

2-Nonanone

14.90.6a

15.60.6a

16.30.7ac

1-Pentanol

11.40.5b

9.60.4c

18.30.8a

1-Hexanol

14.50.6b

15.80.7b

24.01.0d

1-Octanol

4.90.2a

8.30.3c

5.70.2bd

ab, Values in the same row with different superscript letters differ significantly (P <0.01). nd: not found.

POTENTIAL APPLICATIONS OF EVOO BY-PRODUCTS

OLIVE VEGETATION WATERS (OVWs) crude phenolic concentrates (OVW CPC)

and purified extracts (OVW PEs) for:

feed industries as integrators.

Production of functional foods enriched with phenolic compounds.

In food industries, as an additive of natural origin with antioxidant and antimicrobial
activities.

OLIVE VEGETATION WATER PURIFIED

COMPOSITION (mg/g). Esposto et al., 2014.

EXTRACT

(OVW

PE)

Phenol Compounds
3,4-DHPEA

56.5

1.1

3,4-DHPEA-EDA

532.5

9.8

p-HPEA

12.3

0.4

Verbascoside

80.0

4.1

Total phenols

681.3

10.7

Purity

68 %

PHENOLIC

EXPERIMENTAL PROTOCOL.
Amount of
antioxidant added
to renfined olive oil
(OR)

Olive
vegetation
water purified
extract (OVW
PE)

BHT

100

(mg/kg)

ORPE(1)

ORBHT(1)

200

(mg/kg)

ORPE(2)

ORBHT(2)

400

(mg/kg)

ORPE(3)

Esposto et al., 2014.

Oxidative stability of the oils used in the experiment.

Esposto

et al., 2014

9.00

4.0

induction time (h)

peroxide value (meq of O2/kg of oil
acidity (g of oleic acid /100 g of oil
K232
K270

8.00
7.00

3.5
3.0

6.00

2.5

5.00

2.0

4.00

1.5

3.00

1.0

2.00

0.5

1.00

0.0

0.00

-0.5

OR

ORBHT (1)

ORBHT (2)

OREF (1)

OREF (2)

OREF (3)

OR: refined oil ORBHT (1): refined oil added with 100 mg/kg of BHT; ORBHT (2): refined oil added with 200 mg/kg of
BHT; ROPE (1): refined oil added with 100 mg/kg of OVWPE; ROPE (2): refined oil added with 200 mg/kg of OVWPE;
ROPE (3): refined oil added with 400 mg/kg of OVWPE.

Evolution of the sum of Acrolein and - - dimethylacrolein (g/kg) in

OR, ORBHT (1), ORBHT (2), ROPE (1), ROPE (2) and ROPE (3) during
frying. Esposto et al., 2014.
350

300

250

200

150

100

50

The vertical line represents the precision of the analysis, expressed as the standard deviation of three samples. OR: refined oil ORBHT (1): refined oil added
with 100 mg/kg of BHT; ORBHT (2): refined oil added with 200 mg/kg of BHT; ROPE (1): refined oil added with 100 mg/kg of OVWPE; ROPE (2): refined oil
added with 200 mg/kg of OVWPE; ROPE (3): refined oil added with 400 mg/kg of OVWPE.

Evolution of the sum of aldehydes (C7C11) (g/kg) in OR, ORBHT(1), ORBHT

(2), ROPE (1), ROPE (2) and ROPE (3) during frying. Esposto et al., 2014.
40000

35000

30000

25000

20000

15000

10000

5000

The vertical line represents the precision of the analysis, expressed as the standard deviation of three samples. OR: refined oil ORBHT (1): refined oil added
with 100 mg/kg of BHT; ORBHT (2): refined oil added with 200 mg/kg of BHT; ROPE (1): refined oil added with 100 mg/kg of OVWPE; ROPE (2): refined oil
added with 200 mg/kg of OVWPE; ROPE (3): refined oil added with 400 mg/kg of OVWPE; ROPE

SAUSAGES ADDED WITH OLIVE VEGETATION WATER PURIFIED EXTRACT (OVW

PE).

OVWPE
750 mg/kg
1500 mg/kg

ANTIOXIDANT EFFECT

EVOLUTION OF PHENOLIC COMPOUNDS

ON THE UNCOOKED AND COOKED
PRODUCT TO DIFFERENT TIMES OF
CONSERVATION (0, 7, 15 day)

ANTIOXIDANT AND ANTIMICROBIAL

EFFECTS

EVOLUTION OF PHENOLIC COMPOUNDS

AND OXIDATIVE STABILITY AT DIFFERENT
SEASONING TIMES (0, 3, 10 e 20 day)

Antioxidant effects: Peroxide value (meq O2/kg of fat) of cooked sausages.

unpublished data.

aA
abA
bA

aB

Control

bB aB

L 1 (1500 mg/kg of OVWPE)

aB

bB

L 1 (1500
OVWPE)

aB

mg/kg

of

Antioxidant
effects:
evolution
the
phenolic
composition
uncooked
and
cooked
Antioxidant
effects:
evolution
of of
the
phenolic
composition
in in
uncooked
and
cooked
sausage
sausage
added
added
with
with
1500
750mg/kg
mg/kgofofOVWPE.
OVWPE.unpublished
unpublished
data.
data.
a
a

aa
b
b

bb

dc

cd
c

d
e
d

d
c

c
c

e
f

a
a

ac
a
aa

b
b

bc

ac

b
cc

b
c
c
b

ee
d
d

bc
bc
e
d

Data are the mean of three analytical evaluations, standard deviation is reported in bracket. ae, values in the same row with different superscript
letters differ significantly (P< 0.01)

SAUSAGES ADDED WITH OLIVE VEGETATION WATER PURIFIED EXTRACT (OVW

PE).

OVWPE
750 mg/kg
1500 mg/kg

ANTIOXIDANT AND ANTIMICROBIAL

EFFECTS

EVOLUTION OF PHENOLIC COMPOUNDS AND OXIDATIVE STABILITY AT DIFFERENT

SEASONING TIMES (0, 3, 10 e 20 day)

Antioxidant and antimicrobial effects: evolution of the peroxide value (Meq. O 2/Kg di
salami) in salami control and with addition of OVWPE (L1 = 750 mg/kg; L 2 = 1500
mg/kg) at different seasoning times. unpublished data.

aD

aC

aB

B
cC
aA

bA

b
B

b
C

bB
bA

bA

The data significance was evaluated by one way ANOVA using the Fisher LSD test (p 0.05). The lowercase letters indicate the
differences between the various thesis (control, L1 and L2), while the uppercase letters indicate the differences between the different
seasoning time (0, 3, 10 and 20 days)

Antioxidant and antimicrobial effects: evolution of the phenolic composition (mg/kg) in

salami added with OVWPE (L1 = 750 mg/kg; L2 = 1500 mg/kg) at different seasoning
times. unpublished data.

A
B

B
C

C
C

B
C
B
A A

C
A

D
A

D
B

The data significance was evaluated by one way ANOVA using the Fisher LSD test (p 0.05) while the uppercase letters indicate the
differences between the different seasoning time (0, 3, 10 and 20 days)

OLIVES

HARVESTED AT A SUITABLE DEGREE OF MATURATION

ACCORDING TO THE CULTIVAR.

Crushing

WORKED AT AN OPTIMAL SANITARY STATE.

DIFFERENTIATED EFFECT ON THE CONSTITUTIVE

PARTS OF THE DRUPES BY:

Malaxation

TEETH CRUSHER

Control of
BLADE
TIME,
CRUSHER
O2 LEVEL
and TEMPERATURE.
CRUSHER
WITH A LOW TURNS NUMBER

Extraction

STONING

DOUBLE STOKER

THREE-PHASES CENTRIFUGATION
WITH REDUCED WATER ADDITION
concentrates
EVOO dried
anddestoned
purified extracts
pomaces:
for:

OVWs

HIGH QUALITY
EVOO
EVOO

bio-energy
feed industries
and compost
as integrators;
production;
cosmetic
zootechnic
industries
industries
as as
natural
integrative
antioxidants;
feeding:
EVOOofindustries
supplement
high biological
for increasing;
value compounds
polyphenols
acids and
concentration;
polyphenols).
POMACES the(fatty
food industries for producing functional products

THANKS FOR YOUR

ATTENTION !