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Foods: Bioactive Compounds and Quality of Extra Virgin Olive Oil
Foods: Bioactive Compounds and Quality of Extra Virgin Olive Oil
Review
Bioactive Compounds and Quality of Extra Virgin
Olive Oil
Cecilia Jimenez-Lopez 1,2 , Maria Carpena 1 , Catarina Lourenço-Lopes 1 ,
Maria Gallardo-Gomez 3 , Jose M. Lorenzo 4 , Francisco J. Barba 5 , Miguel A. Prieto 1, * and
Jesus Simal-Gandara 1, *
1 Nutrition and Bromatology Group, Department of Analytical and Food Chemistry, Faculty of Food Science
and Technology, University of Vigo, Ourense Campus, E32004 Ourense, Spain;
[email protected] (C.J.-L.); [email protected] (M.C.);
[email protected] (C.L.-L.)
2 Centro de Investigação de Montanha (CIMO), Instituto Politécnico de Bragança, Campus de Santa Apolonia,
5300-253 Bragança, Portugal
3 CINBIO, Universidade de Vigo, Department of Biochemistry, Genetics and Immunology,
Campus Universitario Lagoas Marcosende, 36310 Vigo, Spain; [email protected]
4 Meat Technology Centre Foundation, 32900 San Cibrao das Viñas, Spain; [email protected]
5 Nutrition and Food Science Area, Preventive Medicine and Public Health, Food Science, Toxicology and
Forensic Medicine Department, Universitat de València, Faculty of Pharmacy, Avda, Vicent Andrés Estellés,
s/n, Burjassot, 46100 València, Spain; [email protected]
* Correspondence: [email protected] (M.A.P.); [email protected] (J.S.-G.)
Received: 30 May 2020; Accepted: 24 July 2020; Published: 28 July 2020
Abstract: (1) Background: Extra virgin olive oil (EVOO) is responsible for a large part of many health
benefits associated to Mediterranean diet as it is a fundamental ingredient of this diet. The peculiarities
of this golden, highly valued product are in part due to the requirements that must be met to achieve
this title, namely, it has to be obtained using exclusively mechanical procedures, its free acidity
cannot be greater than 0.8%, it must not show sensory defects, and it has to possess a fruity taste.
(2) Methods: All these characteristics are key factors to EVOO quality, thus the chemical composition
of these many health-promoting compounds, such as unsaturated fatty acids (which are also the
major compounds, especially oleic acid), as well as minor components such as tocopherols or phenolic
compounds (which behave as natural antioxidants) must be preserved. (3) Results: Due to the
presence of all these compounds, the daily consumption of EVOO entails health benefits such as
cardioprotective, antioxidant, anti-inflammatory, anti-tumor properties or acting as regulator of
the intestinal microbiota, among others. (4) Conclusions: Taking all together, conserving EVOO
chemical composition is essential to preserve its properties, so it is worth to control certain factors
during storage like exposure to light, temperature, oxygen presence or the chosen packaging material,
to maintain its quality and extend its shelf-life until its consumption.
Keywords: extra virgin olive oil; chemical composition; bioactive substances; EVOO
quality; applications
1. Introduction
Olea europaea L., commonly known as the olive tree, is a small tree species typically distributed
by the Mediterranean countries [1]. Olive oil, its main derived product, has experienced an increase
in its popularity due to its organoleptic characteristics and its associated beneficial health effects [2].
The olive tree is one of the species that was first cultivated. In fact, there are records for the first
cultivated trees which date from 6000 years in Asia Minor regions according to the International
Olive Council (IOC) [3]. Its origin is linked to Mediterranean civilizations and regions, characterized
by soft and rainy winters and hot dry summers [4]. Although its industry uses large areas for the
cultivation of trees, nowadays, its cultivation has been spread to regions of all continents (excluding
Antarctica). Still, Mediterranean countries remain the main olive producers, led by Spain, Italy, and
Greece. In consequence, the European Union (EU) is responsible for the 70% of the global olive
production, generating a production value of 7000 million euros each year, becoming a key factor for
the agro-industrial sector development and thus, a social and economic engine for the EU southern
regions [5].
The French writer Georges Duhamel once wrote “There, where the olive tree gives up, is where the
Mediterranean ends. The tree of light is the nature and culture of the Mediterranean” [6]. This statement
corroborates the joining and simultaneous development of a culture-bound to olive products and
particularly olive oil. So much, that olive oil is a crucial part of the commonly known as Mediterranean
diet (MED). This diet consists of a balanced combination of low animal protein consumption with
a high intake of fruits, vegetables and cereals and olive oil as the principal source of fat in many
foods [3]. MED has been usually associated with a low incidence of cardiovascular diseases (CVD)
risk in EU southern regions, which likewise, show higher life expectancy when compared to EU
northern countries and the United States of America (USA) [7,8]. Several studies have been conducted
to evaluate and prove MED as the main protective agent in the primary prevention of chronic diseases.
Research has pointed out that the beneficial health effects of MED are attributed to a large extent to
olive oil.
Some of those beneficial properties are gathered up in Figure 1. Its unique composition and
biological properties are largely responsible for this association. Olive oil composition is mainly
formed by triglycerides and a variety of several compounds in small quantities. Among the glyceride
fraction, olive oil shows a high content of fatty acids and particularly, an elevated proportion of
monounsaturated fatty acids (MUFA). Unsaturated acids are up to 85% of its composition, due to its
high content in oleic acid (C18:1), which might range between 70–85% and other fatty acids as linoleic
or palmitoleic acid. In the case of saturated fats, they entail around de 14% of oil composition, basically
due to palmitic and stearic acids. Regarding minor compounds, they signify less than 2% of olive
oil composition, and the best representatives of this group are phenolic compounds, although this
minor group also includes some lipophilic compounds like α-tocopherol (vitamin E). Likewise, there
are several hydrophilic phenolic compounds, among which the simple phenol hydroxytyrosol and
the secoiridoid oleuropein must be highlighted [1,8]. Besides, olive oil is also a source of pigments
like carotenoids [3]. Phenolic compounds are known for their biological properties. In particular,
these olive oil compounds have shown potential as antioxidant, anti-inflammatory and antimicrobial
agents [9]. However, their concentration is related to different factors: olive varieties, environmental
factors, time of harvest and extraction, and storage conditions [1]. To sum up, olive oil is considered as
a superfood due to its health properties derived from its unique composition; this is, its lipid profile
and its bioactive compounds content [10]. Moreover, its singular composition has attracted so much
attention that olive oil has been studied as a source of these bioactive compounds but also their derived
residues have shown high concentrations of these molecules and thus a great potential for industrial
recovery and related applications, such as formulation of new high added value products [10].
Foods 2020, 9, x
Foods 2020, 9, 1014 3 of 31
3 of 31
Figure 1. Scheme of the beneficial and healthy effects of the olive oil inclusion in the Mediterranean
Figure 1. Scheme of the beneficial and healthy effects of the olive oil inclusion in the Mediterranean
diet. Information adapted from [11,12].
diet. Information adapted from [11,12].
Figure 2 shows a detailed procedure of the extraction of olive oil from Olea europaea L. of different
Figure 2 shows a detailed procedure of the extraction of olive oil from Olea europaea L. of different
varieties, whose extracting process follows a common line formed by different steps: collection, grading,
varieties, whose extracting
washing, crushing, malaxing,process follows
separation anda centrifugation,
common line storage
formed and
by different
packaging. steps:
Thiscollection,
process is
grading, washing, crushing, malaxing, separation and centrifugation, storage and packaging. This
not aimed at improving oil quality; however, attention must be paid to its development to not cause a
process is not aimed at improving oil quality; however, attention must be paid to its development to
loss of quality [13,14]. Depending on the modifications of this process and the resultant organoleptic
not cause a loss of quality [13,14]. Depending on the modifications of this process and the resultant
characteristics of the product, among others, the EU defines on the Article 118 of European Commission
organoleptic
(EC) Regulation characteristics of the product,
(EC) No 1234/2007 among
six main types of others,
oils, whichthe are
EU delineated
defines on alsothe by
Article
the IOC 118 of
[15].
European Commission (EC) Regulation (EC) No 1234/2007 six main types of oils,
The specific attributes of each type of oil are shown in Table 1. Regarding all these groups, virgin olive oil which are
delineated
(VOO) mustalso by the IOC due
be highlighted [15].
toThe
theirspecific attributes
organoleptic of each type
characteristics and of oil are shown
beneficial in Table
properties. Among 1.
Regarding all these groups, virgin olive oil (VOO) must be highlighted due
VOOs, the EU establishes three types of oils: extra virgin olive oil (EVOO), VOO, and lampante oliveto their organoleptic
characteristics and beneficial properties. Among VOOs, the EU establishes three types of oils: extra
oil. Furthermore, IOC adds an extra subgroup called ordinary virgin oil. All VOOs are characterized
virgin olive
for being oil (EVOO),
obtained VOO, and
by mechanical lampante
processes (onlyolive oil. Furthermore,
washing, decantation,IOC adds an extra
centrifugation andsubgroup
filtration)
called ordinary virgin oil. All VOOs are characterized for being obtained by mechanical processes
under specific thermal conditions which do not cause any alteration. Afterward, they are divided
(only washing, decantation, centrifugation and filtration) under specific thermal conditions which do
according to their acidity, which gives an idea of the content in free fatty acids based on the percentage
not cause any alteration. Afterward, they are divided according to their acidity, which gives an idea
of oleic acid. Lower acidity values guarantee a high-quality oil, showing it has been obtained from
of the content in free fatty acids based on the percentage of oleic acid. Lower acidity values guarantee
healthy olives and under ideal conditions. Moreover, they are submitted to sensory analysis to asses
a high‐quality
some requirementsoil, showing
[16]. it has been obtained from healthy olives and under ideal conditions.
Moreover, they are submitted to sensory analysis to asses some requirements [16].
Foods 2020, 9, 1014 4 of 31
Foods 2020, 9, x 4 of 31
Figure 2. Summary of the oil extraction process focused on extra virgin olive oil (EVOO) production.
Figure 2. Summary of the oil extraction process focused on extra virgin olive oil (EVOO) production.
The boxes with discontinuous outline refer to the obtainment of other type of oils. Modified from
The boxes with discontinuous outline refer to the obtainment of other type of oils. Modified from
different schemes of [10,17].
different schemes of [10,17].
Table 1. Types of olive oils. Modified from EU Council Regulation (EC) No 1234/2007 [15].
Free
Type of Oil Characteristics
Acidity
They are characterized for being obtained by mechanical or other physical <0.8 g per
EVOO
processes under specific thermal conditions that do not cause alterations in 100 g
Virgin olive the oil and have not suffered any treatment other than washing, decantation, ≤2 g per
Virgin olive oils
oil centrifugation or filtration. Excluded are oils obtained using solvents or 100 g
adjuvants with chemical actions, by re-esterification process or any mixture
Lampante >2 g per
with oils of other types.
olive oil 100 g
≤0.3 g per
Refined olive oil In this case, virgin olive oil is submitted to a refining process.
100 g
Olive oil (composed of refined It is the result of the blending of the two previous oils: virgin olive oils (not ≤1 g per
olive oils and virgin olive oils) lampante oil) with refined olive oil. 100 g
This type refers to oil obtained from olive pomace by using solvents, physical
treatments or oil corresponding to lampante olive oil type, except for certain
Crude olive pomace oil specified characteristics.
As well as in the case of virgin olive oils, excluded are oils obtained by means
of re-esterification and mixtures with oils of other types.
≤0.3 g per
Refined olive pomace oil This type is obtained from refining crude olive pomace oil.
10 g
It if the resultant oil from mixing refined olive pomace oil and virgin olive oil ≤1 g per
Olive pomace oil
different than lampante oil. 100 g
NOTES: (1) The International Olive Council (IOC) establishes olive oil standards in terms of sensory analysis and
chemical composition for each category. (2) In the case of Lampante oil, it is also referred as “virgin olive oil not fit
for consumption” by the IOC and with a free acidity higher than 3.3 g per 100 g.
Foods 2020, 9, 1014 5 of 31
Particularly, EVOO is obtained directly from olives, that is, pure olive juice. It is considered
the highest quality oil, and in general, it is characterized for having a low acidity, up to 0.8% and
a sensory grade higher than 6.5 points, thus, having perfect aroma and flavor [3,10]. Regarding
sensory aspects, EVOO must show a fruity note higher than 0 and, more importantly, a median of
zero defects [18]. In addition, and to offer the most possible information to the consumer, these oils
are usually labeled as intense, medium or light depending on their positive attributes [19]. However,
sensory analysis is always more difficult to unify. On the other hand, EVOO is susceptible to different
chemical alterations during its production. Hydrolytic and oxidative degradation are the main causes
of undesirable product formation so that water plays an important role since lipases (naturally present
at olive pulp or seeds) are active in an aqueous phase. Lipolytic reactions lead to the formation of
free fatty acids, thus increasing the acidity of the oil. To avoid these reactions, the filtration step is
of high importance, as well as paying attention to other factors such as avoiding microorganisms
or mechanical crushing, among others. Moreover, water may affect the transformations of different
phenolic compounds [18,20]. Regarding oxygen, EVOO must have a peroxide index (mEqO2 /kg) ≤20.
Oxygen is crucial for the lipoxygenase cascade reactions and its consequential positive sensory notes,
but its excess can cause defects [20]. Moreover, EVOO must also show specific spectrophotometric
values: absorption λ = 233 nm, ≤ 2.50 and λ = 270 nm, ≤ 0.22, which are quality and authenticity
indicators, respectively [18]. Besides, contaminants (i.e., mycotoxins or pesticides, among others)
are another crucial factor in avoiding associated risks [21]. Considering all the characteristics that
EVOO is required to fulfill, quality control is a must. Both IOC and EC have approved different
standards in matters of purity (oil composition) and quality (organoleptic profile, acidity, peroxide
value, among others) as well as sensory analysis to ensure that alterations have not occurred during
the process [10]. Additionally, another remarkable aspect is traceability. Traceability must be assured
during all the obtaining process of EVOO, including the four basic steps (harvesting, milling, storage,
and packaging batches) of the procedure to guarantee the identity of the product and its production
chain. This approach also leads to the next step, this is certification, carried out by normalization
and certification quality organisms in order to prove the rigorous process system employed at EVOO
production [22,23].
Regarding its composition, EVOO is mainly composed of triglycerides (97–99%) and minor
compounds (1–3%), which are the principal responsible for its biological properties and sensory
attributes. It has a high content of MUFA (65–83%), especially oleic acid, and some polyunsaturated
fatty acids (PUFA) such as linoleic acid, which is considered a potent fatty acid on the reduction of
low-density lipoprotein (LDL) cholesterol. This lipid profile and also high ω6/ω3 ratio have been linked
to protective effects on coronary, autoimmune and inflammatory disorders but also as anti-thrombotic and
regulators of blood pressure [24–26]. Concerning bioactive compounds, their main representatives are the
same of oil in general, namely phenolic compounds such as hydroxytyrosol and derivatives (oleuropein
and tyrosol), tocopherols but also other compounds as hydrocarbons (i.e., squalene) or pigments like
provitamin A compounds [3,26]. However, it must be mentioned that some of these compounds such as
squalene might be lost during refinery, so they can only be found on this type of oil [25].
As mentioned, all these bioactive compounds are known for their biological properties and
positive effects on human health. EVOO inclusion in the diet and its bioactive molecules have been
studied to identify its effects. EVOO is known for having a high content of antioxidant compounds with
protective properties against free radicals. Therefore, it has been pointed out that its high consumption
is related to a generally low risk of suffering colon, breast or skin cancer as well as beneficial effects on
aging and coronary diseases [1]. It has also been proposed as a preventing tool of Alzheimer’s and
other neurodegenerative diseases [3], as anti-inflammatory [7] and also as immune-stimulating [24].
Another study proved that rats fed with EVOO in substitution of lipids and complemented with
physical exercise, could avert cartilage diseases as osteoarthritis [9]. Additionally, EVOO consumption
has shown positive effects on gut microbiota [27]. Some studies have researched the bioavailability
of phenolic compounds of EVOO and found that 55–60% of them can be absorbed, most of them at
Foods 2020, 9, 1014 6 of 31
small intestine [11]. Moreover, their different compounds have shown other beneficial properties like
antimicrobial, antitumor or protective agents against cellular damage [26,28]. More specific studies
have also related EVOO treatment with positive gene regulation and with micro ribonucleic acid
(miRNA) modulation of target genes associated with synaptic plasticity as well as to motor and
cognitive behavior [29]. There are multiple bibliographic references directed towards proving all these
promoting effects. Nevertheless, a specific study must be highlighted, the PREDIMED trial (prevention
through MED, in Spanish). This is one of the largest nutritional studies ever conducted in Spain,
which evaluated the effects on primary prevention of CVD when following a MED supplemented
by EVOO or nuts mix [30]. This project groups together different studies, which have brought to
light several positive consequences: reduction of CVD risk, reduction of C-reactive protein, reduction
of atrial fibrillation, prevention of diabetes and metabolic syndrome, reduction of diastolic blood
pressure, higher protection against breast cancer or lower prevalence of non-alcoholic fatty acid
liver disease [25,31–33]. However, more epidemiologic studies and controlled trials are necessary
to better validate and understand the beneficial effects of EVOO consumption. At last, it is worth
mentioning that new disciplines (encompassed as nutrigenomics) are also working on new approaches
for evaluating the health-promoting effects, characterizing new markers, and understanding their
action mechanisms [29,34].
Regarding all the presented information, this article aims to review the current knowledge
about the unique EVOO composition and its related bioactivities. Therefore, a revision focused
on its beneficial and health-promoting properties, its chemical composition as well as some quality
parameters is presented. To achieve this purpose, a systematic search in several databases was carried
out based on the topic described, all in all, ~5000 documents have been published. Most of the
publications correspond to research articles and book chapters, 56.3% and 15.6%, respectively, followed
by reviews (12.5%). Among them, bibliographic references were chosen according to their concordance
with the covered topics. The extensive information compiled was further classified into the different
sections presented in this review. After analyzing the considerable amount of data available on this
field, it can be said that EVOO is a current topic; although it has been known and used for centuries, it is
still under study, which means EVOO has potential properties and/or applications yet to be discovered.
The concentration of the minor and major fruit components changes and depends on all those variables.
Apart from that, the olives ripening process lasts a few months in which the atmospheric, environmental
and agricultural conditions play a very important role despite the strict genetic control that can be
applied [43,44]. During the maturation and ripening process, the photosynthetic activity decreases
as the oil content in the fruits increases [45]. In the first stage of ripening, denominated green stage,
the ripe fruits have already acquired their final size, so the maturation proceeds, and the chlorophylls
in the skin are slowly swapped by anthocyanins, turning the olives from green to dark violet or purple
until the end of the ripping process. These changes in color define the spotted, purple and black
stages [43,46]. Olives have the highest phenolic compound content at the phase between green and
darker skin, and therefore, the degree of maturation is an important factor to define the right harvest
time that will originate the best quality olive oil [47]. Figure 3 shows a summary of representative
chemical structures of some relevant compounds present in EVOO.
Foods 2020, 9, x 8 of 31
Lipids
Carbohydrate
Pigments
Vitamins
Figure 3. Representative chemical structures of some relevant compounds present in EVOO.
Figure 3. Representative chemical structures of some relevant compounds present in EVOO.
2.1. Primary Metabolites
2.1. Primary Metabolites
2.1.1. Lipids
2.1.1. Lipids
Lipids are one of the principal sources of energy for all living beings and additionally, they are also
involvedLipids are one of the principal sources of energy for all living beings and additionally, they are
in many physiological functions, as their role as a structural component of cell membranes,
also involved in many physiological functions, as their role as a structural component of cell
membranes, nervous system, the production of hormones, brain development and also on the
promotion of liposoluble vitamins absorption.
EVOO is widely used in the human diet, especially in the MED and has been long renowned for
its many health‐promoting properties. Its consumption is associated with reduced risk of several
chronic illnesses, like diabetes, hypertension, obesity and CVD [48,49]. These health properties are
Foods 2020, 9, 1014 8 of 31
nervous system, the production of hormones, brain development and also on the promotion of
liposoluble vitamins absorption.
EVOO is widely used in the human diet, especially in the MED and has been long renowned
for its many health-promoting properties. Its consumption is associated with reduced risk of several
chronic illnesses, like diabetes, hypertension, obesity and CVD [48,49]. These health properties are
related to the presence of bioactive compounds like phenolic compounds but also with the high content
in MUFA. Olive oil has a high content in oleic acid transforming it into a healthy fat, especially when
compared with other vegetable oils [40]. This lipid can decrease the risk of CVD due to its effects on
the lipids present in the blood vessels [50]. According to available data, there is 65.2–80.8% of MUFA
in the lipidic fraction of olive oil [51]. Other fatty acids found in the total fatty acids composition of
olive oils are palmitic acid, palmitoleic acid, stearic acid, linoleic acid, α-linolenic acid, and other minor
ones that are listed in Table 2.
Triacylglycerols constitute a big part of the edible oil and a high percentage of the saponifiable
fraction is constituted by MUFA [27]. The principal triacylglycerol detected in olive oil is
oleic–oleic–oleic (OOO), representing about half of the total triacylglycerol portion found in
EVOO. Other triacyclglycerols also present are palmitic–oleic–oleic (POO), oleic–oleic–linoleic
(OOL), palmitic–oleic–linoleic (POL) and stearic–oleic–oleic (SOO) [43,52]. Diacylglycerols and
monoacylglycerols have been identified in VOO at concentrations of 1–2.8% and 0.25, respectively [53].
Four classes of sterols also take place in olive oil and are commonly used to check its
genuineness because their presence is linked to the quality of the oil. These four classes are
common sterols (4-Desmethylsterols), 4α-Methylsterols, triterpene alcohols (4, 4-Dimethylsterols)
and triterpene dialcohols [43]. Common sterols in EVOO are present in both free and esterified
forms [59]. The leading components of this sterol fraction are campesterol, β-Sitosterol and
∆5-Avenasterol [60,61], and in smaller amounts, it is also possible to find stigmasterol, cholesterol,
cholesterol, brassicasterol, sitostanol, ergosterol, campestanol, ∆7-Cholestenol, ∆7-Avenasterol,
∆7-Stigmasterol, ∆7-Campesterol, ∆5,24-Stigmastadienol, ∆5,23-Stigmastadienol, ∆7,24-Ergostadienol,
∆7,22-Ergostadienol, 22,23-Dihydrobrassicasterol and 24-Methylene-cholesterol [62,63]. The total sterol
content of EVOO varies between 1000 and 2000 mg/kg, being the first value the inferior limit set by the
EU Commission [43]. β-Sitosterol is the main compound in the sterol fraction with values between 75%
and 90% of the total sterol fraction, while ∆5-Avenasterol has values between 5% and 20% [62]. Crop year,
cultivar, ripeness of the fruit, storage time of the olives before oil extraction and geographic influences all
contribute to sterol composition of the final EVOO obtained [64–66]. At the same time, storage time and
conditions of the final product are also factors that can originate several important changes particularly
in the concentrations of each individual sterol [43]. 4-Monomethylsterols are present in smaller amounts
and signify part of sterol biosynthesis as intermediates. They can be found in their free and esterified
forms [67]. The most common are gramisterol, obtusifoliol, cycloeucalenol and citrostadienol [60,62],
and their concentrations vary between 50 and 360 mg/kg of oil [60,68]. Triterpene alcohols, also identified
as 4,4-Dimethylsterol, are a very complex fraction that can be in free and esterified form, and whose
main compounds are butyrospermol, β-Amyrin, cycloartenol and 24-Methylenecycloartanol. In smaller
amounts or trace quantities, cyclosadol, cyclobranol, dammaradienol, germanicol, 24-Tirucalladienol,
24-Methylene-24-Dihydroparkeol, α-Amyrin, taraxerol, 7, parkeol and tirucallol can also be found [62].
Total triterpene alcohol levels range from values of 350 to 1500 mg/kg [68,69]. Lastly, among the
triterpene dialcohols class, erythrodiol (5α-olean-12-ene-3β, 28-diol, homo-olestranol) in free and
esterified form and uvaol (∆12-Ursen-3β,28-diol) are the major triterpene dialcohols found in EVOO [70],
and their presence is mainly affected by cultivation characteristics [68]. EVOO contains levels of total
erythrodiol from 19 to 69 mg/kg of oil, and the free form is inferior to 50 mg/kg [59,68].
2.1.2. Tocopherols
Three isoforms of tocopherols are present in EVOO: α-, β- and γ-tocopherol. α-Tocopherol can be
found in its free form and represents more than 90% of the identified section with ranges from 206.5
to 270.9 mg/kg of oil to 191.5 to 292.7 mg/kg of oil, values that fluctuate with variables as the year of
harvest and spacing between olive trees [71]. Both, the distance between plants and the crop year
influenced statistically tocopherols amount [43,71]. Besides, the high levels of this type of tocopherol
may be linked to the high levels of chlorophyll pigments and the simultaneous necessity for singlet
oxygen deactivation [72].
Lower quantities of β-Tocopherol (~10 mg/kg), γ-Tocopherol (~20 mg/kg) and δ-Tocopherol
(~10 mg/kg) can also be found on EVOO. The total tocopherol concentration seems to decrease in the
ripping of the fruits, and the refining or the hydrogenation process causes their degradation, so they
are only found in the EVOO and VOO [73].
2.1.3. Carbohydrates
There are two hydrocarbons mainly present in olive oil, 2,6,10,15,19,23-Hexamethyl-2,6,10,14,18,22-
Tetracosahexaene also known as squalene and β-Carotene, which will be addressed in the pigments
section of this review. Squalene is the last metabolite synthesized before the sterol ring formation.
Some of the beneficial health effects of olive oil are partially linked with the presence of squalene, and
it has also demonstrated antitumoral effects against certain types of cancer [74,75]. This compound
constitutes more than 90% of the hydrocarbon fraction and is the most abundant compound in the
unsaponifiable matter, with concentrations ranging from 200 to 7500 mg/kg oil [57]. In a different
Foods 2020, 9, 1014 10 of 31
study, squalene was reported in even higher concentrations, up to 12,000 mg/kg oil. Squalene content
depends on several variables like the type of olive cultivation and the oil extraction technique applied,
and it decreases largely during the refining process so it is present in larger quantities in EVOO and
VOO [76].
The remaining fraction of carbohydrates in EVOO is composed of triterpene and diterpene,
isoprenoid polyolefins, hydrocarbons and n-paraffins [43,76].
2.2.2. Pigments
The lipophilic carotenoid and chlorophyll pigments occurring in olive oil are responsible for its
characteristic color [95]. The coloration of EVOO is greener in the presence of green olives that have
higher chlorophyll content whereas using mature olives with higher carotenoid content we obtain a
more yellowish oil, so the final color is a result of the proportions of these pigments [96]. EVOO has a
large variety of carotenoids and chlorophylls, from β-Carotene, violaxanthin, neoxanthin, lutein and
other xanthophylls to chlorophyll a and b, pheophytin a and b and other minor derivatives [97,98].
These pigments can be found in amounts up to 100 ppm of total carotenoids and major pigments like
pheophytin up to 25 ppm, β-carotene up to 15 ppm and lutein up to 10 ppm [96], although these values
depend on various factors. The final concentration of each pigment in the final EVOO relies on the
physicochemical characteristics of the fruit, the geographic origin, climate and irrigation conditions
and the mechanic extraction process used. Storage conditions and final packaging also play a role in
pigment concentration and type [96,99,100].
Quality and adulteration of EVOO are sometimes analyzed through the measuring of pigment
compounds because they are correlated with EVOO nutritional value, freshness and antioxidant
properties [99,101]. In addition, pigments can also be used for the authentication of EVOO, by
measuring the chlorophyll and carotenoid pigments of EVOO and comparing them through a quality
index, in which the total chlorophylls to total carotenoids ratio must be around 1 and the ratio of minor
carotenoids to lutein must be around 0.5, to declare it an authentic olive oil [102]. These parameters are
valid for any olive oil regardless of the studied variety. Furthermore, other pigments like violaxanthin,
lutein and total pigment content can be useful as a tool to identify a monovarietal EVOO [102].
Chlorophylls, carotenoids and other minor pigments like lutein and violaxanthin can be stable for
more than one year in storage regardless of the degree of ripeness and variety of the olives used to
produce that oil [103].
The degradation of chlorophylls occurs as a consequence of a pheophytinization reaction that starts
from the malaxation step during the extraction of the EVOO and increases throughout storage time [104].
During that process, the chlorophylls naturally present (a and b) are slowly but irreversibly converted
into pheophytins a and b, where the central Mg+2 ion of the porphyrin ring is exchanged with two
hydrogen atoms making the molecules more stable. These eventually turn to pyropheophytins by the
removal of the carboxymethyl group, which are the ultimate products of chlorophyll degradation [105].
More recently, the cardioprotective benefits of a MED enriched with EVOO have been proven by
the PREDIMED study. This multicenter, randomized, controlled trial involved ~ 7500 subjects with
potential cardiovascular risk, showing no CVD at enrolment. The PREDIMED trial resulted in a 30%
decrease of a major CVD development, such as stroke or myocardial infarction, in comparison to a
control group that followed a low-fat diet [30,107]. An observational study based on the PREDIMED
cohort indicated that consumptions of 10g EVOO/day are related to CVD risk diminutions up to
10% [112]. A recent systematic review evaluating clinical trials reported that diets enriched with
10–50 mL/day of EVOO (but not diets supplemented with EVOO capsules) significantly decreased
diastolic blood pressure by 0.73mm Hg [113].
In another meta-analysis of randomized controlled trials, case-control and prospective cohort
studies including ~40,000 cases of stroke and ~100,000 cases of coronary heart disease (CHD), it was
reported that for each increase of 25 g of olive oil intake, stroke and CHD risk was reduced by 26% and
4%, respectively. When combined stroke and CHD, olive oil consumption also showed preventing
effects, decreasing the risk of a CVD event by approximately 18% [114].
The preventive role of EVOO polyphenols against CVD was also documented in a meta-analysis
of controlled trials that evaluated the effect of low versus high polyphenol olive oil on markers of CVD
risk. Olive oil consumption ranged from 25–75 mL/day. High polyphenol olive oil significantly reduced
the CVD-risk markers malondialdehyde, oxidized LDL, total cholesterol, high-density lipoprotein
(HDL) cholesterol and also some inflammatory indicators like C-reactive protein (CRP) or interleukin-6
(IL-6) [115].
among unhealthy groups (with type 2 diabetes mellitus or at risk of CVD). Another meta-analysis
comprising 3106 participants also showed a significant reduction of IL-6 and CRP levels, when olive
oil was consumed as a supplementary or natural intake [123]. The adherence to a high-phenol
VOO breakfast decreased the postprandial inflammatory response, reducing the levels of plasma
lipopolysaccharides in patients with metabolic syndrome [124]. The higher polyphenol content of
EVOO may mediate the mentioned favorable effect as it has demonstrated anti-inflammatory effects
in vitro [133]. The anti-inflammatory effect of phenolic compounds-enriched EVOO has also been
reported in the adipose tissue in mice, with anti-atherosclerotic effects [134].
Due to these mentioned capacities, EVOO has also been proposed as a potential therapeutic product,
reducing inflammation in inflammatory bowel diseases, including ulcerative colitis and Crohn’s
disease, being both related to chronic inflammation of the intestinal mucosa [135,136]. The benefits
of EVOO consumption were evaluated in other autoimmune and chronic inflammatory diseases
such as rheumatoid arthritis [137], systemic lupus erythematosus [138] or multiple sclerosis [139,140]
with promising results in murine models. Besides, both in vitro and in vivo studies outline that the
anti-inflammatory activity of EVOO provides a neuroprotective effects that could prevent cognitive
decline and, therefore, the development of Alzheimer’s disease or elderly dementia [3,141].
phenolic compounds intake is not absorbed in the small intestine; therefore, they remain in the large
intestinal lumen where they are subjected to gut microbiota metabolic activities. As a consequence,
polyphenols are converted to low-molecular-weight compounds that are absorbed and responsible for
the health benefits derived from polyphenol-rich food, such as EVOO [132,146,147].
A recent review and meta-analysis of randomized controlled trials supports the prebiotic action
of polyphenols, capable of modulating and improving intestinal microbe populations, which affects to
CVD and colorectal cancer markers [132]. Furthermore, another randomized controlled trial showed
that the ingestion of VOO enriched with phenolic compounds decreases the serum levels of oxidized
LDL in hypercholesterolemic participants as well as increases the presence of Bifidobacterium spp in
feces. Slight changes in the profile of fecal microbial metabolites were also reported. These data
suggest that the cardioprotective effect of phenolic compounds could be mediated by the populations
of bifidobacteria present in the gut microbiota [131]. The possible modulation of gut microbiota by
olive oil and its role in cancer prevention, especially colorectal cancer, has also been suggested [148].
However, the complexity of the human diet, the lack of accuracy in the measurement of dietary
intake and the extensive variation on microbiota between individuals challenges the evaluation of how
diet changes modulate gut microbiota and its metabolic activity [149].
Figure 4. Parameters and factors concerning EVOO quality and its preservation.
with optical image detection from the smartphone’s camera, resulting in on-site analysis of samples
without requiring any pretreatment and without the need for a laboratory environment [155,161].
40 ◦ C cause greater adulterations in the composition of pigments than low temperatures, such as 3 ◦ C,
although the ideal temperature for its conservation is around 23 ◦ C [154,172].
As a curious fact, when EVOO is stored at low temperatures, a crystallization process caused by
freezing can be observed. In this process, triacylglycerols start to freeze through the methyl fraction,
which is just the part that melts first in the opposite process. Through the analysis of the freezing and
melting kinetics, information related to the geographical origin of the EVOO can be obtained [160,173].
Moreover, low temperatures application during EVOO processing has shown to be an effective strategy
to improve its yield and quality [174].
6. Conclusions
EVOO, also colloquially known as “liquid gold” is a natural product of unquestionable value
and not only in the monetary sense of the term but also for its recognized properties and advantages
on health. It is considered food for its nutritional value and is practically a requirement of the MED,
contributing to the benefits associated with it. Its chemical composition allows EVOO to be classified
as a lipophilic product since lipids are the main compounds, especially MUFA, followed by PUFA.
This lipid fraction is responsible for protective properties on coronary, autoimmune and inflammatory
disorders, granting anti-thrombotic and regulation effects of blood pressure Although in a smaller
quantity, other compounds such as tocopherols or polyphenols are also present, which are associated
with the powerful antioxidant and inflammatory activity of EVOO, among other qualities. For all
these reasons, the inclusion of this golden ingredient in the diet, in addition to offering characteristic
organoleptic properties, provides substances capable of preventing the appearance or development
Foods 2020, 9, 1014 20 of 31
of diseases of various nature, from heart and circulatory diseases to metabolic disorders, including
carcinogenic processes. Compared to other types of olive oil, EVOO must meet more stringent
requirements that give it the right to possess that nomenclature. However, it is a shame that, during
distribution and storage, the previously achieved quality is corrupted, leaving some of its most
distinguished and desirable properties on the way. Therefore, the factors that promote the degradation
of its components must be known, intending to reduce the negative impact that may originate in
EVOO and thus increasing its shelf-life. These factors can be divided into intrinsic factors, such as
the variety of olives and their cultivation conditions, about which little can be done once EVOO has
been produced; and external factors, which include exposure to light, temperature, time or the type
of material used for packaging. Due to the functional properties it presents, other applications for
EVOO could also be considered, such as its use in therapies in which other treatments are not very
effective, like certain neurodegenerative diseases or as a vehicle for the administration of certain
pharmacological compounds in a comfortable way, by simply adding them to the diet. Future studies
aimed at optimizing and maximizing the capabilities and applications of this product will, without a
doubt, be welcomed and well received by the industry, not only food and agriculture but also by the
pharmaceutical and cosmetics industries.
Author Contributions: Formal analysis, M.A.P., J.S.-G., C.J.-L., M.C., C.L.-L., M.G.-G., J.M.L. and F.J.B.;
investigation, C.J.-L., M.C., C.L.-L., M.G.G. and F.J.B.; methodology, C.J.-L., M.C., C.L.-L., M.G.-G., J.M.L.
and F.J.B.; supervision, M.A.P. and J.S.G.; validation, M.A.P. and J.S.G.; writing—original draft, M.A.P., J.S.-G.,
C.J.-L., M.C., C.L.-L., M.G.-G. and F.J.B.; writing—review and editing, M.A.P. and J.S.-G. All authors have read
and agreed to the published version of the manuscript.
Funding: The research leading to these results received financial support from Programa de Cooperación
Interreg V-A España—Portugal (POCTEP) 2014–2020 (projects Ref.: 0181_NANOEATERS_01_E and Ref:
0377_IBERPHENOL_6_E) supporting the pre-doctoral work of C.J.L. To MICINN supporting the Ramón&Cajal
grant for M.A.P. (RYC-2017-22891); to the company AlgaMar (www.algamar.com) and Axudas Conecta Peme
(Xunta de Galicia) supporting the IN852A 2018/58 NeuroFood Project that financed the pre-doctoral grant for
C.L.L.; to EcoChestnut Project (Erasmus+ KA202) for supporting the work of M.C.R.; to Ibero-American Program
on Science and Technology (CYTED-AQUA-CIBUS, P317RT0003) for financial support. This project has received
funding from the Bio Based Industries Joint Undertaking (JU) under grant agreement No 888003 UP4HEALTH
Project (H2020-BBI-JTI-2019), the JU receives support from the European Union’s Horizon 2020 research and
innovation program and the Bio Based Industries Consortium.
Conflicts of Interest: The authors declare no conflict of interest.
Abbreviations List
Generic
CHD Coronary Heart Disease
CVD Cardiovascular Diseases
DSC Differential Scanning Calorimetry
DPPH 2,2-Diphenyl-1-picryl-hydrazyl-hydrate free radical assay
EFSA European Food Safety Authority
EU European Union
EVOO Extra Virgin Olive Oil
GC Gas Chromatography
HPLC High Performance Liquid Chromatography
HIV Human Immunodeficiency Viruses
IC50 Half Maximal Inhibitory Concentration
IOC International Olive Council
MED Mediterranean Diet
NMR Nuclear Magnetic Resonance
PREDIMED Prevention Through Mediterranean Diet
USA United States of America
VOO Virgin Olive Oil
Foods 2020, 9, 1014 21 of 31
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