THESIS PROPOSAL

FOR
AVOCADO OIL AND ITS ANALYSIS

INTRODUCTION:
Avocado (Persea Americana) is a fruit native to Central America, grown in warm temperate
and subtropical climates throughout the world. The pulp of this fruit contains about 60% oil,
7% skin, and approximately 2% seed. %). Popularly known as “vegetable butter” or “butter
pear”, the fruit contains a substantial amount of triglycerides (TGs) along with a high content
of unsaturated fatty acids. Avocado fruit is rich in monounsaturated fat and contains
relatively high level of important lipid soluble compounds such as vitamin E, β-sitosterol and
carotenoids. The consumption of avocado fruit is highly related to its potential benefits. It is
also rich in many other bioactive phytochemicals such as tocopherols, phytosterols, aliphatic
alcohols and hydrocarbons. TGs are the most important nutritive group of compounds in
avocado oil and represent a significant amount (~90%) of the entire oil composition.
Chemically, TGs are complex hydrophobic molecular species formed by the esterification of
three fatty acids (FAs) with a glycerol backbone under enzymatic catalysis. The complexity
of TGs is due to a large number of possible FA combinations attached to the glycerol
skeleton, which can differ in the number of acyl carbon atoms (CNs), the degree of
unsaturation, and the position and configuration (cis/trans) of the double bonds (DBs) in each
FA.

Unlike oil extracted from other fruits, the oil from avocado fruit is often extracted from the
mature fruit flesh, and its lipid content has been reported as the highest among all known fruit
and vegetable varieties. This has sparked a growing interest of human nutrition, food
industry, and cosmetics in Avocado oil. Avocado oil has a multitude of applications such as
culinary oil and as an ingredient in healthcare products, cosmetics, pharmaceuticals and
nutraceuticals. The consumption of avocado oil has become popular owing to its high
nutritional value and potential benefit to human health, including the management of
hypercholesterolemia, hypertension, diabetes and fatty liver disease etc. The lipid content,
mainly of monounsaturated fatty acids, is associated with cardiovascular system benefits and
anti-inflammatory effects and possesses anti-cancer and antimicrobial properties. There are
no internationally defined parameters for avocado oil. The values that are commonly used are
those recommended for olive oil. The quality standard for olive oil is available in the Codex
Alimentarius and the International Olive Oil Council (IOC).
Classification of avocado oil based on its extraction method and fruit quality.

Avocado oil of a higher quality, “extra virgin”, corresponds to that produced from high-
quality fruit, extracted only with mechanical methods, using a temperature below 50 ◦C and
without the use of chemical solvents. “Virgin” avocado oil is produced with fruit of a lower
quality (with small areas of rot and physical alterations), extracted by mechanical methods,
using a temperature below 50 ◦ C and without the use of chemical solvents. “Pure” avocado
oil is a type of oil for the production of which the quality of the fruit is not important; it is
bleached and deodorized oil, infused with the natural flavor of herbs or fruits. Finally,
“mixed” avocado oil is combined with olive, macadamia, and other oils. Therefore, it
presents sensory and chemical characteristics that are variable. The Mexican norm states that
the “crude oil of avocado” is a slightly amber-colored fatty liquid, obtained by physical
extraction of the pulp and the seed of the fruit (Persea americana). “Pure” edible avocado oil
is a product with at least 98.5% refined avocado oil. In this work, a systematic review of the
literature was carried out to collect, select, evaluate, and summarize all available evidence
regarding the processes and properties of avocado oil.

The research question was: What are the most published topics on avocado oil? The answer
to this question allowed us to include topics, such as: extraction methods (i.e., cold pressed
method, ultrasound-assisted aqueous extraction method, supercritical CO2 method, CO2
subcritical method, enzymatic extraction, and solvent extraction), procedures of conservation,
contamination/adulteration, technological applications, composition (characteristics
according to the variety and origin of the fruit, physicochemical characterization, avocado
seed oil, and comparison with other oils), and biological effects (human health effects and
experimental studies in animals).

The purpose of this research is to produce a complete profile on avocado oil, including
extraction methods, physicochemical characteristics, nutritional properties, conservation, as
well as various applications in the food and medicine industries. Avocado oil has proven to
be a vegetable oil with a composition of major and minor components that are highly
appreciated by the population, either at low or high temperatures, with multiple technological
applications.

BACKGROUNG AND SIGNIFICANCE:

The avocado tree (Persea Americana) is a member of the Lauraceae family and originated in
Mexico and Central America. Its thick-skinned fruit is botanically considered a berry and
contains a single large seed. Earliest archaeological records of the existence of avocados
came from Coxcatlan in Mexico in approximately 10,000 BC. Evidence suggests they were
cultivated as a food source since at least 5000 BC by Mesoamerican people. The first
published description of avocados, by a Spanish explorer to the New World, was made in
1519 by Martin Fernandez de Enciso in the book Suma de Geografia. During the subsequent
Spanish colonization of Mexico, Central America and parts of South America in the 16th
century, avocado trees were introduced across the region and flourished in the warm climates
and fertile soils. The Spanish also brought avocados across the Atlantic Ocean to Europe and
sold them to other countries like France and England. Europe’s primarily temperate climates
were not ideal for growing avocados though.

While avocados have been eaten by people for thousands of years, avocado oil is a relatively
new innovation, particularly as culinary oil. In 1918 the British Imperial Institute first drew
attention to the possibility of extracting the high oil content from avocado pulp, though there
is no record of avocado oil being produced at this time. In 1934 the California State Chamber
of Commerce noted that some companies were using blemished avocado fruit, unfit for sale,
for oil extraction. Early methods for extracting avocado oil involved drying avocado pulp and
then squeezing out the oil with a hydraulic press. The process was laborious and did not
produce significant quantities of usable oil. In 1942 a solvent extraction method of avocado
oil production was first described by Howard T. Love of the United States Department of
Agriculture. Around this time experiments were conducted for large-scale production of
avocado oil due to shortages of fats and cooking oils during wartime. Solvent extraction of
avocado oil became popular for producing refined avocado oil, used as a lubricant and
particularly in the cosmetics industry. However, the solvent extraction method required
significant further refinement and heating before the oil was ready for commercial use.
Additionally, much of the nutritional value of the avocado was lost in the process. Avocado
oil produced by chemical solvents is still produced today, mainly for use in face creams, hair
products, and other cosmetics. This clear and highly refined avocado oil is not considered
suitable for cooking with.

The primary reason for the increase in avocado oil use as culinary oil throughout the world in
recent years is its nutritional properties and health benefits. Cold pressed avocado oil is high
in vitamin E, an antioxidant with protective effects on the cardiovascular system. It also
contains good concentrations of beta-sitosterol, a phytosterol that reduces cholesterol
absorption during digestion. Lutein is another antioxidant found in avocado oil produced
without excessive heat or chemical solvents. Dietary lutein is associated with improved
vision and a lower risk of age-related macular degeneration. The fatty acid profile of avocado
oil produced by cold pressing is between 72% and 76% monounsaturated fats, with saturated
fats at around 13%. A higher intake of monounsaturated fatty acids to saturated ones is a
central part of the highly regarded Mediterranean diet and the main reason why olive oil is
considered healthy by nutritionists. However, olive oil has a lower ratio of monounsaturates
and a higher percentage of saturated fat than avocado oil. Comparing the nutritional profiles
of the two, avocado oil is superior to olive oil in both antioxidants and fats. Another factor
which makes avocado oil more versatile than olive oil is its significantly higher smoke point.
Smoke point is the temperature at which the structure of cooking oil starts to break down and
begins smoking. Extra-virgin olive oil has a very low smoke point, often listed as low as
220°F (105°C). This makes it unsuitable for frying and cooking at high temperatures. By
comparison, avocado oil has a smoke point as high as 482°F (250°C), making it much better
high temperature cooking oil. Avocado oil also has a flavor which many consumers say they
prefer to the taste of olive oil. It is often recommended as a salad dressing and other culinary
purposes where olive oil is usually used.
RESEARCH DESIGN AND METHODS:

PROCESSING OF AVOCADO OIL:

To produce a quality product requires high quality raw materials. For this reason it is
imperative that the fruits used are of high grade in particular must contain high proportion of
pulp, have high oil content, free of diseases and must not be physically damaged as evident in
pulp discoloration and/or off-flavors. The fruits have to be carefully matured and ripened to
allow for maximum oil development before they are used. Various methods have been used
in the extraction of oil from avocado fruits. The methods vary in their degree of effectiveness
for oil extraction and also subsequent effect on the resultant oil quality. Heating and chemical
extraction have been the traditional methods used and now a new method has been developed
in New Zealand which allows oil to be expressed from the fruits with very minimal
processing. Various methods used in the past for the extraction of avocado oil involving
steam pressure, hydraulic pressing, solvent extraction, centrifugation, freeze-drying,
rendering process and the use of a tube press plant. However the specific application of the
oil should determine the process and method chosen. Only the most practical and suitable
methods are discussed below.

1. Pre-Process Treatment:

Avocado fruits destined for oil production must be firstly inspected for physical damage and
other abnormalities. They are then washed before being processed to remove the skin and
seed. The means used for washing, de-stoning and de-skinning of the fruits vary but all
processes involve this first critical step.

2. Extraction Methods:

Considering the high humidity percentage of avocado (around 70 to 80%), the influence of
the pulp drying method prior to oil extraction has been studied. The quality parameters
(peroxide value, iodine value, amount of oleic acid, refractive index, electrical conductivity,
content of carotenoids, chlorophyll, phenolic compounds, and antioxidant activity) have
shown better results when the pulp is dried at 60 ◦C under vacuum, and the extraction is
performed by the Soxhlet method. Meanwhile, the bioactive compounds were best preserved
when the avocado pulp was dried at 60 ◦C with air ventilation and mechanical pressing. On
the other hand, avocado pulp oil, pressed and dried in a microwave, presented a better quality
—determined by the acidity index, peroxide index, and oxidative stability—when compared
with oil obtained by extraction with ethanol. The composition of fatty acids did not differ
significantly when analyzing oil obtained by drying under microwaves or in a drying oven
with forced air circulation. According to Chimsook and Assawarachan, the studied drying
method of the avocado pulp, prior to the extraction of the oil, does not significantly influence
the composition of fatty acids. However, changes were determined in the antioxidant activity
and vitamin E content of cold-pressed avocado oil. Higher antioxidant activities and higher
vitamin E content were observed in oil, the pulp of which was dried with hot air, when
compared to oils obtained by an air-dried and vacuum process. This study is consistent with
the fact that oils from the fortune avocado variety, obtained by the pulp drying lyophilization
method, resulted in lower concentrations of α-tocopherol, squalene and β-sitosterol, as well as
higher relative concentrations of campesterol and cycloartenol acetate, compared to oils
obtained through hot air-drying processes.

2.1. Aqueous separation:

The water or aqueous extraction is a traditional process used to recover oil from plant
resources. Water separation of avocado oil is performed by enzymatic or mechanical
destruction of tissue cells contained lipids followed by centrifugation or gravity setting to
separate the oil from the oil-water emulsion.

One of outstanding advantages of aqueous separation technique is that it does not need to
remove a large quantity of water from fresh pulp, compared to solvent and pressing
extraction process. The aqueous separation technique is practicable due to its simple
operation and low cost, compared to other oil extraction methods (like organic solvent
extraction). Besides, the benefit of utilizing centrifugal force to separate lies in yielding a
higher quality product with higher purity, which greatly simplifies the subsequent refining
process.

I. Centrifugation Method:

After the pre-processing treatment, the fruits are fed into a mill where it takes the form of a
guacamole. The malaxation takes up to several hours until the release of the fine emulsion of
oil. The paste is then fed to a centrifugal decanter where the oil is separated from the
guacamole. Extraction of oil was most efficient using centrifugal force 12,300 xg, 5:1 water
to avocado ratio, temperature 75oC, with a pH 5.5 and a 5% concentration of either NaCl,
CaCO3 or CaSO4.

I. Hot water Separation Method:

According to the mechanically-assisted hot water separation method, avocado fruits are
subjected to mechanically mashed, then oil is released from slurries by boiling water and
subsequent pressing, the oil layer is then separated by gravity setting.

The fine grounded slurries are diluted or treated with hot water to allow easy release of oil
from the slurries and inactivation of lipolytic enzymes. In this method, the diluted slurries are
generally heated at a higher temperature (100–105℃) to separate the oil part. The diluted
slurries also can be homogenized using a blender prior to heating. Proteins and carbohydrates
existed in avocado fruit can improve the emulsion stability, leading to a harder break-down of
the linkage between oil and these components during heating. To make it easier to break
down the emulsion, homogenization pressure is employed prior to heating.
 III. Ultrasound-Assisted Aqueous Extraction Method (UAAE):

This method uses the cavitation forces produced by acoustic waves to break down the cell
walls of the oil-containing cells. This process allows for the generation of an emulsion, which
facilitates oil extraction. This method can be carried out using an ultrasonic bath or an
ultrasonic horn transducer. The high frequency ultrasound conditioning (0.4, 0.6, and 2 MHz,
5 min, 90 kJ/kg) of the avocado puree can improve the oil separation and potentially reduce
the beating time in industrial processes, without affecting the quality of the oil. If this
treatment is applied after shaking, the extractability of the oil increases by between 2% and
5%. The oils obtained from sonicated purees showed free fatty acids (FFA) and peroxide
values below the levels of industrial specification (peroxide less than 20 meqO2/kg) and an
increase in total phenolic compounds after a 2 MHz treatment. The ultrasound-assisted
aqueous extraction (UAAE) of low virgin avocado oil in FFA, considered as virgin avocado
oil, is that obtained by mechanical or natural means at low temperatures.

2.2. Pressing extraction:

Pressing extraction refer to oils extracted by pressing or squeezing oily materials with screw
press or hydraulic press. Pressing technology commonly used to squeeze oil from oilseed
materials (like sesame) with relatively high oil content. Compared with oilseeds, avocado
pulp contains higher moisture (about 77％) and its cellular contents are different. Water
content of fruit pulp can significantly affect the oil yield. Pretreatment methods of avocado
pulp, thus, can be different prior to pressing. The pre-treatment approaches include (1)
slicing and dying of avocado flesh, (2) microwave-oven drying and (3) the addition of
solid additives. Traditional dying procedures such as oven-drying and sun-drying are time
consuming to dry the slices to 4％ –5％ water content, accompanying with a relatively high
risk of poor oil quality. A high temperature (＞100℃) is accompanied by this high
microwave energy, resulting in severe transforming the structure of idioblastic oil cells. Such
transformed structure has a negative effect on the oil extraction yield. When the highest oil
extraction yield by microwave-assisted squeezing was obtained at the optimized energy
(1.89kJ/g), the idioblastic cells became empty with no major changes.

The addition of solid additives is another way to reduce avocado pulp’s moisture and
viscosity and to increase the oil extraction yield. Solid additives are supposed to have a
certain hardness and granularity and to be non-toxic, insoluble in oil or water. After sufficient
mixing avocado pulp with solid additives, appropriate heating is required for subsequent
squeezing. Such heating favors accelerating the disruption of cell walls during the extrusion
process, making the decrease of the viscosity of cellular oil to increase the oil extraction
yield. Besides, moderate heating favors the inactivity of lipases, which reduces or eliminates
the hydrolysis of avocado oil during the squeezing and storage period.

I. Cold Pressing Method:

In the cold pressing method, oil recovery is only obtained from the parenchyma cells of the
pulp; its rupture begins in the first stages of grinding and it can be seen that the idioblastic
cells (oil carriers) remain intact during the process of extraction. The extraction yield
increases when the pulp is beaten at 45.5 ◦C for 2 h. When this process is complete, a three
phase decanter then separates the mixture into oil, water and solids before polishing takes
place with a multi-cone centrifuge. Extra virgin oil is produced after the first press. The
extraction efficiency is dependent on such things like pH, centrifugation rate, salt, mixing
temperature (<50oC) and duration of pressing. The extraction rates vary from 10-22%. In this
method, a lower extraction yield is obtained, although with higher concentrations of α-
tocopherol and squalene, as well as lower contents of campesterol and cycloartenol acetate
compared to the Soxhlet method. Drying by lyophilization and subsequent extraction by the
Soxhlet method allows for a better extraction performance. However, when drying by
lyophilization and extracting by cold pressing, oils with a greater concentration of
antioxidants, and other bioactive compounds were obtained.

This method involves low temperatures and minimal processing and as a result the oil retains
all its natural flavor, nutrients and healthy properties. Thus the resultant oil is of high quality
and is considered” virgin oil” because of very minimal processes involved. This virgin oil has
brilliant emerald green color when extracted; the color is attributed to high levels of
chlorophyll and carotenoids extracted into the oil (Cold-pressed avocado oil has been
described as having an avocado flavor, with grassy and butter/mushroom like flavors), thus
making it more unstable. This makes packaging in dark bottles or tins and total avoidance of
oxygen a must for a longer shelf life for the product. This oil must be stored in a dark, cool
cupboard where the temperature never rises above 30oC, but should never be refrigerated or
it will solidify. Virgin oil only has a shelf life of two years if stored correctly. More stable
refined cold processed oil can also be produced after it has undergone further processes of
refining, bleaching and deodorized (RBD). The production of cold pressed oil requires little
investment and the process itself is simple. However the process has inefficiencies in that
around 6-15% of the oil remains in the pressed residue.

2.3. Solvent Extraction:

Solvent Extraction is one of the traditional methods commonly used. In this extraction
method various solvents could be used (organic solvents have mostly been utilized). Solvent
extractions using hexane and a Soxhlet extractor on a 10g dried sample for 8 hours and
subsequent removal of the solvent followed by vacuum evaporation and drying until constant
mass is obtained. The resultant oil has a high chlorophyll content meaning the chlorophyll is
co-extracted along with the oil. Chlorophyll levels as high as 192.9 ppm has been extracted
by ethanolic extraction. This method has the highest yield but industrial equipment is very
expensive to install plus the highly flammable solvents used are very dangerous. The
recovery and total removal of the solvent is also an issue which requires a highly
sophisticated plant. Thus because the raw material is an expensive fruit and the oil yield is at
maximum around 22% of the whole fruit it is not economical to extract avocado oil in this
way. This method does have other disadvantages like loss of volatile compounds, long
extraction times, toxic solvent residues and degradation of valuable oil compounds.

2.4. Supercritical CO2 Method:

This method of extraction is based on the use of supercritical fluids, substances that are, in
certain circumstances, in a state in which they have intermediate properties between liquid
and gas. Supercritical CO2 (SC-CO2) is a totally innocuous gas, which becomes a powerful
solvent under conditions of pressure and at a temperature above its critical point. Extraction
with SC-CO2 presents a higher performance at a pressure of 400bars. The use of ethanol as a
co-solvent favors the extraction of residual oil, benefiting the extraction of a fraction enriched
in tocopherols .

Supercritical fluid extraction (SFE) is an environmentally friendly and cost-effective

extraction method with a multitude of applications in the food, pharmaceutical and fine
chemical industries. The supercritical carbon dioxide（SC-CO2）as a green solvent is
biological safety with no solvent residue in the final product, compared with organic solvents
used in oil extraction. In the present study, SFE was used for the extraction of oils from
avocado peel, pulp and seed. The color of the oils extracted from the SFE method was much
lighter than that of the solvent extraction, indicating a smaller amount of chlorophyll and
carotenoids being extracted. An important outcome of extracting with SFE method was that
the unsaponifiable fraction of the oil was found to be higher for the first fractional extraction
(20 minute intervals) and tend to decrease with subsequent extractions. Thus the
unsaponifiable fraction of the oil can be enriched by extracting at time intervals.

The use of supercritical fluid is proven to be a cost-effective technique for laboratory scale
while large scale units still require experimentation for accurate economic valuations. This
method has advantages such as low operating temperatures, shorter extraction periods, high
selectivity in the extraction of compounds and no undesirable solvent residue. It also uses a
safe, readily available gas.

2.5. CO2 Subcritical Method:

Extraction with S-CO2 operates under the same principle as the SC-CO2 extraction, but with
a temperature below 31.1 ◦C and CO2 pressure of 72.9bars. Extraction with S-CO2 was
performed at 27 ◦C and 68 bar CO2. Oils extracted using S-CO2 had higher iodine index
values but lower melting points, determined by slip, free fatty acid content, and
saponification index values. The oils extracted by S-CO2 have a clear color and higher levels
of unsaturated fatty acids than the oil extracted with hexane. Regardless of the extraction
method, the main fatty acids in avocado oils were oleic and palmitic acids, while the main
triacylglycerols in avocado oils were palmitoyl-dioleoyl-glycerol and palmitoyl- Oleoyl-
linoleoyl-glycerol.

2.6. Enzymatic Extraction:

In order to improve the performance of extraction by centrifugation, the incorporation of

enzymes, such as pectinases, α-amylase, proteases, and cellulase, to avocado paste have been
considered. The yield varies depending on the concentration and type of enzyme used and the
reaction time and percentage of water used. It is emphasized that this method improves oil by
up to 25 times, in comparison with the performance of a non-enzymatic centrifugation
 2.7. Rendering process:

This method involves heating of the avocado pulp in avocado oil. The moisture evaporates
off leaving the oil and the dry matter behind. The oil can then be decanted off while the
remaining slurry is subjected to hydraulic pressure to press out more or the remaining oil. It is
reported that laboratory experiments resulted in 94% recovery of the oil using this method.

2.8. Tube Press Plant:

This consists of either one or two tubes. The tubes are filled with avocado pulp and then
subjected to hydraulic pressure to press out the oil through perforations within the inner
tubes. This method aims at extracting out most of the oil with minimal damage to the oil
quality.

REFINING OF AVOCADO OIL:

Virgin and crude oil can be further processed when it does not meet the virgin oil quality
standards. The crude oil with high chlorophyll and carotenoid content is a dark green
transparent oil with brown or yellow tints. This highly unstable oil could be refined using the
following steps:

1. Bleaching:

Bleaching removes color pigments like chlorophyll, carotenoid and lutein using acidified
activated earth at an elevated temperature. This is followed by filtration and a
spectrophotometer may be used to monitor the bleaching process by measuring the optical
density of the oil. Losses as high as 5% may be encountered here.

2. Deodorizing:

Deodorizing is required to remove objectionable flavor/odour which may develop during

bleaching by using steam distillation. This is done under vacuum at elevated temperatures.
Again losses as high as 7% may be incurred here.

3. Winterizing:

The presence of high melting components in the oil makes it cloudy at low temperatures.
These components tend to crystallize at low temperatures and must be removed. Winterizing
aid like oxystearin is used to help the formation of larger crystals which could then be
removed via decanting and filtration.

4. Alkali Refining:

The refining process involves the use of a strong alkali (NaOH) to remove free fatty acids and
peroxides. Both compounds tend to reduce the shelf life of the oil and also give it a rancid
smell. This tedious process may result in losses as high as 7% - 8%. The replacement of
NaOH with Na2CO3 or NaHCO3 has resulted in better refining process that retained more
that 85% of the phenolic compounds. These healthy compounds are usually lost with the
unsaponifiable fraction of the oil during the refining process when NaOH is used.

Another method for refining is by heating the oil under vacuum at elevated temperatures, and
sparged with live steam causing the free fatty acids to be distilled off. Refined oil is pale
yellow, bland, and highly stable. It is highly suitable for general purpose cooking because of
its high smoke point and has a healthy fatty acid composition. It is most suited for barbeques
as most common vegetable oil polymerize and oxidize readily on hot surfaces.

PROCEDURES FOR THE CONSERVATION OF AVOCADO OIL:

The conservation of oils is a necessary issue to address, since it allows for increasing the
useful life of the products. One of the efforts made to improve the conservation of avocado
oil has been the use of physical techniques, such as the electric field.

The electric field (voltage 9 kV cm−1, frequency 720 Hz, time of 5 and 25 min) allows the
polyphenol oxidase enzyme present in the avocado pulp to be inactivated, preserving the
components present in the avocado oil. The modifications in the quality of the refined oil
(established according to the acidity index, peroxides, and iodine) are minimal, considering
the electric field method as an alternative for the addition of synthetic antioxidants.

The oxidative stability (determined by finding the antioxidant activity reducing ferric ion,
(ferric reducing antioxidant power) FRAP), during the storage of cold-extracted avocado oil
in the presence of the oleoresins of Capsicum annuum L. (vegetable material rich in
carotenoids), was assessed. It was determined that the optimal extraction of carotenoids was
at a concentration of 1:3 (w/v: Capsicum annuum L/avocado oil) for 48 h in darkness at room
temperature. The behavior of the oil under stronger conditions (45 ◦C, 30 days) showed the
following characteristics: (1) the extracts were stable to lipid oxidation, with a Totox index
total value of 27.34, (2) 85.6% of carotenoids were conserved, (3) 80.66% of the antioxidant
activity was retained, and (4) there was a color change (∆E) of 1.783. The oleoresins obtained
by extraction with avocado oil can be considered as an economic and sustainable alternative
for the extraction of carotenoids, with a good oxidative stability, compared with organic
solvents.
CHEMICAL AND PHYSICAL PROPERTIES:

1. Oil content:

The weight of oil extracted from 40 g of avocado pulp was determined to calculate the lipid
content. Result was expressed as the percentage of lipids in the dry matter of avocado pulp.

2. Protein content:

Total protein was determined by the Kjeldahl method. Protein was calculated using the
general factor (6.25) (AOAC). Data were expressed in percentage of dry weight.

3. Water and Dry matter content:

This was determined according to the (American Oil Chemist Society (AOAC)).

4. Ash and mineral contents:

To remove carbon, about 2 g (powdered) of avocado pulp, in a porcelain container, was

ignited and incinerated in a muffle furnace at about 550°C for 8 h. The total ash was
expressed in percentage of dry weight.

5. Lipid indices:

Standard procedures of AOAC were used for indices values (AOAC); procedures were
applied for acidic value, iodine value, peroxide value, saponification value.

6. Fatty acid composition:

The oils were converted into methyl esters using a KOH/MeOH method (Standard FIL,
182:1999). The extracted fatty acid methyl esters (FAME) were dissolved in hexane for GC
analysis. The identification of the peaks was achieved by retention times and by comparing
them with authentic standards analyzed under the same conditions.

7. Viscosity determination (V):

Viscosity was followed at 25°C or at other temperature with a Stress Tech Rheologica
Rheometer (Rheologica Instruments) conducted with a steel cone-plate (C40/4) under a shear
stress increased from 7 to 20 Pa.

8. Triacylglycerols (TAG) analysis:

Triacylglycerols were analysed by HPLC. Analysis condition: Column temperature: 30°C;

eluent is MeCN/CH2Cl2 (67/33); 7, 5 L of sample injected in 5 ml of MeCN/CH2Cl2:25.
The peaks were recorded and the under peaks areas measured.

9. Flash point:

Flash point is determined experimentally by heating the liquid in a container and then
introducing a small flame just above the liquid surface. The temperature at which there is
flash/ignition is recorded as the flash point. Standard procedures of AOAC are followed.

10. Relative Density (RD):

Relative density (RD) is determined using the pycometer method according to CODEX;
Refractive index (RI) was carried out following the method AOCS-Cc 7-25.

11. Melting point:

Melting point of fats is used to characterize oils and fats and is related to their physical
properties, such as hardness & thermal behavior. Usually Differential scanning calorimetry
(DSC) technique and softening point (AOCS method Cc 3-25) are used for determination of
melting point of fats/oils.

12. Oxidation stability:

Oxidative stability of avocado oil was determined by the rancimat method, which is
considered as an accelerated determination of oxidation. The rancimat method evaluates the
stability by measuring the oxidation induction time, with the Rancimat apparatus. The
oxidation induction time (OIT) of the oil sample was then automatically recorded at 100, 110,
120, 130 and 140oC.

REFERENCES:
1. Dos Santos, M.; Alicieo, T.V.; Pereira, C.M.; Ramis-Ramos, G.; Mendonça, C.R.
Profile of bioactive compounds in avocado pulp oil: Influence of the drying processes
and extraction methods. J. Am. Oil Chem. Soc. 2014, 91, 19–27.
2. Woolf, A.; Wong, M.; Eyres, L.; McGhie, T.; Lund, C.; Olsson, S.; Wang, Y.; Bulley,
C.; Wang, M.; Friel, E.; et al. Avocado oil. From cosmetic to culinary oil. In Gourmet
and Health-Promoting Specialty Oils; Moreau, R., Kamal-Eldin, A., Eds.; AOCS
Press: Urbana, IL, USA, 2009; pp. 73–125.
3. Tan, C.; Tan, S.; Tan, S. Influence of Geographical Origins on the Physicochemical
Properties of Hass Avocado Oil. J. Am. Oil Chem. Soc. 2017, 94, 1431–1437
4. Wong, M.; Requejo-Jackman, C.; Woolf, A. What is unrefined, extra virgin cold-
pressed avocado oil? Inform 2010, 21, 189–260.
5. Costagli, G.; Betti, M. Avocado oil extraction processes: Method for cold-pressed
high-quality edible oil production versus traditional production.
6. Dos Santos, M.; Alicieo, T.V.; Pereira, C.M.; Ramis-Ramos, G.; Mendonça, C.R.
Profile of bioactive compounds in avocado pulp oil: Influence of the drying processes
and extraction methods. J. Am. Oil Chem. Soc. 2014, 91, 19–27.
7. Marie Wong; Cecilia Requejo-Jackman; Allan Woolf (April 2010). "What is
unrefined, extra virgin cold-pressed avocado oil?” Aocs.org. Retrieved 26 December
2019.
8. "Avocado oil”. Purdue University Center for New Crops and Plants Products. 1999-
02-18. Retrieved 2013-07-13