Innovative Food Science and Emerging Technologies 41 (2017) 109–116

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Innovative Food Science and Emerging Technologies

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Stability of the lipid fraction of avocado puree obtained by flash

vacuum-expansion process
Manuel Vargas-Ortiz a,b, Adrien Servent b, Marco Salgado-Cervantes c,⁎, Dominique Pallet b
a
CONACYT-CIAD (Centro de Investigación en Alimentación y Desarrollo), Coordinación Culiacán, Carretera El dorado Km 5.5, Col. Campo El Diez, Culiacán CP 80110, Sinaloa, Mexico
b
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, F-34398 Montpellier Cedex 5, France
c
Unidad de Investigación y Desarrollo de Alimentos, Instituto Tecnológico de Veracruz, M.A. de Quevedo 2779, Col. Formando Hogar, Veracruz Ver CP 91897, Mexico

a r t i c l e i n f o

Article history:
Received 23 November 2016
Received in revised form 2 February 2017
Accepted 27 February 2017
Available online 02 March 2017

© 2017 Published by Elsevier Ltd.

1. Introduction avocado proteins are oxidative enzymes such as polyphenoloxidase,

The avocado is a tropical fruit of great economic importance, rich in
vitamins A and B, minerals and antioxidants. It is also a source of bene-
ficial compounds such as carotenoids, phytosterols and tocopherols. The
lipid fraction of the avocado fruit comprises approximately 71% mono-
unsaturated fatty acids and 16% saturated fatty acids. This proportion
helps healthy blood profiles in consumers, and boosts bioavailability
of fat soluble vitamins present in the avocado itself. The unsaturated
fatty acids do not promote the formation of cholesterol and may be a
way for the mobility of fat-soluble vitamins. Investigative studies sug-
gest that avocado consumption reduces the risks of cardiovascular dis-
eases and helps in weight control and healthy aging (Dreher &
Davenport, 2013). One of the major problems of avocado is its high sus-
ceptibility to qualitative and quantitative quality losses. Postharvest
processing of the avocado is complex comparing to other fruits and
entails appropriate handling of the fruits at the right stage of maturity,
specific packaging, storage atmospheres and temperatures, transport
distance and time, and pest management among other factors. Inappro-
priate postharvest management leads to physical damage and/or phys-
iological disorders which degrade the nutritional and commercial
quality of the fruits (Bill, Sivakumar, Thompson, & Korsten, 2014). Due
to the difficulty of avocado postharvest management, various industri-
alization methods have been sought; yet this has proven to be an even
greater challenge than postharvest management, since the avocado,
unlike other fruits, has a high fat and protein content. The majority of
⁎ Corresponding author.
E-mail address: msalgadocervantes@yahoo.com.mx (M. Salgado-Cervantes).
lipoxygenase and lipases which rapidly spoil the pulp when the avocado
is processed (Elez-Martínez, Soliva-Fortuny, Gorinstein, & Martin-
Belloso, 2005). Currently, various chemicals are employed to prevent
spoilage of avocado products. However consumers are tending to reject
additives due to their association with potentially adverse effects on
human health (Jacobo-Velázquez, Castellanos-Dohnal, Caballero-Mata,
& Hernández-Brenes, 2013). At present, the worldwide economic
context favors trade of food products between countries, with a trend
toward production of natural foods with minimal quantities of
chemicals involved in their production and preservation (Castañeda-
Saucedo et al., 2014). Hence the necessity to evaluate emerging food
processing alternatives which include, as far as possible, only physical
processes for industrialization and preservation purposes. Flash vacu-
um-expansion (FVE) is a physical transformation process, which pro-
motes breakdown of plant tissues by means of temperature and
pressure differences. This process comprises two stages, in the first
(thermal stage) the raw material is placed in a chamber (at atmospheric
pressure, 101,325 kPa) and is heated by a flow of water vapor (the plant
material gradually increases its temperature, depending on the time of
exposure to the vapor); The second stage consists of introducing the
plant material into an expansion chamber with vacuum pressure (2 to
5 KPa), the pressure difference generates the instantaneous evaporation
of a part of the water constitutive of the plant tissues. When the water
changes from liquid to gas, it increases 1000 times its volume (it ex-
pands), so it requires more space to be contained, this causes the cellular
rupture and a puree of the vegetal material is obtained; the FVE has
been used to obtain guava, mango and passion fruit purees, obtaining
higher yield, soluble solids content, antioxidant capacity and better

http://dx.doi.org/10.1016/j.ifset.2017.02.016
1466-8564/© 2017 Published by Elsevier Ltd.
110 M. Vargas-Ortiz et al. / Innovative Food Science and Emerging Technologies 41 (2017) 109–116
coloration (Cogat, 1994; Brat, Olle, Reynes, Alter, & Cogat, 2002). Partic-
ularly in avocado, Vargas-Ortiz, Rodríguez-Jimenes, et al. (2016) and
Vargas-Ortiz, Servent, et al. (2016) reported that the use of the FVE pro-
cess allowed to obtain puree with the best characteristics (tissue disin-
tegration, enzymatic inactivation, antioxidant capacity and color
retention). Avocado has a high proportion of lipids compared to other
evaluated fruits; the stability of the lipid fraction may be determinant
in the quality of the mash even when it retains a good appearance.
The FVE process can modify various factors that affect lipid stability,
for example cell disruption, reduction in enzyme activity, heat treat-
ment. Therefore the present investigation referring with the evaluation
of the processing time effect on the stability of the lipid fraction of avo-
cado puree. For this purpose, oil quality were evaluated through the
peroxide index, the iodine index, the K270 extinction coefficient and
the tocopherol concentrations (α and γ). The fatty acid profile of the
lipid fraction of the purees during refrigerated storage and the
lipoxygenase activity were also determined.
2. Materials and methods
2.1. Plant material
Hass variety avocados grown in Mexico, mature for consumption
(sensitive to the touch; opaque black skin) were bought from a local
market. The fruits were of homogeneous color and size.
2.2. Flash vacuum-expansion equipment
It is a pilot equipment (manufactured by Aurore developpement, La
Reunion, France), it consisted of a cylindrical stainless steel steam-
heating chamber (radio = 12 cm; height = 24 cm; v = 2.7 L), fed at
normal pressure with a water-steam generator and coupled through a
pneumatic valve (103 kPa; opening time 0.5 s) to a cylindrical vacuum
vessel (radio = 30 cm; height = 48 cm; volume = 34 L) where a vacu-
um (3 kPa) is generated by a vacuum pump cooled by a closed water cir-
cuit connected to a condenser.
2.3. Flash vacuum-expansion treatment
For each treatment, four intact avocados (approximately 750 g)
were placed in the steam-heating chamber. They underwent different
steam-heating times for each treatment: 15 min (T1), 20 min (T2)
and 25 min (T3); a fourth batch of fruit had its peel removed without
making cuts on the pulp, and was heated for 20 min (T4). The internal
temperature of the avocado was measured with a thermocouple
ZA9021-FST (Thermofisher, Massachusetts, USA) coupled with an ac-
quisition database Almemo 2690-8 (Ahlborn, Holzkirchen, Germany),
inserted as far as the seed. The temperature of the puree in the expan-
sion chamber was determined with the same equipment. Processing
times were determined after preliminary trials, by selecting those that
showed better results in the disintegration of the fruits, by selecting as
a central point 20 min. After each treatment time had elapsed, each
batch was instantaneously inserted into the vacuum vessel. The puree
was recovered, and the fragments of peel and seed were manually re-
moved. The puree was placed in hermetically sealable containers,
which were stored at 5 °C. Each treatment was applied in triplicate,
and the analyses of each parameter evaluated were performed in dupli-
cate at intervals of 5 days. The purees were stored without adding any
chemical or natural additives. Four avocados in consumption maturity
was withdrawn peel, seed and were processed for 3 min using a food
processor Thermomix Tm31 (Vorwerk, Wuppertal, Germany) equipped
with a temperature sensor, the puree obtained was used as a control
and was given the same storage which the treatments.
2.4. Oil extraction
Sampling was performed on days 0, 7 and 14 of storage. 200 g sam-
ples of puree were taken of each treatment (in triplicate), they were
placed in trays and subjected to dehydration using a lyophilizer model
SMH 15 (Usifroid, France). Lyophilized samples were stored at −18 °C
until the extraction of oil. The extraction of oil present in the mash
was performed as reported by Santana, dos Reis, Torres, Cabral, and
Freitas (2015) with minor modifications. Extraction was conducted
using petroleum ether as solvent; the lyophilized puree was extracted
for 1 h at 45 °C in a hot water bath with orbital agitation, with a ratio
sample: solvent 1: 3 (w/w). The extracts were vacuum-filtered and
the solvent evaporated using a rotary evaporator (37 °C, 50 mbar). Oil
was stored at −18 °C until analysis.
2.5. Peroxide value
The peroxide index was determined according to Mexican official
norm NMX-F-154-1987 (1987) and Fiebig (2003). Two grams of oil
from each treatment were placed in an Erlenmeyer flask and 50 mL of
an acetic acid-chloroform solution (3:2, v/v) were added. The mixture
was agitated until the oil were totally dissolved. Then 1 mL of saturated
potassium iodide solution (KI) was added. The mixture was agitated
gently for 1 min. Thereafter, 75 mL of distilled water was added and
the mixture was titrated with a 0.1 N solution of sodium thiosulfate, ag-
itating vigorously after each addition, until a slightly yellow coloration
was obtained. 0.5 mL of 1% (M/V) starch solution was added as an indi-
cator, and titration was continued with constant agitation, until the blue
color disappeared. The results were expressed as milliequivalents of ox-
ygen per kilogram of oil.
2.6. Iodine value
The iodine index was determined using the Wijs method, as per
Mexican official norm NMX-F-152-S-1981 and ISO 3961 (2013); with
minor modifications. 200 mg of oil was diluted with 15 mL of petroleum
ether, and 25 mL of a solution of Wijs reagent was added. The mixture
was agitated and placed in the dark for 30 min at 25 °C. Thereafter
20 ml of 10% (m/v) potassium iodide was added as well as 150 ml of dis-
tilled water. The mixture was titrated with 0.1 N sodium thiosulfate
until a yellow coloration was obtained and then 1 mL of a 1% (M/Vv)
starch indicator solution was added. The titration was continued until
the blue color disappeared. The results were expressed in grams of I2
per 100 g of oil.
2.7. K270
The extinction coefficient at 270 (K270) was determined according
to Elez-Martínez, Soliva-Fortuny, and Martín-Belloso (2007). 100 mg
of oil was diluted with 10 mL of cyclohexane (spectrophotometric
grade). The mixture was homogenized and the absorbance was deter-
mined with a spectrophotometer specord 6000 (AnalytikJena, Jena, Ger-
many) in a cell with 1 cm wide, using pure cyclohexane as blank.
2.8. Tocopherol: saponification and quantification
The saponification and tocopherol quantification procedures were
performed as per Insani et al. (2008) and Messina et al. (2009). Around
500 mg of avocado oil was placed in a test tube and 2 mL of pyrogallol
(1% in absolute ethanol) was added. The mixture was vortexed for
30 s, and heated to 70 °C in a hot water bath for 2 min (in darkness).
Thereafter it was cooled by placing the tubes in an ice bath, 1.5 mL of
saturated KOH (12 N) was added, then the tubes were agitated and in-
cubated at 70 °C for 30 min (in darkness). After this time, the tubes were
cooled in an ice bath, 1 mL of distilled water and 5 mL of pure hexane
were added. The mixture was agitated and left to rest for 2 min to
 M. Vargas-Ortiz et al. / Innovative Food Science and Emerging Technologies 41 (2017) 109–116
separate the phases. The organic phase was collected and the aqueous
phase extracted once again. The organic phase was dried out in a rotary
evaporator (37 °C, 50 mbar) and re-suspended in 1 mL of ethanol. The
quantification was performed by HPLC using an Agilent 1100 series
chromatograph (Agilent, Santa Clara, USA), equipped with a C18 Alltima
column (5 μm ∗ 4.6 ∗ 250 mm). The fluorescence detector was set at
296 nm for excitation and 330 nm for emission. 20 μL of the sample
was injected. The mobile phase was ethanol/methanol (60/40), at a
flowrate of 1 mL/min. Calibration curves of DL-α tocopherol and γ-to-
copherol diluted in absolute ethanol, at concentrations of 0.1 to
50 g/mL, were plotted. The results were expressed in mg α tocopherol
or γ-tocopherol per kg oil.
2.9. Fatty acid profile
The fatty acid methylation and methyl ester quantification proce-
dures were performed as set out by standard ISO 12966-2 (2011). Pre-
cisely 50 mg of oil was mixed with 2 mL of 0.8% sodium methylate in
methanol. The mixture was refluxed for 15 min, and two drops of phe-
nolphthalein were added. Then, it was neutralized with 1 M sulfuric acid
until the color disappeared and then was refluxed again for 5 min.
Thereafter, 8 mL of hexane and 10 mL of distilled water were added.
The organic phase was collected for gas chromatography analysis.
Methyl ester quantification was performed in a model GC7890A chro-
matograph (Agilent, Santa Clara, USA) equipped with a flame ionization
detector and a DB-WAX apolar column, 30 m ∗ 250 μm ∗ 0.25 μm
(Agilent, Santa Clara, USA). The carrier gas was helium a flow of
1.2 mL/min. The following temperature gradient was employed in the
oven: initial temperature 150 °C for 3 min, followed by an increase of
3 °C/min until 220 °C, and a holding stage at 220 °C for 10 min. The con-
centrations were calculated taking the calibration curves for each fatty
acid as a reference, and the results were expressed in % of total fatty
acids.
2.10. Lipoxygenase (LOX) enzyme activity
The LOX extraction and activity determination procedure was per-
formed as per Vargas-Ortiz, Servent, et al. (2016). 5 g of puree contain-
ing 1% Triton X100 were homogenized with 15 mL of 0.1 M, pH 6
sodium phosphate buffer (4 °C) for 1 min using a homogenizer Ultra-
Turrax T25 Digital, (IKA Works Inc., Staufen, Germany). Then the mix-
ture was centrifuged at 16000 ×g for 5 min, in a model 5810R centrifuge
(Eppendorf, Hamburg, Germany). The supernatant was used as the
crude enzyme extract. The substrate used was a solution prepared in a
25 mL graduated flask, mixing 20 μL of linoleic acid with 20 μL of
Tween-20. 5 mL of deionized water and 1 mL of NaOH solution (1 N)
were added, subsequently 24 mL of sodium phosphate buffer (0.1 M;
pH 6) was added. The reaction mixture used was 1 mL of 0.1 M pH 6 so-
dium phosphate, 20 μL of substrate and 25 μL of enzyme extract. The
mixture was placed in a UV spectrum cell, and the absorbance at
234 nm read in triplicate every 30 s for 5 min at 25 °C. The activity of
the enzyme lipoxygenase (LOX) was recorded as the final absorbance
after 5 min of reaction.
2.11. Statistical analysis
The data were analyzed using one treatment as the experimental
unit, in a completely randomized design. The data were subjected to a
general linear model (GLM) variance analysis by means of the statistical
package SAS (Statistical Analysis System V. 8.0); the averages were
compared by Tukey's test by means of the same software. The graphs
were produced using the software KaleidaGraph v.5 (Values represent
the mean of 3 replications with their standard deviation).
111
Fig. 1. Peroxide index in the lipid fraction of avocado puree obtained by FVE and puree
control, stored at 5 °C.
3. Results and discussion
3.1. Evolution of the peroxide value
The peroxide index determines the hydrogen peroxide content and
provides a measure of the oxidation of the unsaturated fatty acids. The
initial values of the control and the treatments (duration of the thermal
stage: T1 = 15 min; T2 = 20 min; T3 = 25 min; T4 = fruits without
shell 20 min) did not present any significant differences, with respec-
tively 11 meq O2/kg oil and 14 meq O2/kg oil (Fig. 1). These values
agree with the findings of Yousef and Hassaneine (2010) who reported
an initial value of 12 meq O2/kg oil in avocado oil. However they are dif-
ferent from the findings of Prabath, Sekozawa, Sugaya, and Gemma
(2013), who reported a range from 1.66 to 7.33 meq O2/kg oil in fresh
pulp and 5.85 meq O2/kg in oil. These authors proved that the initial var-
iations in the peroxide value can be attributed to many factors, includ-
ing the variety, agricultural practices and the action of lipolytic
enzymes released from the cells of the fruit during the process. The con-
trol puree exhibited a constant increase during storage, always main-
taining a higher value (P b 0.05) than those presented by treatments
T1 to T4. The increase in peroxide index of the control puree can be at-
tributed to the action of the enzyme lipoxygenase, which causes acid
oxidation of compounds which in turn promote oxidation of the fatty
acids (Jacobo-Velázquez & Hernández-Brenes, 2010). Furthermore,
treatments T1 to T4 exhibited an increase during storage, with no signif-
icant differences between them at any point in the study. Averaging be-
tween 12 and 16 meq O2/kg oil, these values are above the value of
10 meq O2/kg oil permitted by the specific Mexican standard for avoca-
do oil (Nmx-F-052, 2008). It was found that the thermal stage and ex-
pansion stage did not generate significant differences in peroxide
value. All the treatments presented lower values than the control. It is
clear that none of the treatments fully prevented fatty acid oxidation.
However it must also be considered that avocado puree is a mixture of
mainly aqueous and lipid fractions, which may be subjected to oxidative
processes caused by various means, and the resulting products may trig-
ger the oxidation of compounds in either fraction. Hydrolytic enzymes,
such as lipases, may also be active in the puree, causing the hydrolysis of
the ester-fatty acid bonds in triglycerides, to produce free fatty acids and
glycerol. Release of the fatty acids facilitates and accelerates auto-oxida-
tion reactions because they are more readily oxidized by free radicals
than are triglycerides (Gutteridge, 1995). Furthermore, it has also
112 M. Vargas-Ortiz et al. / Innovative Food Science and Emerging Technologies 41 (2017) 109–116
been reported the auto-oxidative process in avocado oil is more intense
than in other plant oils, since there are natural antioxidants present in
small quantities, whereas chlorophyll is present in large quantities,
which promotes auto-oxidation (Werman & Neeman, 1986). The lipid
fraction was influenced solely by the treatments and the storage tem-
perature, the rates of the enzymatic oxidation reactions were possibly
reduced because of the temperature (5 °C) of the purees, yet in treat-
ments T1 to T4 the oxidative processes were less severe than in the con-
trol. The flash vacuum-expansion process, significantly reduced
(P b 0.05) lipid peroxidation process, this favors the stability of the
puree because oxidation reactions can trigger other processes of decay
and generate unpleasant flavors.
3.2. Changes in the iodine value
The iodine index provides information on the secondary products
formed during lipid oxidation. In avocado oil it can be affected by vari-
ous factors, from the processing of the pulp, the extraction method or
storage conditions. Hence values have been reported within a range
from 77 to 160 g I2/100 g oil (Santana et al., 2015; Elez-Martínez et al.,
2007; Yousef & Hassaneine, 2010). The initial iodine index did not ex-
hibit any significant differences between the treatments, or compared
to the control puree (Fig. 2, duration of the thermal stage: T1 =
15 min; T2 = 20 min; T3 = 25 min; T4 = fruits without shell
20 min). Average values between 87 and 92 g I2/100 g oil were found.
During storage the values of the control puree exhibited a marked de-
creasing trend, and were consistently significantly lower (P b 0.05)
than those presented by the treatments. On day 7 of storage, the control
puree presented an average value of 75 g I2/100 g oil, and 69 g I2/100 g
oil on day 14. Furthermore, the iodine index exhibited no significant dif-
ference between the treatments T1 to T4 throughout the study. It pre-
senting average values of between 78 and 83 g I2/100 g oil by day 7 of
refrigerated storage, and of 74 to 77 g I2/100 g oil on day 14. The iodine
index was a direct measure of the double bonds present which is why a
decrease in this parameter would relates to a decrease in double bonds
in unsaturated fatty acids due to oxidation. The final values in all the
treatments were lower than the minimum value of 85 g I2/100 g oil ac-
cepted by the specific official Mexican norm for avocado oil. However, it
must be clarified that there is no specific official legislation for avocado
puree. A reduction in the iodine index due to the FVE process was ob-
served, although the final values were greater than the control. Neither
the regulation of thermal stage nor the expansion stage promoted the
Fig. 2. Iodine index in the lipid fraction of avocado puree obtained by FVE and puree
control, stored at 5 °C.
stability of the oil contained in the puree. However, oxidation of the
lipid fraction of the puree may also be influenced by the atmospheric
conditions in the package during storage, given that oxidative processes
may also be multi-factor (presence of oxygen, humidity, light, tempera-
ture). The heat treatment in the flash vacuum-expansion process and
the storage temperature may have reduced the enzymatic effects. How-
ever the oxygen present in the containers could have triggered the oxi-
dation reactions in the puree, as reported by Elez-Martínez et al. (2007)
who observed a greater decrease in the iodine index in avocado purees
stored in an air atmosphere, than in those purees which were stored in a
nitrogen atmosphere. The oxidation of compounds in the aqueous frac-
tion can promote the oxidation process in the lipid fraction, and once
the oxidative process of the fatty acids has begun, auto-oxidation reac-
tions will lead to lipid breakdown. Thus, oxidative processes occurring
in the lipid fraction of the puree limit the shelf life of product.
3.3. Change in extinction coefficient (K270)
Changes in the UV extinction coefficient at 270 are a good indicator
of secondary lipid oxidation, because this parameter is related to the
quantity of trienes or compounds such as aldehydes and ketones,
which are produced in the final steps of lipid oxidation (Gertz &
Klostermann, 2000). There were no significant differences in the initial
K270 values between any of the purees, which registered values within
a range of 0.26 to 0.31 (Fig. 3). T2 and T4 had the lowest values, while T3
and the control presented the highest values. All the purees presented
an increase in the value of K270 during storage. The control registered
the highest values; 0.37 by day 7 and 0.41 on day 14. Furthermore, treat-
ments T1 to T4 did not exhibit a significant increase on day 7 (values of
between 0.28 and 0.31) above the initial values. However by day 14 the
biggest increase was observed in the values of K270 for treatments T1 to
T4; possibly this behavior is due to the fact that there was less formation
of secondary oxidation products in T1 to T4 than in the control puree.
The significant increase observed up to day 14 may be due to the fact
that not all the primary oxidation products were subjected to secondary
oxidation, hence the compounds detected at 270 nm were not forming.
The increase in the K270 values in the lipid fraction of avocado puree
treated at high hydrostatic pressures and with added antioxidants was
also reported by Elez-Martínez et al. (2005), authors recorded values
of between 0.4 and 0.9 in the second week of storage in avocado purees
under refrigeration (4 ± 1 °C) with added natural antioxidants. The
values obtained in the present investigation were lower than those
Fig. 3. Extinction coefficient at 270 (K270) in the lipid fraction of avocado puree obtained
by FVE and puree control, stored at 5 °C.
 M. Vargas-Ortiz et al. / Innovative Food Science and Emerging Technologies 41 (2017) 109–116 113

Table 1 values within a range from 0.42 to 0.73 mg/kg. T2 had the lowest con-
Concentration of α-tocopherol (mg/kg oil). centration and the control the highest concentration. These values are
Treatment Day 0 Day 7 Day 14 very close to the minimum limit following Mexican standard Nmx-F-
Control 23.39 ± 5.25a 16.37 ± 2.91a 13.89 ± 3.06d
052-Scfi (2008), which stipulates a minimum γ-tocopherol concentra-
T1 17.42 ± 5.55a 24.40 ± 6.50a 17.91 ± 1.70cd tion of 0, and a maximum of 19 mg/kg oil, to ensure that the oil is
T2 23.39 ± 3.53a 19.83 ± 5.48a 22.34 ± 4.92bc good quality. The values recorded in this investigation are lower than
T3 22.25 ± 3.76a 22.12 ± 4.11a 30.83 ± 1.32a those reported by Chun et al. (2006), who evaluated the γ-tocopherol
T4 22.17 ± 4.31a 21.83 ± 2.24a 18.86 ± 0.94bcd
content in avocado pulp, and found values of 3.9 mg/kg oil. γ-tocopherol
Different letters in the same column represent significant difference considering concentration decreased during storage in the control sample. The other
(ANOVAA with Tukey's test, P b 0.05). treatments did not exhibit a definite trend. No significant differences
between the treatments were generated in α-tocopherol or γ-tocoph-
reported by these authors. Possibly the thermal stage of the FVE process erol concentration due to the thermal stage or expansion stage; howev-
reduces the activity of lipolytic enzymes and lipoxygenase, thereby re- er, it was possible to observe that the concentration of these compounds
ducing the formation of precursor compounds of the secondary oxida- was more stable during storage, unlike in the control puree.
tive process.

3.4. Tocopherols content 3.5. Fatty acid profile

The Mexican standard specifies that avocado oil must have a mini-
mum α-tocopherol concentration of 64 mg/kg oil, and a maximum of
100 mg/kg oil to be considered good quality. α-tocopherol concentra-
tions in the avocado purees analyzed were within a range of between
14 and 31 mg/kg oil, as Table 1 shows. All the values registered are
less than the minimum value admitted by the Mexican standard.
Flores, Perez-camino, and Troca (2014) reported concentrations of be-
tween 40 and 130 mg/kg in avocado oil produced and sold in Chile. Fur-
thermore, Lozano, Mayer, Bannon, and Gaydou (1993) reported a range
of between 57 and 103 mg/kg in the oil of various avocado varieties.
However the values found in the present investigation coincide with
the values reported by Hu et al. (2009), and Chun, Lee, Ye, Exler, and
Eitenmiller (2006) of 1.8 mg/100 g oil and 2.66 mg/100 g of edible
part of avocado respectively. The initial α-tocopherol concentration
values did not present any significant differences between the treat-
ments or compared to the control (Table 1, duration of the thermal
stage: T1 = 15 min; T2 = 20 min; T3 = 25 min; T4 = fruits without
shell 20 min). The lowest average value was presented by T2,
16.7 mg/kg, while the highest average values were presented by the
control 23.40 mg/kg and T3 23.40 mg/kg. By day 7 of storage, no signif-
icant differences were found between any of the purees. The values pre-
sented a range of between 16.4 and 27.7. On day 14 tocopherols did
present significant differences; the control 13.9 mg/kg presented the
lowest value and T4 30.8 mg/kg the highest value. The variations in α-
tocopherol concentration may be due to the oxidative processes in
each puree, since it has been reported that they may be involved in re-
ducing lipid oxidation (Isnardy, Wagner, & Elmadfa, 2003). The varia-
tion in tocopherol concentration may be influenced by various factors,
such as the fruit degree of maturity, processing, storage time and avoca-
do variety (Lozano et al., 1993; Chun et al., 2006). Only the control puree
presented a direct relationship with storage time, with α-tocopherol
concentration decreasing as storage time increased. There was any di-
rect relationship found with other parameters evaluated by this
investigation.
The γ-tocopherol values in the lipid fraction of the purees evaluated
were much lower than the α-tocopherol values. On day 0, none of the
purees presented any significant differences (Table 2), registering
Table 2
Concentration of γ-tocopherol (mg/kg oil).
Treatment Day 0 Day 7 Day 14
Control 0.72 ± 0.13a 0.47 ± 0.03b 0.40
T1 0.50 ± 0.13a 0.52 ± 0.09ab 0.50
T2 0.59 ± 0.08a 0.466 ± 0.01b 0.58
T3 0.64 ± 0.07a 0.61 ± 0.09ab 0.65
T4 0.53 ± 0.04a 0.50 ± 0.03ab 0.48
Different letters in the same column represent significant difference (ANOVAA with
Tukey's test, P b 0.05).
The fatty acids profile showed that in both the control and treated
purees evaluated, on day 0 of storage (Table 3, duration of the thermal
stage: T1 = 15 min; T2 = 20 min; T3 = 25 min; T4 = fruits without
shell 20 min), the fatty acid with the highest percentage concentration
was oleic acid (C18:1). T2 presented high outlier value which may be
due to a esterification which conduct to the oleic acid concentration
overestimation. T1 had the lowest concentration and T2 the highest
concentration, whereas the control presented an oleic acid concentra-
tion of 42.43%. In second place came palmitic fatty acid (C16:0), with
concentrations between 12.7 and 23.43%; T1 presented the lowest con-
centration and T2 the highest concentration, while the control puree
registered a palmitic acid value of 15.45%. In third place came
palmitoleic acid (C16:1), which varied between 5.65 and 9.2%; again
T1 presented the lowest concentration and T2 the highest concentra-
tion, while the control puree registered a palmitoleic acid value of
6.8%. In fourth place came linoleic fatty acid (C18:2), the concentration
of which varied between 3.45 and 8.95%. T1 had the lowest concentra-
tion and T2 the highest concentration, while the control presented a
linoleic fatty acid value of 4.89%. Linolenic acid (C18:3) was detected
in a smaller proportion, within a range from 0.39 to 0.73%; T3 had the
lowest concentration and T4 the highest concentration. The control reg-
istered a linolenic acid value of 0.70%. The results obtained by this inves-
tigation agree with the values reported by Yousef and Hassaneine
(2010), for the lipid fraction of fresh avocado pulp. Meyer and Terry
(2010) reported similar concentrations in the lipid fraction of avocado
which had undergone minimal processing and storage in modified at-
mospheres (at 5 °C). Neither the stages nor the processing times evalu-
ated affected the composition of the lipid fraction of the purees
evaluated, possibly because the FVE process comprises applying only
physical phenomena, which can reduce their impact on the proportion
of fatty acids. The very high standard deviation may be due to the fact
that the distribution of fatty acids in the various parts of the pulp (yel-
low center and green exterior) changes significantly (Villa-Rodríguez,
Molina-Corral, Ayala-Zavala, Olivas, & González-Aguilar, 2011). Further-
more, it has also been reported that the appearance of fatty acids which
make up the lipid fraction of Hass avocado is erratic and changeable,
hence the composition of the oil is too, even between fruits harvested
in the same year (Olaeta, Undurraga, & Schwartz, 1999). Moreover, oil
properties were not throughout in this study which deals with the opti-
mization of FVE to obtain a product which present less oxidation than
untreated sample. During storage, minor changes were observed in
the concentration of the various fatty acids, mainly in oleic acid, which
± 0.07b
presented increases on day seven. However on day 14 the concentration
± 0.06ab
± 0.10ab of this acid decreased, whereas other fatty acids presented less substan-
± 0.04a tial increases and decreases. These increases or releases of fatty acids
± 0.05ab may be due to the action of lipases, but could also be attributed to the
reversibility of the enzymatic esterification reaction. The variation in be-
havior may also be due to the fact that as the fatty acids are released,
114 M. Vargas-Ortiz et al. / Innovative Food Science and Emerging Technologies 41 (2017) 109–116

Table 3
Fatty acids (%) in the lipid fraction of avocado puree stored at 4 °C.

Day 0 C16:0 palmitic C16:1 palmitoleic C18:1 oleic C18:2 linoleic C18:3 linolenic

Control 15.45 ± 6.18ab 6.80 ± 3.05ab 42.43 ± 18.10ab 4.89 ± 3.08ab 0.70 ± 0.11ab
T1 12.79 ± 2.77b 5.65 ± 1.20ab 33.04 ± 7.56b 3.45 ± 1.00b 0.54 ± 0.16b
T2 23.43 ± 1.80a 9.92 ± 1.21a 61.09 ± 4.36a 8.95 ± 1.64a 0.63 ± 0.13ab
T3 16.23 ± 2.52ab 7.02 ± 1.29ab 43.11 ± 7.88ab 4.70 ± 1.36ab 0.39 ± 0.15b
T4 14.14 ± 2.86ab 5.77 ± 1.44ab 38.15 ± 8.15ab 5.17 ± 2.76ab 0.73 ± 0.30ab

Day 7

Control 17.97 ± 5.44ab 8.02 ± 2.90a 48.10 ± 16.24a 6.02 ± 2.59a 0.39 ± 0.11b
T1 16.90 ± 1.89ab 7.79 ± 0.85a 46.31 ± 5.68a 5.18 ± 0.91a 0.43 ± 0.12b
T2 21.66 ± 1.28a 8.77 ± 0.52a 54.63 ± 1.04a 7.31 ± 1.09a 0.57 ± 0.16b
T3 18.59 ± 5.39ab 8.36 ± 1.64a 49.91 ± 13.90a 8.02 ± 1.82a 0.63 ± 0.15ab
T4 12.71 ± 3.70ab 4.95 ± 1.61a 33.82 ± 9.38a 3.45 ± 1.23a 0.74 ± 0.08ab

Day 14

Control 18.66 ± 7.38a 8.01 ± 3.22a 47.41 ± 20.84a 5.70 ± 3.08a 0.67 ± 0.15a
T1 16.40 ± 8.18a 9.95 ± 5.04a 57.99 ± 33.88a 8.63 ± 6.06a 1.06 ± 0.72a
T2 19.65 ± 3.15a 8.64 ± 1.37a 52.53 ± 7.80a 6.75 ± 1.35a 0.51 ± 0.12a
T3 12.42 ± 0.60 5.53 ± 0.73a 32.68 ± 2.42a 4.52 ± 1.62a 0.45 ± 0.22a
T4 13.43 ± 2.76a 5.44 ± 1.26a 33.90 ± 7.28a 3.95 ± 1.43a 0.69 ± 0.21a

Different letters in the same column represent significant difference (ANOVAA with Tukey's test, P b 0.05).

especially linoleic and oleic acid, they are oxidized by the action of LOX
and auto-oxidation, as reported by Jacobo-Velázquez et al. (2013).
3.6. Lipoxygenase activity
The lipoxygenase activity was highest (P b 0.05) in the control puree
throughout storage (Fig. 4 duration of the thermal stage: T1 = 15 min;
T2 = 20 min; T3 = 25 min; T4 = fruits without shell 20 min). On day
five of storage, the LOX activity in the control mash decreased. However
by day 10, an increase in LOX activity was observed, returning to the
same value as that presented on day 0. Jacobo-Velázquez and
Hernández-Brenes (2010) observed similar behavior of LOX in avocado
puree subjected to high hydrostatic pressure treatments and refrigerat-
ed at 4 °C. This behavior can be attributed to the ability of the LOX en-
zyme to remain active even in crops subjected to freezing or scalding
(Bahçeci, Serpen, Gökmen, & Acar, 2005). Treatments T1 to T4 regis-
tered initial values significantly lower (P b 0.05) than the control, and
even with the reduction in activity of this enzyme during storage, T2
and T4 presented values between 69 and 79% of relative activity without
ever equaling the values presented by the control. The reduction in ac-
tivity of the enzyme can be attributed initially to the effect of the
thermal phase in the FVE process. T1 registered the second lowest
value at both the beginning and end of storage. On the other hand, T3
presented the lowest activity values during storage (54 and 36% on
day 0 and 10 respectively), possibly due to the effect of the heat treat-
ment, given that the avocado fruits remained in the heating chamber
for a longer time, reaching 95 °C (see Fig. 5), which could have resulted
in partial deactivation of the LOX. The optimum temperature for LOX ac-
tivity in avocado fruits is between 30 and 40 °C according to Jacobo-
Velázquez et al. (2013). However, the same authors report that LOX
has registered activity of up to 40% at 65 °C, and activity of between 5
and 10% at 75 °C. In the present investigation the fruits were exposed
to a steam current for periods of 15, 20 and 25 min, which is sufficient
to considerably reduce the relative activity of LOX, considering that on
average a temperature of 70 °C was reached inside the avocado upon
10 min (see Fig. 5). T4 (20 min heating of peeled avocado fruits) reached
an internal temperature of above 85 °C upon 10 min, while in the other
treatments (T1, T2, T3) this same condition was reached upon 15 min of
steam exposure; the absence of shell in T4 enabled a quicker heat trans-
fer to the fruit core. Despite the temperature/time conditions, total LOX
deactivation was not achieved in any case. The thermal stability of LOX
has been evaluated in various crops, since it is directly related to

Fig. 4. Lipoxygenase activity in avocado puree obtained by FVE and puree control, stored at
5 °C. Fig. 5. Temperature in the avocado seed during the FVE processes.
 M. Vargas-Ortiz et al. / Innovative Food Science and Emerging Technologies 41 (2017) 109–116
spoilage processes of stored foods; for example a 3 min scalding treat-
ment with water at 100 °C achieved reductions of 75% in the bitter
gourd, of 70% in green beans, of 70% in snap peas and of 80% in carrots
(Kaur, Kumar, & Kapoor, 1999). LOX activity was present in all treat-
ments, however in all purées obtained by FVE the activity of this enzyme
was lower than in the control.
3.7. Changes in temperature during treatments
Fig. 5 shows the changes of temperature in the avocado fruit during
the thermal stage. In treatments T1 to T4 the fruits showed values of tem-
perature/time with which it is possible to inactivate lipoxygenase enzyme
as discussed in the previous paragraphs. The temperature increase is
greatest during the first 10 min, subsequently the temperature increases
in less proportion. The temperature of the fruits showed a tendency to-
ward thermal equilibrium with the steam temperature (100 °C) which
it is used in the thermal stage. The temperature increase is faster in T4,
due to the absence of peel heat is transmitted faster by the pulp to seed.
To the pass quickly from the heating chamber (at atmospheric pressure
101.3 kPa) to the expansion chamber (between 2 and 5 kPa), by pressure
difference a part of the constitutive water of the fruits it evaporates in-
stantly, this causes a rapid decrease in the temperature of puree. Similarly
puree contact with the walls of the expansion chamber generates accu-
mulated heat dissipation in the puree. On the other hand, puree control
maintained a relatively constant temperature in a range of 19–24 °C dur-
ing the time of preparation (3 min). The temperature reached by the fruits
during the thermal stage of each treatment is below the temperature
(180 °C) that has been used for the analysis of thermal stability of avocado
oil (Berasategi, Barriuso, Ansorena, & Astiasarán, 2012), however, it could
affect the lipoxygenase enzyme as discussed above.
4. Conclusion
The FVE process does not affect the parameters evaluated in this
study at day 0. The FVE process does not affect the proportion of fatty
acids in the lipid fraction of avocado puree, however, with high standard
deviation were observed in the content of each fatty acid, probably due
to the fact that individual variation were high between fruits and oil ex-
tractions. The content of tocopherols is less than that reported by other
authors. Oxidative process was significantly less strong in the lipid frac-
tion of purees obtained by FVE. Considering our results, T3 FVE (25 min
of heat treatment) was the best parameter to avoid puree spoilage dur-
ing storage. In all case lipoxygenase enzyme was not completely inacti-
vate. Even those results suggested that the best treatment was T3,
because of its lowest values in the lipoxygenase activity. Therefore the
FVE is a potential interesting tool for avocado pulp processing option,
as this will gives stability during refrigerated storage without the need
for additives. Also this storage can be test in the future with control at-
mosphere storage after FVE to test the combination of those modality on
pulp and oil quality. It is also necessary to do more studies on the effects
of FVE process on sensory e.g. aromatic and hedonic evolution and tex-
tural aspects of the final product.
Acknowledgments
This work was supported by PCP France-Mexico (PCP-002-2011).
The authors thank all the technical and administrative staff of CIRAD,
Montpellier, France for its help for the realization of this research.
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