Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161

Contents lists available at ScienceDirect

Food and Bioproducts Processing

journal homepage: www.elsevier.com/locate/fbp

Recovery of bioactive components from avocado

peels using microwave-assisted extraction

Rafael G. Araujo a , Rosa M. Rodríguez-Jasso a , Héctor A. Ruíz a ,

Mayela Govea-Salas a , Manuela Pintado b , Cristobal N. Aguilar a,∗
a DIA – Departamento de Investigación en Alimentos, Facultad de Ciencias Químicas, Universidad Autónoma de
Coahuila, 25280 Saltillo, Coahuila, Mexico
b CBQF – Centro de Biotecnologia e Química Fina, Escola Superior de Biotecnologia, Universidade Católica

Portuguesa/Porto, 4202-401 Porto, Portugal

a r t i c l e i n f o a b s t r a c t

Article history: The avocado processing industry generates large amounts of avocado peels, which contain
Received 7 June 2020 large amounts of high value-added compounds with antioxidant capabilities that have not
Received in revised form 18 yet been explored. Optimization of microwave-assisted extraction (MAE) of bioactive com-
February 2021 pounds from avocado peels with high antioxidant capacity was the objective of present
Accepted 22 February 2021 work. Two experimental designs were implemented, using acetone 70% and ethanol at dif-
Available online 2 March 2021 ferent concentrations, finding at 74.48 ◦ C by 14.32 min and 66.37 ◦ C by 0.97 min with 42.58% of
ethanol as optimal conditions to obtain the highest antioxidant capacity, respectively. Opti-
Keywords: mized extracts with acetone and ethanol showed a high polyphenolic content (379.28 ± 19.35
Avocado peel and 354.43 ± 16.85 mg GAE/g dry extract, respectively); high antioxidant activity measured
Microwave-assisted extraction by DPPH (268.04 ± 25.11 and 233.85 ± 13.58 mgET/g dry extract, respectively); ABTS (895.19
Optimization ± 30.41 and 949.41 ± 7.42 mgET/g dry extract, respectively); ORAC (648.88 ± 28.66 and 692.06
Antioxidant capacity ± 28.80 mgET/g dry extract, respectively), results reported for the first time. Fiber bonded
Bioactive compounds phenolics compounds was extracted of MAE residues show a significant antioxidant activ-
Free and bound phenolics ity. HPLC-MS analysis of optimized extracts showed the present of diverse phenolic acids,
numerous procyanidins dimer A and B in different isomer shapes, catechin, epicatechin and
perseitol.
© 2021 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

1. Introduction vitamins (Alvarez et al., 2012; Cowan and Wolstenholme, 2016; Melgar
et al., 2018; Tremocoldi et al., 2018).
Avocado (Persea americana Mill.) is an important fruit from Lauraceae Avocado industrialization only uses avocado pulp to produce oils,
family, native of Mexico and Central America, cultivated in tropical sauces and guacamole generating a large quantity of residues, peels
and subtropical regions of world. Avocado Hass is a hybrid variety from and seeds, which are an important environmental issue due to the
Mexican and Guatemalan cultivars most popular and commercialized large quantities produced and the lack of uses for avocado residues
(Kosińska et al., 2012; Wang et al., 2010). Mexico is the main avocado (Figueroa et al., 2018a). These residues correspond to 25% of the fruit
producer with more 2 million tons in 2018, evaluated in more of 5 bil- and the peels about 11–17% (Rodríguez-Carpena et al., 2011; Wang et al.,
lion dollars that represents more than 30% of worldwide production 2010). Food and cosmetic industries have a great interest to find and
(FAOSTAT, 2018; SIAP, 2020). Avocado is a very valuable fruit that have a replace the synthetic compounds, as BHA (butylated hydroxyanisole)
complete nutritional content associated to many health benefits due to and BHT (butylated hydroxytoluene), which have been associated with
the high levels of unsaturated fatty acids, minerals, fiber, proteins and negative health effects, by safe bioactive compounds from natural

∗
Corresponding author.
E-mail address: cristobal.aguilar@uadec.edu.mx (C.N. Aguilar).
https://doi.org/10.1016/j.fbp.2021.02.015
0960-3085/© 2021 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161 153

sources as agroindustrial residues and plants (Figueroa et al., 2018b; 2.2. Standards and reagents
Rodríguez-Carpena et al., 2011).
Avocado peels have high content of extractable bioactive com- Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic
pounds such as condensed tannins, phenolic acids and flavonoids
acid), DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2 -azino-
as catechin and various procyanidins, flavonols hydroxybenzoic and
bis(3-ethylbenothiazoline-6-sulphonic acid) diammonium
hydroxycinnamic acids (Figueroa et al., 2018a; Kosińska et al., 2012;
salt), Folin & Ciocalteu’s phenol, AAPH (2,2 -azobis(2-
López-Cobo et al., 2016; Wang et al., 2010). Avocado peels extract have
many biological activities associated such as antioxidant, anticancer,
amidinopropane) dihydrochloride), fluorescein, and gallic
antibacterial, anti-inflammatory and anti-hypertensive proprieties, acid were products purchased from Sigma-Aldrich (St. Louis,
many dermatological uses and additionally not contain potentially MO, USA). Chromatography-grade formic acid and methanol
toxic or harmful compounds, making them a promising natural source were produced by Tedia Company (Fairfield, OH, USA). All
of bioactive compounds to many applications in food, cosmetic and the other regents, such as sodium carbonate anhydrous and
pharmaceutical industry (Chia and Dykes, 2010; Hurtado-Fernández potassium persulfate were of analytical grade and acetone
et al., 2011; Kosińska et al., 2012; Segovia et al., 2016; Tremocoldi et al., and ethanol were purchased from Jalmek (Nuevo León,
2018). Mexico).
The most important bioactivities of avocado extracts to food indus-
try are the antioxidant and antibacterial activity to prevent the food
2.3. Microwave-assisted extraction of bioactive
oxidation and bacterial contamination and obtain more time of con-
compounds
servation with high quality of products (Calder and Iztapalapa, 2016).
The use of avocado peels as source of natural compounds is a way
to obtain added value ingredients for replacement of the synthetic The bioactive polyphenols were extracted by MAE (CEM Mars
compounds and at the same time generate a significant impact in 6, USA) with a temperature control in Teflon vessels of 70 mL
the avocado industry and environmental benefits (Saavedra et al., (Xpress). The extractions were performed in a reactor with
2017; Wang et al., 2010). A sustainable strategy for the future is new working volume of 20 mL in order to obtain a 1:20 (w/v, 1
practices and balanced relationships between the environment and g of dry avocado peel/20 mL of solvent), 2.45 GHz and 600
human industrialization to reduce the environmental impact through W (Aguilar-Reynosa et al., 2017), using two statistic designs,
the application of a circular economy.
one with acetone/water (70:30 v/v) and other with ethanol as
The microwave-assisted extraction (MAE) is a green and safe tech-
solvent. Each extraction by MAE has 3 stages, heating time,
nology to obtain added-value compounds with high extraction yields
extraction time and cooling time, in which the heating time
using short times of extraction, low quantities of solvent and replace-
ment of organic solvents by safe solvent as water or ethanol, when
was set to 7 min (the microwave equipment automatically
compared with conventional methods of extraction (Zhang et al., adjusts the power to reach the desired temperature in each
2011). MAE is applied to extract many natural compounds from treatment), the extraction time depends on each treatment
agroindustrial residues from fruits or vegetables to obtain bioactive and the cooling time in 10 min. After each MAE extraction, the
compounds, mainly antioxidants, oils rich in carotenoids or polyphe- extracts were cooler for 10 min in cold water for subsequent
nol extracts from plants or residues (Chemat and Cravotto, 2013; Pap filtered under vacuum to separate the liquid extract and later
et al., 2013). MAE uses the principle of moisture heating inside the it was stored at −20 ◦ C until further analysis. Each treatment
cells, caused by the direct effect of microwaves on water molecules was performed by triplicated.
by dipole polarization and ionic conduction, which improve the poros-
ity or destruction of cell wall, increasing the compound extraction
2.4. Determination total polyphenol content (TPC)
(Chan et al., 2016; Chimsook, 2017; Flórez et al., 2015; Kala et al.,
2016).
The objective of present study was the optimization of the MAE con-
The total polyphenol content (TPC) of the samples was mea-
ditions to obtain avocado peels extracts with high antioxidant capacity sured using a modified Folin-Ciocalteu colorimetric method
using two statistical designs, one with acetone and other with ethanol described previously by Magalhães et al. (2010) with some
and identify the bioactive compounds of avocado peels extract to modifications. Twenty ␮L of samples and control (distilled
demonstrate the feasibility of MAE uses and the potential of avocado water) were mixed in a tub test with 30 ␮L of Folin-Ciocalteu
peel extract in food applications. reagent (0.25 N), 120 ␮L of sodium carbonate (15% (w/v)) and
400 ␮L of distilled water, added by this exact order. The mixture
was heated at 50 ◦ C for 5 min and cooled to room tempera-
ture (25 ◦ C). The total polyphenol content was determined by
2. Materials and methods absorbance of the resulting blue color that was measured at
700 nm in a microplate of 96 wells by colorimetry using a Tecan
2.1. Material Sunrise RC spectrophotometer (Austria). A standard curve was
performed using different concentrations of gallic acid. All
The avocado peels were obtained from Hass variety in a determinations were performed by triplicate. The results were
ready-to-eat ripening stage from a local restaurant, in Saltillo, expressed as milligram gallic acid equivalents (GAE) per g of
Mexico. The peels were washed to remove pulp residues dry peel (mg GAE/g dry peel).
and cut into small pieces with a kitchen knife for further
freeze-drying. Once lyophilized, the peels were ground using 2.5. Determination of the antioxidant capacity in vitro
a blender (Oster) for 30 s and sieved with mesh number 18 by DPPH
and 35 to obtain a particle size between 0.3–1 mm. The flours
were stored at room temperature protected of light and air. The quantification of the antioxidant capacity by DPPH was
The determination of the proximal composition of the avo- described by Kähkönen and Heinonen (2003), which is based
cado peel was determined by the standards Official Methods on the quantification of the antioxidant capacity of the phe-
of Analysis (AOAC, 1995) to lipids, protein and ashes, 932.06, nolic compounds of the extracts by reducing the free radical of
925.09, 923.03 methods, respectively, and 991.43 for insoluble DPPH, the determination was carried out by spectrophotom-
and soluble fiber. etry in microplate, since the free radical in ethanolic solution
154 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161

has a purple coloration and the effect of the reduction due 2.8. Determination of bound phenolics in avocado peel
to an antioxidant compound is measured by the loss of col-
oration, so that indirectly, the antioxidant capacity of the The bound phenolics were obtained by using the previous
obtained extracts can be quantified. The procedure was car- procedure described by Xie et al. (2015). Briefly, 0.5 g of dry avo-
ried out by mixing directly in a microplate, 193 ␮L of the DPPH cado peel after microwave extractions was dissolved with 20
free radical prepared in ethanol at a concentration of 60 ␮M, mL of 80% ethanol to remove residual soluble phenolic acids,
with 7 ␮L of the extract to be analyzed and allowing it to and then defatted with 6 mL hexane. The extract was cen-
react for 30 min at room temperature and in the dark. The trifuged at 8000 rpm for 10 min, and the bound phenolics
microplate was analyzed at 517 nm in the spectrophotome- were then released by hydrolysing the leftover solid residue
ter Tecan Sunrise RC spectrophotometer. Each extract was with 20 mL of 4 M NaOH at room temperature with stirring
analyzed by triplicate and a calibration curve was used with for 4 h. The transparent mixture was acidified to pH 1.5–2.0
Trolox. The results were expressed in mg of Trolox equivalents by gradual addition of 6 M HCl. After centrifugation at 8000
per gram of dry peel (mg TE/g dry peel). rpm for 30 min, the supernatant was extracted five times with
30 mL ethyl acetate. The fraction of ethyl acetate was dried
through anhydrous Na2 SO4 and evaporated to dryness using
2.6. Determination of the antioxidant capacity in vitro a rotary vacuum evaporator at 30 ◦ C. The resulting residue was
by ABTS then dissolved in 80% ethanol to 10 mL. The extract obtained
was stored at −20 ◦ C until used within a week. The detec-
For ABTS radical cation decolorization assay, the free radical- tion method of the bound phenolics in avocado peel was the
scavenging capacity was determined as described by Re et al. same previously described in Folin-Ciocalteu and antioxidant
(1999). ABTS was dissolved in water at a final concentra- activities.
tion of 7 mM. ABTS radical cation (ABTS·+ ) was produced by
reacting ABTS stock solution with 2.45 mM potassium per- 2.9. Optimization of bioactive compounds extraction
sulfate (final concentration) and kept in the dark at room by MAE
temperature (25 ± 2 ◦ C) for 12−16 h before use. The radi-
cal maintains a stable form for more than two days when For optimization of bioactive polyphenols extraction with
stored in the dark. Before analysis, ABTS·+ was filtered using better antioxidant activity measured by DPPH a Central Com-
a 0.22 ␮m (Orange Scientific, Braine-l’Alleud, Belgium) and posite Design (CCD) was applied with two factors and three
diluted with distilled water to an absorbance of 0.700 (± 0.02) replicates of the center point, using acetone at 70% (v/v)
at 734 nm with an UVmini 1240 UV–vis spectrophotometer as solvent, based in Rodríguez-Carpena et al. (2011), which
(Shimadzu, Washington, USA). A blank was taken with dis- shows better results in total phenol content and antioxidant
tilled water (A0 ). After addition of 1.0 mL of diluted ABTS·+ activities, when compared with methanol and ethyl acetate
solution (A734 nm = 0.700 ± 0.02) 10 ␮L of sample was added extracts. Table 1 shows the independent variables of temper-
and the absorbance was read exactly 6 min after initial mix- ature (X1, ◦ C) and time (X2, min) for three variation levels
ture. Since the inhibition percentage (IP) must be between on DPPH activity (Eq. mg Trolox/g dry peel) and total phenol
20 and 80%, the samples were diluted when needed. A cal- content (mg GAE/g dry peel). In order to obtain a process of
ibration curve prepared using standard solutions of Trolox. extraction of bioactive compounds by a greener process, a sta-
All assays were performed by triplicate. The results were tistical design with ethanol/water as a solvent was designed,
expressed as mg of Trolox equivalent/g dry extract (mg TE/g with a concentration range between 0 to 100 % (v/v) of ethanol,
dry extract). since there was no previous report. A Box-Behnken Design
(BBD) was applied to optimize the extraction of bioactive
polyphenols with better antioxidant activity measured by
2.7. Determination of oxygen radical absorbance DPPH, with three factors and three replicates of the center
capacity (ORAC) point. Low and high factors were coded as −1 and +1; the cen-
ter point was coded as 0. The Table 2 shows the independent
The oxygen radical absorbance capacity (ORAC) assay was variables of temperature (X1, ◦ C), time (X2, min) and ethanol
based on that proposed by Contreras et al. (2011). Briefly, concentration (X3, %) for three variation levels on DPPH activ-
the reaction was carried out at 40 ◦ C in 75 mM phosphate ity (mg ET/g dry peel) and total phenol content (mg GAE/g dry
buffer (pH 7.4) and the final assay mixture (200 ␮L) contained peel).
fluorescein (70 nM), AAPH (14 mM), and either antioxidant The data were analyzed by statistical software STATISTICA
(Trolox (0.2–1.6 nmol/mL) or sample at different concentra- 7® to obtain the Optimal Condition (OC) to each statistical
tions. The fluorescence was recorded during 137 min (104 design and later validated the OC by extraction with the condi-
cycles). A FLUOstar OPTIMA plate reader (BMG Labtech, Offen- tions estimated by the software. The validation was performed
burg, Germany) with 485 nm excitation and 520 nm emission by triplicated. The results were analyzed by analysis of vari-
filters was used. The equipment was controlled by the FLU- ance (ANOVA), and the responses and variables (in coded unit)
Ostar Control software version (1.32 R2) for fluorescence were correlated by response surface analysis to obtain the
measurement. Black polystyrene 96-well microplates (Nunc, coefficients of Eqs. (1) and (2) to CCD and BBD, respectively:
Denmark) were used. AAPH and Trolox solutions were pre-
pared daily and fluorescein was diluted from a stock solution
Y = ˇ0 + ˇ1 X1 + ˇ2 X2 + ˇ1,2 X1 X2 + +ˇ1,1 X12 + ˇ2,2 X22 (1)
(1.17 mM) in 75 mM phosphate buffer (pH 7.4). All reaction
mixtures were prepared by triplicate and at least three inde-
pendent runs were performed for each sample. Final ORAC
values were expressed as mg of Trolox equivalent/g dry extract Y = ˇ0 + ˇ1 X1 + ˇ2 X2 + ˇ3 X3 + ˇ1,2 X1 X2 + ˇ1,3 X1 X3
(mg TE/g dry extract). + ˇ2,3 X2 X3 + ˇ1,1 X12 + ˇ2,2 X22 + ˇ3,3 X32 (2)
 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161 155

Table 1 – Experimental conditions used for bioactive polyphenols extraction from avocado peel with acetone 70%
according to CCD with two variables, temperature (X1 ) and time (X2 ), and results obtained for the total polyphenol
content and antioxidant activity by DPPH.
Assay Variables TPC DPPH
(mg GAE/g dry (mg TE/g
peel) dry peel)
X1 X2

1 60 (−1) 1.0 (−1) 123.68 ± 13.00 173.82 ± 5.97

2 60 (−1) 15.5 (1) 125.41 ± 8.23 164.10 ± 7.29
3 60 (−1) 30.0 (1) 123.33 ± 3.59 157.75 ± 3.30
4 75 (1) 1.0 (−1) 126.15 ± 4.88 155.90 ± 12.06
5 75 (0) 15.5 (0) 119.00 ± 3.78 217.26 ± 12.89
6 75 (0) 15.5 (0) 120.61 ± 8.60 180.82 ± 9.17
7 75 (0) 15.5 (0) 133.13 ± 4.39 190.17 ± 5.50
8 75 (0) 30.0 (1) 127.72 ± 10.55 150.08 ± 8.82
9 90 (1) 1.0 (−1) 129.42 ± 3.44 160.54 ± 9.62
10 90 (1) 15.5 (0) 130.32 ± 6.86 173.26 ± 8.19
11 90 (1) 30.0 (1) 127.05 ± 7.92 160.71 ± 10.82

Table 2 – Experimental conditions used for bioactive polyphenols extraction from avocado peel according to BBD with
three variables, temperature (X1 ), time (X2 ) and ethanol concentration (X3 ), and results obtained for the total polyphenol
content and antioxidant activity by DPPH.
Assay Variables TPC DPPH
(mg GAE/g (mg TE/g
dry peel) dry peel)
X1 X2 X3

1 50 (−1) 0 (−1) 50 (0) 81.50 ± 1.40 186.59 ± 9.41

2 90 (1) 0 (−1) 50 (0) 83.16 ± 2.54 170.75 ± 8.02
3 50 (−1) 30 (1) 50 (0) 88.99 ± 4.37 164.36 ± 8.04
4 90 (1) 30 (1) 50 (0) 85.08 ± 3.55 155.41 ± 5.35
5 50 (−1) 15 (0) 0 (−1) 70.24 ± 2.99 108.30 ± 10.57
6 90 (1) 15 (0) 0 (−1) 63.16 ± 0.62 103.20 ± 7.59
7 50 (−1) 15 (0) 100 (1) 43.21 ± 3.10 85.94 ± 2.12
8 90 (1) 15 (0) 100 (1) 55.45 ± 3.02 101.85 ± 5.49
9 70 (0) 0 (−1) 0 (−1) 74.60 ± 0.35 139.47 ± 3.54
10 70 (0) 30 (1) 0 (−1) 70.00 ± 2.67 116.92 ± 3.89
11 70 (0) 0 (−1) 100 (1) 48.82 ± 2.68 81.40 ± 6.04
12 70 (0) 30 (1) 100 (1) 53.65 ± 2.29 90.27 ± 1.75
13 70 (0) 15 (0) 50 (0) 83.93 ± 0.47 215.51 ± 7.79
14 70 (0) 15 (0) 50 (0) 81.10 ± 1.96 184.24 ± 6.13
15 70 (0) 15 (0) 50 (0) 85.31 ± 1.64 154.95 ± 6.18

Where Y is the measured response; ␤0 is the intercept (con- 30 ◦ C. The mobile phases were water containing 0.2% (v/v)
stant); ␤1 to ␤3,3 are the coefficients associate with linear, of formic acid (solvent A) and acetonitrile (solvent B). The
quadratic and interaction effects, respectively, of variables X1 , following gradient was applied: initial, 3% B; 0–5 min, 9% B
X2 and X3 , respectively. The models study the effect of each linear; 5–15 min, 16% B linear; 15–45 min, 50% B linear. The
independent variable and all interactions between them on a column was then washed and reconditioned for 5 min with
particular response (antioxidant activity). After obtaining the the initial concentration of solvents. The flow rate was main-
extracts under the ideal extraction conditions with each sol- tained at 0.2 mL/min and elution was monitored at 245, 280,
vent, they were evaporated using a rotary vacuum evaporator 320 and 550 nm. The whole effluent (0.2 mL/min) was injected
at 40 ◦ C to remove the solvent and then lyophilized to obtain into the source of the mass spectrometer, without splitting.
the powdered extracts for further antioxidant activities. All MS experiments were carried out in the negative mode
[M–H]−1 . Nitrogen was used as nebulizing gas and helium as
damping gas. The ion source parameters were spray voltage
2.10. Analytical RP-HPLC-ESI-MS
5.0 kV, capillary voltage and temperature were 90.0 V and 350
◦ C, respectively. Data were collected and processed using MS
The analyses by Reverse Phase-High Performance Liquid
Workstation software (V6.9). Samples were firstly analyzed in
Chromatography were performed on a Varian HPLC system
full scan mode acquired in the m/z range 50–2000. MS/MS anal-
including an autosampler (Varian ProStar 410, USA), a ternary
yses were performed on a series of selected precursor ions.
pump (Varian ProStar 230I, USA) and a PDA detector (Varian
ProStar 330, USA). A liquid chromatograph ion trap mass spec-
trometer (Varian 500-MS IT Mass Spectrometer, USA) equipped 2.11. Statistical analyses
with an electrospray ion source also was used. Samples (5 ␮L)
were injected into a Denali C18 column (150 mm × 2.1 mm, 3 Statistical analyses were performed to calculate average ten-
␮m, Grace, USA). The oven temperature was maintained at dency and deviation. To compare the polyphenol content and
156 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161

Table 3 – Analysis of variance (ANOVA) for optimization of extraction of bioactive compounds with better antioxidant
activity by DPPH from avocado peels with a CCD, model as a function of temperature (X1 ) and time (X2 ).
Source Sum of squares df Mean square F-value p-Value

Model 2040.13 5 408.03 1.21 0.4183

x1 0.23 1 146.47 0.4358 0.5383
x1 2 146.47 1 78.65 0.2340 0.6490
x2 78.65 1 1374.80 4.0907 0.0990
x2 2 1374.80 1 65.87 0.1960 0.6765
x1 x2 65.87 1 146.469 0.4358 0.5383
Residual 1680.40 5 336.08
Lack of fit 963.93 3 321.31 0.90 0.5655
Pure error 716.47 2 358.23
Cor total 3720.53 10
R2 0.55
Adj R2 0.10
C.V. 10.70

p-Value < 0.05 indicate model terms are significant.

DPPH antioxidant activity of avocado peel extracts and opti- fore larger variations in antioxidant activity. Moreover, the
mize the conditions of extraction an analysis of variance responses of temperature and time had not significant effect
(ANOVA) and Tukey test with a 95% confidence level were run (p-value > 0.05) in linear or quadratic effects, which shows that
in the STATISTICA 7® software (StatSoft Inc., Tulsa, OK, USA). using acetone 70%, time and temperature have not significant
effect between all treatments of the CCD, since the greatest
3. Results and discussion effect on the extraction of bioactive compounds is associated
with acetone. The ANOVA results, showed in Table 3, revealed
3.1. Microwave-assisted extraction of bioactive that second-order polynomial model was found to establish
compounds from avocado peel the prediction of bioactive compounds extraction with high
antioxidant activity response within the range of experimen-
The avocado peels used in this project contain in dry basis tal variables. The determination coefficient of the model was
3.57 ± 0.11 % of ashes, 2.87 ± 0.22 % of lipids, 4.47 ± 0.55 % R2 = 0.54, indicating a low adjustment of the model and 46% of
of proteins, 67.80 ± 0.42% of insoluble fiber and 21.01 ± 0.57 the total variations were not explained by the proposed model.
% of soluble fiber. Tables 1 and 2, show the values of phenol The TPC showed no statistical difference between the 11
content and antioxidant activity by DPPH of each design, CCD treatments however for antioxidant activity statistical differ-
with acetone and BBD with ethanol. Solvent polarity plays ence was observed, that indicates existence of no correlation
an important role in increasing phenolic compounds solubil- between polyphenolic compounds with antioxidant activity.
ity and extractability. Other important influence of bioactivity At central point (75 ◦ C by 15.5 min) in the 3 replicates values
natural extracts of plants or fruits are the synergism of all com- of TPC between 119.0 and 133.13 mg GAE/g dry peel and val-
pounds that can be created or modified by the solvent. Chavan ues of 217.26 and 190.17 were obtained, respectively. A similar
et al. (2001) reported that acetone at 70% was more efficient effect was reported by Pandey et al. (Pandey et al., 2018; Rasera
that absolute acetone for recovery a maximum amount of con- et al., 2019) in a process of optimizing of extraction of bioac-
densed tannins from peas. However, the chemical structure tive compounds from Rheum moorcroftianum rhizomes and
and polarity of phenolic compounds of each material deter- mustard grains, respectively, showing no correlation between
mines their extractability and efficiency of solvent, and the phenolic content and antioxidant activity. The second-order
effects of temperature and time of extraction changes the polynomial model of the optimization of bioactive compounds
efficiency of extraction, therefore, an optimization study of extraction to better antioxidant activity with acetone (BCEa) as
phenolic compounds extraction for each material is required. function of temperature (X1 ) and time (X2 ) is expressed in the
The results show that acetone is more effective to extract next Eq. (3):
polyphenolic compounds reaching value of 133.13 mg GAE/g
dry peel while with ethanol reaches to 88.99 mg GAE/g dry peel, BCEa(mgET/gdrypeel) = 188.17–0.19X1 –3.62X2 –7.60X1 2
but similar maximum values of antioxidant capacity, 217.26 –23.30X2 2 +4.06X1 X2 (3)
and 215.51 mg TE/g dry peel, respectively.

3.1.1. Statistical analysis and optimization of bioactive With the second-order polynomial of equation 3 was plot-
compounds with CCD ted a three-dimensional response surface in order to estimate
Table 1 shows the experimental data of the treatments of the the optimum condition to bioactive compounds extraction
experimental design. The analysis of variance (ANOVA) was with high antioxidant activity from avocado peel using ace-
carried out to obtain, F-value, p-value and lack of fit of the tone 70% as solvent (Fig. 1). The optimal condition estimated
model and the independent variables, temperature and time, to obtain bioactive compounds with higher antioxidant activ-
X1 and X2 , respectively. This error in model fit can be explained ity was close of central point, 74.48 ◦ C by 14.32 min, estimating
by the high degree of volatility of acetone, which may have led 188.31 mg TE/g dry peel.
to solvent losses in vacuum filtration process and the photo- After validation of optimal conditions, performed by tripli-
sensitivity of bioactive compounds that consequently lead to cate, was obtained 262.09 ± 9.96 mg TE/g dry peel, finding an
variations in bioactive compound concentrations and there- antioxidant activity much higher than estimated, highlight-
 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161 157

with high antioxidant activity response within the range of

experimental variables. The determination coefficient of the
model of 0.93 and p-value, indicating a high adjustment of the
model, only with 7% of the total variations was not explained
by the proposed model. However, only ethanol concentration
has been shown to have a significant quadratic effect on the
extraction of compounds with high antioxidant activity. With
50% of ethanol was found the highest contents of phenolic
compounds and antioxidant activity was found. At concen-
trations of 0 and 100%, the content of phenolic compounds
and antioxidant activity was significantly decreased, which
agrees with Rodríguez-Carpena et al. (Rodríguez-Carpena
et al., 2011), that using a solvent mixed with water had greater
solubilization of phenolic compounds and higher antioxi-
dant activities than pure solvent. The conditions of central
point of BBD was the treatment where the highest con-
tent of phenolic compounds and antioxidant activity were
Fig. 1 – Response surface and contour plot showing the
found.
optimization of bioactive compounds extraction with CCD
The second-order polynomial model of the optimization of
by effects of temperature (X1 ) and time (X2 ) using acetone
bioactive compounds extraction to better antioxidant activity
as solvent.
with ethanol (BCEe) is expressed in the next Eq. (4) solely with
the significant effect of ethanol (X3 ):
ing the variation of the optimization model found. The yield
extraction of polyphenol compounds with acetone was 10.69%
(g/g of avocado peel).
BCEe(mgET/gdrypeel) = 125.37 + 36.83X3 2 (4)

3.1.2. Statistical analysis and optimization of bioactive

compounds with BBD
Table 2 shows the experimental data of the treatments of With the second-order polynomial of Eq. (4) was plotted the
the experimental design. The data show that ethanol con- three-dimensional response surfaces in order to estimate the
centration was the independent variable with more impact in optimum condition to bioactive compounds extraction with
phenolic compounds extraction and more antioxidant activ- high antioxidant activity from avocado peel using ethanol as
ity. solvent (Fig. 2). The optimal condition estimated to obtain
The analysis of variance (ANOVA) was carried out to obtain, bioactive compounds with higher antioxidant activity was
F-value, p-value and lack of fit of the model and the indepen- 66.37 ◦ C by 0.97 min with 42.58% of ethanol, estimating 189.06
dent variables, temperature, time and ethanol concentration, mg TE/g dry peel. After validation of optimal conditions, per-
X1 , X2 , and X3 , respectively. The ANOVA results, showed in formed by triplicate, it was obtained 214.03 ± 4.38 mg TE/g
Table 4, revealed that second-order polynomial model was dry peel with a yield of polyphenols of 9.80% (g/g of avocado
found to be for prediction of bioactive compounds extraction peel).

Table 4 – Analysis of variance (ANOVA) for optimization of extraction of bioactive compounds with better antioxidant
activity by DPPH from avocado peels with a BBD using ethanol as solvent, model as a function of temperature (X1 ), time
(X2 ) and ethanol concentration (X3 ).
Source Sum of squares df Mean square F-value p-Value

Model 22365.53 9 2485.06 4.91 0.0472*

X1 75.18 1 75.18 0.0819 0.8016
X1 2 480.39 1 480.39 0.5237 0.5445
X2 394.44 1 394.44 0.4300 0.5793
X2 2 65.62 1 65.62 0.0715 0.8141
X3 873.28 1 873.28 0.9520 0.4321
X3 2 20038.48 1 20038.48 21.8455 0.0429*
X1 X2 11.89 1 11.89 0.0130 0.9198
X1 X2 2 158.50 1 158.50 0.1728 0.7180
X1 2 X2 71.36 1 71.36 0.0778 0.8065
X1 X3 110.41 1 110.41 0.1204 0.7617
X1 2 X3 465.24 1 465.24 0.5072 0.5502
X2 X3 246.80 1 246.80 0.2691 0.6557
Residual 2529.67 5 505.93
Lack of fit 695.11 3 231.70 0.25 0.8560
Pure error 1834.56 2 917.28
Cor total 24895.20 14
R2 0.93
Adj R2 0.48
C.V. 16.39

p-Value < 0.05 indicate model terms are significant.

158 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161

Fig. 2 – Response surface and contour plot showing the optimization of bioactive compounds extraction from avocado peels
with BBC by effects of temperature (X1 ), time (X2 ) and ethanol concentration (X3 ) using ethanol as solvent.

Table 5 – TPC and antioxidant activities by DPPH, ABTS and ORAC of dry peel extracts obtained with optimal conditions
with acetone and ethanol.
Extract TPC DPPH ATBS ORAC
(mg GAE/g dry (mg ET/g dry (mg ET/g dry (mg ET/g dry
extract) extract) extract) extract)

Acetone 379.28 ± 19.35 268.04 ± 25.11 895.19 ± 30.41 648.88 ± 28.66

Ethanol 354.43 ± 16.85 233.85 ± 13.58 949.41 ± 7.42 692.06 ± 28.80

3.2. Total polyphenol content and antioxidant activity 3.3. Bound phenolics in avocado peel

Antioxidant compounds are substances with the ability of The TPC of fiber obtained from ethanol extraction was higher
free-radical scavenging and inhibition of oxidizing enzymes than in the fiber obtained from acetone extraction, was more
such as reactive oxygen species (ROS), which are associated than double (Table 6). This difference emphasizes that ace-
with many diseases, especially cancer (Ramos et al., 2019). tone has the higher power to extract the phenolic compounds
Kosińska et al. (2012) reported that avocado residues (seeds present in avocado peel than ethanol, removing some fiber-
and peels) have several-fold greater phenolic content and bound phenolic compounds, which is in accordance with the
antioxidant capacity than raw blueberry, which are fruits TPC results obtained in the phenolic extracts since acetone
known for its high antioxidant capacity. The TPC and antiox- extracts contain more phenolic content. Consequently, the
idant capacities of dry peel extracts obtained with optimal antioxidant capacity of the fiber-bound phenolic compounds
condition with acetone and ethanol by MAE are shown in obtained after extraction with ethanol is higher since the con-
Table 5. Other authors reported TPC and antioxidant capac- tent of phenolic compounds is superior. The content of bound
ities of avocado peel extracts obtained only by conventional phenolic compounds in avocado peel may be higher than
extraction with solvents. Wang et al. (2010) by sonication that obtained because a hydrothermal microwave process was
with acetone/water/acetic acid (70:29.7:0.3, v/v/v) as solvent used which may have favored the extraction of phenolic com-
reported 47.5 and 158.4 mg ET/g dry peel for DPPH and ORAC pounds bound to the fiber. Although for the avocado peel is the
activities with a TPC of 12.6 mg GAE/g dry peel. Rodríguez- first time reported, many reports showed the TPC bounded to
Carpena et al. (2011) with conventional extraction using fiber for others fruits. Xie et al. (2015) reported 1.89 and 0.92
acetone 70% reported 89.97 mg GAE/g dry peel and antiox- mg/g of phenolic bound in olive leaves and fruit, respectively.
idant activities DPPH and ORAC of 22.26 and 25.96 mg TE/g Chen et al. (2019) reported about 4 mg/g dry rice bran. Peng
fresh avocado peel. Kosińska et al. (2012) reported 25.32 mg et al. (2017) reported 9.33 mg/g of phenolic bound in seed coat
catequin equivalents/g dry peel and 117.64 and 40.3 mg TE/g soybean. Ajila and Rao (2013) reported 29.52 mg/g dry fiber of
dry peel for ORAC and ABTS activities, respectively. Saavedra mango peel. Quatrin et al. (2019) reported 2.47 mg/ g of dry
et al. (2017) reported 31.1 mg GAE/g dry peel and 383.7 mg jaboticaba peel, which demonstrates the reported results of
TE/g dry peel. Rotta et al. (2016) with a methanolic extrac- bounded phenolic compounds in other fruits are similar to
tion reported 10.85 mg GAE/g of dehydrated peel and 190.98 those obtained in avocado peels.
mg TE/g of dehydrated peel. Trujillo-Mayol and Alarcónenos
(2019) used microwave technology and microwave coupled
3.4. Bioactive compounds identification by analytical
with ultrasound with water ethanol (80:20 v/v) to extract
RP-HPLC-ESI-MS
bioactive compounds from avocado peel, however in the best
condition of extraction showed values for phenolic content
A comprehensive analytical characterization of bioactive com-
and antioxidant activity by DPPH of 281.4 mg GAE/g dry extract
pounds present in avocado peel extracts was performed. The
and 779.1 ␮g TE/g dry extract, which are lower than obtained
analysis allowed the identification of 15 major compounds.
in the present work. The results shown in the present work
The identification was performed on basis their retention time
show that MAE and the optimal operating conditions allows
values, UV–vis spectra and mass information determined,
to extract larger amount of bioactive compounds with higher
together with information previously reported in literature
antioxidant activities from avocado peel, and on the other
(Figueroa et al., 2018a; Kosińska et al., 2012; López-Cobo
hand reducing the extraction times and the use of toxic
et al., 2016). Table 7 summarize the proposed identified com-
organic solvents.
pounds in avocado peel extracts (Fig. 3). Figueroa et al. (2018a)
 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161 159

Table 6 – TPC and antioxidant activity of bound phenolic compounds in avocado peel residues obtained after MAE with
acetone and ethanol.
TPC DPPH ATBS
(mg GAE/g residue) (mg ET/g residue) (mg ET/g residue)

Residue acetone peel 6.64 ± 0.52 2.61 ± 0.17 8.71 ± 0.79

Residue ethanol peel 7.73 ± 0.15 5.64 ± 0.32 18.45 ± 0.20

Table 7 – Identification of phenolic and other polar compounds by RP-HPLC-ESI-MS of ethanol avocado peel extract
obtained with MAE optimal condition.
Peak Proposed compound RT (min) m/z Molecular formula

1 Quinic acid 2.886 191.0 C7 H12 O6

2 Procyanidin dimer B 18.494 577.0 C30 H26 O12
3 3-O-caffeoylquinic acid 20.842 353.0 C16 H18 O9
4 Procyanidin trimer B 22.342 864.9 C45 H38 O18
5 Procyanidin dimer B 22.686 577.0 C30 H26 O12
6 Procyanidin dimer B 23.272 577.0 C30 H26 O12
7 (+)-Catechin 24.775 289.0 C15 H14 O6
8 Procyanidin trimer B 25.824 864.0 C45 H38 O18
9 Procyanidin trimer B 26.322 865.0 C45 H38 O18
10 Procyanidin tetramer B 27.184 1152.9 C60 H50 O24
11 Procyanidin trimer B 27.513 864.9 C45 H38 O18
12 Procyanidin trimer B 28.723 864.0 C45 H38 O18
13 Procyanidin trimer B 29.379 864.0 C45 H38 O18
14 Procyanidin dimer B 30.319 577.0 C30 H26 O12
15 Multinoside A 31.391 609.0 C27 H30 O16

4. Conclusions

Avocado peels are agroindustrial residues with high content

of bioactive compounds with potential use in food, cosmetic
and pharmaceutical application as natural ingredients. The
optimization of MAE conditions allowed to determine the con-
ditions of temperature, time and ethanol concentration to
extract bioactive compounds with high antioxidant capaci-
ties, values never reported, using a safe and green method of
extraction. Procyanidins, catechin and phenolic acids are the
major bioactive compounds found in avocado peel extracts
Fig. 3 – Chromatogram of MAE optimal condition of ethanol that are compounds reported with high antioxidant capacity.
avocado peel extract; a: ethanol interference; 1: Quinic acid; The results obtained in this work encourage the use of avo-
2, 5, 6: Procyanidin dimer B; 3: 3-O-caffeoylquinic acid; 4, 8, cado peels as a source of bioactive compounds by MAE soon
9, 11, 12: Procyanidin trimer B; 7: (+)-Catechin; 10: to scale the process and apply them in cosmetic, pharmaceu-
Procyanidin tetramer B; 15: Multinoside A at 280 nm. tical and food industries in order to reduce the use of synthetic
compounds.

Conflict of interest
with a HPLC-DAD-ESI-QTOF-MS was able to identify 61 com-
pounds in avocado peel extract, showing different family The authors declare no conflict of interest.
of compounds: procyanidins, flavonols, hydroxybenzoic and
hydroxycinnamic acids were the most representative groups. Declaration of Competing Interest
The condensed tannins were most numerous groups with 17
procyanidins with different polymerization degree, where pro-
The authors report no declarations of interest.
cyanidins dimeric A and B appear in different isomeric shapes.
In present study with RP-HPLC-ESI-MS was possible identify
the compounds that are present in higher concentration, such Acknowledgments
as 11 procyanidins, dimers and trimers in different isomer
shapes and one procyanidin tetramer B, organic acids as 3- Author Rafael G. Araújo would like to thank the National Coun-
O-caffeoylquinic and quinic acid, catechin and multinoside cil of Science and Technology of Mexicofor the scholarship
A (quercetin 3-(4-glucosylrhamnoside)). These identified com- received for this project in the PhD program in Food Science
pounds are compounds that have been associated with high and Technology at the Autonomous University of Coahuila,
antioxidant activities due to the large number of hydroxyl México. Also, thanks the scientific collaboration of CBQF under
groups capable of reducing free radical (Kosińska et al., 2012; the FCT – Fundação para a Ciência e a Tecnologiathrough
Spranger et al., 2008; Wang et al., 2010; Zhou et al., 2018). project UID/Multi/50016/2013.
160 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161

References Hurtado-Fernández, E., Carrasco-Pancorbo, A.,

Fernández-Gutiérrez, A., 2011. Profiling LC-DAD-ESI-TOF MS
method for the determination of phenolic metabolites from
Aguilar-Reynosa, A., Romaní, A., Rodríguez-jasso, R.M., Aguilar,
avocado (Persea americana). J. Agric. Food Chem. 59, 2255–2267,
C.N., Garrote, G., Ruiz, H.A., 2017. Comparison of microwave
http://dx.doi.org/10.1021/jf104276a.
and conduction-convection heating autohydrolysis
Kähkönen, M.P., Heinonen, M., 2003. Antioxidant activity of
pretreatment for bioethanol production. Bioresour. Technol.
anthocyanins and their aglycons. J. Agric. Food Chem. 51,
243, 273–283, http://dx.doi.org/10.1016/j.biortech.2017.06.096.
628–633, http://dx.doi.org/10.1021/jf025551i.
Ajila, C.M., Rao, U.J.S.P., 2013. Mango peel dietary fibre:
Kala, H.K., Mehta, R., Sen, K.K., Tandey, R., Mandal, V., 2016.
composition and associated bound phenolics. J. Funct. Foods
Critical analysis of research trends and issues in microwave
5, 444–450, http://dx.doi.org/10.1016/j.jff.2012.11.017.
assisted extraction of phenolics: have we really done enough.
Alvarez, L.D., Moreno, A.O., Ochoa, F.G., 2012. Avocado. In:
Trends Anal. Chem. 85, 140–152,
Tropical and Subtropical Fruits: Postharvest Physiology,
http://dx.doi.org/10.1016/j.trac.2016.09.007.
Processing and Packaging., pp. 437–454.
Kosińska, A., Karamac, M., Estrella, I., Hernandez, T., Bartolomé,
AOAC, 1995. Official methods of analysis. In: Methods 932.06,
B., Dykes, G.A., 2012. phenolic compound profiles and
925.09, 923.03, 991.43, 15th ed. Association of Official
antioxidant capacity of Persea Americana Mill. peels and seeds
Analytical Chemists, Gaithesburg, MD.
of two varieties. J. Agric. Food Chem. 60, 4613–4619,
Calder, M., Iztapalapa, U., 2016. Optimization of the antioxidant
http://dx.doi.org/10.1021/jf300090p.
and antimicrobial response of the combined effect of nisin
López-Cobo, A., Gómez-Caravaca, A.M., Pasini, F., Caboni, M.F.,
and avocado byproducts. LWT - Food Sci. Technol. 65, 46–52,
Segura-Carretero, A., Fernández-Gutiérrez, A., 2016.
http://dx.doi.org/10.1016/j.lwt.2015.07.048.
HPLC-DAD-ESI-QTOF-MS and HPLC-FLD-MS as valuable tools
Chan, C.H., Yeoh, H.K., Yusoff, R., Ngoh, G.C., 2016. A
for the determination of phenolic and other polar compounds
first-principles model for plant cell rupture in
in the edible part and by-products of avocado. LWT - Food Sci.
microwave-assisted extraction of bioactive compounds. J.
Technol. 73, 505–513,
Food Eng. 188, 98–107,
http://dx.doi.org/10.1016/j.lwt.2016.06.049.
http://dx.doi.org/10.1016/j.jfoodeng.2016.05.017.
Magalhães, L.M., Santos, F., Segundo, M.A., Reis, S., Lima, J.L.F.C.,
Chavan, U.D., Shahidi, F., Naczk, M., 2001. Extraction of
2010. Rapid microplate high-throughput methodology for
condensed tannins from beach pea (Lathyrus maritimus L.) as
assessment of Folin-Ciocalteu reducing capacity. Talanta 83,
affected by different solvents. Food Chem. 75, 509–512,
441–447, http://dx.doi.org/10.1016/j.talanta.2010.09.042.
http://dx.doi.org/10.1016/S0308-8146(01)00234-5.
Melgar, B., Dias, M.I., Ciric, A., Sokovic, M., Garcia-Castello, E.M.,
Chemat, F., Cravotto, G., 2013. Microwave-assisted Extraction for
Rodriguez-Lopez, A.D., Barros, L., Ferreira, I.C.R.F., 2018.
Bioactive Compounds.,
Bioactive characterization of Persea americana Mill.
http://dx.doi.org/10.1007/978-1-4614-4830-3.
by-products: a rich source of inherent antioxidants. Ind. Crops
Chen, Y., Ma, Y., Dong, L., Jia, X., Liu, L., Huang, F., Chi, J., Xiao, J.,
Prod. 111, 212–218,
Zhang, M., Zhang, R., 2019. Extrusion and fungal fermentation
http://dx.doi.org/10.1016/j.indcrop.2017.10.024.
change the profile and antioxidant activity of free and bound
Pandey, A., Belwal, T., Sekar, K.C., Bhatt, I.D., Rawal, R.S., 2018.
phenolics in rice bran together with the phenolic
Optimization of ultrasonic-assisted extraction (UAE) of
bioaccessibility. LWT - Food Sci. Technol. 115, 108461,
phenolics and antioxidant compounds from rhizomes of
http://dx.doi.org/10.1016/j.lwt.2019.108461.
Rheum moorcroftianum using response surface methodology
Chia, T.W., Dykes, G.A., 2010. Antimicrobial activity of crude
(RSM). Ind. Crop. Prod. 119, 218–225,
epicarp and seed extracts from mature avocado fruit (Persea
http://dx.doi.org/10.1016/j.indcrop.2018.04.019.
americana) of three cultivars. Pharm. Biol. 48, 753–756,
Pap, N., Beszédes, S., Pongrácz, E., Myllykoski, L., Gábor, M.,
http://dx.doi.org/10.3109/13880200903273922.
Gyimes, E., Hodúr, C., Keiski, R.L., 2013. Microwave-assisted
Chimsook, T., 2017. Microwave assisted extraction of avocado oil
extraction of anthocyanins from black currant marc. Food
from avocado skin and encapsulation using spray drying. Key
Bioprocess Technol. 6, 2666–2674,
Eng. Mater. 737, 341–346,
http://dx.doi.org/10.1007/s11947-012-0964-9.
http://dx.doi.org/10.4028/www.scientific.net/KEM.737.341.
Peng, H., Li, W., Li, H., Deng, Z., Zhang, B., 2017. Extractable and
Contreras, M., Hernández-ledesma, B., Amigo, L., Martín-álvarez,
non-extractable bound phenolic compositions and their
P.J., Recio, I., 2011. LWT - Food Science and Technology
antioxidant properties in seed coat and cotyledon of black
Production of antioxidant hydrolyzates from a whey protein
soybean (Glycinemax (L.) merr). J. Funct. Foods 32, 296–312,
concentrate with thermolysin: optimization by response
http://dx.doi.org/10.1016/j.jff.2017.03.003.
surface methodology. LWT - Food Sci. Technol. 44, 9–15,
Quatrin, A., Pauletto, R., Maurer, L.H., Minuzzi, N., Nichelle, S.M.,
http://dx.doi.org/10.1016/j.lwt.2010.06.017.
Carvalho, J.F.C., Maróstica, M.R., Rodrigues, E., Bochi, V.C.,
Cowan, A.K., Wolstenholme, B.N., 2016. Avocado. In: Caballero, B.,
Emanuelli, T., 2019. Characterization and quantification of
Finglas, P.M., Toldrá, F. (Eds.), Encyclopedia of Food and
tannins, flavonols, anthocyanins and matrix-bound
Health. Academic Press, Oxford, pp. 294–300.
polyphenols from jaboticaba fruit peel: a comparison between
FAOSTAT, 2018. Food and Agriculture Organization of the United
Myrciaria trunciflora and M. Jaboticaba. J. Food Anal. 78, 59–74,
Nations. Agriculture Database.
http://dx.doi.org/10.1016/j.jfca.2019.01.018.
http://www.fao.org/faostat/en/#data/QC.
Ramos, B., Maziero, G., Leone, A., Monteiro, E., Assis, D., Victor, J.,
Figueroa, J.G., Borrás-Linares, I., Lozano-Sánchez, J.,
Gomes, D., Pereira, R., Prandi, B., Valentim, B., Silveira, D.,
Segura-Carretero, A., 2018a. Comprehensive identification of
Machado, R., Melo, T., Pereira, C., Rezende, R., De Cássia, R.,
bioactive compounds of avocado peel by liquid
Gonçalves, R., 2019. Avocado seeds (Persea americana Mill.)
chromatography coupled to ultra-high-definition
prevents indomethacin-induced gastric ulcer in mice. Food
accurate-mass Q-TOF. Food Chem. 245, 707–716,
Res. Int. 119, 751–760,
http://dx.doi.org/10.1016/j.foodchem.2017.12.011.
http://dx.doi.org/10.1016/j.foodres.2018.10.057.
Figueroa, J.G., Borrás-Linares, I., Lozano-Sánchez, J.,
Rasera, G., Hilkner, M., Alencar, S., Castro, R., 2019. Biologically
Segura-Carretero, A., 2018b. Comprehensive characterization
active compounds from white and black mustard grains: an
of phenolic and other polar compounds in the seed and seed
optimization study for recovery and identification of phenolic
coat of avocado by HPLC-DAD-ESI-QTOF-MS. Food Res. Int.
antioxidants. Ind. Crop. Prod. 135, 294–300,
105, 752–763, http://dx.doi.org/10.1016/j.foodres.2017.11.082.
http://dx.doi.org/10.1016/j.indcrop.2019.04.059.
Flórez, N., Conde, E., Domínguez, H., 2015. Microwave assisted
Re, R., Nicoletta, P., Anna, P., Ananth, P., Min, Y., Catherine, E.,
water extraction of plant compounds. J. Chem. Technol.
1999. Antioxidant activity applying an improved ABTS radical.
Biotechnol. 90, 590–607, http://dx.doi.org/10.1002/jctb.4519.
 Food and Bioproducts Processing 1 2 7 ( 2 0 2 1 ) 152–161 161

Free Radic. Biol. Med. 26, 1231–1237, Tremocoldi, M.A., Rosalen, P.L., Franchin, M., Daiuto, R., Augusto,
http://dx.doi.org/10.1016/S0891-5849(98)00315-3. J., Massarioli, P., Denny, C., Paschoal, R., Melo, P.S., De Alencar,
Rodríguez-Carpena, J., Morcuende, D., Kylli, P., Andrande, M.-J., S.M., 2018. Exploration of avocado by-products as natural
Est, M., 2011. Avocado (Persea americana Mill.) phenolics, sources of bioactive compounds. PLoS One 13, 1–12,
in vitro antioxidant and antimicrobial activities, and http://dx.doi.org/10.1371/journal.pone.0192577.
inhibition of lipid and protein oxidation in porcine patties. J. Trujillo-Mayol, I., Alarcónenos, J., 2019. Improvement of the
Agric. Food Chem. 59, 5625–5635, polyphenol extraction from avocado peel by assisted
http://dx.doi.org/10.1021/jf1048832. ultrasound and microwaves. J. Food Process Eng. 42, e13197,
Rotta, E.M., de Morais, D.R., Biondo, P.B.F., dos Santos, V.J., http://dx.doi.org/10.1111/jfpe.13197.
Matsushita, M., Visentainer, J.V., 2016. Use of avocado peel Wang, W., Bostic, T.R., Gu, L., 2010. Antioxidant capacities,
(Persea americana) in tea formulation: a functional product procyanidins and pigments in avocados of different strains
containing phenolic compounds with antioxidant activity. and cultivars. Food Chem. 122, 1193–1198,
Acta Sci. - Technol 38, 23–29, http://dx.doi.org/10.1016/j.foodchem.2010.03.114.
http://dx.doi.org/10.4025/actascitechnol.v38i1.27397. Xie, P., Huang, L., Zhang, C., Zhang, Y., 2015. Phenolic
Saavedra, J., Córdova, A., Navarro, R., Díaz-Calderón, P., compositions, and antioxidant performance of olive leaf and
Fuentealba, C., Astudillo-Castro, C., Toledo, L., Enrione, J., fruit (Olea europaea L.) extracts and their structure-activity
Galvez, L., 2017. Industrial avocado waste: functional relationships. J. Funct. Foods 16, 460–471,
compounds preservation by convective drying process. J. Food http://dx.doi.org/10.1016/j.jff.2015.05.005.
Eng. 198, 81–90, Zhang, H.F., Yang, X.H., Wang, Y., 2011. Microwave assisted
http://dx.doi.org/10.1016/j.jfoodeng.2016.11.018. extraction of secondary metabolites from plants: current
Segovia, F.J., Corral-Pérez, J.J., Almajano, M.P., 2016. Avocado seed: status and future directions. Trends Food Sci. Technol. 22,
modeling extraction of bioactive compounds. Ind. Crops Prod. 672–688, http://dx.doi.org/10.1016/j.tifs.2011.07.003.
85, 213–220, http://dx.doi.org/10.1016/j.indcrop.2016.03.005. Zhou, P., Zhang, L., Li, W., Zhang, S., Luo, L., Wang, J., 2018. In vitro
SIAP, 2020. Servicio de Información Agroalimentaria y Pesquera. evaluation of the anti-digestion and antioxidant effects of
http://infosiap.siap.gob.mx/aagricola siap gb/ientidad/index. grape seed procyanidins according to their degrees of
jsp. polymerization. J. Funct. Foods 49, 85–95,
Spranger, I., Sun, B., Mateus, A., Freitas, V., Silva, J., 2008. http://dx.doi.org/10.1016/j.jff.2018.08.001.
Chemical characterization and antioxidant activities of
oligomeric and polymeric procyanidin fractions from grape
seeds. Food Chem. 108, 519–532,
http://dx.doi.org/10.1016/j.foodchem.2007.11.004.