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Avocado seed contains elevated levels of phenolic compounds and exhibits antioxidant properties. We investigated the effect of Avocado Seed Flour (ASF) on the lipid levels in mice on a hyperlipidemic diet. The concentration of phenols was determined by high-performance liquid chromatography, antioxidant activity was evaluated using the Trolox equivalent antioxidant capacity method, and dietary fiber was measured using the Association of Official Analytical Chemists (AOAC) method. The LD50 of ASF was determined using Lorke's method and hypolipidemic activity was evaluated in a hypercholesterolemic model in mice. Protocatechuic acid was the main phenolic compound found in ASF, followed by kaempferide and vanillic acid. The total phenolic content in the methanolic extract of ASF was 292.00 ± 9.81 mg gallic acid equivalents/g seed dry weight and the antioxidant activity resulted in 173.3 μmol Trolox equivalents/g DW. In addition, a high content of dietary fiber was found (34.8%). The oral LD50 for ASF was 1767 mg/kg body weight, and treatment with ASF significantly reduced the levels of total cholesterol, LDL-C, and prediction of the atherogenic index. Therefore, the antioxidant activity of phenolic compounds and dietary fiber in ASF may be responsible for the hypocholesterolemic activity of ASF in a hyperlipidemic model of mice.
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ORIGINAL PAPER
Hypolipidemic Effect of Avocado (Persea americana Mill)
Seed in a Hypercholesterolemic Mouse Model
María Elena Pahua-Ramos & Alicia Ortiz-Moreno &
Germán Chamorro-Cevallos &
María Dolores Hernández-Navarro &
Leticia Garduño-Siciliano &
Hugo Necoechea-Mondragón &
Marcela Hernández-Ortega
Published online: 2 March 2012
#
Springer Science+Business Media, Inc. 2012
Abstract Avocado seed contains elevated levels of pheno-
lic compounds and exhibits antioxidant properties. We in-
vestigated the effect of Avocado Seed Flour (ASF) on the
lipid levels in mice on a hyperlipidemic diet. The concen-
tration of phenols was determined by high-performance
liquid chromatography, antioxidan t activity was evaluated
using the Trolox equivalent antioxidant capacity method,
and dietary fiber was measured using the Association of
Official Analytical Chemists (AOAC) method. The LD
50
of ASF was determined using Lorkes method and hypolipi-
demic activity was evaluated in a hypercholesterolem ic
model in mice. Protocatechuic acid was the main phenolic
compound found in ASF, followed by kaempferide and
vanillic acid. The total phenolic content in the methanolic
extract of ASF was 292.00±9.81 mg gallic acid equivalents/
g seed dry weight and the antioxidant activity resulted in
173.3 μmol Trolox equivalents/g DW. In addition, a high
content of dietary fiber was found (34.8%). The oral LD
50
for ASF was 1767 mg/kg body weight, and treatment with
ASF significantly reduced the levels of total cholesterol,
LDL-C, and prediction of the atherogenic index. Therefore,
the antioxidant activity of phenolic compounds and dietary
fiber in ASF may be responsible for the hypocholesterole-
mic activity of ASF in a hyperlipidemic model of mice.
Keywords Avocado
.
Dietary
.
Fiber
.
Hypolipidemic
.
Phenolic compounds
.
Seed
Abbreviations
ABTS 2, 2-azino-bis (3-ethylbenzthiazoline-6-
sulphonic acid)
AI Atherogenic Index
ASF Avocado Seed Flour
BW Body Weight
Plant Foods Hum Nutr (2012) 67:1016
DOI 10.1007/s11130-012-0280-6
M. E. Pahua-Ramos
:
A. Ortiz-Moreno (*)
:
M. Hernández-Ortega
Departamento de Ingeniería Bioquímica,
Escuela Nacional de Ciencias Biológicas,
Instituto Politécnico Nacional,
Plan de Ayala y Prol. Carpio s/n, Col. Casco de Santo Tomás,
C.P. 11340 México, D.F., Mexico
e-mail: ortizalicia@hotmail.com
G. Chamorro-Cevallos
:
L. Garduño-Siciliano
Departamento de Farmacia,
Escuela Nacional de Ciencias Biológicas,
Instituto Politécnico Nacional,
Av. Wilfrido Massieu s/n, esq. Manuel L. Stampa,
Col. Unidad Profesional Adolfo López Mateos,
Del. Gustavo A. Madero,
C.P. 07738 México, D.F., Mexico
M. D. Hernández-Navarro
Departamento de Farmacia, Facultad de Química,
Universidad Autónoma del Estado de México,
Paseo Colón esq. Paseo Tollocan, Col. Residencial Colón,
C.P. 50120 Toluca, Estado de México, Mexico
H. Necoechea-Mondragón
Coordinación de Operación de Redes de Investigación,
Instituto Politécnico Nacional,
Edificio Secretaría Académica. Av. Miguel Othón
de Mendizábal s/n, Col. La Escalera,
C.P. 07738 México, D.F., Mexico
GAE Gallic Acid Equivalents
HDL-C High-Density Lipoprotein Cholesterol
LDL-C Low-Density Lipoprotein Cholesterol
LD
50
Median Lethal Dose
TC Total Cholesterol
TG Triglycerides
Trolox 6-hydroxy-2, 5, 7, 8 tetramethylchroman-
2-carboxylic acid
Introduction
The treatment of hypercholesterolemia and related car-
diovascular diseases with medicinal plants has increased
in recent years [1]. Reasons for the increased populari ty
of these herbal medicines may include their relatively
low cost compared to orthodox medicines, availability
(since they are almost always d erived from available
plants in the local region), and efficacy. Although poisonous
plants are ubiquitous throughout the world, herbal medicine is
still used by up to 80% of the total population in developing
countries. Despite widespread use, few scientific studies have
explored the safety and efficacy of traditional herbal remedies
[2] Substances such as chlortalidone and propranolol, which
are used in the treatment of hypercholesterolemia, can have
adverse effects that affect therapy compliance and total quality
of life of the patient [3].
Cardiovascular disease is a growing health problem
throughout the world and represents a leading cause of
mortality a nd morbidity in humans [4]. Several factors,
such as a high caloric diet, age, lack of exercise, smok-
ing, alcohol consumption, and genetic predisposition
have been linked with cardiovascular disease [1]. Elevat-
ed cholest erol levels predispose patients to a condition
known as hypercholesterolemia [5], which in cre ases th e
risk of fatal and nonfatal coronary heart disease in people
over the age of 50 [6].
Several beneficial medicinal proper ties o f compoun ds
present in the avocado seed and peel have been reported,
which are related to the elevated levels of phenolic com-
pounds (64% in seed, 23% in peel, and 13% in pulp). In
addition, the seeds and peels of avocado also contribute
57% and 38% of the antioxidant capacities of the entire
fruit, respectively [7].
Several studies on the hypolipidemic effects of the
avocado seed have been focused on methanolic extracts
[1] and aqueous extracts [8]; howe ver, the study of the
hypolipidemic effects of the entire avocado seed provides
an interesting alternati ve, since the seed represents 13-18%
[9] of the av oc ado fruit and is disca rd ed during avocado
pulp processing. Therefore, the aim of this study was to
investigate the effect of ASF on the lipid levels of mice
on a hyperlipidemic diet.
Materials and Methods
Seed Flour Preparation
Fresh seeds of Persea americana Mill were obtained from an
orchard in Uruapan, Michoacán, Mexico. The seeds were
identified and authenticated with the number 15658 at the
herbarium of the Centro Médico Nacional of the Instituto
Mexicano del Seguro Social. The seeds were washed, cut into
small pieces, and dried in an oven (Fisher Scientific, Isotemp
model 718F, USA) at 40 °C for 48 h until achieving a moisture
level of 4%. The small pieces were then crushed into powder
with a hammer (Wiley Mill standard model No. 3, Arthur
Thomas Co., USA) until the flour passed through US 20 mesh
(0.844 mm).
Phenol Extraction
Phenol extraction was performed according to the method
described by Asaolu et al. [1]. One hundred grams of seed
flour were sloped into a beaker and then extracted with 75%
methanol overnight in a soxhlet extractor (Electro thermal
model ME466, England). The methanolic extract was con-
centrated and allowed to evaporate to dryness at 50 °C using
a rotary evaporator (Büchi 461, Brinkmann Instruments,
Switzerland). The extract was then dissolved in water at a
concentration of 4 g/100 ml.
Total Phenolic Content
HPLC-PDA Analysis of Phenolic Compounds
Twenty microlitres of the methanolic extract were analyzed
using an HPLC-PDA system (Varian 920-LC) and a C18
column (Onmisher 5; 150×4.6 mm i.d.). The solvent mixture
system contained a mixture of 5% formic acid in water (A) and
100% methanol (B), with a flow rate of 0.9 ml/min, and the
gradient flow was as follows: 0 min5% B; 3 min15% B;
13 min25% B; 25 min
30%B;35min3
5
%B;39min
45% B; 42 min45%B;44min50% B; 45 min70% B;
50 min70%B;56min75% B; and 61 min80% B.
Detection was achieved with a photo diode array detector.
Spectrophotometric data from all peaks were monitored at
220280 nm and chromatograms were recorded at 340 nm.
The data were processed with Varian Galaxie Chromatogra-
phy Data Software version 1.9.302.952 (Agilent Technologies
USA). Phenolic compound quantification was determined
using the retention times and absorbance recorded in the
chromatograms relative to external standards [10].
Total phenol content was quantified using the Folin
Ciocalteu [11]. G allic acid was used as a standard for the
calibratio n c ur ve. The result s w ere expressed as gallic
acid eq uiva lents (GAE) m g/g seed dry weight (D W) .
Plant Foods Hum Nutr (2012) 67:1016 11
Antioxidant Activity
Total antioxidant capacity was evaluated using the method
reported by Re et al. [12] with Trolox as the standard. A
stock solution was prepared by reacting 7 mM ABTS with
2.45 mM potassium persulfate to generate the ABTS cation
chromophore. This solution was diluted with absolute
ethanol until reaching an absorbance of 0.7±0.02 at
734 nm. A sample of 10 μl of ASF extract was added
to 990 μL of ABTS solution and the reaction was followed
over a 7 min time course. Total antioxidant activity or capacity
was calculated relative to the reactivity of Trolox as the
standard under the same conditions. The results were
expressed as μmol Trolox equivalent/g of seed DW.
Proximate Analysis
Moisture, protein, ether extract, ash, and crude fiber
content in ASF were determined in triplicate following
the standard methods from the Association of Official
Analytical Chemists International (AOAC) [13], and total
carbohydrate content was calculated as the difference to a
total of 100%. Dietary fiber was also determined in
triplicate using AOAC methods 997.08 and 999.03 [13].
Animals
For the hypolipidemic activity assay, 40 8-10-week-ol d
adult male CD-1 mice with an average weight of 28±2 g
were obtained from the Centro de Investigac ión y Estudios
Avanzados (CINVESTAV) at the Instituto Politécnico Nacional
(IPN-México). Each mouse was weighed and random ly
assigned to groups by body weight (BW). Each group of mice
was housed in cages with wooden chip bedding and maintained
under a 12 h light/dark cycle. Mice were fed with standard
laboratory chow (5001 Lab Rodent Diet, PMI Nutrition
International, Inc., Bienwood, MO) and provided water
ad libitum. The animal experiments and study design
were approved by the Laborato ry Animal Care Commit-
tee of IPN and wer e conducted in compliance with the
Official Mexican standard NOM-062-ZOO-1999 regarding
technical specifications for production, care, and use of labo-
ratory animals [14].
Oral Acute Toxicity
Acute toxicity tests were conducted according to Lorkes
methodology [15].
Hypolipidemic Activity of ASF
Five groups of CD-1 mice (8 mice per group) were formed,
with group 1 serving as the control. A diet rich in cholesterol
was supplied to the animals ad libitum for six days in
order to induce hypercholesterolemia [ 16, 17]. The diet
formula u sed is shown i n Table 1.Group25 recei ve d
the hypercholesterolemic diet, which was administered
with distilled wate r (group 2 ) or differen t dose s of ASF
(groups 3, 4 and 5 received 125, 250, and 500 mg ASF/kg
BW, respectively, once a day) by gavage. The doses were
chosen according to the acute toxicological study divided by a
security factor of 10.
At the end of six days, the TC, HDL-C, LDL-C and TG
concentrations were determined according to the methodology
described by Argüelles et al. [17].
Statistical Analysis
All data for acute toxicity were statistically analyzed by the
Students t-test using Sigma-Stat version 3.5 (Jandel San
Raphael, CA) and P <0.001 was considered statistically
significant. Hypolipide mic activity data were s tatisticall y
analyzed by one-way analysis of varia nce (ANOVA) and
Tuk
eys test using Sigma-Stat version 3.5 (Jandel San
Raphael, CA). The data were reported as mean ± standard
deviation (SD). A P <0.05 was considered statistically
significant for hypolipidemic activity data.
Results and Discussion
Identification of Phenolic Compounds
Analysis of methanolic extract from avocado seed by
HPLC-PDA identified eleven major peaks. Seven phe nolic
compounds were identified using external standards, spectra
characteristics, and retention time. Protocatechuic acid
(128.18±0.01 μg/g DW) was the main phenolic compound
identified, followed by kaempferide (107.42±0.04 μg/g
DW) and vanillic acid (28.67 ±0.001 μg/g DW). In addition,
clorogenic acid, syringic acid, rutin, and kaempferol were
present in small amounts (Table 2). Recen tl y, Rodrígu ez-
Carpena et al. [18] analyzed and classified phenolic
compounds from two avocado varieties as ca techins,
(sum of catechin and ep icate c hin) , hydroxybenzoi c acids
Table 1 Formulation
for the hypercholesterol-
emic diet
Ingredients % of total weight
Lab rodent diet
5001
53.5
Sucrose 30
Casein 10
Butter 5
Cholesterol 1
Sodium cholate 0.5
12 Plant Foods Hum Nutr (2012) 67:1016
(p -coumaric, caffeic, ferulic, and sinapic), hydroxycin-
namic acids (p-hydroxybenzoic, protocatechuic, vanillic,
syringic, and gallic), flavonols, and procyanidins (sum of
dimers, oligomers, and polymers). These authors r eported
that the seed and peel contained the highest amount of
phenols in the entire fruit. Moreover, Terpinc et al. [19]
reported that flavonoids, rutin, catechin, and quercetin
are widespread in nature and may act as powerful
antioxidants.
These findings and our results provide evidence for the
importance of phenols present in avocado seed, since
phenolic compounds have been shown to reduce plasma
lipid levels in human body through the upregulation of
LDL receptor expression, inhibition of hepatic lipid synthesis
and lipoprotein se cretion, a nd incre ase i n c holesterol
elimination through bile acids [20].
Total Phenolic Content and Antioxidant Activity of ASF
The total phenolic content and antioxidant activity of the
methanolic extract of ASF was determined to be 292.00±
9.81 mg GAE/g seed DW and 173.3 μmol Trolox equiv-
alents/g seed DW, respectively. Rodríguez-Carpena et al.
[18] previously reported a total phenolic content of 351.1±
9.88 mg GAE/g seed for t he Hass variety and 416.4±
10.48 mg GAE/g seed dry matter for the Fuerte variety.
According to Wang et al. [7], seeds contain the strongest
antioxidant properties and highest phenol and procyanidin
content compared to the pulp. Soong & Barlow [21 ]
reported a s ignificantly higher total antioxidant capacity
and phenolic content of fruit seeds than the edible portions.
In most fruits, the contribution of the fruit seed fraction
compared to the total antioxidant activity and phenolic
content was more than 95%, and therefore these authors
suggested that the fruit seeds should be further utilized
rather than just discarded as waste [21]. Importantly, our
results are in agreement with the findings by Wang et al.
[17] and Soong & Barlow [21] showing that the antioxidant
activity of fruit seeds components may be responsible for
the hypocholesterolemic activity observed.
Proximate Analysis and Dietary Fiber
We found that the ASF preparation contained 4.0±0.8
moisture, 2. 2±0.14 ash, 4.75±0.01 protein, 6.39±0.5
crude fiber, 4.38±0.8 ether extract, and 79.10±0.8 car-
bohydrates (data expressed as mean ± standard deviation
g/100 g sa mple fresh weight; n0 3). The low oil content
of the ASF suggested that oil and its fatty acids could
have a minimal effect on cholesterol and LDL-C reduc-
tion, since i t only represented a small portion of t he total
daily oil intake of the treated mice. Nijjar et al. [22]
found t hat nuts a re a good source of mono and pol y-
unsaturated fatty acids and also contain dietary fiber,
phytosterols, and polyphenols. These components likely
combine to a reduction in LDL-C levels beyond the
effects predicted by equations based solely on fatty acid
profiles. Nevertheless, the high crude fiber content
(6.39 g/100 g DW sample) of ASF could have a beneficial
effect on total cholesterol and LDL-C reduction in the
plasma of the g roups of mice treated [23].
The ASF preparation was found to contain 34.8±3.4 g
dietary fiber/100 g DW samp le, which is relevant in this
study, since the natural gel-forming or viscous fibers
(pectin, gums, mucilage, algal polysaccharid es, some
storage polysaccharides, and some hemicelluloses) are
water-soluble and resistant to digestion by human gastro-
intestinal enzymes that are part of the dietary fiber.
Moreover, this content has been shown to be associated
with a cholesterol-lowering effect [24]. The dietary fiber
content of the avocado seed is similar to another Mexican
seed called chia [25
]. Reyes-Caudillo et al. [25]r
e
portedthat
chia seeds from Jalisco and Sinaloa States contain a total
dietary fiber content of 39.9% a nd 36.9%, respectively.
Therefore, the die tary fibe r conte nt in chia see ds is of
sufficient level to promote beneficial health effects,
including a reduction of cholesterolaemia, modification of
the glycemic and insulinaemic responses, changes in intestinal
function, and antioxida nt a ctivity. The high content of
dietary fiber in ASF found in the present study
suggests that dietary fiber could play an important role in
the hypocholesterolemic activity in mice.
Oral Acute Toxicity
In the first stage of the oral acute toxicity study, the
animals did not exhibit any toxicological signs, including
depression, writhing, diarrhea, hypermotility, or aggression
Table 2 Phenolic compounds in methanolic extract from Persea
americana Mill
Peak number Rt (min) UV (nm) μg/g
1 Protocatechuic acid 6.12 243, 322 128.18±0.01
2 Clorogenic acid 7.37 242,278,439 0.516±0.02
3 Syringic acid 8.98 242,314, 443 2.51±0.002
4 Vanillic acid 9.87 242,380,436 28.67±0.001
5 NI 11.00 242,307,446
6 Rutin 11.67 242,277,319,386 9.63±0.008
7 NI 13.28 242,315,363
8 NI 14.48 242,318,446
9 Kaempferol 16.30 242,311,386,429 2.19±0.002
10 Kaempferide 23.81 216,242 107.42±0.04
11 NI 51.71 241,334,380
Data expressed as mean ± standard deviation; n0 3. NI0 not identified
Plant Foods Hum Nutr (2012) 67:1016 13
compared to the control group. No signs of toxicity or death
were observed in any of the animals, and all animals survived
to the end of the 14 day study period. Weight gain in the
control animals was minimal (< 4%), while the treated
animals exhibited a slight increase in w eight, although
there was no significant difference in the percent weight
change between the groups (P<0.05). In the s ec on d stage
of the study, we observed 100% mortality by day six in
the group fed with 2500 mg ASF/kg BW. Table 3 lists
the effects of different doses of ASF on daily food and
water intake and on the weight of the main organs,
which was exp res sed as a ratio of r elati ve w eig ht ( RW)
to total body weight. Mice administered 100 and
1000 mg ASF/kg BW exhibited significant differences
(P 0.001) com pared with the control, where by liver
weight was lower and kidney weight was higher than
control group. An increase in the RW of the kidney has
also been reported by Ozolua et al. [8] in adult rats fed
aqueous seed extract from avocado. In addition, Brai et al.
[26] found that liver weights were significantly increased in
albino rats fed avocado aqueous leaf extract after induction of
a hyperlipid em ic di et compared to normal cont rol r at s,
which was accompanied with a significant increase in
liver cholesterol level. These findings together with the
results of this study sugges t that the comp ound s pre sent
in avocado leaves and seeds are different and have an
opposite influence on the liver.
In the second stage of the acute toxicity study, no significant
differences (P>0.001) were found in daily food and water
intake between mice treated with 1250 mg ASF/kg BW and
control mice; however, significant differences (P 0.001)
were found in daily food and water intake between mice
treated with 2500 mg ASF/kg BW and control mice.
Based on these results, we determined the oral LD
50
for
ASF to be 1767 mg/kg BW by using the geometric mean of
the dose tha t cau sed 100 % m ortality and the dose that
caused no mortality, as suggested by Lorke [15]. The oral
LD
50
of ASF in mice indicated that it exhibited a low
toxicity [17]. It has previously been shown that ether
and aqueous extracts of Persea americana Mill seed
administered by intraperitoneal injection in rats also had
low toxicity, with LD
50
values of 751.6 mg/kg BW an d
10 g/kg BW, respectively [8 , 27].
Hypolipemic Activity of ASF
To determine the hypolipemic activity of ASF, mice were
dosed according to the LD
50
of ASF found in this study. The
Table 3 Daily food and water consumption and relative weight of liver and kidney in CD-1 mice treated with different doses of avocado seed flour
Groups Dose (mg/kg) Daily food consumption (g) Daily fluid consumption (ml) LBW (%) KBW (%)
Control 1st step
a
38.3±10.2 40.8±12.8 6.80±0.15 1.58±0.09
ASF 10 28.4±10.4
a
34.4±14.3 6.75±0.37 2.67±0.02
a
100 31.9±8.3 47.1±11.2 5.48±0.11ª,
b
1.83±0.05
a,b
1000 33.7±7.3 50.2±10.4
a
5.04±0.22ª,
b
1.75±0.09
a,b
Control 2nd step
c
35.5±6.2 53.4±9.7 5.50±0.28 1.54±0.16
ASF 1250 40.8±8.0 46.9±7.6 5.14±0.14 1.44±0.10
2500 21.3±14.5
c
28.1±15.8
c
ND ND
Data expressed as mean ± standard deviation; n0 8. ASF, Avocado Seed Flour; LBW, liver-to-body weight ratio; KBW, kidney-to-body
weight ratio.
a
P<0.001, significant difference with respect to control 1st step,.
b
P<0.001; significant difference with respect to 10 mg/kg of ASF;
c
P<0.001, significant difference with respect to control 2nd step. ND: Not determined.
Table 4 Effect of avocado seed flour on lipid profile of mice
Treatment ASF Dosis (mg/kg) TC (mmol/L) LDL-C (mmol/L) HDL-C (mmol/L) TG (mmol/L) AI
Normocholesterolemic 31.9±7.16 12.50±3.02 18.92±4.25 1.05±0.13 1.6±0.05
Hypercholesterolemic 106.7±9.70 92.1±10.26 15.00±2.1 0.852±0.09 7.9±1.86
ASF 125 70.9±4.49
a
55.8±5.35
a
15.6±1.79 0.872±0.09 3.9±0.59
a
250 69.0±5.15
a
56.1±5.28
a
13.5±0.78 0.914±0.07 4.3±0.58
a
500 67.6±4.92
a
54.4±5.37
a
13.8±1.21 0.930±0.12 4.3±0.72
a
Data expressed as mean ± standard deviation; n0 8, analyzed by ANOVA and Tukey-Kramer test.
a
P 0 .0001; significant difference with
respect to the hypercholesterolemic control. ASF, Avocado Seed Flour. TC, Total Cholesterol; LDL-C, Low-density Lipoprotein
Cholesterol; HDL-C, High-density Lipoprotein Cholesterol; TG, Triglycerides; AI, Ath erogenic Index.
14 Plant Foods Hum Nutr (2012) 67:1016
hypolipidemic effect of ASF was evaluated at doses at 125,
250, and 50 0 mg/kg BW. Acute supplementation of
cholesterol produced a significant (P 0.05) elevation in
plasma cholesterol levels in the hypercholesterolemic
control compared to the normocholesterolemic control.
In addition, the TC increa sed from 31.9±7.16 to 106.7±
9.70 mmol/L, LDL-C increased from 12.5±3.02 to 92.1±
10.26 mmol/L, and the calculated AI increase from 1.6±
0.05 to 7.9±1.86 between the two groups, respectively
(Table 4 ). No significant (P>0.05) changes were found
in the plasma HDL-C (18.92±4.25 vs 15.0±2.1 mmol/L,
respectively) and TG (1.05±0.13 vs. 0.85±0.09 mmol/L,
respective ly) between the two groups. These o bse rvat ions
could be associated with insulin activit y [28].
Similar results were reported by Asaolu et al. [1]in
normocholesterolemic and hypercholesterolemic groups for
TC (3.12±0.83 mmol/L vs 7.52±1.11 mmol/L, respectively)
andLDL-C(0.36mmol/Lvs5.79±2.10mmol/L,respec-
tively) using a methanol extract of avocado seeds. In
addition, t he AI was significantl y increased in the hyper-
cholesterolemic group compared to the normocholestero-
lemic group of that study (4.3 vs 1.3, respectiv ely) .
Treatment of mice with 125 mg ASF/kg BW significantly
(P 0.05) reduced the elevated levels of TC by 33% (106.7±
9.70 to 70.9±4.49 mmol/L) and LDL-C by 39.4% (92.1±
10.26 to 55.8±5.35 mmol/L). In addition, treat ment with
250 mg ASF/kg BW reduced TC and LDL-C by 34 and
39%, respectively, while treatment with 500 mg ASF/kg
BW reduced the TC and LDL-C levels by 36 and 41%,
respectively. A similar effect was reported by Asa olu et al.
[1] in Albino rats administered 200 mg of avocado seed
extract (75% methanol)/kg BW, where they observed a
significant reduction in TC, LDL-C, and TG levels by 47,
69, and 44%, respectively, compared to hypercholesterolemic
control mice. In addition, it was reported that the cholesterol
levels of hype rt ensiv e rats treated with 5 00 mg/kg BW of
avocado aqueous seed extract were reduced by 19.2,
42.5, 47.9, and 13.6% in the plasma, kidney, heart, and
liver, respectively, compared to hyp e rtens ive control mic e
[29]. In add it ion, significant r ed ucti ons in LDL-C and
triglycerides were also observed. These studies together
with our results indicate that aqueous or methanol seed
extract or seed flour of avocado can be used as an
effective supplement in mice and rats for treating
hyperlipidemia.
Conclusion
In this study, we found that ASF has low toxicity and can
significantly reduce the cholesterol and LDL-C levels in
hypercholesterolemic mice. This effect could be attributed
to the phenolic content, antioxidant activity, and/or dietary
and crude fiber content of the seed. Further research is
required in order to identify the components of ASF that
are responsible for the observed hypocholesterolemic
effects.
Acknowledgments This research was partly funded by the Consejo
Nacional de Ciencia y Tecnología (CONACyT) scholarship, Secretaria
de Investigación y Posgra do-IPN Proyect Number. 20100788,
Comisión de Operación y Fomento de Actividades Académicas del
IPN (COFAA-IPN), and Universidad Autónoma del Estado de México.
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16 Plant Foods Hum Nutr (2012) 67:1016
... Avocado fruit has an olive-green peel with a smooth, fatty, thick and almost creamy-textured pulp. Avocado seed makes up 13% of the Nutraceuticals 2024, 4 418 fruit and contains 64% of the phenolic compounds contributing to 57% of the antioxidant activity of the whole fruit [13]. The seed contains higher antioxidant activity, and higher procyanidin content than the pulp [14]. ...
... Phytochemicals found in avocado seed include flavonoids, alkaloids, saponins, glycosides and tannins [15][16][17]. The phenolic compounds in avocado seed include protocatechuic acid, kaempferide, vanillic acid, chlorogenic acid, syringic acid, rutin and kaempferol [13]. Phenolic compounds reduce adipogenesis, inhibit digestive enzymes and possess antiinflammatory and antioxidant effects [18,19]. ...
... Avocado seed has been reported to contain catechins, hydroxybenzoic acids, hydroxycinnamic acids, flavonols and procyanidins [13]. Another study showed the presence of 5-O-caffeoylquinic acid, quercetin 3-O-glucoside, quercetin 3-O-galactoside, quercetin 3-O-rutinoside, quercetin-3,4 ′ -diglucoside and quercetin 3-O-arabinoside [58]. ...
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Avocado seed contains 64% of the phenolic compounds of the whole fruit. This makes avocado seed a potential candidate for the development of treatments for different illnesses, including obesity (the major risk factor for metabolic disorders). The aim of this study was to investigate the effects of avocado seed powder on high-fat diet-induced obesity in rats. Sprague Dawley rats (16 rats) were fed a high-fat diet for 10 weeks. After 10 weeks, the rats were assigned into two groups of eight animals each and were fed either a high-fat diet (HFD; control group) or a high-fat diet containing avocado seed powder (HFD-A; treatment group) for 6 weeks. Animals were weighed weekly, and weekly weight gain was determined. Animals in the treatment (avocado seed) group showed significantly lower body weight gain (7.8 ± 9.63 g) than animals in the control group (33.9 ± 10.84 g) at the end of this study. The treatment group presented with lower triglycerides than the control, with LDL and HDL comparable to the control group. Avocado seed powder showed potential to reduce obesity in rats fed a high-fat diet. Avocado seed can therefore be investigated further as a potential anti-obesity nutraceutical.
... Pahua-Ramos et al. [80] noted that carbohydrates found in avocado may regulate blood cholesterol. However, De la Torre-Carbot et al. [81] propose that the cardioprotective action of avocado may be influenced by the high level of tocopherols in its oil, as observed in rats fed diets with avocado oil. ...
... Interestingly, avocado waste may be an important source of bioactive compounds that can exert beneficial health effects, including cardiovascular health. However, these effects have only been noted on dyslipidemia parameters in animal models [18,75,[80][81][82][83][84][85][86][87][88][89][90][91]. More details about it were described by Pineda-Lozano et al. [18]. ...
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Botanically speaking, avocado (Persea americana) is a fruit. It consists of a single large seed surrounded by a creamy, smooth-textured edible mesocarp or pulp covered by a thick, bumpy skin. Avocado is a nutrient-dense fruit, containing a range of bioactive compounds which have been independently associated with cardiovascular health. These compounds have been obtained from the pulp, peel, and seed. This narrative review summarizes the current understanding of the cardioprotective potential of avocado fruit, especially the pulp and seed, and its food products, and examines the biological mechanism behind it.
... For example, avocado seeds have 15 % higher phenolic content [7] and exhibit more antioxidants (70 %) than the fresh edible parts (pulp) [8], however, their chemical compositions can differ because of the variation in geographical settings, soil chemistry, and agronomic practices [6]. Furthermore, fresh and lyophilized extracts of avocado seeds have been shown to exhibit sizable physiological health and functional properties, including cholesterol-lowering [9], antidiabetic [10] antioxidant, anti-inflammatory [11,12] and anti-microbial activities [13]. These unique qualities are in high demand for valorization such as for commercial use in the production of healthy functional ingredients for food and non-food processing applications, which is critical for minimizing by-product waste. ...
... The use of plants in the treatment of hyperlipidemia and cardiovascular disorders has grown in recent years, and there is a growing consumer demand for fresh, natural, safe, and healthful foods that need little work or time to prepare [9]. Actually, because they have a low to medium energy density and are significant providers of major nutrient deficiencies, dietary guidelines worldwide advocate consuming more fruit and vegetables. ...
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Annually, cardiovascular diseases (CVDs) claim the lives of more than 17 million individuals. In this study, hypolipidemia potential of Persea americana, Moringa oleifera, Chrysophyllum albidum and Capsicum annuum seed oils were evaluated. Fifty adult Wistar rats of both sexes weighing between 120 and 170 g were grouped into ten, with five animals in each group. The animals were stabilized for one week prıor to the induction period. The seed oils were administered in their meal for fourteen days. Blood samples were collected through a cardıac puncture into heparinized tubes centrifuged at 5000 rpm for 10 minutes and were used for haematological and lipid profile assays. Comparatively between the analyte and control groups, the result showed no significant difference (p≥0.05) in haemoglobin (HB) with a significant increase (p≤0.05) in the packed cell volume (PCV) level of the rats. In addition, there were significant reductions in the levels of triglycerides, total cholesterol, and low-density lipoprotein-cholesterol. Among the male rats of the analyte and control group, there was a significant increase (p<0.5) in the level of high-density lipoprotein-cholesterol ın their blood samples. This study reveals that the rich essential fatty acid reported in the seed oils has hypolipidemic effect potential.
... Por outro lado, até um certo ponto, o aumento da temperatura da extração favorece a liberação de metabólitos secundários graças a danificação das paredes celulares que levam ao aumento da permeabilidade da membrana celular além de aumentar a transferência de massa, da solubilidade e da difusão do soluto e solvente[27].A literatura traz informações sobre altos teores de ácido protocatéquico, caempferida, ácido vanílico, ácido clorogênico, ácido seringico e rutina, considerados excelentes agentes antioxidantes além de apresentar atividade hipocolesterolémica[24]. Assim, as suas propriedades como os compostos fenólicos e seus derivados são tem alto potencial e já são amplamente usados nas áreas farmacêutica, nutricional e em aplicações veterinárias.Os resultados de atividade antioxidante dos extratos hexanóico e etanoico obtidos nas diferentes temperaturas de extração (4, 25 e 60 °C) estão apresentados na tabela 5.Tabela 5. Atividade antioxidante dos extratos hexanólico e etanoico obtidos da farinha de semente de abacate em diferentes temperaturas, expressa em valores de IC₅₀. ...
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Introdução: as indústrias de processamento de frutas geram grande quantidade de biomassa residual que poderia ser reaproveitada. Considerando o elevado volume de resíduos produzidos pelo descarte das sementes do abacate e destacando o alto teor de compostos bioativos, é um produto interessante para ser avaliado. Objetivo: avaliar a composição centesimal da farinha das sementes de abacate e as propriedades antioxidantes e antimicrobianos dos seus extratos. Métodos: os extratos foram obtidos por maceração da farinha do caroço do abacate desidratada em diferentes temperaturas (4,25 e 60 °C) utilizando n-hexano e etanol como solventes. Resultados: verificou-se que a farinha é uma excelente fonte de carboidratos, com alto teor de fibras, proteínas e minerais (N, K, Mg e Ca, entre outros). A temperatura da extração influenciou tanto no rendimento como no conteúdo de fenóis totais, atividades antioxidantes e antimicrobianas dos extratos. Os extratos etanoicos obtidos à 60 °C apresentaram maior rendimento (18%) e teor de compostos fenólicos totais (~840 mgEAG/g). Também os extratos etanoicos apresentaram maior atividade antioxidante (IC50= 0,013 mg/mL e 0,018 mg/mL) em temperaturas mais baixas de extração, 4 °C e 25 °C, respectivamente. Já extrato hexanóico obtido à 4 °C apresentou maior atividade antimicrobiana para as quatro bactérias testadas (L. monocytogenes, S. aureus, S. choleraesuis e E. coli). Conclusão: a farinha obtida das sementes de abacate, apresentam alto valor biológico e podem ser usadas como suplementos na alimentação humana.
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Background: Avocado seed (Persea americana Mill.) which is usually thrown away after avocado fruit consumption, contains active compounds that can lower blood triglyceride level. Utilization of avocado seed as a medication to lower triglyceride has potentials that still needs to be discovered before it can be used as an alternative herbal medicine. Objectives: The purpose of this research is to find out the effect of avocado seed’s particle size variation during extraction on the extract effectiveness in lowering blood triglyceride level. Methods: This is an experimental research which is done on Sprague Dawley rats. The rats are propylthiouracil (PTU) for 14 days to reach hyperlipidemic state. The hyperlipidemic rats are divided into 2 groups of intervention. The first group receives avocado seed extract which was extracted from avocado seeds that had been milled for 15 minutes. The second group receives avocado seed extract which was extracted from avocado seeds that had been milled for 30 minutes. Particle size is determined by the average diameter of 10 random avocado seed particle in each group. The avocado seed particle is extracted using remaceration method. Avocado seed extract is given for 14 days with a daily dose of 250 mg/kgBW. Triglyceride level in the rats’ blood is measured after PTU has been given for 14 days and is measured again after 7 days, and 14 days of avocodo seed extract intervention. Unpaired T-test and Paired T-test are used to analyze the data. Results: The average particle size of avocado seed in the second group avocado seed’s extraction the second group is 13.50 µm. Avocado seed extract lowers blood triglyceride level on hyperlipidemic rats from day 0-7, day 0-14 and day 7-14 in the first group (p<0.05). The findings in the second group are similar (p<0.05), except for day 7-14 (p>0.05). There is no significant difference in the decrease of blood triglyceride level between the first group and the second group (p>0.05). Conclusions: Avocado seed extract can lower the level of blood triglyceride in rats. The variation of avocado seed’s particle size on its extraction has no effect on the decrease of blood triglyceride.
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Watermelon (Citrullus lanatus) and Avocado (Persea americana) seeds are often considered as byproducts or waste in the food industry. This study investigated the fatty acid profile of oil derived from the seeds of watermelon (Citrullus lanatus) and avocado. Soxhlet extraction protocol was used to extract the oils, and their fatty acid compositions were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS). The main fatty acids identified and quantified include saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs). The investigation revealed that the oil derived from Watermelon seeds (Citrullus lanatus) contained more unsaturated fatty acids than the oil from Avocado seeds (Persea americana). However, oil extracted from avocado seeds (Persea americana) has a larger percentage of monounsaturated fatty acids than oil extracted from watermelon seeds (Citrullus lanatus). As a result of its high concentration of essential oils, the oil derived from these two seed sources has the potential to be used in the food and cosmetics industries. The study's findings also shed light on the nutritional significance of these oils, particularly critical fatty acids like omega-3 and omega-6. In conclusion, the study on the fatty acid profile of oils extracted from watermelon and avocado seeds holds promise for uncovering valuable information with implications for nutrition, health, agriculture, and industry.
Chapter
Nutraceutical potential of avocado waste
Article
A comprehensive investigation of extraction methods on the efficiency of the total polyphenol contents (TPC) and antioxidant activities for both avocado seed groups with and without removing antinutrients (signed as groups A and B) and assessing acute toxicity on the extraction products. The Soxhlet extraction showed the most effective method for producing an avocado seed extract possessing the highest TPC and strong antioxidant activities, followed by the maceration, hot extraction, and percolation extraction. Interestingly, it was found that antinutrients containing in the seed can reduce TPC. For instances, sample 4B (Soxhlet method) exhibited the largest TPC (189.67±3.58 mgGAE/g DW) and DPPH inhibitory activity (IC 50 = 19.24±1.23 μg/mL), while those of the sample 2A (percolation method) were 68.95±3.79 mgGAE/g DW and 67.48±0.98 μg/mL, respectively. Additionally, sample 4B did not exhibit acute oral toxicity (D max = 21.05 g/kg mice weight), indicating a safe and high antioxidant activity of avocado seed extract for clinical applications. A high correlation analyzed by Pearson's correlation was found for the TPC to DPPH assay and ferric reducing/ antioxidant assay, showing their correlation parameters of ‐0.898 and 0.847 (p < 0.05), respectively.
Article
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The effects of aqueous seed extract of Persea americana Mill. (avocado), var. Fuerte, on blood pressures, plasma and tissue lipids of albino rats were investigated. Twenty-five rats were divided into 5 groups of 5 rats each. Group 1 (normal), group 2 (hypertensive), group 3 (hypertensive + 200 mg/kg b .wt of extract), group 4 (hypertensive + 500 mg/kg b. wt of extract) and group 5 (hypertensive + 700 mg/kg b. wt of extract). Except for group 1, which received 100% growers mash, all other groups received 92% growers mash and 8% NaCl as their daily meal for 4 weeks. The different dose of P. americana aqueous extract, significantly (P<0.05) reduced blood pressures of the hypertensive rats. Reduction in total cholesterol, LDL and triacylglycerol levels were observed at the 500 mg/kg b. wt of seed extract in the plasma, kidney, liver and heart. These results suggest that the use of aqueous seed extract of this plant in the treatment of hypertension may produce a favourable lipid profile at the 500 mg/kg dose level.
Article
A method for the screening of antioxidant activity is reported as a decolorization assay applicable to both lipophilic and hydrophilic antioxidants, including flavonoids, hydroxycinnamates, carotenoids, and plasma antioxidants. The pre-formed radical monocation of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS*+) is generated by oxidation of ABTS with potassium persulfate and is reduced in the presence of such hydrogen-donating antioxidants. The influences of both the concentration of antioxidant and duration of reaction on the inhibition of the radical cation absorption are taken into account when determining the antioxidant activity. This assay clearly improves the original TEAC assay (the ferryl myoglobin/ABTS assay) for the determination of antioxidant activity in a number of ways. First, the chemistry involves the direct generation of the ABTS radical monocation with no involvement of an intermediary radical. Second, it is a decolorization assay; thus the radical cation is pre-formed prior to addition of antioxidant test systems, rather than the generation of the radical taking place continually in the presence of the antioxidant. Hence the results obtained with the improved system may not always be directly comparable with those obtained using the original TEAC assay. Third, it is applicable to both aqueous and lipophilic systems.
Article
This study was carried out to determine the acute and subacute toxicities of ether extract of Persea americana seed in rats. Acute toxicity study was conducted by intraperitoneal (i.p.) administration of graded doses of seed extract of the plant. The i.p. LD50 was found to be 751.6 ± 98.6 (S.E) mg/kg body weight. For subacute toxicity study , a daily i.p. treatments by P. americana seed extract for 14 days with doses of 75 and 150 mg/kg b.w. caused a significant (P < 0.05 ) decrease in food consumption , body weight, blood glucose, hemoglobin and hepatic cholesterol. Whereas, serum creatinine, uric acid, total protein and total bilirubin were not significantly altered. Histopathological examinations of the liver and kidney at the end of the study (14 days) showed normal architecture suggesting no morphological disturbances induced by the seed extract in these organs. In conclusion, acute toxicity study showed a relatively low LD50 for P. americana seed extract. A daily treatment of rats with seed extract for 14 days decreased food consumption, body weight, blood glucose, hemoglobin and hepatic cholesterol levels. Serum creatinine, uric acid, total protein and total bilirubin were not significantly altered.
Article
A method for the screening of antioxidant activity is reported as a decolorization assay applicable to both lipophilic and hydrophilic antioxidants, including flavonoids, hydroxycinnamates, carotenoids, and plasma antioxidants. The pre-formed radical monocation of 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) is generated by oxidation of ABTS with potassium persulfate and is reduced in the presence of such hydrogen-donating antioxidants. The influences of both the concentration of antioxidant and duration of reaction on the inhibition of the radical cation absorption are taken into account when determining the antioxidant activity. This assay clearly improves the original TEAC assay (the ferryl myoglobin/ABTS assay) for the determination of antioxidant activity in a number of ways. First, the chemistry involves the direct generation of the ABTS radical monocation with no involvement of an intermediary radical. Second, it is a decolorization assay; thus the radical cation is pre-formed prior to addition of antioxidant test systems, rather than the generation of the radical taking place continually in the presence of the antioxidant. Hence the results obtained with the improved system may not always be directly comparable with those obtained using the original TEAC assay. Third, it is applicable to both aqueous and lipophilic systems.
Article
In this study, the antioxidant activities of Camelina sativa methanolic extracts were evaluated by different chemical assays: reducing power, 2,2-diphenyl-1-picrylhydrazyl (DPPH·) assay, the β-carotene bleaching method and the metal chelating activity assay. An LC–MS profiling method was used for a comprehensive study of the phenolic compounds and their representation in camelina seeds, cake and oil. For this purpose, 4-vinyl derivatives of hydroxycinnamic acids were synthesized by thermal decarboxylation of the corresponding phenolic acids and sinapine was isolated from kale (Brassica oleracea) applying a new method and confirmed by NMR. The results revealed that besides the total phenolic content and antioxidant activity, seeds and cake also possess a similar phenolic profile. In addition to sinapine and 4-vinyl derivatives, other antioxidants were successfully identified: ellagic acid, protocatechuic acid, p-hydroxybenzoic acid, sinapic acid, salicylic acid, catechin, rutin, quercetin and quercetin glucoside. Since after oil pressing most of the phenolic compounds remain in the seed residues, only a few compounds were identified in the oil. Camelina cake was found to have the best reducing power and radical scavenging activity, whereas camelina oil, with a relatively low phenolic content, exhibited the highest iron-chelating capacity and the best inhibitory action against β-carotene discolouration in an emulsified system.
Article
The total antioxidant capacity and phenolic content of edible portions and seeds of avocado, jackfruit, longan, mango and tamarind were studied. In addition, the relationship between antioxidant activity, phenolic content and the different degrees of heating of mango seed kernel was investigated. The seeds showed a much higher antioxidant activity and phenolic content than the edible portions. The contribution of all the fruit seed fractions to the total antioxidant activity and phenolic content was always >70%. ABTS cation radical-scavenging and FRAP assays were employed for the determination of antioxidant activity; FCR assay was used to measure the total phenolic content. The AEAC and FRAP of ethanolic extracts of MSKP products increased to a maximum after heating to 160 °C. The total phenolic content in extracts of MSKP products increased from 50.3 to 160 mg/g GAE with an increase in heating temperature to 160 °C.
Article
In this study, our objective was to investigate the possible hypolipemic effect of Persea americana (PA) on hypercholestrolemic rats. Acute administration of cholesterol resulted in the elevation of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDLC), very low density lipoprotein cholesterol (VLDLC) and reduction in high density lipoprotein cholesterol (HDLC). However, treatment with various doses of the methanolic extract of the seeds of Persea americana caused a significant reduction in the levels of TC, TG, LDLC and VLDLC while the levels of HDLC increased significantly. These effects were dose dependent as marked changes were observed at the highest concentration (300mg/kg) of the methanolic extract of Persea americana seeds. It was concluded that Persea americana seeds showed an hypolipemic effect and may serve as possible alternative treatment for hyperlipemia and hypertension.