Physicochemical characterization of propolis collected by Apis mellifera L
(Hymenoptera: Apidae) in Pacho and Bogotá Cundinamarca.
Caracterización fisicoquímica de propóleo colectado por Apis mellífera L (Hymenoptera:
Apidae) en Pacho y Bogotá Cundinamarca
Zharick Tatiana Cortes Zapata
Engineer in Agroecology. Member of the research group
MAEC. E-mail: zcortezapa@uniminuto.edu.co, ORCID:
https://orcid.org/0000-0001-9161-5526
Daniel Augusto Acosta-Leal
D. in Education. Professor at Corporación Universitaria Minuto
de Dios - UNIMINUTO. E-mail:
daniel.acosta@uniminuto.edu, ORCID: https://orcid.org/0000-
0002-6135-7439
Adriana Mireya Zamudio Sanchez
Assistant Professor Jorge Tadeo Lozano University. E-mail:
adriana.zamudio@utadeo.edu.co . ORCID:
https://orcid.org/0000-0002-6449-5095
Abstract
The objective of this work was to characterize and implement a reliable, viable and effective
physicochemical methodology for the quality of Apis mellifera propolis collected in two apiaries
located in Pacho and Suba-Bogotá, Cundinamarca, Colombia. A qualitative analysis was designed and
validated to diagnose and evaluate the quality of propolis for laboratory analysis. Samples were
collected by the scraping method, ethanolic extracts of propolis (EEP) were obtained and results were
compared under several Latin American standards (Cuban, Brazilian and Salvadoran standards). The
presence of phenols and flavonoids was determined by means of solubility in lead acetate, ferric
chloride, sodium hydroxide, pW, humidity, oxidizing property, UV-vis spectroscopy profiles, pH,
mechanical masses, dry mass, percentage of phenolic compounds, ashes, determination of major and
minor minerals and quantity of flavonoids in quercetin. The results showed that the solubility in lead
acetate and sodium hydroxide were positive for the 10 samples in duplicate, plus the 3 validation
samples; the results showed significant differences between locations and compared with current
standards.
Key words: Antioxidants, Phenols, Flavonoids, bee products.
zcortezapa@uniminuto.edu.co
http://centrosuragraria.com/index.php/revista, Published by: Edwards Deming Institute,
Quito - Ecuador, October - December vol. 1. Num. 15. 2022, This work is licensed under
a Creative Commons License, Attribution-NonCommercial-ShareAlike 4.0 International.
https://creativecommons.org/licenses/by-nc-sa/4.0/deed.es
Received February 17, 2022
Approved: May 11, 2022
124
Resumen
El objetivo de este trabajo fue caracterizar e implementar una metodología fisicoquímica confiable,
viable y eficaz para la calidad del propóleo de Apis mellifera recolectado en dos apiarios ubicados en
Pacho y Suba-Bogotá Cundinamarca Colombia. Se diseñó y validó el análisis cualitativo para
diagnosticar y evaluar la calidad de propóleos para analizarlo en el laboratorio. Las muestras fueron
colectadas por el método de raspado, se obtuvieron extractos etanólicos de propóleos (EEP) y
resultados que fueron comparados bajo varias normas a nivel latinoamericana (Normas cubana,
brasilera y salvadoreña). Se determinó la presencia de fenoles y flavonoides por medio de solubilidad
en acetato de plomo, cloruro férrico, hidróxido de sodio, pW, humedad, propiedad oxidante, perfiles
de espectroscopio UV-vis, pH, masas mecánicas, masa seca, porcentaje de compuestos fenólicos,
cenizas, determinación de minerales mayores y menores y cantidad de flavonoides en quercetina. Los
resultados evidenciaron que la solubilidad en acetato de plomo e hidróxido de sodio fueron positivos
para las 10 muestras por duplicado, más las 3 muestras de validación; los resultados presentaron
diferencias significativas entre las localidades y comparados con las normas vigentes.
Palabras Clave: Antioxidantes, Fenoles, Flavonoides, productos apícolas.
Introduction
The name propolis comes from the Greek propolis Pro "front or defense" from and polis
"city" in this case the beehive, this product is highly appreciated for its anti-
inflammatory, antitoxic, anesthetic, antioxidant, bactericidal, antiseptic and healing
properties, in addition to its wide use in human medicine, veterinary, agriculture, joinery
and food concentration (Rodríguez Martín, 2014).
The production of propolis in Colombia and in the world is scarce because the volumes
produced are small and the beekeepers are unaware of the benefits of propolis and its
characteristics, but because of the profit they can receive from its production, it could
be a product of commercial interest. However, there is no standardized technology that
allows it to be collected efficiently in order to strengthen the increase in production and
consolidate the subsector in the face of future demand for the product. (Patiño, 2001).
The main producers of royal jelly, pollen and propolis in the world are Japan, Poland,
France, Netherlands, Germany and the United States; The world's largest importer of
propolis is Japan with 7,100 tons and the largest consumer of propolis in the planet is
Germany with 4,600 tons and the main exporters are Brazil, Argentina, China, Mexico
(Rodríguez & Nuñez, 2014).
Flavonoids are the main characteristic of propolis for its antioxidant properties for
disease prevention, in the early twentieth century medicine officially recognized
Cortes, Acosta-Leal, Zamudio 2022
October - December vol. 1. Num. 15, 2022
propolis with therapeutic properties, English surgeons proved the healing action of
propolis and decreasing mortality from gangrene. (Morales, 2012).
In Colombia the use and exploitation of propolis has had little disclosure, this hinders
the study to implement a characterization of propolis and avoids establishing quality
measures and a classification of raw propolis, to have initiative it is necessary to have
quality protocols required to characterize it and thus obtain an added value that allows
beekeepers to be encouraged and have an alternative productive activity for an
apiculture in crisis. (Gil, Viloria, Durango, & Garcia, 2012)..
The composition of propolis depends on the botanical origin, bees collect raw material
from plant resins, buds, branches, young fruits, bark and mechanical impurities that they
collect from various plants and certain trees in the collection area. From the resinous
substances they collect together with secretions from their glands they obtain propolis,
their scent can be from tree species, honey and waxes. (Núñez R. et al., 2018)..
Bees disinfect the inside of the hive with propolis, close cracks and reduce access routes,
cover the corpses of enemies that try to enter the hive to prevent their decomposition.
This property of propolis was already known to the Egyptians and the priests who used
it to mummify the dead. (Carrillo, Castillo, & Mauricio, 2011).
Propolis in agriculture is used to treat diseases such as fungal diseases in fruit trees,
vegetables and viticulture; propolis acts as a systemic treatment, i.e. it does not act only
in contact areas but penetrates the plant sap and distributes it, it is not preventive but
curative when the fungus is present. Propolis can be combined with other preventive
treatments such as nettle slurry, Bordeaux mixture for downy mildew and powdered
sulfur for powdery mildew. (Mercedes & Modroño, 2019).
Phenolic compounds originate from the plant kingdom, as they are one of the main
secondary metabolites in plants and in the animal kingdom by ingestion. Phenols are
synthesized by plants and are genetically regulated.
Bactericidal properties were demonstrated with staphylococci, streptococci and
salmonellae, for fungal properties it was shown that they cause parasitic conditions or
mycosis and shows that Apis mellifera has a total of seven types of antibiotics that
protects bees. Its food, propolis venom and beeswax, for anesthetic properties is linked
by its volatile oil content that is granted effects superior to those of cocaine and
novocaine.(Muñoz, Linares and Narváez, 2011)..
In the agroecological-social area, the objective was to analyze the quality of propolis to
avoid losses of propolis in the production system and to have it as an efficient cycle.
Agroecology has tools available for the service of agricultural production, incorporating
bees because this improves the quality of the fruits due to the effect of pollination and
small producers benefit from a small amount of income from the use of honey and in
126
some cases pollen. However, propolis can contribute from the economic factor due to
the small amount of income as products for sale. From the social component it can be
used for self-consumption, from the environmental part it will avoid generating waste,
which is why the benefit of this raw material closes the cycle of beekeeping production
from the agroecological vision.
Materials and methods
The propolis samples were collected by the scraping method, 5 from Suba-Bogotá
Cundinamarca and 5 from Pacho Cundinamarca and one more for the validation of the
methodology.
The propolis samples were taken in Suba - Bogotá 4°44'38.4 "N 74°04'22.8 "W with an
altitude of 2560 masl, temperatures from 7°C to 19°C, an annual precipitation of
797mm, wind speed of 4.1km/h-7.9km/h, relative humidity 77- 83%, the second
location was in Pacho-Cundinamarca in the village of llano la hacienda 5°09'29.8 "N
74°07'44.2 "W at 2134 masl with temperatures from 12°C to 23°C, wind speed 3.5 km/h
to 5.7km/h, relative humidity 57%, precipitation 51mm - 205mm monthly and 1440mm
annually.
In this research the parameters of pH, pW, dry extract, oxidant property, determination
of flavonoids and phenols, mechanical masses were evaluated according to the
methodology described by Tagliacollo (2010) and from the book "Origin, nature,
physicochemical properties and therapeutic value of propolis" of (Salamanca, 2017) ash
and moisture were determined, an adaptation of the methodology was made for reading
in uv-vis spectrophotometry and the results of all parameters were compared under the
Cuban agricultural branch standard NRAG 1994, Russian (Joint Resolution SPRI No
94/2008 and SAGPA No. 357/2008), Brazilian, Mexican, Salvadorian, Salvadorian
(Joint Resolution SPRI No 94/2008 and SAGPA No. 357/2008), Brazilian, Mexican,
Salvadorian, Salvadorian and Cuban standards.Salvadoran (Lemus, Rodriguez, &
Orellana, 1995)Argentina (Kozlowski, 2002)For the quantification of flavonoids, it was
carried out under the methodology of (Palomino G, García P, Gil G, Rojano, & Durango
R, 2009) Finally, for the determination of minerals, the following elements were
determined: manganese, zinc, copper, iron, magnesium, calcium, potassium, sodium by
means of flame atomic absorption spectrophotometry, and 5 g of raw propolis ash, since
this represents the mineral content of the propolis sample.
Propolis was stored in amber jars, marked and deposited in cool, dark and dry places,
kept at 4°C for 24 hours, the following analyses were performed applying a validation:
reproducibility, accuracy or trueness, average. (Tagliacollo, 2014)
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STATISTICAL DESIGN: A completely randomized design was used in which the
following statistical measures were employed: mean, reproducibility, coefficient of
variation, accuracy or trueness.
The data obtained were analyzed using SPSS ® Statistics 25 software, where their
normal distribution was checked and the Student's t-test was applied to accept or reject
the null hypothesis for each of the parameters evaluated, and to confirm the assumption
of normal distribution, the Shapiro Wilk test was applied due to the fact that there is a
group of less than 30 samples.
According to the statistical analysis performed with SPSS ® Statistics 25R software, the
hypothesis of equality of variances for phenols, oxidative property, dry extract,
moisture, mechanical masses and pW is rejected, so that it is concluded that with a
significance level of 95% there are significant differences between the samples taken in
Pacho and the samples taken in Suba-Bogota, but for pH and ash for the results obtained
in the laboratory there is a significance level between each of the variables for each
zone.
Result
Results and comparison under the regulations A parameterization was elaborated for
the qualitative tests depending on the aspect of the result of the final solution, which
was applied for the determination of phenolic compounds with sodium hydroxide, ferric
chloride; and for flavonoids the reaction of the propolis extract with lead acetate
according to the regulations in force.
The first comparative result is the mineral content in each of the duplicate samples from
the two study zones (see Table 1).
Table 1 Mineral content in propolis
Minerals
Pacho (mg/g)
Potassium
738,424136
Sodium
16,7900821
Calcium
607,207669
128
Magnesium
125,185616
Iron
53,7146915
Copper
1,43265741
Zinc
24,3871311
Manganese
7,85367682
A table was prepared to represent the qualitative tests of major and minor minerals in
crude propolis ash (see Table 4).
Determination Of Phenolic Compounds:
Reaction With Sodium Hydroxide:
Table 2 Parameters for the determination of phenolic compounds in samples of
ethanoic extract of propolis
Solution appearance
Qualitative test result
Slightly granular
+
Granular
++
Highly granular
+++
The reaction of the propolis extract with sodium hydroxide must be positive in order for
the propolis to determine if phenolic compounds are present and thus be accepted
according to the regulations. The reaction and the basis is positive when the sodium
hydroxide breaks the C ring of a flavonoid which can be a flavonone or a flavone and
this rupture causes the formation of a chalcone, evidenced by the yellow coloration that
varies its intensity in the reaction (See Table 3).
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Table 3 Determination of phenols location 1
NaOH reaction
analysis
Sample Suba -
Bogota
Qualitative test
result
Coloring
1
10%
++
Yellow
5%
++
10%
++
Yellow
5%
++
10%
+
Yellow
5%
+
10%
+++
Yellow
5%
+++
5
10%
++
Yellow
5%
++
The following table shows the results of the sodium hydroxide analysis in Pacho (see
Table 4).
Table 4 Determination of phenols location 2
NaOH reaction
analysis
Pacho Sample
Qualitative test
result
Coloring
1
10%
+++
Brownish green
5%
++
10%
+
Brownish green
5%
+
10%
++
Green
5%
++
10%
+
Green
5%
+
5
10%
+++
Brownish green
5%
+++
Determination Of Flavonoids - Reaction With Lead Acetate
The reaction is positive when it presents a yellow precipitate or a cloudy solution to be
accepted by the standards. It is based on the formation of a colored complex between
lead and dihydroxyphenols and polyhydroxyphenols such as flavones, flavanones and
130
anthocyanins contained in propolis and it is necessary to have these compounds in order
to obtain its antimicrobial, antimicrobial and antiviral characteristics (see Table 5).
Table 5 Determination of flavonoids location 1
Sample
Suba -
Bogota
Coloring
Qualitative
result
1
Yellow
-
Yellow
-
Yellow
-
Yellow
-
5
Yellow
-
The following table shows the results for the determination of flavonoids in lead
acetate in Pacho (see Table 6).
Table 6 Determination of flavonoids location 2
Pacho
Sample
Collation
Qualitative
Result
1
Green
+
Brownish
green
+++
Green
+
Green
+
5
Green
+++
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The flavonoid content was determined by the colimetric method with lead acetate and
for the quantification of the total flavonoid content it was determined by the method of
Kumazawa et al. (Kumazawa, Hamasaka, & Nakayama, 2004). with minor
modifications, to 0.5 ml of ethanolic propolis extract solution diluted 1:1000 at 5%
ALCL3 and after 1 hour at room temperature and avoiding light, the absorbance was
measured at 430nm.
CALIBRATION CURVE: For the calibration curve standards, 2.5mg of quercetin was
used and dissolved in 25ml of 96o methanol, the total flavonoid content was calculated
as quercetin (mg/g), subsequently this stock solution was diluted to 5, 10, 15, 15, 20 and
25μg/mL as follows.
To read the samples a dilution of the ethanolic extract of propolis was made, 0.1ml of
EEP was taken and was taken to a volume of 25ml of ethanol to each sample, already
having the dilutions 2ml and 2ml of aluminum chloride 5% were taken from each one,
they were shaken with a bortex, the samples were left in a place with room temperature
and dark for 1 hour and we proceeded to read each one of the samples in triplicate on
the calibration curve. Finally, an average of the weight of 0.1ml of EEP was taken from
each region.
Flavonoid Quantification
The quantification of flavonoids were few compared to other international propolis, but
it was higher than the norm, the highest amount of flavonoids was obtained in Pacho
with 6.52 mg/g and the highest amount in Bogota was 4.3 mg/g, being very significant
the difference.
Ferric Chloride Reaction
This reaction of propolis extract with ferric chloride is used to identify the presence of
phenols in the propolis extract sample, colored compounds are formed as the hydroxyl
groups act to activate the benzene ring by donating density, the aromaticity of the ring
is weakened and it acquires more the characteristics of a conjugated polyene, the iron
(III) ion is a moderate oxidant that induces the conversion of a pyrogallol or a
hydroxylated orthoquinone, with higher concentration of pyrogallol it promotes the
formation of a polymer of the quinone formed and a very long pi system is produced so
that absorption is obtained in the whole spectrum. (Penadillos & Antonio, 2013)
Dry Extract
For the propolis samples from the municipality of Pacho, it was found that most of the
samples were below the range established by the Latin American legislative norm,
giving a range of 3.026% - 12.496%, therefore 40% (4 samples) of the 10 samples meet
the Latin American norms, these being from Suba-Bogotá. It is defined that the range
132
of the results is high and may be related to the botanical origin and the area where the
bees collected the resin.
Oxidizing Property
The samples analyzed were below the standards, which suggest a maximum reaction
time of 22 seconds to have an optimal oxidation to identify a good quality of propolis
due to its high oxidation rate, the time range for the 10 samples was 9.96 - 21.01 seconds.
These results can be affected by the relative humidity that favors the natural oxidation
of the product during its production, a high percentage of humidity is higher than the 22
seconds established by the regulations is due to a bad storage of the product.
(Tagliacollo, 2014).
Solubility Of Eep Particles In Lead Acetate And Sodium Hydroxide
For both analyses, the objective is to verify the homogenization of the extract particles
in a saline solution to identify phenols and flavonoids. All the samples from the studied
areas were positive.
pH
The range of the sample results is between 3.69 - 4.8 which indicates that the samples
tend to be acidic. The standards do not establish a value, but the literature on pH in
alcoholic media reports that the pH value is close to 5.0.
Mechanical Masses
The mechanical masses are wood shavings, wax, plant remains, the regulations show
that they constitute 40 - 50% of the total mass, the rest is mass that is biologically active.
In the samples analyzed a range of 59.79 - 26.68% was obtained, the quality of propolis
is inversely proportional to the content of mechanical masses since if the propolis is
harvested in traps it is purer in terms of propolis content since if it is harvested with the
scraping system more mechanical masses are obtained.
Ash In Dry Mass
In the locality of Suba-Bogotá, high ash indices were obtained with 1.5% and in Pacho
with 0.8%. The propolis from the city of Bogotá complies with Brazilian standards, with
100% of the 5 samples collected in Suba-Bogotá complying with the standards and 10%
complying with the standards for the 5 samples collected in Pacho.
Humidity
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October - December vol. 1. Num. 15, 2022
The results obtained do not exceed the percentage of Brazilian standards. A range of
3.6% to 4.9% was obtained for Pacho and Bogotá with respect to the regulations, which
indicate that the maximum result can be 8%, and the dry base (pW) did not exceed 8%
because the two locations had a range of 3.7% to 5.2% for moisture on a dry base.
Uv-vis spectrophotometry
A sweep of each of the samples diluted 1:1000 was made to read them in the
spectrophotometer, a base line with deionized water was needed for the spectrum to read
it based on that line and compare it with the propolis samples and proceed to read the
10 samples by washing and purging the cuvette to avoid contaminating the sample with
the other samples. Next, one of the sweeps that the spectrophotometer marked is
observed, being 230 the maximum absorbance peak in Pacho.
According to the methodology reported by (Palomino G et al., 2009) adjustments were
made to the calibration curve adapted from the methodology of (Lemus et al., 1995).
Determination of minerals by atomic absorption spectrophotometry
For the determination of minerals there are regulations and methodology that establishes
some percentage of certain minerals in propolis, the results obtained between 0.5 - 728.4
mg/g of propolis were made in duplicate and an average was made for each of the
samples, Pacho has the highest amount of minerals than in Bogota, the highest amount
that has the crude propolis is in potassium and the lowest amount of minerals was
obtained in copper as follows:
Potassium<Calcium<Magnesium<Iron<Zinc<Sodium<Manganese<Copper.
In the extraction process of propolis by centrifugation 60% of the samples presented a
milky and opaque turbidity because the water contained in the crude propolis causes
insolubility of waxes and less hydrophilic components (Grosso, 2017).
Regarding the mechanical masses the result may vary due to the collection method, for
the determination of ashes the results of Suba-Bogotá were very low with respect to the
legislation established for it to be a quality propolis, but in comparison at national level
the articles reviewed (Bastos, Guzman, Figueroa, Tello, & Scoaris, 2011) in Caldas it
was found that the percentage of ashes with the scraping methodology is 0.22 - 0.78 on
the other hand for the studied zones (Pacho and Suba-Bogotá) with the same
methodology it is 0.8% for Suba-Bogotá and 1.5% for Pacho. (Julián Martínez, Carlos
Garcia, Diego Durango, & Jesús Gil, 2012). According to the (Salamanca Grosso, 2017)
the amount of ash in the samples is the amount of total fixed solids that represents the
mineral content of the propolis, since the muffle incinerates the organic matter of the
propolis, i.e. the more ash content the sample has the better because it fixes a nutritional
rate in the product.
134
Regarding moisture and pW, it was determined in the samples and considers the water
loss of the sample after having removed moisture in the field and pW is the percentage
of moisture that the propolis sample can retain at room temperature without alteration
from an external factor. In this case, the samples from Suba-Bogotá contain more
moisture than those from Pacho due to the environmental conditions in Pacho, which
also affects other propolis characteristics such as the oxidizing property. The levels of
oxidizing activity for the alcoholic extracts of propolis are 22 seconds, Pacho has a less
oxidizing activity than Suba-Bogota since Pacho had 12.37 seconds as opposed to Suba-
Bogota that obtained 18.93 seconds, these results favor the product during its
production, if it has any alteration to the maximum limits established by the regulations
it is due to a bad storage of the product, this would also be an indication of possible
fermentations generating unwanted products or contaminants being toxic and harmful
to health.(Delgado Aceves, Andrade Ortega, & Ramírez Barragán, 2018).
The results obtained in Pacho contain a higher level of acidity than Suba-Bogotá.
The samples analyzed with respect to the dry extract varied from 5.8% to 9.8%, 11%
being the value established by Brazilian regulations, therefore 40% of the samples of 10
samples analyzed comply with the current Brazilian legislation, these samples from the
town of Pacho being more in accordance with the regulations.
The content of phenols present in the extract varied in 2.6 for Bogota and 6.14 for Pacho
being 0.5% the ideal content of phenols according to Brazilian regulations therefore all
samples exceed and are consistent with the regulations in terms of quality, the high
content of phenols that has the propolis increases the value of apitherapy in the
international market for its antimicrobial effect, as well as other substances present in
the propolis (Tagliacollo, 2014)The phenolic compounds are recognized for their high
biological, anticarcinogenic, anti-inflammatory, analgesic and oxidant activity. (Bastos
et al., 2011).
The components of propolis are different depending on the flora and climatic conditions,
according to the results obtained, the less disturbed natural environment has a higher
concentration of phenols as in Pacho Cundinamarca improves the quality of propolis,
from the statistical analysis it is concluded that apart from the fact that there are
significant differences there is the possibility of having a higher quality propolis, The
hypothesis under the established parameters the propolis of the two zones are of the
same quality and statistically if there are differences between the two and the social
benefit for small producers at an economic level and own consumption for their health,
bronchodilator benefits of propolis by high contents of phenols, aromatic acids, esters,
aldehydes, camarinas and triglycerides.
Conclusions
Cortes, Acosta-Leal, Zamudio 2022
October - December vol. 1. Num. 15, 2022
The difference in color in the reactions for the determination of phenols and flavonoids
is due to the difference in flora and climatic conditions in which each of the samples
was collected. The most important characteristic that gives the difference in quality of
propolis is the amount of phenolic compounds, because the one from Suba-Bogota is of
better quality due to its content of phenolic compounds.
Propolis does not have a chemical formula, it is a resin made up of several elements
from different plants collected by Apis mellifera bees. Propolis contains 2.6% - 6.14%
phenols in the areas of Pacho and Suba - Bogota respectively, favoring human health
due to its pharmaceutical properties and its biological potential, mainly its antioxidant
activity.
Environmental benefit, the use of tree resins by bees and continuous visits to the flora,
promote forest conservation and protection of natural resources, use for their hives for
their antimicrobial properties.
Most of the ethanolic extracts analyzed meet the regulations of the Brazilian legislation,
the ash content obtained in the propolis of Suba-Bogotá make them promising as
nutritional additives for the human diet, the ash content also indicates the mineral
content since when calcining a sample evaporates much of the amount of organic matter
and remains only the minerals, highlighting that minerals are very important for
biological activity and cellular metabolism, for the human being can prevent
arteriosclerosis and the increase of the immune capacity, the mineral content is used to
characterize geographical areas where propolis is produced since it can be an indicator
of environmental pollution of the environment where Apis mellifera collects its raw
material to produce its propolis, the presence of iron, chromium, nickel, copper and lead
are considered to be stable complexes that form the same flavonoids. (Betances, 2018).
The propolis environments were different, the environment of Pacho was rural, a
eucalyptus forest and the environment of Bogota was more urban, full of bushes, flowers
and various trees, according to the literature, eucalyptus trees contain more phenols and
flavonoids, therefore it may affect the content of flavonoids and phenols is higher in
Pacho, also depends on the climate affects the oxidative property which is an index of
the amount of phenols and flavonoids in propolis.
Acknowledgments
Thanks to the research direction of the Bio-systems Center of the Jorge Tadeo Lozano
University and the research direction of the MAEC research group of Corporación
Universitaria Minuto de Dios for carrying out this research.
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