Revista Brasileira de Farmacognosia (2023) 33:288–299

https://doi.org/10.1007/s43450-023-00374-x

REVIEW

Brazilian Brown Propolis: an Overview About Its Chemical

Composition, Botanical Sources, Quality Control, and Pharmacological
Properties
Victor Pena Ribeiro1 · Jennyfer Andrea Aldana Mejia2 · Debora Munhoz Rodrigues2 ·
Gabriel Rocha Alves2 · Ana Maria de Freitas Pinheiro2 · Matheus Hikaru Tanimoto2 · Jairo Kenupp Bastos2 ·
Sérgio Ricardo Ambrósio1

Received: 22 November 2022 / Accepted: 7 February 2023 / Published online: 24 February 2023
© The Author(s) under exclusive licence to Sociedade Brasileira de Farmacognosia 2023

Abstract
Brazil is one of the largest propolis producers in the world. Propolis is produced by bees from plant exudates and tissues,
leading to many variations in the types of propolis. Generally, Brazilian propolis types are green, brown, and red. Despite
not being the main research focus as the green and red propolis, brown propolis is the second most produced propolis type in
Brazil and has tremendous economic and medicinal importance. Propolis has drawn attention with the rise in the search for
healthier lifestyles, functional foods, biocosmetics, and natural products as therapeutic sources. This review covers the main
chemical constituents identified in different types of Brazilian brown propolis, and their botanical sources, chemistry, and
biological activities. The economic aspect of brown propolis is also presented. There are many gaps to be filled for brown
propolis regarding the development of analytical methods, and quality control to allow its standardization, limiting its appli-
cability in the food and pharmaceutical industries. Future perspectives regarding brown propolis research were discussed,
especially biological activities, to support the medicinal uses of different types of brown propolis.

Keywords Propolis · Chemical composition · Botanical source · Biological activity

Introduction with other medicinal plants, playing an important role in main-

taining community health for disease prevention and treatment
Propolis is a resinous and complex product produced by hon- (Rivera-Yañez et al. 2020). These medicinal properties reported
eybees (Apis mellifera) through the collection of different for Brazilian brown propolis have become the target of intense
parts of plants and exudates mixed with bees’ wax and sali- research mainly due to their activities, such as, antioxidant (Sar-
vary secretions in the hive (Salas et al. 2016). This mixture tori et al. 2012; Calegari et al. 2017), antibacterial (Fernandes
forms a robust adhesive material used for structural repairs to et al. 2015; Pimenta et al. 2015), anti-inflammatory (Sartori et al.
maintain sealing, humidity, and internal hive temperature. It 2012; Ribeiro et al. 2018), cytotoxic (Lima et al. 2019), anti-
is also important as a defense against invading microorgan- leishmanial (Santana et al. 2014), antigenotoxicity (Fernandes
isms that may threaten the community, essential for the hive’s et al. 2014, 2019), and antimycoplasma (Araújo et al. 2020).
survival (Simone-Finstrom et al. 2017; Bankova et al. 2018). The chemical composition of propolis is directly linked
Propolis has been used since ancient times in different civi- to the botanical sources surrounding beehives in each region
lizations in traditional medicine, either alone or in combination (Simões-Ambrosio et al. 2010). Considering Brazil’s conti-
nental dimension and its conspicuous plant diversity, several
* Jairo Kenupp Bastos Brazilian types of propolis have been classified, including
[email protected] color, botanical source, and phytochemical profile. Green,
1
red, and brown propolis are the main types (Park et al. 2002).
Núcleo de Pesquisa Em Ciências Exatas E Tecnológicas,
The green propolis has one primary botanical source,
Universidade de Franca, Franca, SP 14404‑600, Brazil
2
Baccharis dracunculifolia DC., Asteraceae (Arruda et al.
Faculdade de Ciências Farmacêuticas de Ribeirão Preto,
2020a, b). Red propolis has two botanical sources: Dalbergia
Universidade de São Paulo, Av. Do Café, Ribeirão Preto,
SP 14040‑930, Brazil ecastaphyllum (L.) Taub., Fabaceae, rich in isoflavonoids,

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pterocarpans, and chalcones; and Symphonia globulifera these two propolis types (Fonseca et al. 2011). According
L.f., Clusiaceae, rich in polyprenylated benzophenone to Rodrigues et al. (2016), besides those traditional green
(Ccana-Ccapatinta et al. 2020). propolis chemical markers, BP can also present lower con-
The different types of Brazilian brown propolis (BBP) bear centrations of propenoic and cinnamic acid. Other phenolic
different chemical profiles, leading to different chemical com- compounds like chrysin, pinocembrin, galangin, caffeic acid
positions and, consequently, different biological activities. This phenylethyl ester, and pinobanksin-3-O-acetate have also
represents a great challenge in the case of BBP, since it is pro- been detected by HPLC–UV-ESI–MS analysis in Brazilian
duced in different regions of Brazil, from the northeast to the propolis samples (Fabio et al. 2019).
south of the country, thus having several biomes and vast plant An RP-HPLC–DAD–ESI–MS/MS was used to chemically
diversity as possible botanical sources. (Tazawa et al. 2016). characterize a BP sample from Paraná (Araújo et al. 2020).
In this review, we discuss botanical sources, the impor- The raw material was submitted to extraction and partition
tance of analytical methods development, quality control, with different solvents in a Soxhlet system, revealing the preva-
pharmacological properties, and the economic aspects. lence of some classes of compounds in the fractions. In all
fractions, artepillin C, baccharin, and 3-hydroxy-2,2-dimethyl-
8-prenyl chromane-6-propenoic acid were identified, although
Search Strategies the hexane fraction was enriched with propenoic and cinnamic
acid derivatives like drupanin; the hexane:ethyl acetate frac-
A search in the literature was undertaken by using the fol- tion presented propenoic and cinnamic acid derivatives, and
lowing platforms: Web of Science, SciFinder, Pub-Med, flavonoids as kaempferol and quercetin. In comparison, the
Science Direct, and Google Scholar. The terms “Brown ethyl acetate fraction was rich in flavonoids, chlorogenic acids,
Propolis” and “Brazilian propolis” were used as keywords and quinic acid esterified by one or more units of cinnamic,
in search engines. The inclusion criteria of the articles were p-coumaric, caffeic, or ferulic acids. The methanol and aque-
that propolis must be classified as brown and contained at ous fractions contained quinic and caffeoyl acid derivatives.
least one of the following points: chemical composition, Furthermore, diterpenes as isocupressic acid (5), (E)-
identification of the botanical origin, and/or evaluation communic acid (6), (Z)-communic acid (7), and abietic
of some biological activity. Brown propolis articles from acid (8) have been isolated from samples of BP from Par-
regions other than Brazil were not considered in this review. aná State (Tazawa et al. 2016). The presence of the 15-ace-
No time filter was used. Articles published in Portuguese, toxyisocupressic acid and an unreported diterpene, the
Spanish, and English were considered. rel-(5S,6S,8R,9R,10S,18R,19S)-18,19-epoxy-2-oxocleroda-
3,12(E),14-triene-6,18,19-triol 18,19-diacetate 6-benzoate,
were reported on BP ethanolic extract (Santos et al. 2021).
Discussion Capillartemisin A and caffeoylquinic acid derivatives like
3,4-di-O-E-caffeoylquinic acid, the 3,5-di-O-E-caffeoylquinic
Chemical Composition acid, O-hexosyl-caffeoyl dihydrocaffeate, 4,5-di-O-E-
caffeoylquinic acid, and O-E-coumaroyl-caffeoylquinic acid
The chemical composition of propolis is highly dependent were detected in Minas Gerais (Southeast region) BP by LC-
on the conditions of the location and the chemical constit- DAD-MS analysis (Dembogurski et al. 2018). Brown propo-
uents of the botanical source (Bankova et al. 2018). One lis from Mato Grosso also furnished acetylisocupressic acid,
production center of BP is the southern region of Brazil. dihydro-p-coumaric acid (9), caffeic acid (10), and aroma-
Forty-four samples of BP from Paraná and the Santa Cata- dendrin (Fernandes et al. 2019). Analysis by UPLC-MS of
rina States were analyzed to identify the regional identity of the hydroalcoholic extract of BP from Rio Grande do Sul
these locations. Their compounds were identified and quan- (South region) revealed the presence of rutin, chlorogenic
tified by comparing their chemical profile with authentic acid, ferulic acid, and caffeic acid (Waller et al. 2017).
standards (Machado et al. 2021). According to this study, Another area of production of BP is the northeast region.
the BP from Southern Brazil is chemically characterized A sample from Bahia State was submitted to extraction with
by caffeoyl-quinic acids (range, 11.14 − 21.45 mg/g), hexane, methanol, and dichloromethane, and the chemical
p-coumaric acid derivatives (6.27 − 12.17 mg/g), flavonols profile was assessed by CG-MS (Santos et al. 2017). The
(9.35 − 23.55 mg/g), followed by benzoic acid derivatives hexane fraction was composed of pentadecane, hexadecane,
(3.18 − 7.45 mg/g), and dihydroflavonols (0.17 − 4.25 mg/g). heptadecane, and tricosane. Methyl cinnamate and sitosterol
Phenolic compounds already reported in Brazilian green cinnamate were isolated from the hexane fraction, and ana-
propolis have been found on standardized BP extracts, like nixanthone was isolated from the dichloromethane fraction.
p-coumaric acid (1), drupanin (2), artepillin C (3), and bac- The GC–MS chromatographic analysis of BP from Piauí
charin (4), revealing similarity on phenolic profile between (Northeast region) displayed the triterpenes lupeol, germanicol,

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β-amyrin, hop-22-(29)-en-3-one, and lupenone for hexanic The sesquiterpenes β-caryophyllene (11) and humulene
fraction, and for dichloromethane fraction, the compounds were the most abundant compounds in the BP sample from
2,3-dihydroxybenzofurane, lupeol, and dodecanoic acid. In Bahia state (Olegário et al. 2019); acetophenone, (R)-α-
ethyl acetate fraction, p-coumaric acid and 3,5-dihydroxyben- pinene, and ( +)-δ-cadinene (12) were predominant in the
zoic acid were identified (Santana et al. 2014). Paraná sample; d -limonene and nonanal were the major
Samples from Paraná (South) and Ceará (Northeast) state compounds in the Sergipe state sample (Northeast region).
were submitted to the acid/n-butanol hydrolysis method to Brown propolis from Minas Gerais sample displayed ses-
detect proanthocyanidins and their quantification by pre- quiterpenes (33.62%), oxygenated sesquiterpenes (26.98%), and
cipitation with BSA (bovine serum albumin). All the sam- oxygenated monoterpenes (18.99%) (Ribeiro et al. 2021a, b, c).
ples had a positive reaction for proanthocyanidins with low Quantification differences were observed for samples from this
tannin content values (between 0.6 and 1%) but without a state, although the sesquiterpenes β-caryophyllene and α-copaene
complete chemical characterization (Mayworm et al. 2014). (13) were predominant. 1,8-Cineol (14), terpineol-4-ol (15), nero-
The volatile oil from BP has been characterized by sev- lidol (16), spathulenol (17), δ-cadinene, aromadendrene (18),
eral studies, showing differences in the chemical profile of γ-muurolene (19), and the alkyl-phenylketone, acetophenone
propolis from different locations. Headspace solid-phase have also been reported (Lima et al. 2019; Olegário et al.
micro-extraction (HS-SPME) and GF-MS analysis helped 2019; Ribeiro et al. 2021a, b, c; Símaro, et al. 2021).
identify more than 315 volatile compounds in BP from The volatile oil of BP from Mato Grosso do Sul displayed
Bahia, Minas Gerais, Paraná, and Sergipe States (Olegário (E)-caryophyllene, δ-cadinene, spathulenol, α-copaene, (E)-
et al. 2019). Terpenes were the predominant class of com- nerolidol, and aromadendrene, with the prevalence of vir-
pounds in all samples, followed by the aldehydes. idiflorene and trans-α-bergamotene (Fernandes et al. 2015).

Botanical Sources sources, and even its chemical composition, making it dif-
ficult to standardize this medicinal product.
One of the major bottlenecks in propolis studies is the eluci- Several approaches are aiming to find the probable botan-
dation of its botanical sources. The botanical source visited ical source of propolis under study in the literature. One of
by bees is directly related to the chemical composition of the approaches is to observe bees in the field, as Apis mel-
propolis, impacting its pharmacological properties. Many lifera bees collect red exudates from Dalbergia ecastaphyl-
publications on propolis do not describe its type, botanical lum to produce red propolis, which was confirmed by the

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chemical similarity between the plant exudate and propolis

(Daugsch et al. 2008). Another approach is the identifica-
tion of chemotaxonomic markers and correlating them with
botanical species near the hives, which led Ccana-Ccapatinta
et al. (2020) to describe Symphonia globulifera as the source
of benzophenones in Brazilian red propolis. Some research-
ers use palynology for botanical identification of propolis,
but bees visit many plants for nectar collection and a few
plants for resin collection, making it challenging to identify
the botanical source for propolis production (Freitas et al.
2011). Currently, the metabolomics associated with tech-
niques such as UPLC-MS/MS has shown to be an essential
tool in propolis’s chemical prospection to identify its origin.
The botanical origins of some Brazilian propolis were well-
established, as Baccharis dracunculifolia for green propolis
Fig. 1  Collection of exudates of Araucaria angustifolia by Apis mellifera
and Dalbergia ecastaphyllum and Symphonia globulifera for bee in União da Vitória (Paraná state, Brazil)
red propolis. Several plants have been described as responsi-
ble for their composition regarding brown propolis, as Pinus
spp., B. dracunculifolia, Eucalyptus spp., and Araucaria Araucaria’s participation was confirmed later through the
angustifolia (Freitas et al. 2011; Frota et al. 2021; Ribeiro phytochemical study of this propolis (Santos et al. 2021).
et al. 2021a, b, c; Santos et al. 2021; Serafim et al. 2022). The participation of A. angustifolia in the chemical com-
Baccharis dracunculifolia, popularly known as position of propolis is mainly due to the presence of acid
“alecrim-do-campo,” is largely distributed in South Amer- diterpenes, such as 13-epi-cupressic acid, abietic acid, and
ica from Southeastern Brazil to Argentina and Uruguay communic acid (Santos et al. 2021; Tazawa et al. 2016).
(Ribeiro et al. 2018). It is the primary plant source of From volatile compounds, A. angustifolia presents the ses-
Southeastern Brazilian propolis, called green propolis, quiterpene germacrene-D and the diterpenes hibaene and
because of its color. Green propolis contains high lev- phyllocladene as significant components of its volatile oil
els of prenylated p-coumaric acids, mainly artepillin C (Brophy et al. 2000). The anti-inflammatory and antimicro-
and baccharin, and the volatile compounds nerolidol and bial biological properties are attributed to diterpenes. Many
spathulenol, all found in B. dracunculifolia (Beserra et al. diterpenes isolated from BP and A. angustifolia possessed
2021; Bernardes et al. 2022). antimicrobial activity (Bankova et al. 1999; Ribeiroet al.
Due to its geographic location, B. dracunculifolia is found 2021a, b, c).
mainly in BP samples collected in the southeastern region Diterpenes from Pinus spp. and Eucalyptus spp. are also
of Brazil, and there are other possible associated botanical found in BP. Almost all isolated diterpenes from a Brazilian
sources, making it brown. The phenolic acids of B. dracuncu- Southeast BP sample were reported in Pinus resin, as 19-ace-
lifolia and its volatile components nerolidol and spathulenol toxy-13-hydroxyabda-8(17),14-diene, totarol, 7-oxodehydro-
are described in several BP (Dembogurski et al. 2018; Araújo abietic acid, dehydroabietic acid, communic acid, and isopi-
et al. 2020; Ribeiro et al. 2022). Compounds not described maric acid. Pinoresinol and matairesinol lignans were also
for Baccharis are usually identified in phytochemical stud- isolated from the same propolis sample (Ribeiro et al. 2021a,
ies, thus evidencing the participation of other plants in the b, c). These lignans are described as major compounds in
production of brown propolis (Beserra et al. 2021). A. angustifolia knots resin and are also described as major
Diterpenic acids reported in Brazilian Southeast BP are compounds in Pinus taeda resin (Eberhardt et al. 1993).
also found in conifers species as Araucaria angustifolia. Many BBP contain α-pinene and β-pinene as major com-
Some phytochemical studies of Brazilian brown propo- pounds of their volatile fraction. These compounds are chem-
lis confirmed A. angustifolia as the primary plant source ical markers of Pinus volatile oil, corroborating this plant as a
(Sartori et al. 2021; Ribeiro et al. 2021a, b, c; Santos et al. botanical source for some BBP (Ioannou et al. 2014). Euca-
2021). In a previous work published by our research group, lyptus spp. also contributes with flavonoids and glucopyra-
we reported the isolation of diterpenes from Araucaria sp. noside compounds for BP composition (Freitas et al. 2008).
Brazilian brown propolis. During the collection of propolis
samples in the field, bees collected the exudates from the Analytical Methods and Quality Control
A. angustifolia trunk for propolis production (Santos et al.
2021). The bees collect A. angustifolia exudate, store it in Determination of total phenolic and flavonoid contents
the corbicula of the left leg, and take it to the hive (Fig. 1). in propolis samples has been widely used to determine

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biological properties (Sawaya et al. 2011), mainly using analyzer apparatus to determine the chemical composition of
spectrophotometric assays. Usually, total phenolic content ethanol extracts from different types of Brazilian propolis,
is measured by Folin–Ciocalteau’s method, and flavonoid considering their predominant botanical origin (Salomão et al.
content is measured by the ­AlCl3 complexation method 2008). Fonseca et al. (2011) quantified phenolic compounds
(Machado et al. 2021). However, these methods do not by HPLC–MS in brown and green Brazilian propolis from São
specify each compound in the class of phenolic and flavo- Paulo. Tazawa et al. (2016) discovered a novel diterpene in BP
noids, being not selective. It is crucial to identify the com- from the state of Paraná, using 1D- and 2D-NMR analyses and
pounds responsible for the biological activities to guarantee identified five more diterpenes. All the six compounds were
the quality of propolis and its products. The development quantified in the sample by ultra-performance liquid chroma-
of validated analytical methods is mandatory to guarantee tography (UPLC) using calibration curves without validation.
selectivity, accuracy, and precision in quantifying com- Rodrigues et al. (2016) used HPLC–DAD to quantify
pounds (Machado et al. 2021). prenylated phenolic acids and phenolic acids in brown and
Several analytical methods have been developed to ana- green Brazilian propolis samples from Paraná and Minas
lyze raw propolis and its commercial products. Many of Gerais, respectively. Waller et al. (2017) detected 17 com-
them aim to identify the chemical components with biologi- pounds in a BP sample from the state of Rio Grande Sul
cal activities, mainly phenolic compounds used as biomark- using HPLC–MS, in which the compounds were character-
ers/standards (Fabio et al. 2019). It is challenging to develop ized by their UV and mass spectra. They used external stand-
analytical methods for propolis analysis because it bears a ards calibration curves to quantify p-coumaric acid, rutin,
complex matrix demanding different methods’ approaches chlorogenic acid, ferulic acid, and caffeic acid.
to analyze all the compound classes present in propolis Mayworm et al. (2014) quantified tannins in different
(Pavlovic et al. 2020). For propolis’ polyphenol analysis, types of Brazilian propolis. Tannic acid was used as a ref-
thin-layer chromatography (TLC), gas chromatography erence for determining tannin content using the precipita-
(GC), high-performance liquid chromatography (HPLC), tion method with bovine serum albumin (BSA). Brown
and capillary electrophoresis (CE) are the most used meth- propolis samples contained tannins in a low quantity com-
ods. HPLC coupled with photodiode array detector (DAD) is pared with other propolis types.
beneficial for polyphenol analysis, but HPLC coupled with a
mass spectrometer (HPLC–MS) has gained space in propolis
analysis, and it allows the identification of compounds in Validated Methods
complex matrices (Fabio et al. 2019).
Besides phenolic analysis, propolis volatile com- Currently, there are few articles with details reporting
pound analysis is essential, and solid-phase microextrac- validated methods for BP analysis. Machado et al. (2016a,
tion (SPME) with GC coupled to the mass spectrometer b) identified and quantified phenolic compounds in dif-
(GC–MS) is a good choice for this class of compounds ferent types of propolis, including samples of BBP from
(Pavlovic et al. 2020). Headspace solid-phase microextrac- the southern states Santa Catarina, Rio Grande do Sul,
tion (HS-SPME) coupled with gas chromatography-mass and Paraná. The analyses were made in HPLC–DAD, and
spectrometry (HS-SPME GC–MS) allows the study of sev- the article mentions that the method was validated based
eral samples. Furthermore, HS-SPME has the advantage of on National Health Surveillance Agency (Anvisa 2017)
avoiding using solvents, except when the matrix effect inter- and National Institute of Metrology, Standardization, and
feres in analysis (Burzynski-Chang et al. 2018). Industrial Quality (Inmetro) guidelines.
There are some qualitative and quantitative methods Ribeiro et al. (2021a, b, c) validated a method to quan-
reported in the literature for BP analysis, using the above- tify volatile compounds from Brazilian southeast brown
cited techniques. The qualitative methods are used only for propolis using GC-FID. The authors identified 56 com-
chemical characterization, and there are reported quantify- pounds by GC–MS in a volatile oil extracted from BP.
ing method samples, even without validation. Most of the Eight major compounds were isolated and used in the vali-
methods are not validated and usually show incomplete dated method. The chemical structures were determined
information about compound quantification. This review did by GC–MS and NMR analyses. The method was validated
not include the qualitative and classical spectrophotometric following International Conference on Harmonization
methods for phenolic and flavonoid analysis. (ICH) and Anvisa guidelines and can be used for quality
control to quantify these compounds in different types of
Quantitative Methods propolis.
Santos et al. (2021) also validated a method for BP dit-
HPLC has been the most used equipment for quantitative erpene analysis using HPLC–DAD. The major diterpenes
method development, varying the type of detector and mass from a BBP from Paraná state were isolated and identified

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by 1H and 13C-NMR. The method was validated based on above, with inhibition values between 50 and 100% of cell
Anvisa guidelines, and all the parameters were analyzed to growth and with low cytotoxicity against normal cells,
guarantee the method’s reliability to identify and quantify showing excellent activity and selectivity against PC3 and
the six diterpenes in BP samples. Machado et al. (2021) HL60 cancer cells (Frota et al. 2021).
analyzed 44 BBP samples from Paraná by HPLC–DAD. The BP from Paraná also showed moderate activ-
Analytical standards were used to quantify the total content ity against human ovarian cancer cells (OVCAR-8) and
of caffeoylquinic acid, phenylpropanoids (p-coumaric acid colorectal carcinoma (HCT-116), with 75 and 50% of cell
derivates), benzoic acids, flavonols, and dihydroflavonols growth inhibition, respectively. It is interesting to report
in the samples. that Machado et al. (2021) tested 44 types of BP from the
Based on the literature review, developing more validated southern region with different levels of antitumor activi-
analytical methods is still necessary for BP analysis around ties, ranging from inactive extracts to highly active extracts
the world. It is still indispensable to develop validated against three human cancer cells (OVCAR-8, HCT-116, and
methods using HPLC–MS and GC–MS, which are already SF-295).
broadly used in quality control as they give structural infor- Other BP samples from Santa Catarina and the Rio
mation and allow the quantification of many compounds. Grande do Sul were also evaluated against different cancer
HPLC–MS is a powerful technique and should be more cell lines, but they did not show in vitro antiproliferative
explored as it provides a complete characterization of the effect against the tested cells (Silva et al. 2017).
biomarker compounds of propolis. BP composition varies Overall, little is known about the antitumor activities
depending on several factors, and it is increasingly impor- of brown propolis since there is a wide range of possibili-
tant to construct a database with the already characterized ties to be tested. Furthermore, chemical variations directly
biomarkers and develop validated methods to analyze them. influence biological results, and each Brazilian region has
specific BP that could display antitumor activities. Scien-
Pharmacological Properties tific studies on BBP should be encouraged to discover new
molecules with antiproliferative actions.
Several diseases do not have available medicines in the mar-
ket for their treatment, and sometimes the available medi- Mutagenic and Antimutagenic Activity
cines present relevant side effects. Therefore, novel drugs
must be developed for treating these illnesses. Propolis has A series of in vivo experiments with Drosophila mela-
been used since ancient times as a healing agent. Its isolated nogaster were performed to evaluate the mutagenic potential
compounds have been tested against many pathologies and of Brazilian brown propolis. In this test, the genotoxicity was
parasites. The biological effects of BBP and its chemical evaluated through the somatic mutation and recombination
composition are displayed in Table S1. test (SMART), in which different mutant strains of D. mela-
nogaster are used to be crossed and exposed to the samples
Cytotoxicity Assays (propolis extract, isolated compounds, or volatile oil). The
offspring obtained from the emerging adults of the nonlethal
The cytotoxicity studies on BBP are still scarce; the litera- concentrations are analyzed, observing spots in the wings
ture reports especially the cytotoxicity of the ethanolic brown that indicate mutagenic treatment activity.
propolis extract from the different geographic origins of Bra- Ethanolic extract of brown propolis from Paraná state in
zil, such as the northeastern (Frota et al. 2021), the southeast- concentrations between 0.5 and 7.5 mg/ml showed no induc-
ern (Ribeiro et al. 2021a, b, c), and the southern (Machado tion of genetic and chromosomal mutations or the somatic
et al. 2021) regions. As propolis activities are related to the recombination indicating safety in use at low concentrations
environmental conditions prevailing in each region, the cyto- (Rodrigues et al. 2016). This was confirmed by another
toxicity reported from BP of different regions in Brazil also study, where BP from Mato Grosso do Sul State, at 1, 2,
shows variations in the antiproliferative activities. and 4 mg/ml, did not indicate mutagenicity (Fernandes et al.
The ethanol extract of BP from Ceará inhibited more 2019). However, the isolated compounds caffeic acid and
than 75% of the proliferation of human cancer cells, such as p-coumaric acid presented toxicity against D. melanogaster
colorectal (HCT116), leukemia (HL60), prostate (PC3), and larvae at 40 mmol/l. Similarly, the acetylisocupressic acid
murine melanoma cancer cells (B16F10). Despite observ- was mutagenic at 2.76 mmol/l on the individuals of the HB
ing an excellent inhibition against these cancer cell lines, cross (crossing with the P450 cytochrome bioactivation),
inhibition values in normal cells also exhibited remarkable indicating a promutagen effect. On the other hand, the essen-
cytotoxicity, demonstrating the non-selectivity of the tested tial oil of BP from this location showed a lack of mutagenic
extract. This study also evaluated the hexane extract of BP activity and somatic mutations at concentrations of 0.05, 0.1,
against the same human and murine cancer cells reported and 0.2% (Fernandes et al. 2015).

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The ethanolic extract of Mato Grosso do Sul state propo- (Paraná, Minas Gerais, and Mato Grosso do Sul states),
lis was tested in vivo on D. melanogaster larvae. Concen- extracted with ethanol 96%–water (7:3, v/v) and ethanol
trations of 0.375, 0.75, and 1.5 mg/ml, administered with 96%, were previously submitted to wax and non-polar com-
doxorubicin hydrochloride (DRX), a mutagenic agent, ponent removal prior to being tested on the DPPH assay
indicated inhibition of mutation frequencies in ST descend- (Dembogurski et al. 2018).
ants, between 32.5 and 51.8%. Despite the HB crosses, with BP potential of preventing oxidative stress in the skin was
metabolic activation of procarcinogens and promutagens by tested using a UVB-induced oxidative model (Fonseca et al.
CYP450 enzymes, no inhibition of mutation was observed. 2011). Previously, the extract displayed significant antioxi-
Instead, the highest concentration presented a marked co- dant activity against superoxide radicals on the xanthine/
mutagenic activity between the extract and the DXR. The luminol/XOD system, with an ­IC50 of free radicals’ inhibition
authors attribute this result to the presence of compounds value of 0.005 μl/ml in an in vivo topical pretreatment (2.5%)
like flavonoids that can potentiate the effect of some chemo- and oral treatment (100 mg/kg) of BP extract, tested on hair-
therapeutic agents. Considering that BBP did not show a less mice of the HRS/J strain exposed to UV irradiation. The
mutagenic effect, its potential antigenotoxicity effect was oral treatment showed recovery of glutathione (GSH) levels.
assessed (Fernandes et al. 2014). Furthermore, topical pretreatment and oral treatment inhib-
Fractions from the ethanolic extract showed no genotoxic- ited the cutaneous metalloproteinase activity induced by UV
ity against D. melanogaster larvae. Instead, treatment with irradiation exposure. However, the treatments did not exhibit
butanolic (n-BuOH) fraction decreased the mutations caused inhibition of metalloproteinases (MMP-2 and MMP-9) in the
by DXR on the adults of the crosses, with inhibition of the irradiated animals. These enzymes are involved in elastin
mutagenic effect higher than 90% on the HB descendants in degradation and aging processes.
a dose–response manner. It indicates a possible metabolic The antioxidant capacity of the essential oil of BP from
activation of n-BuOH fraction constituents and CYP450 Minas Gerais was also tested on the DPPH and ABTS
enzymes (Fernandes et al. 2014). assays by a spectrophotometric method (Lima et al. 2019).
The volatile oil displayed I­ C50 of 30.1 μg/ml on the ABTS
assays and 25 μg/ml on the DPPH test. The last value was
Antioxidant Effect close to the positive control, butylated hydroxytoluene
(BHT), indicating an antioxidant potential of the volatile
Several researchers have tested the antioxidant potential of constituents of BBP.
BP from different locations. Compared to other propolis,
brown types showed low values of flavonoids and phenolic
compounds, decreasing their antioxidant potential (Machado Toxicity
et al. 2016a, b; Silva et al. 2017). However, some papers
reported an excellent antioxidant capacity of ethanolic The toxic potential of BBP is poorly studied, and as men-
extracts, fractions, and essential oil of brown propolis. tioned, it was focused on the mutagenic activity. Only one
According to Mohafez et al. (2010), the BP ethanolic study focuses on the acute toxicity assays of BBP extracts
extract at 50 μg/ml displayed antioxidant activity compa- on Artemia salina (Santos et al. 2017). According to their
rable to the hydroxyl ammonium chloride, an antioxidant results, the hexane extract enriched with hydrocarbons did
compound. By increasing the reaction time and the concen- not exhibit toxicity with a ­DL50 of 1000 mg/ml, although
tration, the scavenging effect reached 47.5% at 100 μg/ml on the dichloromethane extract enriched with ananixathone
the DPPH assay. A Mato Grosso do Sul propolis ethanolic presented ­DL50 toxicity value of 68.99 mg/ml. The metha-
extract and its fractions were tested on the DPPH assay to nolic extract without a chemical characterization presented
calculate the I­ C50 value (Fernandes et al. 2014). The extract a ­DL50 value of 118.1 mg/ml.
displayed an ­IC50 of 532.2 μg/ml, while their ethyl acetate
and n-butanol fractions presented the lowest I­ C50 values
(109.3 and 38.8 μg/ml, respectively), which were the most Antifungal Activity
potent ones. Even with potential, these fractions did not dis-
play the caffeic acid antioxidant potential ­(IC50 3.47 μg/ml). The antifungal activity of BP was evaluated by Waller
Hexane and dichloromethane fractions presented a poor et al. (2017) using Sporothrix brasiliensis. The BP could
antioxidant potential due to the significant presence of non- inhibit the growth of 100% of the S. brasiliensis and pre-
polar compounds (Fernandes et al. 2014; Santos et al. 2017). sented a minimum inhibitory concentration (MIC) range
It revealed that the solvent and the extraction method have a from 0.19 to 1.56 mg/ml. The minimum fungicidal concen-
remarkable influence on the biological effect. The antioxi- tration (MFC) was also evaluated, and the values ranged
dant capacities of propolis samples from different locations from 0.78 to 3.125 mg/ml. More than 70% of the strains

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Revista Brasileira de Farmacognosia (2023) 33:288–299 295

used in this study were itraconazole resistant. Deegan concentration ­(CC50) to macrophages for the amastigote
et al. (2019) compared the activity of BP ethanolic extract forms. The hydroalcoholic fraction displayed the highest
against Malassezia pachydermatis and some commercially activity ­(CC50 21.69 μg/ml), and the ethyl acetate fraction,
antifungal agents. BP displayed a MIC value > 16 mg/ml, which contains p-coumaric acid as the major compound
but it did not exhibit lethality (MFC) at the highest tested (17.16%), displayed the weaker activity (­ CC50 > 400 μg/ml).
concentrations. The MIC and MFC values of amphotericin The antileishmanial activity of VOBP was reported by
B, itraconazole, and ketoconazole ranged from 0.0625 to Ribeiro et al. (2021a, b, c). The in vitro assay was accom-
16 μg/ml. plished with promastigote and amastigote forms of the
Candida is a genus comprising more than 300 species, parasite, and it was noticed that VOBP inhibited flagellar
and some can cause human infections that significantly motility in the promastigote forms in a dose-dependent man-
affect immunocompromised people (Ruhnke 2006). Differ- ner with I­ C50 21.3 μg/ml and also increased the death of
ent types of Brazilian BP were not active against Candida amastigote forms with I­ C50 25.1 μg/ml. Amphotericin B was
albicans (Silva et al. 2017). Salomão et al. (2008) reported the positive control used at 50 μg/ml, and VOBP was more
the results of BP extract against C. albicans and Paracoc- active against amastigote forms than the positive control.
cidioides brasiliensis, and it was more active against P. Nerolidol was the major compound of VOBP, which has
brasiliensis, but still with weak activity. The volatile oil of already demonstrated leishmanicidal activity against L. bra-
BP (VOBP) was tested against three strains of Candida: C. ziliensis (Ceole et al. 2017).
krusei, C. glabrata, and C. parapsilosis. The best activity One study reported the activity of BP extract and its frac-
was against C. parapsilosis with MIC of 100 μg/ml (Ribeiro tions against trophozoite forms of T. vaginallis with MIC of
et al. 2021a, b, c). 400 μg/ml. Furthermore, one dichloromethane fraction and
one ethyl acetate fraction inhibit 100% of the trophozoite
Antiparasitical Activity viability at 500 μg/ml (Dembogurski et al. 2018). Different
types of BP extract were used, and the ethanolic extracts
There are 20 neglected tropical diseases (NTDs) listed by the were the most active.
World Health Organization (WHO 2019) including Leishma-
niasis and Chagas disease. These NTDs affect people who Antiviral Activity
live in great poverty, corresponding to 1.4 billion people
worldwide. (Holmes et al. 2017). Hocchein et al. (2019) reported the antiviral activity of BP
Trypanosoma cruzi is a protist hemoflagellate responsible hydroalcoholic extract and its dichloromethane and aque-
for causing Chagas disease. Salomão et al. (2008) evalu- ous fractions against herpes simplex virus type-1 (HSV-1).
ated the in vitro activity of some BP extracts against tripo- Dichloromethane fraction displayed cytotoxicity activity
mastigote forms of Trypanossoma cruzi. Crystal violet, the over the cell with C
­ C50 88.4 μg/ml, followed by the crude
positive control, displayed ­IC50 187 μg/ml, and BP samples hydroalcoholic extract ­(CC50 143.7 μg/ml), and aqueous
displayed ­IC50’s ranging from 200 to 2000 μg/ml. BP con- fraction with weak activity ­(CC50 1240 μg/ml). They also
taining 24.74% of benzoic acid displayed the smaller ­IC50. evaluated the percentage of an inhibitory concentration
Another 96 h in vitro study with different extracts of BP of the fractions and extract, finding a range of I­ C50 from
against epimastigote forms of T. cruzi with concentrations 58.5 μg/ml (dichloromethane fraction) to 294 μg/ml (butanol
between 75 and 300 mg/ inhibited the growth of T. cruzi by fraction). Likewise, two fractions inhibited 90% of the virus
90% (Silva et al. 2017). replication at concentrations above 200 μg/ml.
Considering other studies where BP showed potential
against Plasmodium, Trypanosoma, and Leishmania genus, Antibacterial Activity
it became an exciting alternative for discovering new com-
pounds with antileishmanial activities. Santana et al. (2014) The antimicrobial activity of BP has been studied worldwide
reported the activity of BP against promastigote and amas- against gram-positive and gram-negative bacteria, drawing
tigote forms of L. amazonensis, comparing different frac- the attention of pharmaceutical and food companies. Moreo-
tions obtained by the partition of the crude extract. The in ver, the need to develop new drugs with antimicrobial poten-
vitro assay of growth inhibition of L. amazonensis promas- tial has increased annually due to the high number of new
tigote form after 72 h with dichloromethane fraction contain- resistance against the usual treatment (Morrill et al. 2015).
ing 2,3-dihydroxybenzofurane as its major compound dis- A good activity of BP extract was observed against gram-
played the best result ­(IC50 3.22 μg/ml). The hydroalcoholic positive bacteria Staphylococcus aureus and Enterococcus spp.
fraction ­(IC50 4.64 μg/ml), hexane fraction ­(IC50 4.79 μg/ (MIC 31.3–500 μg/ml). However, no activity was found against
ml), and ethyl acetate fraction (­ IC50 8.83 μg/ml) were also gram-negative bacteria, like Klebsiella spp. and Escherichia
active. The activity was evaluated by the 50% cytotoxicity coli (Silva et al. 2017). Salomão et al. (2008) reported a similar

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result, by describing the activity of BP extract against Strepto- et al. 2010). A dose of 100 mg/kg of BP ethanolic extract,
coccus pneumonia (MIC 0.2–0.8 μg/ml) and Staphylococcus in an oral pretreatment, showed hepatic protection against
aureus (MIC 1.6–52.4 μg/ml). On the other hand, it was inac- lipid peroxidation effect of C­ Cl4, decreasing the malondial-
tive against gram-negative Klebsiella pneumoniae. dehyde (MDA) level and increasing the glutathione (GSH)
Other studies reported the inactivity of BP extracts concentration. Also, BBP pretreatment reduced hemorrhagic
against strains of Salmonella spp., P. aeruginosa, S. aureus, lesions induced by indomethacin in rat stomachs, reducing
and E. coli, including resistant strains (Bastos et al. 2011). the gastric ulcer index by approximately 65%, and maintain-
Furthermore, forty samples of BP from different origins ing the typical histological structure. Even when indometha-
were tested against S. aureus, E. faecalis, P. aeruginosa, cin causes ROS-generated inflammation, BP pretreatment
and E. coli. A few of them displayed weak MIC values attenuates this injury effect, preventing ulcer damage.
(156–1250 μg/ml), and the majority were inactive. The BP has been evaluated as a food supplement of lambs in
same occurred for the minimum bactericidal concentration comparison with the traditional dietary additive monensin
(MBC), with no extract being active at any concentration (Ítavo et al. 2011). The propolis extract was added to the diet
(Machado et al. 2021). of lambs daily with a calculated dry residual/flavonoid propor-
Dembogurski et al. (2018) evaluated the antibacterial activ- tion of 0.9 g/20.2 mg. The animal group treated with propolis
ity of BP against planktonic cells of Staphylococcus aureus showed good efficiency in weight gain, feed efficiency, and
and biofilm cells. Five different fractions of BP extract were conversion, making BP extract a potential food supplement
tested on seven strains of the gram-positive Mycoplasma to replace monensin sodium, which has been indicated as a
genera, and the less polar fractions displayed the best MIC potential inhibitor of food consumption in sheep.
value (3.9 μg/ml) (Araújo et al. 2020). The result seemed to
be better for planktonic cells. Still, the BP assay against Pseu- Production and Economic Aspects
domonas aeruginosa (gram-negative) did not show activity.
The VOBP also displayed activity against Helicobacter The Brazilian beekeeping activity is a promising national
pylori, Mycobacterium tuberculosis, and M. avium with MIC enterprise and has gained traction in the international market
3.25, 50, and 62.5 μg/ml, respectively. Tetracycline displayed since the enormous Brazilian biodiversity favors the diversi-
MIC of 1 μg/ml against H. pillory and isoniazid displayed MIC fied bee products. Scientific investigations showing Brazilian
of 1.47 μg/ml against both Mycobacterium (Lima et al. 2019). propolis’s antimicrobial and immunological activities have
Fernandes et al. (2015) reported the antibacterial evaluation contributed to increasing propolis’s competitiveness and
of another VOBP sample with MIC > 1000 μg/ml against five demand nationally and internationally (Berretta et al. 2017).
strains of bacteria, including gram-positive and gram-negative The range of biological properties of BP has drawn the
bacteria (Enterococcus fecalis, S. aureus, E. coli, P. aerugi- interest of the pharmaceutical, food, and cosmetic industries,
nosa, and K. pneumonia). As expected, it was noticed that mainly due to its antimicrobial and antioxidant potential with
BP has more significant potential against gram-positive than beneficial health effects for consumers. The characteristic
gram-negative bacteria. Similar results were reported by other organoleptic properties, such as smell, color, and texture of
authors suggesting that the difference occurs due to more com- most propolis, are essential for consumer acceptability and
plexity of the cell wall of gram-negative bacteria. commercialization. Its medicinal and food supplement uses
Nevertheless, the variation of the antimicrobial activity are well-established in several parts of the world, especially
found in samples of BP extract and its volatile oil can be in Eastern Europe, China, and Japan. Brazil is a big player in
explained by the chemical difference among the samples, the international market, moving millions of dollars annually
which leads to different chemical properties. It is known that with bee products. (Hata et al. 2012; Berretta et al. 2017).
the chemical composition of propolis depends on its botani- CONAP-Brazil (National Beekeeping Cooperative)
cal origin, which is directly correlated to the surrounding reported in 2020 that propolis’s national and international
plant biodiversity (Rodrigues et al. 2020). markets increased by approximately 40 and 50%, respec-
tively, devoid of the COVID-19 outbreak. It is estimated that
approximately 90% of CONAP’s revenue comes from the
Other Activities exports of bee products, mainly to Asian countries, including
Japan, South Korea, and Taiwan, registering an increase of
The enzymatic analysis of gastric tissues indicated that prop- 94% in the exports compared to 2019 (Conap 2020).
olis caused restoration of glutathione and superoxide dis- Minas Gerais is the largest Brazilian State propolis pro-
mutase activity to normal levels and reduced thiobarbituric ducer, responsible for 70% of all propolis production in Bra-
acid reactive substances (TBARS) levels. The antioxidant, zil of the total estimated production of 120 tons per year,
lipid peroxidation-inhibitory, and anti-ulcer in vivo activi- with 85% of green propolis and 15% of brown propolis (Gui-
ties of BBP ethanolic extract were also reported (Mohafez marães 2020). As the world demand for propolis has grown,

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Revista Brasileira de Farmacognosia (2023) 33:288–299 297

with exportation to Europe and the USA, other regions of Southern Brazil. J Food Sci Technol 57:4228–4235. https://​doi.​
Brazil are also increasing their productions, such as the org/​10.​1007/​s13197-​020-​04461-y
Arruda C, Ribeiro VP, Almeida MO, Mejía JAA, Casoti R, Bastos
northeast and south regions. However, to consolidate brown JK (2020) Effect of light, oxygen and temperature on the sta-
propolis in the international market, more scientific stud- bility of artepillin C and p-coumaric acid from Brazilian green
ies must be undertaken on propolis from each region, thus propolis. J Pharm Biomed Anal 178:112922. https://​doi.​org/​10.​
adding value to this Brazilian bee product (Nordeste 2021). 1016/j.​jpba.​2019.​112922
Arruda C, Ribeiro VP, Mejía JAA, Almeida MO, Goulart MO, Can-
dido ACBB, dos Santos RA, Magalhães LG, Martins CHG,
Bastos JK (2020) Green propolis: cytotoxic and leishmanicidal
Conclusions activities of artepillin C, p-coumaric acid, and their degradation
products. Rev Bras Farmacogn 30:169–176. https://​doi.​org/​10.​
BP has a greater chemical variety than other types of Brazilian 1007/​s43450-​020-​00043-3
Bankova V, Boudourova-Krasteva G, Sforcin JM, Frete X,
propolis since several possible botanical sources are listed. Kujumgiev A, Maimoni-Rodella R, Popov S (1999) Phytochem-
Thus, further research using BP to assess its pharmacological ical evidence for the plant origin of Brazilian propolis from São
activity should consider its chemical variation and its possible Paulo State. Z Naturforsch C 54:401–405. https://​doi.​org/​10.​
botanical sources. Powerful tools like ultra-efficiency liquid 1515/​ZNC-​1999-5-​616
Bankova V, Popova M, Trusheva B (2018) The phytochemistry of the
chromatography systems coupled with high-resolution mass honeybee. Phytochemistry 155:1–11. https://​doi.​org/​10.​1016/j.​
spectrometry associated with metabolomics studies can be an phyto​chem.​2018.​07.​007
alternative to propose new propolis classification taking into Bastos EMAF, Galbiati C, Loureiro EM, Scoaris DO (2011) Indica-
account their chemical profile. The volatile fractions’ compo- dores físico-químicos e atividade antibacteriana de própolis mar-
rom frente à Escherichia coli. Arq Bras Med Vet Zootec 63:1255–
sition of different types of propolis has been poorly explored, 1259. https://​doi.​org/​10.​1590/​S0102-​09352​01100​05000​32
and it undoubtedly plays an essential role in its biological Bernardes CT, Ribeiro VP, Carvalho TC, Furtado RA, Furtado NAJC,
activities, which have not been fully elucidated. Therefore, Bastos JK (2022) Disinfectant activities of extracts and metabo-
different types of Brazilian brown propolis have great poten- lites from Baccharis dracunculifolia DC. Lett Appl Microbiol
75:261–270. https://​doi.​org/​10.​1111/​lam.​13725
tial and should be further investigated to enable its production Berretta AA, Arruda C, Miguel FG, Baptista N, Nascimento AP,
and commercialization as effective and safe health products. Marquele- Oliveira F, Hori JI, Barud HS, Damaso B, Ramos C,
Ferreira R, Bastos JK (2017) Functional properties of Brazilian
Supplementary Information The online version contains supplemen- propolis: from chemical composition until the market. Superfood
tary material available at https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 43450-0​ 23-0​ 0374-x. and Functional Food - an Overview of Their Processing and Uti-
lization. https://​doi.​org/​10.​5772/​65932
Acknowledgements The authors are thankful to Dr. Rogelio Pereda- Beserra FP, Gushiken LFS, Hussni MF, Ribeiro VP, Bonamin F,
Miranda, Editor-in-Chief of Brazilian Journal of Pharmacognosy, for Jackson CJ, Pellizzon CH, Bastos JK (2021) Artepillin C as an
his suggestions to improve the manuscript. The authors are thankful to outstanding phenolic compound of Brazilian green propolis for
Fundação de amparo à Pesquisa do Estado de São Paulo (FAPESP) for disease treatment: a review on pharmacological aspects. Phytother
financial support, Grants # 2017/04138-8 and 2022/00659-1. Res 35:2274–2286. https://​doi.​org/​10.​1002/​ptr.​6875
Brophy JJ, Goldsack RJ, Wu MZ, Fookes CJR, Forster PI (2000) The
Author Contribution VPR contributed to the conception, design, litera- steam volatile oil of Wollemia nobilis and its comparison with
ture search, and writing of the manuscript. Data compilation, and writ- other members of the Araucariaceae (Agathis and Araucaria).
ing and editing of the manuscript were done by JAAM, DMR, GRA, Biochem Syst Ecol 28:563–578. https://​doi.​org/​10.​1016/​S0305-​
AMFP, and MHT. JKB and SRA supervised and critically revised the 1978(99)​00090-3
manuscript. All the authors have read and approved the manuscript. Burzynski-Chang EA, Ryona I, Reisch BI, Gonda I, Foolad MR, Gio-
vannoni JJ, Sacks GL (2018) HS-SPME-GC-MS analyses of vola-
Funding The authors received financial support, Grants # tiles in plant populations—quantitating compound × individual
2017/04138–8 and 2022/00659–1, from Fundação de amparo à Pes- matrix effects. Molecules 23:2436. https://​doi.​org/​10.​3390/​molec​
quisa do Estado de São Paulo (FAPESP). ules2​31024​36
Calegari MA, Prasniewski A, Silva C, Sado RY, Maia FMC, Tonial
Declarations LMS, Oldoni TLC (2017) Propolis from Southwest of Parana
produced by selected bees: influence of seasonality and food
Conflict of Interest The authors declare no competing interests. supplementation on antioxidant activity and phenolic profile. An
Acad Bras Cienc 89:45–55. https://​doi.​org/​10.​1590/​0001-​37652​
01620​160499
Ccana-Ccapatinta GV, Mejía JAA, Tanimoto MH, Groppo M, Carvalho
References JCAS, Bastos JK (2020) Dalbergia ecastaphyllum (L.) Taub and
Symphonia globulifera L.f.: the botanical sources of isoflavonoids
Anvisa (2017) RDC 166/2017, Dispõe sobre a validação de métodos and benzophenones in Brazilian red propolis. Molecules 25:2060.
analíticos e dá outras providências, Agência Nacional de Vig- https://​doi.​org/​10.​3390/​MOLEC​ULES2​50920​60
ilância Sanitária, Ministério da Saúde, Brazília, DF Ceole LF, Cardoso MDG, Soares MJ (2017) Nerolidol, the main con-
Araújo C, Mayworm MAS, Yatsuda R, Negri G, Salatino MLF, stituent of Piper aduncum essential oil, has anti- Leishmania bra-
Salatino A, Timenetsky J, Campos GB (2020) Chemical com- ziliensis activity. Parasitology 144:1179–1190. https://​doi.​org/​10.​
position and antimycoplasma activity of a brown propolis from 1017/​S0031​18201​70004​52

13
298 Revista Brasileira de Farmacognosia (2023) 33:288–299

Conap (2020) Exports increase in the pandemic. https://​www.​Jorna​ JPC, da Silva Julião MS, Bandeira PN, Pessoa CÓ, dos Santos HS,
lbelv​edere.​Com.​Br/​Index.​Php/​Estilo/​Saude/​Item/​1653-​Conap-​ Zocolo GJ, Rodrigues THS (2021) Inhibitory activity of brown
Expor​tacoes-​Aumen​tam-​Na-​Pande​mia propolis extracts on a norfloxacin-resistant strain of Staphylococ-
Daugsch A, Moraes CS, Fort P, Park YK (2008) Brazilian red propolis cus aureus. Rev Bras Farmacogn 31:249–255. https://​doi.​org/​10.​
- chemical composition and botanical origin. Evid Based Com- 1007/​s43450-​021-​00150-9
plementary Altern Med 5:435–441. https://d​ oi.o​ rg/1​ 0.1​ 093/e​ cam/​ Hata T, Tazawa S, Ohta S, Rhyu MR, Misaka T, Ichihara K (2012)
nem057 Artepillin C, a major ingredient of Brazilian propolis, induces a
Deegan KR, Fonseca MS, Oliveira DCP, Santos LM, Fernandez CC, pungent taste by activating TRPA1 channels. PLoS One 7:e48072.
Hanna SA, Machado BAS, Umsza-Guez MA, Meyer R, Portela https://​doi.​org/​10.​1371/​JOURN​AL.​PONE.​00480​72
RW (2019) Susceptibility of Malassezia pachydermatis clinical Holmes KK, Bertozzi S, Bloom BR, Jha P (2017) Disease Control
isolates to allopathic antifungals and Brazilian red, green, and Priorities, Third Edition (Volume 6): Major Infectious Dis-
brown propolis extracts. Front Vet Sci 6:1–12. https://​doi.​org/​10.​ eases. World Bank, Washington, DC. https://​doi.​org/​10.​1596/​
3389/​fvets.​2019.​00460 978-1-​4648-​0524-0
Dembogurski DS, Silva TD, Boaretto AG, Rigo GV, Silva RC, Tasca Ioannou E, Koutsaviti A, Tzakou O, Roussis V (2014) The genus
T, Macedo AJ, Carollo CA, Silva DB (2018) Brown propolis- Pinus: a comparative study on the needle essential oil composi-
metabolomic innovative approach to determine compounds capa- tion of 46 pine species. Phytochem Rev 13:741–768. https://​doi.​
ble of killing Staphylococcus aureus biofilm and Trichomonas org/​10.​1007/​s11101-​014-​9338-4
vaginalis. Food Res Int 111:661–673. https://​doi.​org/​10.​1016/j.​ Ítavo CCBF, Morais MG, Costa C, Ítavo LCV, Franco GL, Silva
foodr​es.​2018.​05.​033 JA, Reis FA (2011) Addition of propolis or monensin in the
dos Santos DC, David JM, David JP (2017) Composição química, ativi- diet: behavior and productivity of lambs in feedlot. Anim Feed
dade citotóxica e antioxidante de um tipo de própolis da Bahia. Quim Sci Technol 165:161–166. https://​doi.​org/​10.​1016/j.​anife​edsci.​
Nova 40:171–175. https://​doi.​org/​10.​21577/​0100-​4042.​20160​174 2011.​02.​020
Eberhardt TL, Bernards MA, He L, Davin LB, Wooten JB, Lewis NG Lima VHM, Almeida KCR, Alves CCF, Rodrigues ML, Crotti AEM,
(1993) Lignification in cell suspension cultures of Pinus taeda. Souza JM, Ribeiro AB, Squarisi IS, Tavares DC, Martins CHG,
In situ characterization of a gymnosperm lignin. J Biol Chem Miranda MLD (2019) Biological properties of volatile oil from
268:21088–21096. https:// ​ d oi. ​ o rg/ ​ 1 0. ​ 1 016/ ​ S 0021- ​ 9 258(19)​ Brazilian brown propolis. Rev Bras Farmacogn 29:807–810.
36897-8 https://​doi.​org/​10.​1016/j.​bjp.​2019.​07.​004
Elian Guimarães (2020) With therapeutic properties, the search for green Machado BAS, Silva RPD, Barreto GDA, Costa SS, Silva DF,
propolis increases in the pandemic. Jornal Estado de Minas. https://​ HNRocha JLCN, Dellagostin OA, Henriques JAP, Umsza-Guez
www.​em.​com.​br/​app/​notic​ia/​econo​mia/​2021/​06/​26/​inter​nas_​econo​ MA, Padilha FF (2016) Chemical composition and biological
mia,12807​10/​com-​propr​iedad​es-​terap​eutic​as-​busca-​por-​propo​lis-​ activity of extracts obtained by supercritical extraction and etha-
verde-​aumen​ta-​na-​pande​mia.​shtml nolic extraction of brown, green and red propolis derived from
Fabio G, Federica C, Alfredo F, Nicola V (2019) Recent advances in different geographic regions in Brazil. PLoS One 11:e0145954.
analytical approaches for the standardization and quality of poly- https://​doi.​org/​10.​1371/​journ​al.​pone.​01459​54
phenols of propolis. J Med Plant Res 13:487–500. https://​doi.​org/​ Machado CS, Finger D, Lima TC, Modesto T, Monteiro MC, Oliveira
10.​5897/​jmpr2​019.​6849 FCE, Pessoa C, Torres YR (2021) Seeking the regional iden-
Fernandes FH, Rosa GZ, Garcez WS, Lopes SM, Corsino J, Garcez tity of brown propolis produced in southern brazil and linked
FR (2014) Assessment of the (anti)genotoxicity of brown propo- to total levels of bioactive compounds and biological activity.
lis extracts from Brazilian Cerrado biome in a Drosophila mela- ACS Food Sci Technol 1:176–185. https://​doi.​org/​10.​1021/​acsfo​
nogaster model. Food Res Int 62:20–26. https://d​ oi.o​ rg/1​ 0.1​ 016/j.​ odsci​tech.​0c000​52
foodr​es.​2014.​02.​029 Machado CS, Mokochinski JB, Lira TO, Oliveira FDCE, Car-
Fernandes FH, Guterres ZDR, Corsino J, Garcez WS, Garcez FR doso MV, Ferreira RG, Sawaya ACHF, Ferreira AG, Pessoa
(2019) Assessment of the mutagenicity of propolis compounds C, Cuesta-Rubio O, Monteiro MC, Campos MS, Torres YR
from the Brazilian cerrado biome in somatic cells of Drosophila (2016) Comparative study of chemical composition and biologi-
melanogaster. Orbital 11:307–313. https://​doi.o​ rg/1​ 0.​17807/o​ rbit​ cal activity of yellow, green, brown, and red Brazilian propolis.
al.​v11i5.​1418 Evid-Based Complement Altern Med 2016:6057650. https://​doi.​
Fernandes FH, Guterres ZDR, Violante IMP, Lopes TFS, Garcez WS, org/​10.​1155/​2016/​60576​50
Garcez FR (2015) Evaluation of mutagenic and antimicrobial Mayworm MAS, Lima CA, Tomba ACB, Fernandes-Silva CC, Sala-
properties of brown propolis essential oil from the Brazilian Cer- tino MLF, Salatino A (2014) Does propolis contain tannins?
rado biome. Toxicol Rep 2:1482–1488. https://​doi.​org/​10.​1016/j.​ Evid-Based Compl Alt 2014:613647. https://​doi.​org/​10.​1155/​
toxrep.​2015.​11.​007 2014/​613647
Fonseca YM, Marquele-Oliveira F, Vicentini FTMC, Furtado Mohafez OMM, Abdel-Raheem IT, Nafady AM (2010) Antioxidant,
NAJC, Sousa JPB, Lucisano-Valim YM, Fonseca MJV (2011) lipid peroxidation-inhibitory and antiulcer activities of brown
Evaluation of the potential of Brazilian propolis against UV- propolis. Bull Pharm Sc 33:169–177. https://​doi.​org/​10.​21608/​
induced oxidative stress. Evid Based Complement Alternat Med bfsa.​2010.​64741
2011:863917. https://doi.org/10.1155/2011/863917 Morrill HJ, Pogue JM, Kaye KS, LaPlante KL (2015) Treatment
Freitas AS, Barth OM, Sales EO, Matsuda AH, Almeida-Muradian LB options for carbapenem-resistant Enterobacteriaceae infections.
(2011) A palynological analysis of Brazilian propolis samples. Open Forum Infect Dis 2:ofv050. https://​doi.​org/​10.​1093/​ofid/​
JAAS 3:67–74. https://​doi.​org/​10.​3896/​IBRA.4.​03.2.​01 ofv050
Freitas MO, Ponte FAF, Lima MAS, Silveira ER (2008) Flavonoids Nordeste, B (2021) Potential of propolis production in the North-
and triterpenes from the nest of the stingless bee Trigona spinipes. east. Publishing Journal Banco Do Nordeste. https://​bnb.​gov.​
J Braz Chem Soc 19:532–535. https://​doi.​org/​10.​1590/​S0103-​ br/​web/​guest/​agron​egocio/​agroi​nform​a/-/​asset_​publi​sher/​qg5dL​
50532​00800​03000​22 6xAGf​oP/​conte​nt/​poten​cial-​da-​produ​cao-​de-​propo​lis-​no-​norde​
Frota VM, Dias FMF, do Nascimento MF, da Silva Mendonça L, dos ste/​37609​65?​inher​itRed​irect=​false​&​redir​ect=​https%​3A%​2F%​
Santos Moita EC, dos Santos Gomes LC, Leal ALAB, Barreto 2Fbnb.​gov.​br%​2Fweb%​2Fgue​st%​2Fagr​onego​cio%​2Fagr​oinfo​
HM, Silva MFS, dos Santos Fontenelle RO, Gomes GA, do Vale rma%​3Fp_p_​id%​3D

13
Revista Brasileira de Farmacognosia (2023) 33:288–299 299

Olegário LS, Andrade JKS, Andrade GRS, Denadai M, Cavalcanti RL, Evid-Based Complement Altern Med 5:317–324. https://​doi.​org/​
Silva MAAP, Narain N (2019) Chemical characterization of four 10.​1093/​ecam/​nem058
Brazilian brown propolis: an insight in tracking of its geographical Santana LCLR, Carneiro SMP, Caland-Neto LB, Arcanjo DDR, Moita-
location of production and quality control. Food Res Int 123:481– Neto JM, Citó AMGL, Carvalho FAA (2014) Brazilian brown
502. https://​doi.​org/​10.​1016/j.​foodr​es.​2019.​04.​004 propolis elicits antileishmanial effect against promastigote and
Park YK, Alencar SM, Aguiar CL (2002) Botanical origin and amastigote forms of Leishmania amazonensis. Nat Prod Res
chemical composition of Brazilian Propolis. J Agric Food Chem 28:340–343. https://​doi.​org/​10.​1080/​14786​419.​2013.​856904
50:2502–2506. https://​doi.​org/​10.​1021/​JF011​432B Santos MFC, Oliveira LC, Ribeiro VP, Soares MG, Moraes GOI, Sar-
Pavlovic R, Borgonovo G, Leoni V, Giupponi L, Ceciliani G, Sala S, tori AGO, Rosalen PL, Bastos JK, Alencar SM, Veneziani RCS,
Bassoli A, Giorgi A (2020) Effectiveness of different analytical Ambrósio SR (2021) Isolation of diterpenes from Araucaria sp.
methods for the characterization of propolis: a case of study Brazilian brown propolis and development of a validated HPLC
in Northern Italy. Molecules 25:504. https://​doi.​org/​10.​3390/​ method for its analysis. J Sep Sci 44:3089–3097. https://​doi.​org/​
molec​ules2​50305​04 10.​1002/​jssc.​20210​0374
Pimenta HC, Violante IMP, Musis CR, Borges ÁH, Aranha AMF Sartori G, Pesarico AP, Pinton S, Dobrachinski F, Roman SS, Pau-
(2015) In vitro effectiveness of Brazilian brown propolis against letto F, Rodrigues LC, Prigol M (2012) Protective effect of brown
Enterococcus faecalis. Braz Oral Res 29:1–6. https://​doi.​org/​10.​ Brazilian propolis against acute vaginal lesions caused by herpes
1590/​1807-​3107B​OR-​2015.​vol29.​0058 simplex virus type 2 in mice: involvement of antioxidant and anti-
Ribeiro VP, Arruda C, Abd El-Salam M, Bastos JK (2018) Brazilian inflammatory mechanisms. Cell Biochem Funct 30:1–10. https://​
medicinal plants with corroborated anti-inflammatory activities: doi.​org/​10.​1002/​cbf.​1810
a review. Pharm Biol 56:253–268. https://​doi.​org/​10.​1080/​13880​ Sartori GO, Spada FP, Ribeiro VP, Rosalen PL, Ikegaki M, Bastos
209.​2018.​14544​80 JK, Alencar SM (2021) An insight into the botanical origins of
Ribeiro VP, Arruda C, Mejia JAA, Bastos JK, Tripathi SK, Khan SI, propolis from permanent preservation and reforestation areas
Khan IA, Ali Z (2021) Phytochemical, antiplasmodial, cytotoxic of southern Brazil. Sci Rep 11:22043. https://​doi.​org/​10.​1038/​
and antimicrobial evaluation of a southeast Brazilian brown s41598-​021-​01709-1
propolis produced by Apis mellifera bees. Chem Biodivers Sawaya ACHF, Cunha BS, Marcucci MC (2011) Analytical methods
18:e2100288. https://​doi.​org/​10.​1002/​cbdv.​20210​0288 applied to diverse types of Brazilian propolis. Chem Cent J 5:27.
Ribeiro VP, Arruda C, Mejía JAA, Candido ACBB, Santos RA, Magal- https://​doi.​org/​10.​1186/​1752-​153X-5-​27
hães LG, Bastos JK (2021b) Brazilian southeast brown propo- Serafim MS, Rodrigues DM, Ribeiro VP, Ccana-Ccapatinta GV,
lis: gas chromatography method development for its volatile oil Groppo M, Martins CHG, Ambrósio SR, Bastos JK (2022) Euca-
analysis, its antimicrobial and leishmanicidal activities evaluation. lyptus botryoides’ resin and its new 2-O-galloyl-1,6-O-di-trans-
Phytochem Anal 32:404–411. https://​doi.​org/​10.​1002/​pca.​2988 p-coumaroyl-β-D-glycopyranoside compound display good anti-
Ribeiro VP, Ccana-Ccapatinta GV, Mejía JAA, Berretta AA, Moraes microbial activity. Nat Prod Res 5:1–10. https://​doi.​org/​10.​1080/​
LA, Bastos JK (2022) Chemical characterization of Brazilian 14786​419.​2022.​20654​86
propolis using automated direct thermal desorption–gas chroma- Silva RPD, Machado BAS, Barreto GA, Costa SS, Andrade LN,
tography–mass spectrometry. J Sci Food Agric 102:4345–4354. Amaral RG, Carvalho AA, Padilha FF, Barbosa JDV, Umsza-
https://​doi.​org/​10.​1002/​jsfa.​11788 Guez MA (2017) Antioxidant, antimicrobial, antiparasitic, and
Ribeiro VP, Símaro GV, Mejia JAA, Arruda C, Bastos JK (2021) Anti- cytotoxic properties of various Brazilian propolis extracts. PLoS
inflammatory and antinociceptive activities of the hydroalcoholic One 12:e0172585. https://​doi.​org/​10.​1371/​journ​al.​pone.​01725​85
extract and the volatile fraction of southeastern Brazilian brown Simões-Ambrosio LMC, Gregório LE, Sousa JPB, Figueiredo-Rinhel
propolis. Rev Bras Farmacogn 31:59–66. https://d​ oi.​org/1​ 0.​1007/​ ASG, Azzolini AECS, Bastos JK, Lucisano-Valim YM (2010)
s43450-​020-​00122-5 The role of seasonality on the inhibitory effect of Brazilian green
Rivera-Yañez N, Rivera-Yañez CR, Pozo-Molina G, Méndez-Catalá propolis on the oxidative metabolism of neutrophils. Fitoterapia
CF, Méndez-Cruz AR, Nieto-Yañez O (2020) Biomedical prop- 81:1102–1108. https://​doi.​org/​10.​1016/J.​FITOTE.​2010.​07.​008
erties of propolis on diverse chronic diseases and its potential Simone-Finstrom M, Borba RS, Wilson M, Spivak M (2017) Propolis
applications and health benefits. Nutrients 13:78. https://​doi.​org/​ counteracts some threats to honeybee health. Insects 8:46. https://​
10.​3390/​NU130​10078 doi.​org/​10.​3390/​INSEC​TS802​0046
Rodrigues CRF, Plentz LC, Marcucci MC, Dihl RR, Lehmann M Tazawa S, Arai Y, Hotta S, Mitsui T, Nozaki H, Ichihara K (2016)
(2016) In vivo evaluation of mutagenic and recombinagenic Discovery of a novel diterpene in brown propolis from the state
activities of Brazilian propolis. Food Chem Toxicol 96:117–121. of Parana, Brazil. Nat Prod Commun 11:201–205. https://​doi.​org/​
https://​doi.​org/​10.​1016/j.​fct.​2016.​07.​037 10.​1177/​19345​78X16​01100​218
Rodrigues DM, de Souza MC, Arruda C, Pereira RAS, Bastos JK Waller SB, Peter CM, Hoffmann JF, Picoli T, Osório LG, Chaves F,
(2020) The role of Baccharis dracunculifolia and its chemical Zani JL, Faria RO, Mello JRB, Meireles MCA (2017) Chemical
profile on green propolis production by Apis mellifera. J Chem and cytotoxic analyses of brown Brazilian propolis (Apis mellif-
Ecol 46:150–162. https://​doi.​org/​10.​1007/​s10886-​019-​01141-w era) and its in vitro activity against itraconazole-resistant Sporo-
Ruhnke M (2006) Epidemiology of Candida albicans infections and thrix brasiliensis. Microb Pathog 105:117–121. https://d​ oi.o​ rg/1​ 0.​
role of non-Candida albicans yeasts. Curr Drug Targets 7:495– 1016/j.​micpa​th.​2017.​02.​022
504. https://​doi.​org/​10.​2174/​13894​50067​76359​421 WHO (2019) World Health Organization. Control of Neglected Tropi-
Salas AL, Alberto MR, Zampini IC, Cuello AS, Maldonado L, Ríos cal Diseases. https://​www.​who.​int/​teams/​contr​ol-​of-​negle​cted-​
JL, Schmeda-Hirschmann G, Isla MI (2016) Biological activities tropi​cal-​disea​ses/​overv​iew
of polyphenols-enriched propolis from Argentina arid regions.
Phytomedicine 23:27–31. https://​doi.​org/​10.​1016/J.​PHYMED.​ Springer Nature or its licensor (e.g. a society or other partner) holds
2015.​11.​007 exclusive rights to this article under a publishing agreement with the
Salomão K, Pereira PRS, Campos LC, Borba CM, Cabello PH, author(s) or other rightsholder(s); author self-archiving of the accepted
Marcucci MC, Castro SL (2008) Brazilian propolis: correla- manuscript version of this article is solely governed by the terms of
tion between chemical composition and antimicrobial activity. such publishing agreement and applicable law.

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