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. 2021 Jun 7;26(11):3462. doi: 10.3390/molecules26113462

Chemical Composition of Volatile Compounds in Apis mellifera Propolis from the Northeast Region of Pará State, Brazil

Mozaniel Santana de Oliveira 1,2,*, Jorddy Neves Cruz 1, Oberdan Oliveira Ferreira 1, Daniel Santiago Pereira 3, Natanael Santiago Pereira 4, Marcos Enê Chaves Oliveira 3, Giorgio Cristino Venturieri 5, Giselle Maria Skelding Pinheiro Guilhon 6, Antônio Pedro da Silva Souza Filho 3, Eloisa Helena de Aguiar Andrade 1,2,6
Editor: Juraj Majtan
PMCID: PMC8201256  PMID: 34200300

Abstract

Propolis is a balsamic product obtained from vegetable resins by exotic Africanized bees Apis mellifera L., transported and processed by them, originating from the activity that explores and maintains these individuals. Because of its vegetable and natural origins, propolis is a complex mixture of different compound classes; among them are the volatile compounds present in the aroma. In this sense, in the present study we evaluated the volatile fraction of propolis present in the aroma obtained by distillation and simultaneous extraction, and its chemical composition was determined using coupled gas chromatography, mass spectrometry, and flame ionization detection. The majority of compounds were sesquiterpene and hydrocarbons, comprising 8.2–22.19% α-copaene and 6.2–21.7% β-caryophyllene, with additional compounds identified in greater concentrations. Multivariate analysis showed that samples collected from one region may have different chemical compositions, which may be related to the location of the resin’s production. This may be related to other bee products.

Keywords: Amazon, bioproducts, propolis, aroma, bioactive compounds

1. Introduction

Honeybees are among the most studied insects because of their high economic value and fundamental role in agriculture and ecosystems [1]. The species Apis mellifera is known worldwide as an important pollinator of agricultural crops [2]. This species is native to Europe, Africa, the Middle East, and parts of Asia [3,4], and has great potential for adaptation to different biomes and climatic conditions [3]. Apis mellifera is not restricted to honey production; it also produces propolis through the addition of saliva and wax to organic liquids collected from plant sap, resin, gum, and latex [5].

Propolis, also called “bee glue”, is a resinous substance similar in some aspects to natural wax found in hives [6]. This substance has a dark yellow to brown color and is formed from materials collected by bees from flower buds, leaves, and other plant parts [7]. Propolis is sticky and adhesive in nature [8]. For bees, propolis is of paramount importance, as the insects use it as a coating to seal cracks or spaces in the hive, a base for making honey [6], colony protection, and defense against infections and parasites [9].

The protection that propolis offers to bees is related to the pharmacological properties of this bioactive product [10], as indicated by its uses in traditional medicine. Propolis is mainly used for treating diseases of the vascular and blood system (anemia), respiratory infections, ulcers, mycoses, and cancer, along with improving the immune system [11]. The chemical characteristics of propolis are directly related to their biological activity [12,13]. Previous literature has discussed the great potential of propolis as an antimicrobial and antioxidant material [14,15] with immunological, antiparasitic, and cytotoxic properties [16], as well as antiviral activity against the SARS-Cov-2 virus [17].

Several products based on propolis, mainly as drinks and health foods, have been commercialized. The function of propolis as a supplement and bioactive food preservative has caused constant growth in the demand for similar products [9,18]. Users of this bee product have gained great benefits related to the biological activities and volatile components of propolis [8].

Propolis is a phytochemical and complex mixture composed of 50% resin (containing flavonoids and 66 phenolic acids), 30% wax, 10% essential oil, 5% pollen, and 5% other organic compounds [5,19]. Studies have reported that the volatile compound profile of propolis comprises a variety of chemicals and volatile compounds such as 1-methyl-naphthalene, naphthalene, 3-methyl-1-butanol [20], limonene, β-caryophyllene, nerolidol [21], (E)-isoeugenol, linalool, butanoic acid [22], and acetophenone [23]. The chemical composition of propolis is related to the biosynthetic capacity of plants and their secondary metabolites used by bees [9,24]. In this context, the present study aims to evaluate the chemical composition of the volatile fraction of seven samples of propolis collected in the Northeast of Pará, Eastern Amazon.

2. Results and Discussion

The chemical composition of the volatile compounds of the different propolis samples from Apis mellifera, as analyzed by gas chromatography mass spectroscopy (GC-MS) and gas chromatography flame ionization detection (GC-FID) are shown in Table 1. In total, 87 compounds were identified, demonstrating the diverse chemical composition of the volatile compounds in the propolis. Chi et al. [25] identified approximately 406 compounds, mainly comprising monoterpenes, phenol alcohols, sesquiterpenoids, acid esters, aldoketones, and hydrocarbons. However, in the present study, the predominant classes were sesquiterpene hydrocarbons (80.6–89.2%), followed by oxygenated monoterpenes (3.6–8.4%). The class of phenylpropanoids (0.2%), was identified only in sample 3 (Table 1). Phenyl propanoids are the predominant class in red propolis from Brazil, followed by hydrocarbon sesquiterpenes [16]. In propolis from the Cerrado biome (Campo Grande, MS, Brazil), sesquiterpenes, hydrocarbons, and oxygenated compounds are the main components [26].

Table 1.

Chemical composition (%) of volatile compounds identified in different propolis samples of Apis mellifera collected in the city of São João de Pirabas state of Pará.

Constituent RIL RIC Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Sample 7
2-Heptanone 889 a 888 0.4 0.45
α-Pinene 932 a 933 0.2 0.8 0.6 0.4 0.5 0.7 0.86
Benzaldehyde 952 a 953 0.1 1 0.61
6-Methyl-5-hepten-2-one 981 a 985 0.5 0.7 0.7 0.5 1.2 0.8 1.16
p-Cymene 1020 a 1022 0.1 0.1 0.1 0.1 0.1 0.1
Limonene 1024 a 1025 0.1 0.1 0.1 0.1 0.1 0.1
1,8-Cineole 1026 a 1027 0.2 0.1 0.1 0.1
(Z)-Linalool oxide (furanoid) 1067 a 1069 0.1 0.4 0.1 0.1 0.1 0.2
trans-Linalool oxide (furanoid) 1084 a 1090 0.1 0.1 0.1 0.1 0.1 0.33
Linalool 1095 a 1100 0.8 0.6 0.8 0.5 1.3 0.7 1.51
Naphthalene 1178 a 1182 4.3 1.4 1.1 7.4 5.8 5 4.99
Methyl chavicol 1195 a 1197 0.3 0.2 0.8 0.3 0.4 0.3 0.3
β-Cyclocitral 1217 a 1217 0.1
n-Decanal 1201 a 1229 0.3 0.06
Neral 1235 a 1235 0.1
Geranial 1264 a 1266 0.1 0.1
Benzenepropanoic acid. methyl ester 1278 b 1272 0.09
(Z)-Methyl cinnamate 1299 a 1280 0.1
(E)-Anethole 1282 a 1282 5.3
2-Undecanone 1293 a 1292 0.17
Tridecane 1300 a 1300 0.1
α-Cubebene 1345 a 1345 1.3 2.6 3.7 3.4 1.1 2.1 5.01
α-Ylangene 1373 a 1367 1.6 0.9 0.9 1.3 1.5 1.6 0.7
α-Copaene 1374 a 1375 8.2 15.4 17.1 16.2 9.4 14.3 22.19
β-Patchoulene 1379 a 1378 0.5 0.2
2-epi-α-Funebrene 1380 a 1380 0.3 0.6 0.26
α-Duprezianene 1387 a 1387 0.3
β-Bourbonene 1387 a 1387 0.3 0.1 0.3
β-Elemene 1389 a 1389 0.5
7-epi-Sesquithuejene 1390 a 1391 0.7 1.5 0.5 0.6
Cyperene 1398 a 1398 12.6 1.1 3.2 3.6 10 4.6
α-Gurjunene 1409 a 1400 0.8 0.7 0.5 0.7 0.5 7.77
(Z)-α-Bergamotene 1411 a 1411 5.3 4.5 2 2.5 2.3
2-epi-β-Funebrene 1411 a 1412 2 3.3
β-Caryophyllene 1417 a 1418 11.8 9 6.2 7.9 21.7 13.6 17.69
β-Cedrene 1419 a 1421 0.4 0.4 0.5 0.25
β-Copaene 1430 a 1426 0.3 0.1 0.3 0.4 0.4 0.26
(E)-α-Bergamotene 1432 a 1430 1.4 22.1 19.1 4.9 2.8 7.1 1.81
α-Guaiene 1437 a 1434 0.31
6,9-Guaiadiene 1442 a 1437 0.6 0.4 0.7 0.6 0.4 0.13
Aromadendrene 1439 a 1439 1.7 0.1 0.1 0.2
trans-Muurola-3,5-diene 1451 a 1445 0.88
α-Humulene 1452 a 1451 0.9 0.5 0.1 1 3 4.3
(E)-β-Farnesene 1454 a 1454 6.6 1 2.8 7 8 3
Rotundene 1457 a 1456 2 0.7 0.1 1 0.1 0.83
Allo-aromadendrene 1458 a 1458 0.5 0.6 1.7 0.4 0.07
(Z)-cadina-1(6),4-diene 1461 a 1467 0.8 0.9 0.9 0.68
(Z)-Muurola-4(14),5-diene 1465 a 1470 1.7 1.9 2.9 1.49
4,5-di-epi-Aristolechene 1471 a 1471 0.5
β-Acoradiene 1469 a 1474 0.9 0.8 1 0.7 1 0.7
γ-Gurjunene 1475 a 1475 0.5
ar-Curcumene 1479 a 1477 0.4 1 1 1.5 0.7 1.7 0.49
γ-Muurolene 1478 a 1478 1 6.6 3.3
β-Selinene 1489 a 1483 9.9 0.6 0.7 1 2.2 1.6 1.53
(E)-Muurola-4-(14),5-diene and 1493 a 1486 0.5 0.8 0.7 0.4 0.3 0.48
α-Selinene 1498 a 1489 9.8 1.3 1.3 1.5 2.3 2.3 1.64
α-Muurolene 1500 a 1493 0.3 0.6 0.9 0.4 0.7 0.45
Cis-cadina-1,4-diene 1495 a 1495 0.8
(E)-β-guaiene 1502 a 1497 0.2 0.4 0.3 0.19
β-Bisabolene 1505 a 1504 2.3 3.4 7 4 7.8 10 5.46
(E,E)-α-Farnesene 1505 a 1505 1.1
γ-Cadinene 1513 a 1507 1.2 0.1 0.7 1.1 1.3 0.8 0.48
δ-Cadinene 1522 a 1513 5.4 4.4 5.9 12.6 4.3 7 3.76
Sesquicineole 1515 a 1515 0.1
(E)-Calamenene 1521 a 1516 1.9 1.1 2.3 2.4 1.2 1.9 1.64
β-Sesquiphelandrene 1521 a 1521 0.5
(E)-cadina-1,4-diene 1533 a 1527 0.3 0.4 0.3 0.5 0.3 0.25
α-Cadinene 1537 a 1530 0.4 0.1 0.2 0.3 0.6 0.2 0.12
α-Calacorene 1544 a 1535 0.6 0.4 0.6 1 0.6 0.1 0.28
β-Calacorene 1564 a 1544 0.6
Elemicin 1555 a 1555 0.2
(E)-Nerolidol 1561 a 1558 0.2 1.2 0.3 0.1 0.3 0.43
Caryophyllenyl alcohol 1570 a 1570 0.1
Caryolan-8-ol 1571 a 1571 0.1
Caryophyllene oxide 1582 a 1576 0.4 0.1 0.1 0.2 0.5 0.3 0.2
Sphatulenol 1577 a 1577 0.1
Gleenol 1586 a 1590 0.1 0.1
Hexadecane 1600 a 1600 0.1 0.1
Junenol 1618 a 1603 0.2 0.3 0.1 0.3 0.3 0.6 0.18
α-Corocalene 1622 a 1622 0.1 0.1 0.2
1,10-di-epi-Cubenol 1618 a 1623 0.2 0.2 0.2 0.3 0.1 0.3 0.17
Cubenol 1514 a 1638 0.2 0.2 0.2 0.3 0.4 0.19
α-Cadinol 1652 a 1650 0.2 0.1 0.1 0.1 0.1 0.2 0.11
Cadalene 1675 a 1667 0.2 0.09
β-Bisabolol 1674 a 1674 0.4 0.1 0.1
epi-α-Bisabolol 1683 a 1683 0.1 0.1 0.2 0.1 0.2
α-Bisabolol 1685 a 1685 0.1 0.1 0.12
Hydrocarbon monoterpene 0.9 3 1.4 1 1.7 1.6 3.18
Oxygenated monoterpene 5.7 3.6 8.4 8.6 7.5 6.4 7.78
Hydrocarbon sesquiterpene 84.9 80.6 85.5 88.3 87 89.2 85.49
Oxygenated sesquiterpene 1.6 2.5 1 1.8 1.3 2.6 1.5
Phenylpropanoids 0.2
Others 0.1 0.37
Total 93.2 89.7 96.5 99.7 97.5 99.8 98.32
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Org = organic; Min = mineral; Cont. = control; RI(C): Calculated Retention Index; RI(L): Literature Retention Index. (a) Adams [52]; and (b) Nist [53].

Multivariate analyses, principal component analysis (PCA) (Figure 1), and hierarchical cluster analysis (HCA) were performed to analyze the correlation between the classes of compounds identified in the different samples, as shown in Figure 1 and Figure 2. As shown in Figure 1, the principal components (PC) contained the main components analyzed, PC1 and PC2, which accounted for 42.8% and 26.1% of the variables, respectively. In combination, both variables accounted for 68.9% of the variance in the analyzed data. In the HCA analysis, the similarity between the identified classes was evaluated; four groups were observed. Group I, including samples 1, 4, 5, and 6, showed a similarity of 51.04% (Figure 2), and comprised oxygenated monoterpenes and hydrocarbon sesquiterpenes (Figure 1). Groups II, III, and IV contained only one sample each and comprised phenylpropanoids, oxygenated sesquiterpenes, and hydrocarbon monoterpenes, with similarities of 23.02%, 16.32%, and 0%, respectively (Figure 2). Because of the complex chemical composition of propolis, chemometric analysis is widely used in studies to define groups of chemically correlated samples [27,28,29,30,31,32].

Figure 1.

Figure 1

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Biplot (principal component analysis) from the analysis of volatile compound classes identified in the aromas of seven samples of bee propolis from Apis mellifera.

Figure 2.

Figure 2

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Dendrogram presenting the relational similarity of the volatile compound classes identified in the aromas of seven bee propolis samples from Apis mellifera.

The compounds identified at the highest concentrations are listed in Table 1. Sample 1 contained the following: naphthalene (4.3%), α-copaene (8.2%), cyperene (12.6%), β-caryophyllene (11.8%), (E)-β-farnesene (6.6%), β-selinene (9.9%), α-selinene (9.8%), and δ-cadinene (5.4%). The composition of sample 2 is as follows: α-copaene (15.4%), (Z)-α-bergamotene (5.3%), β-caryophyllene (9%), (E)-α-bergamotene (22.1%), and δ-cadinene (4.4%). For sample 3, the components present were: (E)-anethole (5.3%), α-copaene (17.1%), (Z)-α-bergamotene (4.5%), β-caryophyllene (6.2%), (E)-α-bergamotene (19.1%), β-bisabolene (7%), and δ-cadinene (5.9%). Sample 4 contained the following: naphthalene (7.4%), α-copaene (16.2%), β-caryophyllene (7.9%), (E)-α-bergamotene (4.9%), (E)-β-farnesene (7%), γ-muurolene (6.6%), β-bisabolene (4%), and δ-cadinene (12.6%). In sample 5, naphthalene (5.8%), α-copaene (9.4%), cyperene (10%), β-caryophyllene (21.7%), (E)-β-farnesene (8%), β-bisabolene (7.8%), and δ-cadinene (4.3%). Sample 6 contained naphthalene (5%), α-copaene (14.3%), cyperene (4.6%), β-caryophyllene (13.6%), (E)-α-bergamotene (7.1%), β-bisabolene (10%), δ-cadinene (7%), β-bisabolene (10%), and δ-cadinene (7%). For sample 7, naphthalene (4.99%), α-cubebene (5.01%), α-copaene (22.19%), α-gurjunene (7.77%), β-caryophyllene (17.69%), α-humulene (4.3%), and β-bisabolene (5.46%) were present. These results are qualitatively similar to those reported in the literature [19,33].

In other studies, the major compounds were thymol (29.61%), its isomer carvacrol (30.57%) from Kermanshah City in the west of Iran [34], carvone (40.34%), β-bisabolene (10.6%), β-thujone (11.45%), carvone (40.34%) from Tehran Province, Iran [35], carvacrol (20.7%), acetophenone (13.5%), spathulenol (11.0%), (E)-nerolidol (9.7%), β-caryophyllene (6.2%) from Atlantic Forest in São Lourenço MG, Brazil [36], β-pinene (2.0–21.8%), α-pinene (1.2–46.5%), limonene (11.6%), dihydrosabinene (17.8%), 1,8-cineole (0.1–11.0%), p-cymene (0.1–5.3%), 2,7-dimethyl-3-octen-5-yne (trace-11.7%), octanal (12.9%), (E)-β-ocimene (17.8%), α-thujene (trace-11.0%), and styrene (13.5%) from South Africa [37], δ-cadinene (1.29–13.31%), γ-cadinene (1.36–8.85%) and α-muurolene (0.78–6.59%), β-eudesmol (2.33–12.83%), T-cadinol (2.73–9.95%) and α-cadinol (4.84–9.74%) from different Italian regions [19], and α-pinene, β-pinene, γ-terpinene, α-muurolene, γ-cadinene and δ-cadinene from different regions of Croatia [38].

Multivariate analysis, principal component analysis (PCA) (Figure 3), and hierarchical cluster analysis (HCA) (Figure 4) were applied to the chemical compounds identified in the different volatile compounds present in the aroma fractions of propolis samples from Apis mellifera. The first component PC1 accounted for 32.5% of the variation, while PC2 accounted for 23.3% of the variation. Combined, both components comprised 55.8% of the variance (Figure 3). HCA, considering the Euclidean distances and complete bonds, confirmed the formation of two distinct groups, without group I shown in Figure 1. The first of these, formed by samples I, IV, V, and VI, with a similarity of 12.29% (Figure 4), comprised 7-epi-sesquithuejene, allo-aromadendrene, δ-cadinene, sesquicineole, (E)-calamenene, β-sesquiphelandrene, (E)-cadina-1,4-diene, α-cadinene, α-calacorene, γ-muurolene, (E)-calamenene, (E)-β-farnesene, α-langene, naphthalene, γ-cadinene, cyperene, aromadendrene, α-selimene, β-selimene, and rutundene (Figure 3). The second group was formed by grouping samples II, III, VI, and VII, with a similarity of 16.01% (Figure 4). This arose from the consolidation of the following compounds: arcucumene, (Z)-α-bergamotene, β-acoradiene, (E,E)-α-farmasene, (E)-nerolidol, α-copaene, (E)-anthole, (Z)-muurola-4(14),5-diene, β-bisabolene, α-cubebene, 6-methyl-5-hepten-2-one, α-humulene, 2-epi-b-funebrene, linalool, and β-carophyllene (Figure 3).

Figure 3.

Figure 3

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Biplot (PCA) from the analysis of volatile compounds identified in the aromas of seven samples of bee propolis from Apis mellifera.

Figure 4.

Figure 4

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Dendrogram representing the relational similarity of the volatile compounds identified in the aromas of seven bee propolis samples from Apis mellifera.

The difference between the chemical composition of the present samples (Table 1) and those reported in the literature may be related to the geographical origin and the biome in which the bees collected the raw materials to form the propolis [8]. The isolation and analysis techniques [39] can also directly influence the chemical composition of both the volatile compounds and compounds of higher molecular weight, or those with greater polarity [40]. Olegário et al. [41] used PCA to determine the volatile compounds that quantitatively constituted propolis samples collected in different regions of Brazil. The geographic origin of the samples influenced their chemical composition in all the cases analyzed by the authors.

Because propolis is a product of plant origin, its chemical composition depends on factors including local flora, place of collection, and the seasonal and circadian period of collection of raw materials by bees, as the plants producing volatile compounds tend to produce different compounds at different times. The period of the year, climate and temperature, and rainfall index, among other factors, can induce variability in the chemical composition of propolis. Furthermore, the volatile compounds identified in propolis can be added to other analyses of chemical composition and serve as markers to identify their botanical origin [8,42]. This was also observed in propolis samples from Morocco [43], the northeastern states of Brazil [44], Yemen [45], other regions of Brazil, Estonia, China, Uruguay [46], South Africa [37], and Argentina [21]. Volatile compounds constitute a small fraction of propolis and are important for characterizing its botanical origin [47]. In addition, volatile compounds can be used as food preservatives in propolis-based packaging [48] by exploiting their antioxidant [15,25], antifungal [49,50], antibacterial [51], and other biological activities [23].

3. Materials and Methods

3.1. Collection Area

Apis mellifera propolis samples were collected in apiaries located in the city of São João de Pirabas, which is in the northeastern region of the state of Pará-Eastern Amazon (geographic coordinates: 0°46′08″ S 47°10′26″ O). The samples were collected from seven different hives from a producer. The hives were arranged at a distance of 2 m from each other in a forest with different types of plants, as shown in the Supplementary Material S1. Propolis was collected with the aid of sterile spatulas. According to the methodology described by Dutra et al. [54], the samples were placed in sterile plastic bags and kept at a temperature of 5–10 °C after collection, see Supplementary Material S1.

3.2. Aroma Extraction

Before the aroma extraction process, the propolis samples were frozen and crushed. For aroma extraction, 10 g of the sample was mixed with water (20 mL) and subjected to simultaneous distillation–extraction (SDE) for 3 h using a Chrompack Micro-Steam Distillation Extractor (Likens–Nickerson) and pentane (2 mL) as the organic mobile phase, as described in the literature [55,56].

3.3. Analysis of Chemical Composition of Volatile Compounds

The chemical compositions of the volatile fraction of the seven propolis samples was analyzed using GC-MS via a Thermo DSQ-II system equipped with a DB-5MS silica capillary column (30 m × 0.25 mm; 0.25 mm). For this analysis, the following conditions were used: the temperature was increased from 60 to 240 °C at a rate of 3 °C/min; the injector temperature was set to 240 °C; helium was used as the carrier gas (linear velocity of 32 cm/s, measured at 100 °C); aqueous 2:1000 n-hexane was injected in one step (0.1 mL); the temperature of the ion source and other parts was set at 200 °C. The quadrupole filter was swept in the range of 39–500 Da every second. Ionization was achieved by using an electronic impact technique at 70 eV. The volatile components were identified by comparison with the literature [52,53]. The volatile constituents were quantified by peak-area normalization using the FOCUS GC/FID, as previously reported by our research group [42].

3.4. Statistical Analysis

Multivariate analysis was performed according to a previously reported methodology [42,57,58] using Minitab 17® software (free version, Minitab Inc., State College, PA, USA). The chemical constituents of the essential oils were used as the variables. The raw data were first standardized to the same “weight.” PCA was then performed using the matrix type correlation configuration in the software. In the HCA of the samples, the Euclidean distance options were used for distance measurement, and the connection method used was complete. Multivariate analysis was applied to the samples, where the concentration of the compounds was ≥1%.

4. Conclusions

Different volatile compounds present in the aroma were obtained from the analyzed samples of propolis. Compounds belonging to the sesquiterpene class were present in the highest concentrations. Variability of the samples was observed using multivariate analysis. This may be related to the bee collection area. Based on the analyzed data, different groups were delineated, both for the classes of compounds and for the compounds analyzed in the form of a correlation matrix. These data are important because they can provide guidelines for future studies on the botanical origins of propolis.

Acknowledgments

The author Mozaniel Santana de Oliveira thanks PCI-MCTIC/MPEG, as well as CNPq for the scholarship process number: [301194/2021-1]. The authors appreciate the support of the Federal Rural University of Semi-Arid, Empresa Brasileira de pesquisa Agropecuária (Embrapa) Amazônia oriental and Projetos integrados da Amazônia—Fundo-Amazônia BNDS 16.17.01.004.00.00. The authors would like to thank the Universidade Federal do Pará—Propesp/PAPQ—Programa de Apoio à Publicação Qualificada.

Supplementary Materials

The following are available online, Figure S1: Collection of propolis samples.

Click here for additional data file. (1.6MB, zip)

Author Contributions

Conceptualization, M.S.d.O., J.N.C. and O.O.F.; methodology, D.S.P., N.S.P., M.E.C.O., G.C.V., G.M.S.P.G., A.P.d.S.S.F. and E.H.d.A.A.; formal analysis, A.P.d.S.S.F. and E.H.d.A.A.; investigation, M.S.d.O.; original draft preparation, M.S.d.O., J.N.C. and O.O.F.; visualization, A.P.d.S.S.F. and E.H.d.A.A.; supervision, A.P.d.S.S.F. and E.H.d.A.A.; project administration, A.P.d.S.S.F. and E.H.d.A.A. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

Sample Availability

Samples of the compounds in the Museu Paraense Emílio Goeldi are available from the authors.

Footnotes

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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