Propolis Samples Research Articles

Wound Healing, Anti-Inflammatory and Anti-Oxidant Activities, and Chemical Composition of Korean Propolis from Different Sources

Propolis, such as is used as bio-cosmetics and in functional materials, is increasing because of its antioxidant medicinal benefits. However, its pharmacological and chemical composition is highly variable, relative to its geography and botanical origins. Comparative studies on three propolis samples collected from different regions in Korea have been essential for linking its provenance, chemical composition, and biological activity, thereby ensuring the efficient utilization of its beneficial properties. Here, we report the chemical composition and biological activities such as the antioxidant, wound healing, and anti-inflammatory effects of the ethanolic extract of Korean propolis collected from two regions. We compared the chemical constituents of three 70% ethanol-extracted (EE) samples, including the Andong, Gongju field (GF), and Gongju mountain (GM)-sourced propolis using Gas chromatography–mass spectrometry (GC–MS). The major and common components of these EE Korean propolis were flavonoids such as pinocembrin (12.0–17.7%), chrysin (5.2–6.8%), and apigenin (5.30–5.84%). The antioxidant property using a 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay of EEP showed substantial differences among samples with the highest from Andong. The sample 10% GM levigated in simple ointment was found to be the most active in wound healing activity based on the excision, incision, and dead space wound models. The potential of propolis for wound healing is mainly due to its evidenced properties, such as its antimicrobial, anti-inflammatory, analgesic, and angiogenesis promoter effects, which need further study. The anti-inflammatory activity and NO production inhibitory effect were highest in GM samples. However, GM and GF samples demonstrated similar inhibitory effects on the expression of inflammatory mediators such as iNOS, IL-1β, and IL-6. The presence of a higher concentration of flavonoids in Korean EE propolis might be responsible for their promising wound healing, anti-inflammatory, and antioxidant properties.

Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis

Objectives: The objective of this study is to evaluate the wound healing potential of Eurasian propolis by analyzing the phytochemical profile and the biological effects of three representative propolis samples. Methods: Specific colorimetric assays were used to estimate the total phenolic and flavonoid contents and the triterpenoids content. Some of the main components of Eurasian propolis (pinocembrin, pinobanksin, CAPE, chrysin and galangin) were analyzed using HPLC-DAD. Scavenging activity and total antioxidant capacity were assessed through DPPH and ORAC assays, respectively. Human keratinocyte, fibroblast, and monocytic cell lines were used for the biological in vitro analyses. The direct wound healing properties were tested through scratching assays and ELISA kits for the assessment of the production of growth factors (FGF-7, Latency Associated Peptide-LAP), while the indirect effects were evaluated through the estimation of the levels of MMP9, IL-1β, IL-8, and TNF-α using ELISA kits together with a cell-free test on the inhibition capacity on collagenases. Network Pharmacology analysis was employed to further explore possible mechanisms of the action of propolis on the healing process. Results: The analyses confirmed the high phenolic content of Eurasian propolis (142.50–211.30 mg GAE/g), dominated by flavonoids (95.50–196.80 mg Galangin Equivalents/g), and terpenes (431.50–650.00 mg β-sitosterol Equivalents/g), while also verifying the significant antioxidant (4.9–8.9 mM/g Trolox Equivalents) and antiradical (DPPH IC50 26.1–54.4 μg/mL) activities. The samples showed indirect wound healing properties by mitigating inflammation and remodeling (reduced IL-1β and MMP9) and potentially modulating the immune response (upregulated IL-8). In vitro studies confirmed these effects, demonstrating decreased MMP9 production and collagenase inhibition when cells were co-treated with propolis and a stressor. Propolis also suppressed IL-1β release in fibroblasts, although its impact on TNF-α was inconclusive. Notably, co-treatment upregulated IL-8 in monocytes, suggesting a potential immunomodulatory role. Conclusions: Eurasian propolis may not directly stimulate cell proliferation during wound healing. Its anti-inflammatory and immunomodulatory properties could indicate an indirect contribution in helping the process.

Antimicrobial Activity of Honey and Propolis from Alba County, Romania.

Investigating the quality of bee products obtained across different geographical regions and analyzing their antimicrobial activity is of significant interest to various scientific disciplines. This study focuses on comparing the antimicrobial activity of honey and propolis samples from different areas of Alba County, Romania. The quality parameters of five samples of two types of bee products (honey and propolis) were assessed. Then, the samples were tested to comparatively determine their antimicrobial properties against 12 species of bacteria (Escherichia coli, Salmonella typhimurium, Salmonella enteritidis, Salmonella anatum, Salmonella choleraesuis, Pseudomonas aeruginosa, Pseudomonas fluorescens, Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Bacillus subtilis, and Listeria monocytogenes) and 7 fungal strains (Candida albicans, Aspergillus niger, Aspergillus flavus, Penicillium chrysogenum, Rhizopus stolonifer, Fusarium oxysporum, and Alternaria alternata). Of the bacterial strains, the most sensitive to the action of honey samples were the two strains of Staphylococcus followed by P. fluorescens. The two strains of Pseudomonas and L. monocytogenes were the most sensitive to the activity of propolis. Of the fungal strains, F. oxysporum was the most sensitive to the actions of both honey and propolis, followed by P. chrysogenum in the case of honey samples and the two Aspergillus strains in the case of propolis. These findings indicate that bee products are rich sources of bioactive compounds exhibiting strong antimicrobial properties and significant potential for the development of new phytopharmaceutical products.

Antimicrobial activity of propolis agents on some pathogenic microbes

The study aims to investigate the antimicrobial activity of propolis obtained from different regions of Iraq compared with that of propolis obtained from Iran. Samples were investigated for their antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Eschericha coli, Klebsiella pneumoniae, Bacillus cereus , Staphylococcus epidermidis and Candida albicans using standard antimicrobial assays. Marked variations in the antimicrobial activity of the different propolis samples were observed, the method of extraction selected gives the highest antimicrobial activity and the best alcohol concentration using in the extraction of propolis , then the crude extract of propolis showed synergistic effect with some antibiotics in inhibition of pathogenic microorganisms.

Antimicrobial, Anti-biofilm, Anti-swarming, Anti-quorum Sensing Activities, and Cytotoxicity of Propolis Samples from Northeast of Türkiye

Aim of study: Studies on propolis have increased as it has been revealed that it contains biologically active molecules. In the current study, it was aimed to analyze biological activity, and cytotoxicity of ethanolic extract of three different propolis samples from Türkiye. Material and methods: The antibacterial activity of the extracts against 14 microorganisms was assessed using the agar well diffusion method and the microdilution method. Chromobacter violeceum was used in quorum-sensing assay, and Pseudomonas aeruginosa PAO1 strain was used in swarming and biofilm assays. Using the MTT test, the cytotoxic effect of the extracts was examined on the lung adenocarcinoma cell line (A549), pancreatic tumoral cell line (AR42J), breast cancer cell line (MDA-MB-231), and normal epithelial cell line (Vero). Main results: All propolis extracts were effective against 8/14 microorganisms included in the study. While all propolis extracts have shown anti-quorum sensing activity, there was not any anti-swarming and anti-biofilm activity in each sample. It was demonstrated that every propolis sample had a dose-dependent cytotoxic effect on the examined cell lines. Research highlights: Due to the biological activity shown by the propolis samples included in the study, it is considered that it has the potential to influence the creation of novel medications in the future.

Chemical Markers in Italian Propolis: Chrysin, Galangin and CAPE as Indicators of Geographic Origin

Knowledge of the chemical composition of propolis is crucial for understanding the characteristics of products of different origins, but also for quality control and regulatory purposes. To date, official monographs or official analyses that allow researchers to evaluate propolis in a proper way have not yet been released. This study focuses on the characterization of twenty-seven Italian propolis samples and the identification of chemical markers that define its geographical provenance. Total polyphenol (TP) and total flavonoid (TF) content, alongside the quantification of pinocembrin, chrysin, galangin, and caffeic acid phenethyl ester (CAPE), were identified as potential markers. Additionally, DPPH assays were conducted to evaluate the antiradical activity of propolis samples. Our findings demonstrated that TPs, TFs and pinocembrin differed in propolis of different origins, especially in samples from the islands. However, the quantification of the sum of chrysin and galangin and CAPE provided a clearer distinction of the geographical origin of the propolis samples. In contrast, the DPPH assay did not prove useful for this purpose, as most results were similar and, therefore, not significant. This study lays the groundwork for future research on propolis. These findings could contribute to the development of more refined methods for distinguishing propolis origins, enhancing the understanding, valuation and quality control of this natural product in various applications.

Phytochemical profiling and characterization of flavonoid derivatives from propolis sample and investigation of cytotoxic and antiprotozoal activities

Recently, the growth of consumer demand for functional foods with potential nutritional and health benefits led to rapid growth of analytical tools for profiling of bioactive metabolites and assure quality. Bee propolis is one of the most important bee products owing to its myriad health value. As a gummy exudate produced in beehives after harvesting from different plant species, bee propolis contains bioactive secondary metabolites. The current study aims to profiling the chemical composition of propolis samples from Nigeria using HPLC–UV-ELSD and with the aid of NMR-based analysis for assignment of metabolites classes abundant in Nigerian propolis. Red Nigerian propolis samples were subjected to phytochemical analysis using HPLC–UV-ELSD and NMR. Further chromatographic separation of promising fractions was performed by column chromatography and size exclusion chromatography. Screening of the antitrypanosomal and cytotoxic activities against Trypanosoma brucei and human leukemia cell lines (U937), respectively, was performed. The performance of LC–MS permitted identification of the different components from which 13 compound were identified and allowed combination of fractions to afford 9 fractions from which two isoflavonoids were isolated and identified using 1D and 2D NMR analysis with MS as isosativan and Medicarpin. Red Nigerian propolis crude extract showed the highest inhibitory activity at 6.5 µg/ml compared to moderate activity for the isolated compounds with MIC of 7.6 µg/ml and 12.1 µg/ml for medicarpin and isosativan, respectively. Moreover, the fraction RN-6 from the total extract showed the potent cytotoxic effect with IC50 = 26.5 µg/ml compared to standard diminazen which showed IC50 = 29.5 µg/ml.

Phenolic changes in propolis during in vitro digestion and cytotoxic effects on human cancer cell lines

PurposeThis study aimed to evaluate the compositional changes and bioaccessibility of phenolics and antioxidants in propolis during in vitro digestion as well as the cytotoxic effects of digested propolis on various cancer cell lines.Design/methodology/approachSix propolis samples were obtained and subjected to in vitro oral, gastric and intestinal digestion. Both digested and undigested samples were analyzed for their total phenolic, flavonoid and antioxidant activities. Additionally, changes in phenolic composition in the in vitro digestion system were revealed by the HPLC-DAD system. The cytotoxic effects of the digested samples were assessed on lung (A549, H1299), skin (A431), liver (Hep-G2) and colon (Caco-2) cancer cells as well as on fibroblast (Bj) cells.FindingsThe mean bioaccessibility values of phenolic and flavonoid compounds were found to be less than 35 and 24%, respectively, while the TEAC and CUPRAC antioxidant results ranged between 225.08–649.04 and 398.68–1552.28 µmol TE/g, respectively. The release of p-coumaric, ferulic, 3,4-dimethoxycinnamic acids, naringenin, pinocembrin and chrysin increased progressively from the oral to the intestinal stage. The cytotoxic effects of samples on cell lines were ranked, based on IC50 results, as A431 > Hep-G2 > Caco-2 > A549 > H1299 > Bj.Originality/valuePropolis has been recognized for centuries as a natural remedy, and numerous studies have explored its bioactive components. However, no studies have previously examined the changes in the phenolic compositions of propolis samples during digestion or their cytotoxic effects on cancer cells. Therefore, this study provides novel insights and an approach to the existing literature on this topic.Graphical abstract

Investigation of Antimicrobial and Antiviral Effects of Türkiye Propolis Water Extracts: An In Vitro Study

Propolis is a natural bee product that has been used as a therapeutic agent for centuries. Propolis extracts are natural resources that attract the attention of scientists looking for new components due to the insufficiency of existing drugs. In the current study, the antiviral and antimicrobial activity of propolis water extracts prepared from three different raw propolis samples were investigated. The total flavonoid contents (TFC) and total phenolic content (TPC) of the extracts were measured. It was determined that the TPC and TFC ranged from 5.87±0.36 to 20.47±1.46 mg GAE g-1, and 0.48±0.04 to 2.10±0.22 mg QUE g-1, respectively. The antimicrobial activity of the extracts against 14 microorganisms and their effect against the Pseudomonas aeruginosa biofilm were investigated. Additionally, anti-quorum sensing and anti-swarming activities of the extracts were tested. The antiviral activity of the extracts was examined by MTT and qRT-PCR methods. The water extracts of propolis samples included in the study did not show antimicrobial, anti-swarming, anti-quorum sensing, and anti-viral activities. However, the extracts were found to have strong anti-biofilm activities. The results show that aquatic propolis extracts can be evaluated in the treatment of biofilms.

Design of experiment for extraction optimization and antioxidant assay studies of biologically active compounds of propolis

Bees gather propolis from tree sap and combine it with their saliva to seal and sanitize the hive. There are various countries where propolis extracts are used in food and beverage as additives mainly due to the presence of phenolic compounds. Therefore, extraction is the initial and the most vital step in the purification and recovery of phenolic compounds from propolis. The purpose of this study is to enhance propolis extract yield by thoroughly optimizing the extraction process and to evaluate and compare the antioxidant assay of the propolis extracts. Several factors such as the chemical nature of phenolic compounds, composition of solvents, temperature and solvent pH may influence the extraction efficiency and quantity of phenolic compounds. These factors have been extensively studied by univariant optimization; however, the univariant optimization fails to consider the effect of interaction between variables on extraction efficiency. Therefore, the Design of Experiment (DoE) approach is adopted in this study in order to evaluate the correlation between various parameters and their impacts on the extraction of propolis in order to determine TPC, TFC, and DPPH scavenging activity. The DoE was carried out to correctly optimize different extraction parameters including the percentage of ethanol in extraction solvent, temperature and time. The ethanol percentage showed the highest F ratio and minimum p-value for the extraction of 28 total phenolic compounds and flavonoids with a negative correlation showing the presence of more polar compounds in propolis. While in the case of DPPH scavenging assay, the temperature was found to have a significantly positive effect on the extraction efficiency. Optimized extraction parameters obtained by the design were found to be different for TFC, TPC and DPPH, e.g., optimum solvent composition for TFC, TPC and DPPH were 51%, 50% and 90%, respectively. The ultrasonic extracts of propolis showed a higher TPC (574.017 µg GAE/g); in 51% ethanol/water at 25 °C while sonicated for only 2 min. The total flavonoid content (TFC) was found higher (353.16 µg Quercetin/g) in 50% ethanol/water when sonicated for 60 min at 56 °C. DPPH scavenging assay showed higher capacity (1617.27 µg Ascorbic acid/g) in 90% ethanol/water at 62 °C for 60 min sonication. Fourteen propolis samples from Pakistan and Turkey were extracted in order to compare TPC, TFC, and DPPH assay results. In Pakistani propolis, the TPC, TFC, and DPPH scavenging assay varies from 418–1026 µg/g, 264–866 µg/g, and 302–1162 µg/g, respectively. Turkish propolis was demonstrated to have higher TPC, TFC, and DPPH scavenging assay values than Pakistani propolis, with values of 1312–1535 µg/g, 1182–1446 µg/g, and 885–2756 µg/g, respectively. It was found that propolis produced naturally possesses more potential phenolic compounds than propolis purchased from farmer’s markets.

ANALYSIS OF PROPOLIS STINGLESS BEE BIOACTIVE COMPOUNDS FROM SEVERAL REGIONS IN INDONESIA

Objective: Propolis is a bee product collected by honeybees from a resinous substance of various plant sources. Its antioxidant activities are different from various geographic origins. This study aimed to analyze bioactive compounds by lC-MS/MS, compare antioxidant activity total phenolic and flavonoid contents in stingless bee propolis samples from several regions in Indonesia. Methods: The propolis samples were taken from stingless bee hives of Tetragonula clypearis from Sumbawa, Tetragonula laeviceps from Magelang, Tetragonula biroi from Bogor, and Geniotrigona thoracica from South Kalimantan). Analysis of bioactive compounds was identified by lC-MS/MS. The quantification of the chemical compound determined its total phenolics and flavonoid (TPC and TFC) contents. The antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay. Results: The results of this study obtained 10 compounds from G. thoracica, 8 compounds each from T. laeviceps and T. biroi, and 7 compounds from T. clypearis. The results of TPC was ranging from 57.24±5.35 to 139.39±15.79 mg GAE/g. The TFC was ranging from 22.13±0.79 to 37.20±3.63 mg QE/g. With the highest TPC and TFC was propolis of G. thoracica. The IC50 of antioxidant activity was ranging from 11.12 to 162.72 ppm. Conclusion: The compounds contained in propolis T. clypearis, T. laeviceps, G. thoracica and T. biroi have a potential as a new herbal candidate as antioxidant agents.

Identification of volatile compounds and evaluation of certain phytochemical properties of Turkish propolis

AbstractThe purpose of this study was to assess volatile component profiles and the antioxidant activity of propolis samples from eight different locations in Türkiye. α‐Pinene, β‐pinene, 3‐carene, limonene, 2‐acetylfuran, benzaldehyde, acetic acid, benzoic acid, longifolene, isopentyl acetate, m‐cymene, styrene, and δ‐cadinene were the most common volatile components found in the most of propolis samples. Principal component analysis (PCA) and hierarchical cluster analysis were performed on the gas chromatography–mass spectrometry (GC–MS) data to identify trends and clusters in the propolis samples. As a result of the PCA, the common components in all propolis were m‐cymene, decanal, α‐pinene, limonene, pinocarvone, benzaldehyde, and butanoic acid. The samples had 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH) inhibition activity ranging from 19.2% to 92.5% and 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) values ranging from 480 to 1370 μM trolox/g extract. The total phenolic content (TPC) ranged from 10,900 to 34,033 mg GAE/100 g. Compared with butylated hydroxytoluene (BHT) (control), all but one of the propolis samples exhibited higher DPPH activity. In addition, ABTS levels of propolis extracts were higher than those of BHT. These results unequivocally show that Turkish propolis has remarkable antioxidant qualities, which makes it a viable option for addition to food and medicine products.

Exudate and Propolis from Escallonia pulverulenta: Phytochemical Characterization and Antibacterial Activity.

Propolis is a complex mixture formed from exudates that bees collect from plants and then mix with beeswax and their own salivary enzymes. Chilean propolis is characterized by the presence of phenolic compounds, which are considered responsible for the biological activities. The endemic species Escallonia pulverulenta (Ruiz and Pav.) Pers. [Escalloniaceae] is a recognized source of exudate to produce propolis. This study reports for the first time the chemical profile and antibacterial activity of E. pulverulenta exudate and leaves, as well as two samples of Chilean propolis. Palynological and morphological analysis showed the presence of E. pulverulenta as one of the main species in the propolis samples. UPLC-MS/MS analyses enabled the identification of phenolic acids in the leaves and in the propolis. Conversely, flavonoids are mainly present in exudates and propolis. Quercetin is the most abundant flavonol in the exudate, with similar concentrations in the propolis samples. Nevertheless, the main compound present in both samples of propolis was the flavanone pinocembrin. The antibacterial results obtained for exudate and propolis have shown a similar behavior, especially in the inhibition of Streptococcus pyogenes. These results show the importance of the exudates collected by the bees in the chemical composition and antibacterial capacity of propolis.

In Vitro Antimicrobial Potential of Portuguese Propolis Extracts from Gerês against Pathogenic Microorganisms.

Antimicrobial resistance (AMR) is one of humanity's main health problems today. Despite all the breakthroughs and research over the past few years, the number of microbial illnesses that are resistant to the available antibiotics is increasing at an alarming rate. In this article, we estimated the biomedical potential of Portuguese propolis harvested from the Gerês apiary over five years, evaluating the in vitro antimicrobial effect of five hydroalcoholic extracts prepared from five single propolis samples and of a hydroalcoholic extract obtained from the mixture of all samples. The antimicrobial potential was firstly assessed by determining the minimum inhibitory concentration (MIC) of these extracts against a panel of three Gram-positive (Bacillus subtilis, methicillin-sensitive Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus) and one Gram-negative bacteria (Escherichia coli), as well as two yeasts (Candida albicans and Saccharomyces cerevisiae). As MIC values against each bacterium were consistent across all the evaluated propolis extracts, we decided to further conduct a disk diffusion assay, which included three commercial antibiotics-erythromycin, vancomycin, and amoxicillin/clavulanic acid-for comparison purposes. In addition to displaying a concentration-dependent antibacterial effect, the hydroalcoholic extracts prepared with 70% ethanol exhibited stronger antimicrobial capacity than vancomycin against B. subtilis (% of increase ranged between 26 and 59%) and methicillin-sensitive S. aureus (% of increase ranged between 63 and 77%). Moreover, methicillin-resistant S. aureus (MRSA) showed susceptibility to the activity of the same extracts and resistance to all tested antibiotics. These findings support that propolis from Gerês is a promising natural product with promising antimicrobial activity, representing a very stimulating result considering the actual problem with AMR.

Physicochemical Properties, Antioxidant Activity, and High-Performance Thin-Layer Chromatography Profiling of Propolis Samples from Western Australia.

This study reports on the physicochemical and antioxidant properties of propolis samples from various regions across Western Australia and identifies some phenolic constituents using high-performance thin-layer chromatography (HPTLC). Total phenolic content (TPC) was determined using a modified Folin-Ciocalteu assay, and antioxidant activity was investigated with the Ferric Reducing Antioxidant Power (FRAP) assay and also visualised and semi-quantified by HPTLC-DPPH analysis. TPC values ranged from 9.26 to 59.3 mg gallic acid equivalent/g of raw propolis and FRAP assay data from 4.34 to 53.8 mmol Fe2+ mmol/kg of raw propolis, although some of these variations might be related to differences in extraction yields obtained with 70% ethanol. The presence of luteolin, taxifolin, naringenin, and 4-hydroxyphenylacetic acid was confirmed based on a comprehensive, validated matching approach against an HPTLC-derived database. The findings of the study highlight the importance of future research on the chemical composition and bioactivity of Western Australian propolis.

Characterisation of New Zealand Propolis from Different Regions Based on Its Volatile Organic Compounds.

Propolis is a bee product mainly consisting of plant resins and is used by bees to maintain the structural integrity of the colony. Propolis is known to contribute to bee health via its antimicrobial activity and is a valued product for human use owing to its nutritional and medicinal properties. Propolis is often characterised into seven categories depending on the resin source. New Zealand propolis is typically assumed as being poplar-type propolis, but few studies have chemically characterised New Zealand propolis to confirm or reject this assumption. Here, for the first time, we characterise propolis originating from different regions in New Zealand based on its volatile organic compounds, using gas chromatography coupled with mass spectrometry (GC-MS). To support this characterisation, we also collected and analysed resin samples from a variety of resin-producing plants (both native to New Zealand and introduced). Our findings suggest that bees mainly use poplar as a resin source, but also utilize native plant species to produce propolis. While regional variation did not allow for clear separation between samples, some patterns emerged, with samples from some regions having more chemical complexity and a higher contribution from native species (as suggested by a higher number of compounds unique to native species resin). Further studies are needed to accurately identify the botanical sources contributing to these samples. It may be also of interest to explore the biological activity of regional propolis samples and their potential nutritional or medicinal benefits.

Stability of Propolis Phenolics during Ultrasound-Assisted Extraction Procedures.

Propolis has gained popularity in recent years as a potential preventive and therapeutic agent due to its numerous health benefits, which include immune system boosting, blood pressure lowering, allergy treatment, and skin disease treatment. The pharmacological activity of propolis is primarily attributed to phenolics and their interactions with other compounds. Given that phenols account for most of propolis's biological activity, various extraction methods are being developed. The resin-wax composition of the propolis matrix necessitates the development of an extraction procedure capable of breaking matrix-phenol bonds while maintaining phenol stability. Therefore, the aim of this study was to assess the stability of two major groups of phenolic compounds, flavonoids and phenolic acids, in propolis methanol/water 50/50 (v/v) extracts obtained after ultrasound-assisted extraction (USE) under different extraction parameters (extraction time and pH) and heat reflux extraction (HRE). The methodology involved varying the USE parameters, including extraction time (5, 10, and 15 min) and pH (2 and 7), followed by analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify phenolic recoveries. Results revealed that benzoic acid and chlorogenic acid derivatives demonstrated excellent stability across all ultrasound extraction procedures. The recoveries of flavonoids were highly diverse, with luteolin, quercitrin, and hesperetin being the most stable. Overall, neutral pH improved flavonoid recovery, whereas phenolic acids remained more stable at pH = 2. The most important optimization parameter was USE time, and it was discovered that 15 min of ultrasound resulted in the best recoveries for most of the phenols tested, implying that phenols bind strongly to the propolis matrix and require ultrasound to break the bond. However, the high variability in phenol extraction and recovery after spiking the propolis sample shows that no single extraction method can produce the highest yield of all phenols tested. As a result, when working with a complex matrix like propolis, the extraction techniques and procedures for each phenol need to be optimized.

Antibacterial activity of three ethanolic extracts of propolis from the municipalities of Sevilla, Cartago, and Armenia, Colombia, against MRSA (Methicillin-Resistant Staphylococcus aureus)

Introduction: Staphylococcus aureus is a pathogen commonly associated with hospital and community-acquired infections, particularly methicillin-resistant strains that impose limi- tations on treatment options. The resistance of these strains to beta-lactam antibiotics has spurred the search for new antimicrobial sources, and natural products such as propolis extracts have demonstrated efficacy against Methicillin-Resistant Staphylococcus aureus. The an- timicrobial activity of propolis depends to the flora used by the bees to make the propolis. Aim: This study aims to determine the antibacterial activity of three ethanolic extracts of propolis from the municipalities of Sevilla, Cartago, and Armenia, Colombia, against Methicillin-Resistant Staphylococcus aureus. Methodology: The methodology involved obtaining ethanolic extracts of propolis collected from different regions of Colombia, followed by fractionation and identification of the propolis with the highest bactericidal activity. Subsequently, an analysis using gas chromatography coupled with mass spectro- metry (GC-MS) was identified the predominant compound in the selected fractions. Re- sults: All three propolis samples evaluated in this study demonstrated bactericidal activity at concentration of 10 mg/mL, with the Cartago propolis showing concentrations below 5 mg/mL. Chromatographic analysis identified 3-Hexadecyloxycarbonyl-5-(2-hydroxye- thyl) as the predominant compound. Conclusion: In conclusion, the need for additional research to understand the pharmacological potential of Colombian propolis, specifically the bactericidal activity of Cartago propolis against Methicillin-Resistant Staphylococcus aureus.

Analysis and evaluation of quality parameters of commercial propolis products using a new high Performance Liquid Chromatography (HPLC) method and Comparison of antimicrobial properties

Propolis, which has an important place in functional foods, is a natural bee product with high biological active value and is consumed in different formulations and packages worldwide. Thanks to the active compounds it contains, propolis shows biological properties such as antibacterial, antifungal, antiviral, antiprotozoal, antitumoral, antiulcer and anti-inflammatory. However, it is seen that propolis supplements are prepared unconsciously in the market.Such commercial products need to be standardized. Within the scope of this study, characteristic compounds of Turkish propolis were initially determined with support from the literature. Subsequently, bioactive components of 23 commercial propolis products were analyzed using the new HPLC method. Then, the flavonoid content and antimicrobial activity of the samples were determined.According to the results of these analyses, the amounts of Caffeic Acid (2,16–1620,57 ppm, Quercetin (0,19–203,99 ppm), Apigenin (0,30–319,08 ppm), Kaempferol (0,56–251,15 ppm), Chrysin (3,97–2830,93 ppm), and Caffeic Acid Phenethyl Ester (13,08–9666,48 ppm) were found to be quite different. The highest flavonoid content was measured as P2: 20847,43 mg/L and the lowest was 00003,10 mg/L in sample P13. Samples prepared for children were found to be highly diluted in terms of content. Again, it was determined that the samples with high preservation of active ingredients were products with production certificates based on laboratory analyzes. The samples were generally found to have inhibition activity against Gram (+) bacteria. Propolis samples P6 and P22 showed high activity against Gram (+) and Gram (−) bacteria and even fungi. When we evaluated some products that are insufficient in terms of content and antimicrobial activity, it was observed that antimicrobial activity increased depending on the other auxiliary chemicals used in the products. In this context, propolis products, which are very valuable in terms of health, may lose their real activity due to uncontrolled production and will gradually be devalued in the market. It is important that this valuable product should be produced and offered to the market with the necessary analysis methods without losing its effectiveness.

The Influence of Geographical Origin on Poplar Propolis Composition and the Impact of Human Microbiota.

Ethanol extracts obtained from 13 poplar propolis samples originating from various European countries by traditional maceration were tested for total polyphenols, flavonoid content, and antioxidant activity. Moreover, the content of 18 polyphenolic compounds (from the group of phenolic acids and flavonoids) was determined using the HPLC method. The inhibitory effect of six selected extracts with the highest activity was assessed by well-diffusion method against five strains (Bifidobacterium spp., L. rhamnosus, L. acidophilus, E. coli, and Bacteroides spp.) of intestinal bacteria self-isolated from the faeces of obese probands with the use of selective media. It was found that the antioxidant activity of propolis varied depending on geographical origin and even among samples from the same region, which indicates that some other factors also influence propolis quality. The samples of different geographical origin varied mainly in the share of individual phenolic compounds, and it was not possible to find a characteristic marker of origin, excluding the galangin present in the Polish samples only. Assessing the inhibitory activity of propolis (in the range of 70 mg to 10 µg per mL) indicated that the concentration of 100 µg/mL was found as being safe for tested fecal bacteria (Bifidobacterium spp., L. rhamnosus, L. acidophilus, E. coli, and Bacteroides spp.). As no negative effect of low doses of propolis on the intestinal microflora was found, it can be suggested that its use in recommended doses brings only beneficial effects to the body.