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Affiliation(s)

1. Croatian Institute for Experimental and Translation Oncology (CIETO), Koledinečka 03, Zagreb 10040, Croatia
2. Institutes of Biomedical Sciences, Medical Faculty of Maribor, Taborska 8, Maribor 2000, Slovenia
3. Retired, Glavarjeva 19, Ljubljana 1000, Slovenia
4. Research and Developement, Medex D.o.o., Linhartova 049a, Ljubljana 1000, Slovenia
5. Institutes of Microbiology and Immunology, Medical Faculty, Zaloška 4, Ljubljana 1105, Slovenia

ABSTRACT

Propolis is a resinous natural product, produced by bees (Apis mellifera), from vegetable parts and plant secretions. Propolis’ samples A, B, C and D were extracted with phosphate buffer saline (PBS) or with 70% EtOH at pH values 8.0, 7.2 and 6.4 followed by: (1) reverse-phase high-performance liquid chromatography (RP-HPLC) on Purospher® Star RP-18 column, the quantity of caffeic acid, chrysin, pinocembrin and galangin was determined; (2) determination of total flavonoids in both extracts; (3) antimicrobial tests of both extracts against (a) Gram-positive bacteria: methicillin-resistant Staphylococcus aureus (MRSA), St. aureus, Streptococcus pyogenes, Str. agalactiae, (b) Gram-negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Acinetobacter baumanii and (c) yeast: Candida albicans. The antimicrobial activity of propolis’ PBS extracts against Gram-positive bacteria shows the lowest minimal inhibitory concentration (MIC, mg/mL) at pH 8.0 in sample C, followed by A, B and D. In sample C, MICs at pH 8.0 were 0.007 (Str. agalactiae), 0.015 (MRSA), 0.015 (Str. pyogenes) and 0.007 (St. aureus). The polyphenol content of sample C is: flavonoid content 5.47 ± 0.62 mg/mL, caffeic acid 1.33 ± 0.92 mg/mL, chrysin 41.02 ± 4.22 µg/mL, pinocembrin 2.93 ± 0.33 mg/mL and galangin 41.87 ± 4.23 mg/mL. PBS extracts against Gram-negative bacteria show the lowest MIC (mg/mL) at pH 8.0 in sample D, followed by B, C and A. In sample D, MICs at pH 8.0 were 0.003 (Acin. baumanii, Pr. mirabilis, Ps. aeruginosa) and 0.007 (E. coli). Polyphenol content of sample D is: flavonoids 8.28 ± 0.92 mg/mL, caffeic acid 3.56 ± 0.32 mg/mL, chrysin 677.42 ± 68.42 μg/mL, pinocembrin 146.49 ± 13.89 mg/mL and galangin 59.81 ± 5.86 mg/mL. The strongest anti C. albicans activity, with the lowest MIC (mg/mL), at pH 8.0 was in the sample C, followed by samples D, A and B. In sample C, the MIC at pH 8.0 is 0.001 (PBS extract). The antimicrobial activities of selected propolis samples correlate with their polyphenol content, more precisely, flavonoid, caffeic acid, chrysin, pinocembrin and galangin content.

KEYWORDS

Propolis, pH, phosphate buffer saline extract, 70% EtOH extract, antimicrobial activity.

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