Comparison of Total Phenolic Content and Antioxidant Activity of Indonesian Propolis Extracted with Various Solvents.

  • Diah Kartika Pratami Faculty of Pharmacy, Universitas Indonesia, Cluster of Health Sciences Building, Depok, West Java, 16424, Indonesia
  • Nur Elistiani Eksadita Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, West Java, 16425, Indonesia
  • Muhamad Sahlan Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, West Java, 16425, Indonesia
  • Abdul Mun’im National Metabolomics Collaborative Research Center, Faculty of Pharmacy, Universitas Indonesia, Kampus UI Depok, West Java, 16424, Indonesia
  • Asep Bayu Research Center for Vaccine and Drugs, National Research and Innovation Agency, Jalan Raya Jakarta Bogor KM. 46, Cibinong, Bogor 16911, Indonesia
  • Kaysa Faradis Mahira Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan


Propolis is a widely used medicine that may be found in both its pure form and when combined with other
natural ingredients in over-the-counter preparations, cosmetics, and health foods. Some customers develop alcohol sensitivity after using ethanol, which is a common solvent to extract propolis. As propolis solvents, various Natural Deep Eutectic Solvents (NaDES) were examined in this study. Choline chloride, propylene glycol, glycerol, citric acid, and 1,2-propanediol were combined in molar ratios to create some of the NaDES that were employed. Propolis and solvents were used in a kinetic maceration ratio for the extraction process. The Folin-Ciocalteu method was used to measure the total phenolic content (TPC), and the DPPH assay was used to measure antioxidant activity. In ethanol and NaDES solvents, the TPC of propolis extract ranged from 136.52 ± 27.9 to 365.8 ± 20.54 mgGAE/g. The values for antioxidant activity were 45.94 to 183.76 ppm. The propolis extract with the lowest IC50 and the highest TPC content was choline chloride-glycerol (CCG). It was discovered that the optimum NaDES solvent for extracting propolis was CCG-based. NaDES may be a potential solvent for use in Indonesian propolis, both as an extraction medium and as a formulation element.


1. Toreti VC, Sato HH, Pastore GM, Park YK. Recent progress of propolis for its biological and chemical compositions and its botanical origin. Evidence-based Complement Altern Med. 2013;2013:1–13.

2. de Groot AC. Propolis: a review of properties, applications, chemical composition, contact allergy, and other adverse effects. Dermat contact, atopic, Occup drug. 2013;24(6):263–82.

3. Popova M, Trusheva B, Bankova V. Propolis of stingless bees: A phytochemist’s guide through the jungle of tropical biodiversity. Phytomedicine. 2021;86(June 2019):153098.

4. Pratami DK, Mun’im A, Sundowo A SM. Phytochemical profile and antioxidant activity of propolis ethanolic extract from Tetragonula bee. Pharmacogn J. 2018;10(1):73–80.

5. Miyata R, Sahlan M, Ishikawa Y, Hashimoto H, Honda S, Kumazawa S. Propolis components and biological activities from stingless bees collected on South Sulawesi, Indonesia. HAYATI J Biosci. 2020;27(1):82–8.

6. Zulhendri F, Perera CO, Chandrasekaran K, Ghosh A, Tandean S, Abdulah R, et al. Propolis of stingless bees
for the development of novel functional food and nutraceutical ingredients: A systematic scoping review of the experimental evidence. J Funct Foods. 2022;88(December 2021):104902.

7. Pujirahayu N, Suzuki T, Katayama T. Cycloartane-type triterpenes and botanical origin of propolis of stingless Indonesian bee Tetragonula sapiens. Plants. 2019;8(57):1–14.

8. Bankova V, Trusheva B, Popova M. Propolis extraction methods: a review. J Apic Res. 2021;60(5):734–43.

9. Trusheva B, Petkov H, Popova M, Dimitrova L, Zaharieva M, Tsvetkova I, et al. “Green” approach to propolis
extraction: natural deep eutectic solvents. Comptes Rendus L’Academie Bulg des Sci. 2019;72(7):897–905.

10. Picot-Allain C, Mahomoodally MF, Ak G, Zengin G. Conventional versus green extraction techniques — a comparative perspective. Curr Opin Food Sci. 2021;40:144–56.

11. Chemat F, Vian MA, Ravi HK, Khadhraoui B, Hilali S, Perino S, et al. Review of alternative solvents for green extraction of food and natural products: Panorama, principles, applications and prospects. Molecules. 2019;24(16):1–27.

12. Skarpalezos D, Detsi A. Deep eutectic solvents as extraction media for valuable flavonoids from natural sources. Appl Sci. 2019;9(19):1–23.

13. Obluchinskaya ED, Pozharitskaya ON, Zakharova L V, Daurtseva A V, Flisyuk E V, Shikov AN. Efficacy of natural deep eutectic solvents for extraction of hydrophilic and lipophilic compounds from fucus vesiculosus. Molecules. 2021;26(14):4198.

14. Cunha SC, Fernandes JO. Extraction techniques with deep eutectic solvents. TrAC - Trends Anal Chem. 2018;105:225–39.

15. Bankova V, Bertelli D, Borba R, Conti BJ, da Silva Cunha IB, Danert C, et al. Standard methods for Apis mellifera propolis research. J Apic Res. 2019;58(2):1–49.

16. Anjum SI, Ullah A, Khan KA, Attaullah M, Khan H, Ali H, et al. Composition and functional properties of propolis (bee glue): A review. Saudi J Biol Sci. 2019;26(7):1695–703.

17. Margeretha I, Suniarti DF, Herda E, Mas’ud ZA. Optimization and comparative study of different extraction methods of biologically active components of Indonesian propolis Trigona spp. J Nat Prod. 2012;5:233-42.

18. Fikri AM, Sulaeman A, Handharyani E, Marliyati SA, Fahrudin M. The effect of propolis administration on fetal development. Heliyon [Internet]. 2019;5(10):e02672. Available from:

19. Pujirahayu N, Ritonga H, Uslinawaty Z. Properties and flavonoids content in propolis of some extraction method of raw propolis. Int J Pharm Pharm Sci. 2014;6(6):338–40.

20. Benoit C, Virginie C, Boris V. The use of NADES to support innovation in the cosmetic industry. R. V, G.-J. W,
Y.H. C, editors. Vol. 97, Advances in Botanical Research. GATTEFOSSÉ Research & Development Centre, Saint-Priest Cedex, France: Academic Press Inc.; 2021. p. 309–32.

21. Zain MSC, Yeoh JX, Lee SY, Shaari K. Physicochemical properties of choline chloride-based natural deep eutectic solvents (Nades) and their applicability for extracting oil palm flavonoids. Sustain. 2021;13(23):1-18.

22. Liu Y, Friesen JB, McAlpine JB, Lankin DC, Chen S-N, Pauli GF. Natural Deep Eutectic Solvents: Properties, Applications, and Perspectives. J Nat Prod. 2018;81(3):679–90.

23. Funari CS, Sutton AT, Carneiro RL, Fraige K, Cavalheiro AJ, da Silva Bolzani V, et al. Natural deep eutectic solvents and aqueous solutions as an alternative extraction media for propolis. Food Res Int. 2019;125(108559):1-15.

24. Hikmawanti NPE, Ramadon D, Jantan I, Mun’im A. Natural deep eutectic solvents (Nades): Phytochemical extraction performance enhancer for pharmaceutical and nutraceutical product development. Plants. 2021;10(3390):1-18.

25. Yusuf B, Astati SJ, Ardana M, Herman, Ibrahim A, Rijai L, et al. Optimizing natural deep eutectic solvent citric acid-glucose based microwave-assisted extraction of total polyphenols content from Eleutherine bulbosa (Mill.) bulb. Indones J Chem. 2021;21(4):797–805.

26. Herman, Ibrahim A, Rahayu BP, Arifuddin M, Nur Y, Prabowo WC, et al. Single factor effect of natural deep eutectic solvent citric acid-glucose based microwave-assisted extraction on total polyphenols content from
mitragyna speciosa Korth. Havil Leaves. Pharmacogn J. 2021;13(5):1109–15.

27. Costa AG, Yoshida NC, Garcez WS, Perdomo RT, Matos M de FC, Garcez FR. Metabolomics approach expands the classification of propolis samples from Midwest Brazil. J Nat Prod. 2020 Feb 28;83(2):333–43.

28. Pratami DK, Mun’Im A, Yohda M, Hermansyah H, Gozan M, Putri YRP, et al. Total phenolic content and antioxidant activity of spray-dried microcapsules propolis from Tetragonula species. In: AIP Conference Proceedings. 2019.

29. Asem N, Abdul Gapar NA, Abd Hapit NH, Omar EA. Correlation between total phenolic and flavonoid contents with antioxidant activity of Malaysian stingless bee propolis extract. J Apic Res. 2020;59(4):437–42.

30. Ishida Y, Gao R, Shah N, Bhargava P, Furune T, Kaul SC, et al. Anticancer activity in honeybee propolis: functional insights to the role of caffeic acid phenethyl ester and its complex with gamma-cyclodextrin. Integr Cancer Ther. 2018 Sep;17(3):867–73.

31. Kumazawa S, Hamasaka T, Nakayama T. Antioxidant activity of propolis of various geographic origins. Food Chem. 2004;84(3):329–39.

32. Chen J, Yang J, Ma L, Li J, Shahzad N, Kim CK. Structure-antioxidant activity relationship of methoxy, phenolic hydroxyl, and carboxylic acid groups of phenolic acids. Sci Rep. 2020;10(1):2611.

33. Bibi A, Shah T, Sadiq A, Khalid N, Ullah F, Iqbal A. L-isoleucine-catalyzed michael synthesis of N-alkylsuccinimide derivatives and their antioxidant activity assessment. Russ J Org Chem. 2019;55:1749–54.

34. Jiménez-Fernández M, Juárez-Trujillo N, Mendoza-Lopez MR, Monribot-Villanueva JL, Guerrero-Analco JA. Nutraceutical potential, and antioxidant and antibacterial properties of Quararibea funebris flowers. Food Chem. 2023;411(135529):1–8.

35. Sharma N, Biswas S, Al-Dayan N, Alhegaili AS, Sarwat M. Antioxidant role of kaempferol in prevention of hepatocellular carcinoma. Antioxidants. 2021;10(1419):1–17.

36. Falcão SI, Vale N, Gomes P, Domingues MRM, Freire C, Cardoso SM, et al. Phenolic profiling of Portuguese propolis by LC–MS spectrometry: Uncommon propolis rich in flavonoid glycosides. Phytochem Anal. 2013;24(4):309–18.

37. Bratek-Gerej E, Ziembowicz A, Godlewski J, Salinska E. The mechanism of the neuroprotective effect of kynurenic acid in the experimental model of neonatal hypoxia–ischemia: The link to oxidative stress. Antioxidants. 2021;10(1775):1–13.

38. Karmahapatra S, Kientz C, Shetty S, Yalowich JC, Rakotondraibe LH. Capsicodendrin from Cinnamosma fragrans exhibits antiproliferative and cytotoxic activity in human leukemia cells: modulation by glutathione. J Nat Prod. 2018;81(3):625–9.

39. Silva FM da, Ferreira MLS, Cavalcante-Lucena JE, Bicudo ÁJ de A. DL-carnitine as supplementary levocarnitine source in plant-based diets to Nile tilapia (Oreochromis niloticus) fingerlings. Lat Am J Aquat Res. 2018;46(4):825–30.

40. Zhang J, Powell C, Meruvu S, Sonkar R, Choudhury M. Pyrroloquinoline quinone attenuated benzyl butyl phthalate induced metabolic aberration and a hepatic metabolomic analysis. Biochem Pharmacol. 2022;197:114883.

41. Alirezaei M, Khoshdel Z, Dezfoulian O, Rashidipour M, Taghadosi V. Beneficial antioxidant properties of betaine against oxidative stress mediated by levodopa/benserazide in the brain of rats. J Physiol Sci. 2015;65(3):243–52.

42. Cao X, Wu L, Wu M, Zhu C, Jin Q, Zhang J. Abscisic acid mediated proline biosynthesis and antioxidant ability in roots of two different rice genotypes under hypoxic stress. BMC Plant Biol. 2020;20(1):1–14.
How to Cite
PRATAMI, Diah Kartika et al. Comparison of Total Phenolic Content and Antioxidant Activity of Indonesian Propolis Extracted with Various Solvents.. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 21, n. 1, p. 121-129, apr. 2023. ISSN 2614-6495. Available at: <>. Date accessed: 23 june 2024. doi: