• TANTI AZIZAH SUJONO Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Universitas Muhammadiyah Surakarta, 57162, Surakarta, Indonesia
  • IKA TRISHARYANTI DIAN KUSUMOWATI Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Muhammadiyah Surakarta, 57162, Surakarta, Indonesia.
  • RIMA MUNAWAROH Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Muhammadiyah Surakarta, 57162, Surakarta, Indonesia.


Objective: Muntingia calabura L fruits have several advantages for health and can be consumed as alternative medicine. This study aims to evaluate the immunomodulatory activity of methanol extract (ME) and fractions of M. calabura L fruits against non-specific immune response in mice and its total phenolic and flavonoid contents.

Methods: This research using carbon clearance assay or phagocytosis method which was done on mice. Seventy male Swiss mice were divided into 14 groups. Mice were treated with ME, hexane fraction (HF), dichloromethane fraction (DF), and ethyl acetate fraction (EAF) of M. calabura L. orally at a dose of 50, 100, and 200 mg/kg daily for 7 days. Zymosan with a dose of 15 mg/kg was used as a positive control and given intraperitoneally, and CMC Na was used as a negative control. On the 8th day, mice were injected with carbon ink suspension intravenously and blood samples were taken at the minutes of 0, 5, 10, 15, 20, and 30. Furthermore, the transmittance was measured at λ 675 nm. The total phenolic content was measured using the Folin–Ciocalteu method, and flavonoid content was measured using the AlCl3 method.

Results: Zymosan, ME, and EAF of M. calabura L. at a dose of 200 mg/kg showed a strong immunostimulatory effect (phagocytic index >1.5) while the HF and DF groups showed that the higher doses decreased the phagocytic index. The total phenolic contents of ME, HF, DF, and EAF were 27.90, 11.11, 16.72, and 30.11%, respectively, while the flavonoid contents were 4.07, 0.17, 3.07, and 1.86%, respectively.

Conclusion: ME and EAF of M. calabura L. fruits have immunostimulatory activity.

Keywords: Muntingia calabura L., Phagocytic index, Carbon clearance assay, Immunomodulator


1. Mahima, Ingle AM, Verma AK, Tiwari R, Karthik K, Chakraborty S, et al. Immunomodulators in day to day life: A review. Pak J Biol Sci 2013;16:826-43.
2. Jantan I, Ahmad W, Bukhari SN. Plant-derived immunomodulators: An insight on their preclinical evaluation and clinical trials. Front Plant Sci 2015;6:655.
3. Preethi K, Premasudha P, Keerthana K. Anti-inflammatory activity of Muntingia calabura fruits. Pharmacogn J 2012;4:51-6.
4. Pramono VJ, Santoso R. Pengaruh ekstrak buah kersen (Muntingia calabura) terhadap kadar gula darah tikus putih (Rattus norvegicus) yang diinduksi streptozotocin (STZ). J Sain Vet 2014;32:218-23.
5. Sarimanah J, Adnyana IK, Sukandar EY, Kurniati NF. The antirheumatic activity of Muntingia calabura L. Leaves ethanol extract and its fraction. Asian J Pharm Clin Res 2017;10:84-6.
6. Siddiqua A, Premakumari KB, Sultana R, Vithya, Savitha. Antioxidant activity and estimation of total phenolic content of Muntingia calabura by colorimetry. Int J Chem Technol Res 2010;2:205-8.
7. Prakosa AP. Imunomodulasi Ekstrak Buah Talok (Muntingia calabura Linn) Terhadap Makrofag Peritoneum Tikus Wistar Jantan yang Diinduksi Actinobacillus Actionycetemcomitans. Indonesia: Universitas Jendral Soedirman; 2014.
8. Kalaskar MG, Surana SJ. Free radical scavenging, immunomodulatory activity and chemical composition of Luffa acutangula Var. Amara (Cucurbitaceae) pericarp. J Chil Chem Soc 2014;59:2299-302.
9. Patel S, Vajdy M. Induction of cellular and molecular immunomodulatory pathways by Vitamin A and flavonoids. Expert Opin Biol Ther 2015;15:1411-28.
10. Grigore A. Plant phenolic compounds as immunomodulatory agent. In: Phenolic Compound-Biological Activity. London: Intech Open; 2017. p. 75-88.
11. Ganeshpurkar A, Saluja AK. Experimental animal models used for evaluation of potential immunomodulators: A mini review. Bull Fac Pharmacy Cairo Univ 2017;55:211-6.
12. Chun OK, Kim DO, Lee CY. Superoxide radical scavenging activity of the major polyphenols in fresh plums. J Agric Food Chem 2003;51:8067-72.
13. Saleem M, Maqbool M, Musadiq H, Ahmad S, Hassan A, Ahmad S. Pathomorphological effects of alloxan induced acute hypoglycaemia in rabbits. Alex J Med 2013;49:343-53.
14. Ahmad AR, Sakinah, Wisdawati, Asrifa WO. Study of antioxidant activity and determination of phenol and flavonoid content of pepino’s leaf extract (Solanum muricatum Aiton). Int J Pharm Technol Res 2014;6:600-6.
15. Faradilla M, Iwo MI. Efck imunomodulator polisakarida rimpang temu putih immunomodulatory effect of polysaccharide from white turmeric [Curcuma zedoaria (Christm) Roscoe)] rhizome. J Ilmu Kefarmasian Indones 2014;12:273-8.
16. Iwo MI, Soemardji AA, Retnoningrum DS, Sukrasno, U UM. Immunostimulating effect of pule (Alstonia scholaris L. R.Br., Apocynaceae) bark extracts. Clin Hemorheol Microcirc 2000;23:177-83.
17. Shrestha P, Handral M. Evaluation of immunomodulatory activity of extract from rind of Nephelium lappaceum fruit. Int J Pharm Pharm Sci 2016;9:38-43.
18. Wagner H, Kraus S, Jurcic K. Search for Potent Immunostimulating Agents from Plants and Other Natural Sources. Immunomodulatory Agents from Plants. Berlin, Germany: Springer; 1999. p. 1-39.
19. Dillasamola D, Aldi Y, Fakhri M, Diliarosta S, Oktomalio PB, Noverial N. Immunomodulatory effect test from moringa leaf extract (Moringa oleifera L.) with carbon clearance method in male white mice. Asian J Pharm Clin Res 2018;11:241-5.
20. Ghiringhelli F, Rebe C, Hichami A, Delmas D. Immunomodulation and anti-inflammatory roles of polyphenols as anticancer agents. Anticancer Agents Med Chem 2012;12:852-73.
21. Putri DA, Fatmawati S. Metabolit sekunder dari Muntingia calaburadan bioaktivitasnya. Alchemy J Penelit Kim 2019;15:57.
22. Underhill DM. Macrophage recognition of zymosan particles. J Endotoxin Res 2003;9:176-80.
23. Pereira GA, Arruda HS, de Morais DR, Eberlin MN, Pastore GM. Carbohydrates, volatile and phenolic compounds composition, and antioxidant activity of calabura (Muntingia calabura L.) fruit. Food Res Int 2018;108:264-73.
24. Wagle N, Nagarjuna S, Sudheer A, Roopesh C, Sapkota HP, Dangi NB, et al. Evaluation of immunomodulatory activity of petroleum ether extract of seeds of Pithecellobium dulce in wistar rats. Int J Pharm Pharm Sci 2015;7:471-9.
25. Maji AK, Mahapatra S, Banerjee D. In vivo immunomodulatory potential of standardized Pueraria tuberosa extract and its isoflavonoids. Int J Pharm Pharm Sci 2014;6:861-7.
26. Ragasa CY, Tan MC, Chiong ID, Shen CC. Chemical constituents of Muntingia calabura L. Pharma Chem 2015;7:136-41.
27. Mahmood ND, Nasir NL, Rofiee MS, Tohid SF, Ching SM, Teh LK, et al. Muntingia calabura: A review of its traditional uses, chemical properties, and pharmacological observations. Pharm Biol 2014;52:1598-623.
28. Sibi G, Naveen R, Dhananjaya K, Ravikumar KR, Mallesha H. Potential use of Muntingia calabura L. Extracts against human and plant pathogens. Pharmacogn J 2012;4:44-7.
29. Bilia AR, Isacchi B, Righeschi C, Guccione C, Bergonzi MC. Flavonoids loaded in nanocarriers: An opportunity to increase oral bioavailability and bioefficacy. Food Nutr Sci 2014;5:1212-27.
30. Pratiwi G, Martien R, Murwanti R. Chitosan nanoparticle as a delivery system for polyphenols from meniran extract (Phyllanthus niruri L.): Formulation, optimization, and immunomodulatory activity. Int J Appl Pharm 2019;11:50-8.
196 Views | 143 Downloads
How to Cite
SUJONO, T. A., I. T. DIAN KUSUMOWATI, and R. MUNAWAROH. “IMMUNOMODULATORY ACTIVITY OF MUNTINGIA CALABURA L. FRUITS USING CARBON CLEARANCE ASSAY AND THEIR TOTAL FLAVONOID AND PHENOLIC CONTENTS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 13, no. 2, Feb. 2020, pp. 140-5, doi:10.22159/ajpcr.2020.v13i2.36449.
Original Article(s)