• Bosco Lawarence Department of Botany and Biotechnology, Government Arts College, Trivandrum - 695 014, Kerala, India.
  • Murugan K Department of Botany and Biotechnology, Government Arts College, Trivandrum - 695 014, Kerala, India.


Objective: The objective of the present work is to isolate, purify, and fractionate anthocyanin from selected Osbeckia species and also to compare the analgesic and anti-inflammatory potentiality using animal models.

Methods: Methodologies include extraction of anthocyanin from the in vitro callus culture of Osbeckia aspera and Osbeckia reticulata, purification using amberlite column chromatography, and fractionation by liquid chromatography-tandem mass spectrometry. The analgesic activity was determined by tail immersion method, analgesy meter, hot plate, and acetic acid-induced writhing test. Anti-inflammatory activity was evaluated by carrageenan-induced paw inflammation in mice.

Results: Anthocyanin-producing callus cultures were established in MS medium fortified with various combinations of phytohormones and sucrose. Optimal callus formation in O. aspera was initiated on cultures containing 0.5 mg/L of 2, 4-D, and 0.5 mg/L 6-benzylaminopurine (BA). In O. reticulata callus was initiated in the presence of 1.2 mg/L BA and 1.4 mg/L naphthalene acetic acid. The same hormonal combination on extended treatments turned the white friable callus into red compact callus. Anthocyanins obtained from Osbeckia species were purified and fractionated containing malvidin-3-diglucoside, delphinidin, cyanidin aglycone, and peonidin. Purified anthocyanin of O. aspera at the concentrations 50, 100, 150, and 200 mg/kg after 120 min exhibited significant analgesic activity by tail immersion method, in comparison to O. reticulata. However, with hot plate method, anthocyanin of O. reticulata produced a significant analgesic activity even at lower doses (50 and 100 mg/kg) after 120 min. However, in writhing test, theepurified anthocyanin of O. aspera significantly stopped the number of writhes at a dose of 200 mg/kg and also by the anthocyanin of O. reticulata. In the evaluation of anti-inflammatory effect using plethysmometer, anthocyanin at doses of 100,150 and 200 mg/kg started producing anti-inflammatory effect after 30 min, which lasted until 120 min.

Conclusion: It is concluded from the present study that the purified anthocyanin of Osbeckia possesses potent analgesic and anti-inflammatory activities.

Keywords: Osbeckia, Purified anthocyanin, In vitro culture, Analgesic activity, Anti-inflammatory activity, Acetic acid, Carrageenan.

Author Biographies

Bosco Lawarence, Department of Botany and Biotechnology, Government Arts College, Trivandrum - 695 014, Kerala, India.
Assistant Professor
Murugan K, Department of Botany and Biotechnology, Government Arts College, Trivandrum - 695 014, Kerala, India.


1. Lawarence B, Murugan K. Folklore use of Osbeckia species from Munnar hills, Kerala. J Tradit Folk Pract 2017;5:131-41.
2. Jayaweera DM. Medicinal Plants (Indigenous and Exotic) Used in Ceylon. Part I. Colombo: National Science Council of Sri Lanka; 1981. p. 205-6.
3. Aswathy JM, Sumayya SS, Lawarence B, Kavitha CH, Murugan K. Purification, fractionation and characterization of anthocyanin from in vitro culture of Bridelia retusa (L) spreng. Indian J Pharm Sci 2018;80:52-64.
4. Huang W, Yan Z, Li D, Ma Y, Zhou J, Sui Z. Antioxidant and anti-inflammatory effects of blueberry anthocyanins on high glucose-induced human retinal capillary endothelial Cells. Oxid Med Cell Longev 2018;2018:1-10.
5. Bishayee A, Háznagy-Radnai E, Mbimba T, Sipos P, Morazzoni P, Darvesh AS, et al. Anthocyanin-rich black currant extract suppresses the growth of human hepatocellular carcinoma cells. Nat Prod Commun 2010;5:1613-8.
6. Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 1962;15:473-97.
7. Sutharut J, Sudarat J. Total anthocyanin content and antioxidant activity of germinated colored rice. Int Food Res J 2012;19:215-21.
8. OECD. Acute Oral Toxicity-Fixed Dose Procedure (chptr). OECD Guideline Test Chemicals; 2001. p. 1-14.
9. OECD. OECD Guideline for the Testing of Chemicals: Repeated Dose 90-Day Oral Toxicity Study In Rodents; 1998.
10. Meshram GG, Kumar A, Rizvi W, Tripathi CD, Khan RA. Evaluation of the anti-inflammatory activity of the aqueous and ethanolic extracts of the leaves of Albizia lebbeck in rats. J Tradit Complement Med 2016;6:172-5.
11. Vogel HG. Drug Discovery and Evaluation : Pharmacological Assays. Berlin: Springer; 2008.
12. Tasleem F, Azhar I, Ali SN, Perveen S, Mahmood ZA. Analgesic and anti-inflammatory activities of Piper nigrum L. Asian Pac J Trop Med 2014;7S1:S461-8.
13. Liang X, Liu R, Chen C, Ji F, Li T. Opioid system modulates the immune function: A review. Transl Perioper Pain Med 2016;1:5-13.
14. Tamrat Y, Nedi T, Assefa S, Teklehaymanot T, Shibeshi W. Anti-inflammatory and analgesic activities of solvent fractions of the leaves of Moringa stenopetala bak. (Moringaceae) in mice models. BMC Complement Altern Med 2017;17:473.
15. Fan SH, Ali NA, Basri DF. Evaluation of analgesic activity of the methanol extract from the galls of Quercus infectoria (Olivier) in rats. Evid Based Complement Alternat Med 2014;2014:976764.
16. Moniruzzaman M, Rahman MA, Ferdous A. Evaluation of sedative and hypnotic activity of ethanolic extract of Scoparia dulcis Linn. Evid Based Complement Alternat Med 2015;2015:873954.
17. Kuete V, KaraosmanoÄŸlu O, Sivas H. Anticancer activities of african medicinal spices and vegetables. Medicinal Spices and Vegetables from Africa. London: Elsevier; 2017. p. 271-97.
18. Adebisi MI, Abubakar A, Abubakar K, Giaze RT. Analgesic effect and anti-inflammatory activity of aqueous extract of Boswellia dalzielii (burseraceae) stem bark. Int J Pharm Pharm Sci 2018;10:139-42.
19. Agyare C, Spiegler V, Asase A, Scholz M, Hempel G, Hensel A. An ethnopharmacological survey of medicinal plants traditionally used for cancer treatment in the Ashanti Region, Ghana. J Ethnopharmacol 2018;212:137-52.
20. Ammar I, Salem MB, Harrabi B, Mzid M, Bardaa S, Sahnoun Z, et al. Anti-inflammatory activity and phenolic composition of prickly pear (Opuntia ficus-indica) flowers. Ind Crops Prod 2018;112:313-9.
21. Hashmi MA, Khan A, Farooq U, Khan S. Alkaloids as cyclooxygenase inhibitors in anticancer drug discovery. Curr Protein Pept Sci 2018;19:292-301.
22. Patwari BN, Ahmed AB, Das T, Sengupta R. Phytochemical screening and analgesic effects of ethanolic extract of plant Murdania nudiflora (L) Brenan (Commelinaceae) in albino mice using hot plate method. Int J Pharm Pharm Sci 2014;6:512-5.
23. Valenza A, Bonfanti C, Pasini ME, Bellosta P. Anthocyanins function as anti-inflammatory agents in a drosophila model for adipose tissue macrophage infiltration. Biomed Res Int 2018;2018:1-9.
24. Duarte LJ, Chaves VC, Nascimento MV, Calvete E, Li M, Ciraolo E, et al. Molecular mechanism of action of pelargonidin-3-O-glucoside, the main anthocyanin responsible for the anti-inflammatory effect of strawberry fruits. Food Chem 2018;247:56-65.
220 Views | 247 Downloads
How to Cite
Lawarence, B., and M. K. “COMPARISON OF ANALGESIC AND ANTI-INFLAMMATORY ACTIVITIES OF PURIFIED ANTHOCYANIN FROM OSBECKIA ASPERA (L.) BLUME AND OSBECKIA RETICULATA BEDD. USING ANIMAL MODELS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 1, Jan. 2019, pp. 106-11, doi:10.22159/ajpcr.2019.v12i1.27402.
Original Article(s)