• Sarabjit Kaur Department of Pharmaceutical Sciences, Guru Nanak Dev University, Amritsar - 143 005, Punjab, India.
  • Anupam Sharma University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, India
  • Preet Mohinder Singh Bedi Department of Pharmaceutical Sciences, Guru Nanak Dev University, Amritsar - 143 005, Punjab, India




Melilotus officinalis, Aerial parts, Anxiolytic, High-performance thin layer chromatography


Objective: Anxiety is one of the most common and serious mental illness affecting humankind and its extensiveness is on the rise at an alarming rate. Anxiolytic substances are highly acclaimed in the ranking of the most utilized drugs by human. The clinical applications of most widely used anxiolytic agents, that is, benzodiazepines are restricted by their undesirable side effects. Therefore, the development of new pharmacological agents for the treatment of this problem is well justified. Among medicinal plants, Melilotus officinalis (yellow sweet clover) has been recommended for relief of insomnia, convulsions, and as nervine tonic in traditional system of medicine. Nevertheless, no pharmacological studies have thus far evaluated its anxiolytic effect. Therefore, the aim of this study was to evaluate antianxiety effect of different extracts of M. officinalis in mice.

Methods: The extracts of roots and aerial parts of the plant were prepared according to the polarity, that is, petroleum ether, chloroform, ethanol, and water. The anxiolytic effects of petroleum ether, chloroform, ethanol, and aqueous extract of aerial parts and roots of the plant (50, 100, and 200 mg/kg, p.o) were examined in albino mice using elevated plus maze (EPM) and mirror-chamber models of anxiety. High-performance thin layer chromatography (HPTLC) studies were carried out using toluene: Acetone: Formic acid as mobile phase.

Results: Various extracts prepared from roots did not produce significant effect in both the models, whereas the ethanol extract prepared from aerial parts at 100 and 200 mg/kg showed a significant anxiolytic effect as compared to control and standard group. The petroleum ether, chloroform, and water extracts (50, 100, and 200 mg/kg) of the aerial parts of the plant did not produce meaningful effects in this study. HPTLC analysis of the ethanol extract revealed the presence of nine components.

Conclusion: These results suggest that the ethanol extract of aerial parts of M. officinalis plant has statistically significant dose-dependent antianxiety activity which can be attributed to the presence of coumarin, and flavonoid compounds in it.


Download data is not yet available.

Author Biography

Sarabjit Kaur, Department of Pharmaceutical Sciences, Guru Nanak Dev University, Amritsar - 143 005, Punjab, India.


Department of Pharmaceutical Sciences


Anwar MS, Mohtasheem M, Azhar I, Ahmed SW, Bano H. Chemical constituents from Melilotus officinalis. J Basic Appl Sci


Plesca-Manea L, Pârvu AE, Pârvu M, Taamas M, Buia R, Puia M. Effects of Melilotus officinalis on acute inflammation. Phytother Res 2002;16:316-9.

Anwer MS, Shamim S, Ahmed S, Mohtasheemul M, Azhar I. Hypotensive activity of Melilotus officinalis (L.) Pallas. Eur J Med 2015;3(2):80-5.

Braga PC, Sasso MD, Lattuada N, Marabinil L, Calo R, Antonacci R, et al. Antioxidant activity of Melilotus officinalis extract investigated by means of the radical scavenging activity, the chemiluminescence of human neutrophil bursts and lipoperoxidation assay. J Med Plant Res 2013;7(7):358-65.

Nasser NM, Al-Araji M, Widad MK. Antifungal activity of Mellilotus officinalis of Iraq. J Chem Pharm Res 2014;6(11):611-7.

Heidari MR, Najafi F, Asadi PA, Ansari M, Zahedi MJ, Vahedian M. Analgesic and ulcerogenic effect of methanolic extract of Melilotus officinalis. J Kerman Univ Med Sci 2001;8(4):210-9.

Martino E, Ramaiola I, Urbano M, Bracco F, Collina S. Microwave-assisted extraction of coumarin and related compounds from Melilotus officinalis (L.) Pallas as an alternative to soxhlet and ultrasound-assisted extraction. J Chromatogr 2006;1125:147-51.

Kang SS, Lim CH, Lee SY. Soyasapogenols B and E from Melilotus officinalis. Arch Pharm Res 1987;10(1):9.

Kang SS, Won SW. Melilotogenin, a new Sapogenin from Meliltous officnalis. J Nat Prod 1998;51(2):335-8.

Manabu U, Junei K, Naotoshi Y, Toshihiro N. A new oleanene glucuronide having a branched chain sugar from Melilotus officinalis. Chem Pharm Bull 1998;46(3):526-7.

Sutiashvili MG, Alaniya MD. Flavonoids of Melilotus officinalis. Chem Nat Compd 1999;35(5):584.

Duke JA, Godwin MJ, Judi DC, Duke PK. Handbook of Medicinal Herbs. 2nd ed. New York: CRC Press; 2002. p. 806-7.

Kumar M. Ethnobotanical studies on some medicinal plants: A review. World J Pham Res 2014;3(8):342-61.

Evans W. Trease and Evans Pharmacognosy. London: WB Saunders Company Ltd.; 1996. p. 98.

Kulkarni SK. Handbook of Experimental Pharmacology. 3rd ed. Pitampura, New Delhi: Vallabh Prakashan; 2003. p. 135-40.

Malik R, Gilhaotra, NA. Novel non-receptor and non-gabaergic antianxiety-like activity of forskolin: Synergy with diazepam. Int J Pharm Pharm Sci 2015;7(2):200-3.

Lamberty Y. The mirror chamber test for testing anxiolytics: Is there a mirror-induced stimulation? Physiol Behav 1998;64(5):703-5.

Khandelwal KR. Techniques and Experiments in Practical Pharmacognosy. Pune: Nirali Prakashan; 2003. p. 149.

Khanum F, Razack S. Anxiety-herbal treatment: A review. Res Rev Biomed Biotechnol 2010;1:71S.

Ravindran A, Manohar VR, Rai M, Raveendran N, Naik H. Chronic anxiolytic-like activity of aqueous extract of Coriandrum sativum seeds using elevated plus maze test in swiss albino mice. Int J Pharm Pharm Sci 2014;6(2):93-5.

Bloom FE, Kupfer DJ. Psychopharmacology: The Fourth Generation of Progress. New York: Raven Press; 1994. p. 1301.

Bhatacharya SK, Satyan KS. Experimental methods for evaluation of psychotropic agents in rodents: Anti-anxiety agents. Indian J Exp Biol 1997;35(6):565-75.

Kulkarni SK, Reddy DS. Animal behavioural models for testing antianxiety activity. Methods Find Exp Clin Pharmacol 1996;18:219-30.

Siqueira IR, Lara DR, Silva D, Gaieski FS, Nunes DS, Elisabersky E. Psychopharmacological properties of Ptychopetalum olacoides Bentham. Pharm Biol 1998;36(5):327-34.

Dajas F, Rivera-Megret F, Blasina F, Arredondo F, Abin-Carriquiry JA, Costa G. Neuroprotection by flavonoids. Braz J Med Biol Res 2003;136(12):1613-20.

Perez-Ortega G, Guevara FP, Chavez M, Herrera J, Martinez A, Martinez AL, et al. Sedative and anxiolytic efficacy of Tilia americana var. Mexicana inflorescences used traditionally by communities of State of Michoacan. M J Ethnopharmacol 2008;116(3):461-8.

Medina JH, Viola H, Wolfman C, Marder M, Wasowski C, Calvo D, et al. Overview - Flavonoids: A new family of benzodiazepine receptor ligands. Neurochem Res 1997;22(4):419-25.

Marder M, Estiu G, Blanch LB, Viola H, Wasowsk C, Medina JH, et al. Molecular modeling and QSAR analysis of the interaction of flavone derivatives with the benzodiazepine binding site of the GABA(A) receptor complex. Bioorg Med Chem 2001;9(2):323-35.

Marder M, Paladini AC. GABA(A)-receptor ligands of flavonoid structure. Curr Top Med Chem 2002;2(8):853-67.

Seon HJ, Xiang HH, Seong SH, Ji SH, Ji HH, Lee D, et al. Monoamine oxidase inhibitory coumarins from the aerial parts of Dictamnus albus. Arch Pharm Res 2006;29:1119-24.

Singhuber J, Baburin I, Ecker GF, Kopp B, Hering S. Insights into structure-activity relationship of GABAA receptor modulating coumarins and furanocoumarins. Eur J Pharmacol 2011;668(1):57-64.



How to Cite

Kaur, S., A. Sharma, and P. M. S. Bedi. “EVALUATION OF ANXIOLYTIC EFFECT OF MELILOTUS OFFICINALIS EXTRACTS IN MICE”. Asian Journal of Pharmaceutical and Clinical Research, vol. 10, no. 6, June 2017, pp. 396-9, doi:10.22159/ajpcr.2017.v10i6.17183.



Original Article(s)