• B. Satheesh Kumar Bhandary
  • K.P Sharmila
  • N. Suchetha Kumari
  • S. Vadisha Bhat


Punica granatum (L), Ellagic acid, ethanolic extract, acute toxicity, subacute toxicity


Objective: The objective of the present study was to evaluate the acute and sub-acute toxicity profile of ethanolic extracts of Punica granatum (L).whole fruit (EPWF) and seeds (EPS) and synthetic ellagic acid (EA) in Swiss albino mice following OECD guidelines.

 Methods: In acute toxicity study, mice were orally administered 2000 mg/kg body weight of extracts and synthetic compound. In subacute toxicity study, the animals were orally administered with the extracts and synthetic compound (2000 mg/kg body weight) daily for 28 days whereas, control group received normal saline.

Results: There were no behavioral alterations or mortality recorded in the treated groups. The LD50 value was more than 2000 mg/kg body weight. Test groups did not record any significant alterations (p>0.05) in body weight gain, food and water intake. The hematological and biochemical parameters and organ weights did not record any significant alterations (p>0.05) in the treated groups when compared to control. A detailed examination of histoarchitecture of the liver and kidney did not reveal any observable cellular damage in the treated groups compared to control.

Conclusion: The overall finding of this study suggests that Punica granatum (L) whole fruit (EPWF) and seeds (EPS) ethanolic extract and synthetic ellagic acid (EA) is safe up to 2000 mg/kg body weight oral administration and can be considered as non toxic.

Keywords: Punica granatum (L), Ellagic acid, ethanolic extract, acute toxicity, subacute toxicity


Akah PA. Indigenous Knowledge and Medical practice. Ethnopharmacology 2008; 1-13.

Kamboj VP. Herbal Medicine. Current science 2000; 78(1): 35-39.

Jaouad EH, Israilli ZH, Lyoussi B. Acute toxicity and chronic toxicological studies of Ajugaiva in experimental animals. Journal of ethnopharmacology 2004; 91: 43-50.

Kalplowitz N. Hepatotoxicity of herbal remedies: Insight into the intricacies of plant-animal warfare and cell death. Gastroenterol 1997; 113: 1408-1412.

Saad B, Azaizeh H, Abu-Hijleh G, Said O. Safety of traditional Arab herbal. Evid Based Complement Alternat Med 2006; 3: 433–9.

Colson CR, De Broe ME. Kidney injury from alternative medicines. Adv Chronic Kidney Dis 2005; 12:261-75.

Kwan TH, Tong MK, Leung KT, Lai CK, Poon WT, Chan YW et al. Acute renal failure associated with prolonged intake of slimming pills containing anthraquinones. Hong Kong Med J 2006; 12: 394–7.

Zhu YP. Toxicology of the Chinese herb mu tong (Aristolochia manshuriensis). What history tells us. Adverse Drug React Toxicol Rev 2002; 21: 171–7.

Vanherweghem LJ. Misuse of herbal remedies: the case of an outbreak of terminal renal failure in Belgium (Chinese herbs nephropathy). J Altern Complement Med 1998; 4:9– 13.

Horowitz RS, Feldhaus K, Dart RC, Stermitz FR, Beck JJ. The clinical spectrum of Jin Bu Huan toxicity. Arch Intern Med 1996; 156:899–903.

Moritz F, Compagnon P, Kaliszczak IG, Kaliszczak Y, Caliskan V, Girault C. Severe acute poisoning with homemade Aconitum napellus capsules: toxicokinetic and clinical data. Clin Toxicol 2005; 43:873–6.

Gaibazzi N, Gelmini GP, Montresor G, Canel D, Comini T, Fracalossi C et al. Long QRS tachycardia secondary to Aconitum napellus alkaloid ingestion. Ital Heart J Suppl 2002; 3:874–7.

Ernst E. Serious psychiatric and neurological adverse effects of herbal medicines - asystemic review. Acta Psychiatr Scand 2003; 108:83–91.

Jensen WI, Allen JP. Naturally occurring and experimentally induced castor bean (Ricinus communis) poisoning in ducks. Avian Dis1981; 5:184-94.

Langley P. Why a pomegranate? BMJ 2000; 321: 1153–1154.

Gil MI, Tomas-Barberan FA, Hess-Piercd B, Holcroft DM, Kader AA. Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing. Journal of Agricultural and Food Chemistry 2000; 48:4581–4589.

Schubert SY, Lansky EP, Neeman I Antioxidant and eicosanoid enzyme inhibition properties of pomegranate seed oil and fermented juice flavonoids. Journal of Ethnophamacology 1999; 66:11–17.

Aviram M, Dornfeld L, Pomegranate juice consumption inhibit serum angiotensin converting enzyme activity and reduces systolic blood pressure. Atherosclerosis 2001; 158:195–198.

Kaplan M, Hayek T, Raz A, Coleman R, Dornfeld L, Vaya J, et al. Pomegranate juice supplementation to atherosclerotic mice reduces macrophage lipid peroxidation, cellular chlesterol accumulation and development of atherosclerosis. The Journal of Nutrition 2001; 131:2082–2089.

Halvorsen BL, Holte K, Myhrstad MCW, Barikmo I, Hvattum E, Remberg SF, et al. A systematic screening of total antioxidants in dietary plants. The Journal of Nutrition 2002; 132:461–471.

Noda Y, Kaneyuki T, Mori A, Packer L, Antioxidant activities of pomegranate fruit extract and its anthocyanidins, Delphinidin, cyanidin, and pelargonidin. Journal of Agricultural and Food Chemistry 2002; 50:166–171.

Singh RP, Murthy KNC, Jayaprakasha GK, Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extract using in vitro models. Journal of Agricultural and Food Chemistry 2002; 50:81–86.

Kim ND, Mehta B, Yu W, Neeman I, Livney T, Amichay A, et al. Chemopreventive and adjuvant therapeutic potential of pomegranate (Punica granatum) for human breast cancer. Breast Cancer Research and Treatment 2002; 71: 203–217.

Vattem DA, Shetty K. Biological Function of Ellagic Acid: A Review. Journal of Food Biochemistry 2005; 29: 234–266.

Seeram NP, Adams LS, Henning SM, et al. In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. J. Nutr. Biochem 2005; 16(6): 360–7.

Madal, Shivappurkar, Galati, Stoner. Inhibition of N-nitrosobenzymethylamine metabolism and DNA binding in cultured rat esophagus by ellagic acid. Carcinogenesis1988; 9(7): 1313–1316.

Malini P, Kanchana G, Rajadurai M, Antibiabetic Efficacy of Ellagic Acid in Streptozotocin induced Diabetes Mellitus in Albino Wistar Rats. Asian J Pharm Clin Res 2011; 4(3):124-128.

Mandal, Stoner GD. Inhibition of N-nitrosobenzymethylamine-induced esophageal tumorigenesis in rats by ellagic acid.Carcinogenesis1990; 11(1): 55-61.

OECD.Test Guideline 425. Acute Oral Toxicity – Up-and-Down Procedure 2001.

Shah Ayub MA, Garg SK, Garg KM. Subacute toxicity studies on Pendimethalin in rats. Indian J Pharmacol 1997; 29:322-324.

Burger C, Fischer DR, Cordenunzzi DA, Batschauer de Borba AP, Filho VC, Soares dos Santos AR.Acute and subacute toxicity of the hydroalcoholic extract from Wedelia paludosa (Acmela brasilinsis) (Asteraceae) in mice. J. Pharm Sci 2005; 8 (2): 370-373.

OECD (testing guideline, 407), Repeat dose 28 days oral toxicity study in rodents. In-Guidance document for the development of OECD guideline for testing of chemicals. Environmental monogr 1995; No.76.

Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of lowdensity lipoprotein cholesterol in plasma, without the use of preparative centrifuge. Clin. Chem1972; 18,499-502.

Sathya M, Kokilavani R, Ananta TKS. Acute and subacute toxicity studies of ethanolic extract of Acalypha indica Linn in male wistar albino rats. Asian J Pharm Clin Res 2012; 5 (1): 97-100.

Mythilypriya R, Shanthi P, Sachdanandam P. Oral acute and subacute toxicity studies with Kalpaamruthaa, a modified indigenous preparation on rats. J. Health Sci 2007; 53(4): 351-358.

Ogbonnia S, Adekunle AA, Bosa MK, Enwuru VN. Evaluation of acute and subacute toxicity of Alstonia congensis Engler (Apocynaceae) bark and Xylopia aethiopica (Dunal) A. Rich (Annonaceae) fruits mixtures used in the treatment of diabetes. Afr. J. Biotechnol 2008; 7(6): 701-705.

Patel C, Dadhaniya P, Hingorani L, Soni MG .Safety assessment of pomegranate fruit extract: acute and subchronic toxicity studies. Food Chem Toxicol 2008;46(8):2728-35

Tasaki M, Umemura T, Maeda M, Ishii Y, Okamura T, Inoue T, Kuroiwa Y, et al. The Safety assessment of ellagic acid, a food additive, in a subchronic toxicity study using F344 rats. Food Chem Toxicol 2008; 46(3):1119-24.

Ogbonnia SO, Nkemehule FE, Anyika EN. African Journal of Biotechnology 8(9): 1793 -1799.

Walum E. Environmental Health Perspectives 1998; 106 (2): 497-502.



How to Cite

Kumar Bhandary , B. S., K. Sharmila, N. Suchetha Kumari, and S. Vadisha Bhat. “ACUTE AND SUBACUTE TOXICITY STUDY OF THE ETHANOL EXTRACTS OF PUNICA GRANATUM (LINN). WHOLE FRUIT AND SEEDS AND SYNTHETIC ELLAGIC ACID IN SWISS ALBINO MICE”. Asian Journal of Pharmaceutical and Clinical Research, vol. 6, no. 8, Sept. 2013, pp. 192-8,