Int. J. Pharm. Investigation, 2019;9(2):71-75 International Journal of Pharmaceutical Investigation, Vol 9, Issue 2, Apr-Jun, 2019 71 Original Article Acute Toxicity Study of Annona reticulata Leaves Extract in Swiss Albino Mice Lohith Mysuru Shivanna 1 , Halugudde Nagaraja Sarjan 2 , Asna Urooj 1, * 1 Department of Studies in Food Science and Nutrition, Manasagangotri, University of Mysore, Mysuru, Karnataka, INDIA. 2 Department of Studies in Zoology, Manasagangotri, University of Mysore, Mysuru, Karnataka, INDIA. ABSTRACT Background: Annona reticulata (AR) or custard apple belongs to family Annonaceae. The plant is traditionally used to treat various ailments and is also known for its medicinal properties. Objectives: The aim of this re- search work was to evaluate the safety of usage of AR leaf extract in nu- traceutical formulations through acute toxicity study. Materials and Meth- ods: The leaves of AR were cleaned, dried and powdered; and aqueous extract (ARAQ) was prepared. The acute toxicity test was conducted using female swiss albino mice. As per Organization for Economic Co-operation and Development (OECD) 423 guideline, a single dose of 2000 mg/kg body weight of ARAQ was administered via oral gavage for 14 days. Mortality, signs of toxicity, body weight and behavioral changes were observed dur- ing the study period. Following the 14-day treatment, the mice were sacri- fced for hematological, biochemical and histopathology studies. Results: No mortality, signs of toxicity and changes in behavior were observed at 2000 mg/kg body weight. In addition, no signifcant differences (p>0.05) were noticed in body and organ weight between the control and ARAQ treated groups. Also, there were no signifcant elevations observed in he- matological and biochemical blood parameters. Further, histopathological examination revealed normal architecture of liver, kidney and pancreas. No signifcant adverse effects were observed in these organs. Conclusion: Overall, ARAQ did not produce any signifcant toxic effect in mice and the results also indicate the safety of the oral administration of ARAQ at 2000 mg/kg body weight. Hence, ARAQ can be utilized in nutraceutical formula- tions. Key words: Acute Toxicity, Annona reticulata, Nutraceutical, Animal studies, Swiss albino mice, OECD 423. Correspondence Dr. Asna Urooj, Professor, Department of Studies in Food Science and Nutrition, Manasagan- gotri, University of Mysore, Mysuru-570006, Karnataka, INDIA. Phone no: +91 8212419632 Email: asnaurooj@foodsci.uni-mysore.ac.in DOI: 10.5330/ijpi.2019.2.14 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms. INTRODUCTION World Health Organization (WHO) defnes herbal medicine as a plant- derived material or preparation with therapeutic values and other hu- man health benefts, which contains either raw or processed ingredients from one or more plants. 1 Due to lesser side efects caused by herbal medicines, many pharmaceutical companies target herbal plant sources for the synthesis of synthetic compounds. 2 Annona reticulata (AR) is one of the traditionally important herbal medicinal plants used for the treat- ment of various ailments and also possess several medicinal properties such as analgesic, anti-infammatory, anti-hyperglycemic, anthelmintic, anti-ulcer, wound healing and anti-cancer efects. It is commonly known as Bullock’s heart or Custard apple in English and Ramaphala in Kan- nada. 3 It belongs to family Annonaceae. 4 Te plant is indigenous to the West Indies. It is widely cultivated in West Bengal and southern regions of India as a fruit consuming plant and deciduous tree. 3 Traditionally, the plant is used for treating dysentery, cardiac problem, epilepsy, consti- pation, haemorrhage, bacterial infection, dysuria, fever and ulcer. 3 Dif- ferent parts of AR have several phytoconstituents. Stem bark contains alkaloid, tannins and phenolic compounds. Leaves contain favonoids, amino acids, alkaloids, carbohydrates, steroids, tannins, proteins, glyco- sides and phenolics. Root has acetogenin, alkaloid, carbohydrates, pro- teins, favonoids and tannins. Te plant also found to be rich in minerals viz., K, Mg, Fe, Cu, Se, Na, Cl, S, Mn, Zn, Co, Cr, P, Ni and Ca. 3,5,6 Also, in our preliminary screening, the aqueous extract of AR has exhibited the presence of rutin, a favonoid compound known for its apoptosis induc- ing potential. A study reported cytotoxic efect of AR leaves in Caco-2, Hep G2, HEK cell lines. 7 Te roots of AR also exhibited in vivo antican- cer activity against melanoma cells in mice. 8 and in vitro cytotoxic activ- ity on MDA-MB-435 human melanoma cells. 9 Biological activities such as DPPH free radical scavenging activity; antibacterial and antifungal activity of leaf extract of AR has also been reported in a study. 10 Anti-hy- perglycemic efect of AR leaves extract has been reported in Streptozoto- cin (STZ) induced diabetic rat model, proving it to be a potent glucose- lowering agent. 11 As there are no earlier reports on toxicity assessment of aqueous extract of Annona reticulata leaves, the present study aimed to ascertain and establish the safety profle of aqueous extract of AR leaves through acute toxicity study as per OECD guidelines. MATERIALS AND METHODS Chemicals and reagents Clinical diagnostics kits were purchased from M/s. Agappe Diagnostics Ltd, Kerala, India. All other chemicals and reagents used in the study were of analytical grade. Plant Materials Fresh leaf samples of Annona reticulata (AR) were collected during July 2016 and the plant samples were authenticated (Reference no: Tree reg. vol.1. page no.2 annona 10) and supplied by Dr. GSK Swamy from Col- lege of Horticulture, Mysuru, Karnataka, India.