ANTIHYPERURICEMIA ACTIVITY OF THE ETHANOL EXTRACT OF ROSELLE CALYX AND ITS FRACTION (HIBISCUS SABDARIFFA LINN) ON MALE WISTAR RATS Original Article SRI WAHYUNINGSIH a, b ELIN YULINAH SUKANDAR a , SUKRASNO a , DEBBY NADYA LOFIKA b a School of Pharmacy, Bandung Institute of Tecnology, Jalan Ganeca no. 10 Bandung,Indonesia, b Faculty of Pharmacy, Universitas Jenderal Achmad Yani, Jalan Terusan Jenderal Sudirman Po Box 148 Cimahi, Indonesia Email: sri_wahyuningsih40@yahoo.co.id Received: 21 Nov 2015 Revised and Accepted: 03 Feb 2016 ABSTRACT Objective: The objective of this study was to determine anti-hyperuricemia activity of ethanol extract of roselle calyx (Hibiscus sabdariffa L) and its fraction on male Wistar rats. Methods: Two different methods were used. First method induction was done by giving potassium oxonate and diet high purine whereas allopurinol as a comparison. The parameter was level uric acid in serum (uricostatic activity). Second methods induction was done by giving potassium oxonate intraperitoneally and sodium urate orally; with pprobenecid as comparison Parameters used were uric acid level in serum and urine (uricosuric activity). Results: Ethanol extract roselle calyx at a dose of 40 mg/kg bw, water fraction at a dose of 37.5 mg/kg bw and ethyl acetate fraction at a dose of 6.25 mg/kg bw at day 14 showed a significant reduction in serum uric acid level compared to control group (p<0.05). Besides that ethanol extract of Roselle calyx at dose 80 mg/kg bw, water fraction at doses 37.5 and 75 mg/kg bw also ethyl acetate at doses 6.25 and 12.5 mg/kg bw had uricosuric effect that increased the excretion of uric acid in urine significantly compared to control group (p<0.05). Conclusion: The water fraction of roselle calyx at doses 37.5 and 75 mg/kg bw and ethyl acetate at dose 6.25 and 12.5 mg/kg bw was effective as an anti-hyperuricemic agent. The ethanol extract of rosele calyx and its fraction had a uricosuric effect similar to probenecid. The best uricosuric effect was showed by ethyl acetate fraction at dose 6.25 mg/kg bw. Keywords: Hibiscus sabdariffa Linn, Water fraction, Ethyl acetate fraction, Calyx, Uric acid, Antihyperuricemia © 2016 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/ ) INTRODUCTION The prevalence of hyperuricemia has increased in the last five years in the world [1-3]. Uric acid is the end product of purine degradation. A Higher level of blood uric acid is called hyperuricemia and can cause the precipitation of uric acid in the joint inducing pain sensory and the symptom called Gout. Hyperuricemia could be due to the increase of metabolism (overproduction) of uric acid, or reduction secretion in the urine (underexpression) or a combination of both [4]. Hyperuricemia may be an asymptomatic condition with an increased serum uric acid level, if uric acid concentration higher than 7.0 mg/dl, it is associated with an increased risk for Gout [4]. Allopurinol is the only drug approved for use in inhibiting uric acid synthesis through the inhibition of xanthine oxidase. The major side effects of allopurinol are skin rash, leukopenia, and occasional gastrointestinal toxicity [4,5]. Because of that, many types of research are developed for medicinal plants having the effect of lowering the uric acid level that can be therapeutic options for hyperuricemia. Medicinal plants are believed to be important sources of new substances with potential therapeutic effect. The Roselle calyx (Hibiscus sabdariffa Linn) have been used in Indonesian traditional medicine for the hyperuricemia treatment [6]. However data as the hypouricemic agent has not been reported, so this experiment was conducted to find out the activity of roselle calyx as anti hyperuricemic. The aims of this study were to determine the anti hyperuricemic activity of extract ethanol of roselle calyx (Hibiscus sabdariffa) and its fractions on male Wistar rats. MATERIALS AND METHODS Material Oxonic acid potassium salt/Potassium oxonate (Sigma), uric acid (Sigma), Probenecid (Sigma), Allopurinol (Sigma), Uric acid kit reagent (Reagent Human), Gnetum genom seeds. Roselle calyx collection and determination Roselle calyx was collected from Manoko Lembang Indonesia. It was determined in The Herbarium Bandungense School of Life Science Biosciences ITB. Roselle calyx was then dried and ground to powder. Roselle calyx extraction Roselle powder (250 g) was extracted using Soxhlet apparatus with ethanol 96 % as solvent until clear filtrat was obtained. The extract was evaporated until a viscous extract was obtained (yield: 35.8%). Roselle calyx extracts characterization Specific parameters such as organoleptic, water soluble extractive and ethanol soluble extractive, nonspesific parameters were included loss on drying, total ash, and acid insoluble ash, phytochemical screening was measured. Roselle calyx extracts fractionation Fractionation of the extract was performed by dispersing extract in etanol-water (1:1) followed by liquid extraction with hexane. The aqueous fraction was further extracted with ethyl acetate. Hexane, ethyl acetate, and the water fractions obtained were evaporated under reduced pressure to dryness. Yield of n- hexane, ethyl acetate, and water fractions were 46.27%, 15.28 %, and 8.68.%, Antihyperuricemic assay The animals that used were male Wistar rats aged 8-10 w, 200-250 gram weight, from Biosciences and Biotechnology Research Center, ITB. Antihyperuricemic assay was performed in two methods. The first methods were done in order to evaluate uricostatic activity. The second method was employed to evaluate the uricosuric activity. This experiments design was approved by the Animal Research Ethics Committee of ITB (Reg. No. 03/KEPHB-ITB/11-2014). International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 8, Issue 3, 2016