The 5 th International Conference on Pharmaceutical Nanotechnology / Nanomedicine 2021 | 60 ANTIDIARRHEAL ACTIVITY OF EXTRACT ETHANOL MELINJO LEAVES (GNETUM GNEMON L. (LINN.)) IN WISTAR MALE WHITE RATS INDUCED BY ESCHERICHIA COLI AND EXTRACT STANDARDIZATION Original Article HERLINA 1 , INDAH SOLIHAH 1 , DIAN PERMATA WIJAYA 1 , ANISSA NADIA NURRAHMAH 1 1 Department of Pharmacy, Faculty of Mathematics and Sciences Sriwijaya University, Inderalaya Campus, Indonesia * Email: rinaafdil@gmail.com Received: 15 Dec 2021, Revised and Accepted: 18 Mar 2022 ABSTRACT Objective: Determination of the antidiarrheal activity of extract ethanol melinjo leaves (Gnetum gnemon L. (Linn.)) in Wistar male white rats induced by Escherichia coli and extract standardization has been performed. Methods: This research was conducted by in vivo method using white male rats of Wistar strain induced by Escherichia coli. The treatment group was divided into 6 groups: normal, negative control, positive control, and groups with doses 150, 300, and 600 mg/kg BW. Negative control was given Na-CMC and positive control Gentamicin. The initial time diarrhea occurred 24-30 h after administration of Escherichia coli suspension. Results: The results of the phytochemical screening of the ethanolic extract of melinjo contain secondary metabolites of flavonoids, steroids, tannins, and saponins. Standardization of the extract ethanol melinjo leaves (Gnetum gnemon L.) meet the predetermined standards, while the acid insoluble ash content parameters did not meet the predetermined standards. The parameters feces weight, feces consistency, frequency diarrhea, body weight, number of Escherichia coli colonies feces dose 600 mg/kg BW had an effect that was almost close to positive control. The results showed that anti-diarrheal effect dose 150 mg/kg BW 35.75%, dose 300 mg/kg BW 43.02%, dose 600 mg/kg BW 50.14%. This shows that ethanol extract melinjo leaves dose 600 mg/kg BW wasn’t significantly different from the positive control (p<0.05). Effective Dose (ED50) Ethanol Extract Melinjo Leave as antidiarrheal was 578.2468 mg/kg BW. Conclusion: Extract ethanol melinjo leaves (Gnetum gnemon L. (Linn)) dose 600 mg/kg BW has the potential to be used as antidiarrheal and its extract meet the predetermined standards, except the acid insoluble ash content parameters did not meet the predetermined standards. Keywords: Melinjo leaves, Antidiarrheal, Escherichia coli, Effective dose 50 © 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2022.v14s3.12 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Diarrhea is an increase in frequency and decreased consistency of feces when compared to normal individuals, in other words, diarrhea is loose or liquid bowel movements that occur three or more times within 24 h [1]. In Indonesia, diarrhea is the second leading cause of death among children under five and number five for all ages. One of the side effects of diarrhea is the body experiencing dehydration (loss of fluids and electrolytes) so that if it is not addressed it can immediately cause death [2]. One of the causes of diarrhea is a bacterial infection. Escherichia coli bacteria are the bacteria that most often cause diarrhea with a ratio of 19% [3]. Escherichia coli bacteria become pathogenic when the number of these bacteria in the digestive tract increases. Modern medicine in the form of antibiotics to fight Escherichia coli infection has developed rapidly. The irrational use of antibiotics in various fields of medical science is one of the causes of resistance that can be obtained. The use of natural ingredients as traditional medicine can be an alternative to the use of antibiotics [4]. One of the plants that have antibacterial properties is the leaves of Melinjo (Gnetum gnemon L.). Melinjo leaves have many benefits in the health sector, such as preventing cancer, lowering blood sugar, as an antioxidant, high nutritional food, inhibiting the aging process, antibacterial [5]. Melinjo contains active compounds such as alkaloids, flavonoids, saponins, and tannins [6]. Flavonoid compounds as antibacterial can damage bacterial cell membranes, followed by the release of bacterial intracellular compounds [7]. Alkaloid compounds as antibacterials work by disrupting the constituent components of the cell wall layer so that they are not formed completely [8]. Tannin compounds have antibacterial power by inhibiting the reverse transcriptase and DNA topoisomerase enzymes so that bacterial cells cannot be formed. Based on the results of observations, a concentration of 60% was able to inhibit Escherichia coli bacteria in a maximum inhibition zone of 11 mm with the category of strong inhibition [6]. In this study, the researchers wanted of antidiarrheal activity of extract ethanol melinjo leaves (Gnetum gnemon L. (Linn.)) in Wistar male white rats induced by Escherichia coli and extract standardization. MATERIALS AND METHODS Plant material The materials used in this research consisted of melinjo leaves (Gnetum gnemon L.). The sample used was the leaves of the melinjo plant (Gnetum gnemon L.) with wet weight of 3.2 kg obtained from the Inderalaya region, South Sumatra, Indonesia. The plant determination of the melinjo leaves (Gnetum gnemon L.) is carried out with the aim of macroscopically identifying the truth of the plant to be used. Determination was carried out at the Laboratory of the Plant Conversion Center of the Purwodadi Botanical Gardens, Purwodadi, Pasuruan, East Java, Indonesia (No. 0044/IPH.06/HM/I/2019). Chemical and reagent Male white rat Wistar strain (Abduh Tikus Palembang), ethanol 70% (Dira Sonita ® ), concentrated hydrochloric acid (Brataco ® ), sodium acetate (Brataco ® ), filter paper (Whatman ® ), Mayer reagent (Paison ® ), Wagner reagent (Paison ® ), Liebermann-Bourchard reagent (Paison ® ), Dragendrof reagent (Paison ® ), metal magnesium (Brataco ® ), aluminum chloride (Brataco ® ), iron (III) chloride (Merck ® ), aluminum foil, standard rat food, and Gentamicin injection. Preparation of extract ethanol melinjo leaves Extract Ethanol Melinjo leaves were made using the maceration method. A total of 1 kg of Melinjo leaf powder was extracted by the maceration method for 3 x 24 h using 5 L of 70% ethanol solvent. Remaceration was carried out after 72 h by replacing the solvent of 2.5 l 70% ethanol for 3 x 24 h. The next remaceration is carried out in the same way as the first remaseration. Stirring is done 2 times a International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 14, Special Issue 3, 2022