Danni Ramdhani et al. Int. Res. J. Pharm. 2019, 10(5) 56 INTERNATIONAL RESEARCH JOURNAL OF PHARMACY www.irjponline.com ISSN 2230 – 8407 Research Article ANTIMUTAGENIC ACTIVITY OF WATER FRACTION AND ITS SEPARATING FRACTION FROM THE PUSPA PLANT (SCHIMA WALLICHII KORTH) BY THE MICRONUCLEUS METHOD Danni Ramdhani 1 *, Anas Subarnas 2 , Ahmad Muhtadi 2 , Cucu Hadiansyah 3 1 Pharmaceutical Analysis and Medicinal Chemistry Departement, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor, Sumedang, West Java, Indonesia 2 Pharmacology and Clinical Pharmacy Departement, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor, Sumedang, West Java, Indonesia 3 Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jatinangor, Sumedang, West Java, Indonesia Corresponding author email: ramdhani07@gmail.com Article Received on: 22/03/19 Approved for publication: 08/05/19 DOI: 10.7897/2230-8407.1005162 ABSTRACT Objective: An investigation on mutagenic activity of two fraction separated from the water fractions of Schima wallichi Korth has been carried out on Swiss Webster male mice using the micronucleus method. Methods: The simplicia of puspa leaves was extracted by maceration method with methanol solvent for 2 days. The crude extract obtained was fractionated based on the principle of polarity, using n-hexane, ethyl acetate, n-butanol and water solvents. Each fraction was administered orally at the same doses of 350 and 700 mg/ kg of body weight. Results: Antimutagenic activity was slowly decreasing frequency of micronucleus in every 1000 erythrocytes of bone marrow preparation for each test animal. The result of the investigation indicated that the two fractions, at a dose of 700 mg/kg of body weight decreased the frequency of micronucleus significantly as compared with the control group, with the inhibitory of 34.25 % and 44.29 % respectively potency. Phytochemical screening of the fractions indicated that the first fraction contained two main compounds, quinone and polyphenol, and the second one contained saponin in addition to quinone and polyphenol. Conclusion: The antimutagenic activity of the puspa (Schima wallichii Korth) plant from the water fraction and its separation showed significant results. The biggest percentage of inhibition of mutation is indicated by fraction 3 which is 44.29 %. There is an alleged correlation between the presence of saponin compounds in fraction 3 with the ability to inhibit the occurrence of mutations. Keywords: Antimutagenic, Schima wallichii Korth, micronucleus assay, cyclophosphamide, polyphenol, saponin. INTRODUCTION Mutations are the cause of congenital metabolic disorders cellular system that can trigger morbidity and mortality in living organisms 1 . Mutagen compounds are involved in the process of initiation and promotion of several human diseases especially cancer. The ability of new bioactive compounds phytocompounds in counteracting promutagenic and carcinogenic effects get some satisfying results 2 . Antimutagenic and anticarcinogenic properties of a wide variety of dietary constituents and plant secondary metabolites have been reported. The antimutagenic effect is closely related to many classes of phytocompounds, especially flavonoids, phenolic, and saponin compounds present in plants 3 . Puspa (Schima wallichi Korth) plant is an Indonesian tropical plant. Puspa plants are found to grow in groups in tropical forests on Java and Sumatra. Puspa plants belonging to the tribe of Theacea, are widely used empirically in traditional medicinal herbs by the Javanese tribe 4 . The study of genotoxicity through an in vivo micronucleus (MN) test on mouse bone marrow is an important part of the tests used to identify the mutagen hazards 5 . Schmid and Heddle have the initiative that a simpler approach to measuring chromosome damage in vivo is to measure micronucleus 6 . Micronuclei frequencies have been considered to be a reliable index for detecting chromosome breakages and loss 7 . MN is cell nucleus that contains a chromosome fragment that is damaged in part or all of its chromosomes. In the process of cell division, micronucleus is not inserted into the nucleus. MN can be induced by defects incell repair machines and accumulated DNA damage and chromosomal aberrations. Genotoxic agents can cause the formation of MN which causes cell death, genomics instability, or development of cancer 8 . Previous research using the micronucleus method using mice animals showed that methanol extract from the puspa plant had antimutagenic activity 9 . MATERIALS AND METHODS Plant Material and Sample Preparations Plant material used in this study is the leaf part of the puspa plant (Schima wallichii Korth) obtained from the Ciwidey forest, in West Java Province, Indonesia. Puspa leaves obtained are then dried and crushed with a mechanical grinder. The resulting powder material is then extracted by maceration with methanol