Research methods 1. Tool Materials and Methods 1. The Place and Time of Study

Proceedings of MatricesFor IITTEP – ICoMaNSEd 2015 ISBN: 978-602-74204-0-3 Chemistry Page 166 in addition to the use of DDT also can cause health problems and environmental problems, NGuessan, et al., 2009 and Riedel, et al., 2008. The use of abate in Indonesia has been done since 1976 or has been used for more than 30 years, so the continuous use of insecticides may increase the risk of pesticide residue contamination in water, especially drinking water Aradilla, 2009. Syahputra et al. 2006 reports from various regencies in Indonesia, there are more than 40 species of plants that can potentially be used as a botanical insecticide. One of the plants that have been isolated by researchers which contains the active compound of vegetable insecticide is soursop Annona muricata seed with LC50 = 117. 27 ppm Komansilan et al. 2012. Plants that have the potential to be developed as a phyto-insecticide is Barringtonia asiatica Kurz Lecythidaceae with the common name ‘ Pohon Racun Laut ’Sea Poison Tree or in Indonesia known as Bitung Ecology and Evolutionary Biology Greenhouse EEBG, 2006. B. Asiatica is known to have active compounds which are known to cause the death of the insect pests. Methanol extract of B. asiatica seeds are toxic for Crocidolomia pavonana with LC50 values of 0.66 at 7 days after treatment. The application of B. asiatica seed extract also has affected the oviposition with the effective concentration of 0.96 that causes female C. pavonana do not lay eggs on plants. Response of larvae shows that the extract of B. Asiatica, besides having a poisonous character, it also has antifeedant activity Dono and Sujana, 2007. One of the active compounds in the B. Asiatica seeds is a saponin Burton et al. 2003. In some places, B. asiatica is used as a medicine and poison of fish. Active compounds in the B. Asiatica seeds that poison fish is saponin compounds group EEBG, 2006. One of the most toxic compounds to fish from B. asiatica seed extract is ranuncide VIII Burton et al, 2003. Research on seed extract of B. Asiatica has been carried out but their toxicity against Dengue Hemorrhagic Fever vector Aedes aegypti mosquito larva is unknown. Therefore, the use of alternative insecticides that are relatively safe for the environment and have minimal side effect or no adverse effects on non-target organisms, is necessary. An alternative method to control the Aedes aegypti mosquito larvae by phyto-insecticides from the extract of B. Asiatica seeds. More intensive research is expected to be able to extract the Hutun B.asiatica seed to kill Aedes aegypti larvae that could help overcome the problem of Dengue Hemorrhagic Fever in Indonesia, especially Manado city. 2. Materials and Methods 2.1. The Place and Time of Study This study is planned to be conducted in Chemistry and Biology Laboratory, Manado State University, on March 2013- December 2013 2.2. Research methods 2.2.1. Tool The tools used in this study are: knife, blender, analytical balance, beaker glass, funnel, erlenmeyer, micro pipette, rotary vacuum evaporator, desiccators, pipette, test tubes, glass, and wristwatch. Proceedings of MatricesFor IITTEP – ICoMaNSEd 2015 ISBN: 978-602-74204-0-3 Chemistry Page 167

2.2.2. Material

The chemicals used in this study is ethanol, methanol, chloroform, butane, concentrated sulfuric acid, ethyl acetate, acetic acid, dimethylsulfoxide, Hutun Baringtonia asiatica seeds , A. aegypty mosquito larvae .

2.2.3. Work Procedures

Materials used in this study is old Hutun Baringtonia asiatica seed that have been collected from Malalayang coast in Manado. The preparations of materials are crop determination, materials collecting, cleaning, drying by blowing not under direct sun and milling to powder using blender.

2.2.4. Biolarvacide Toxicity Test of Hutun Seed Methanol Extract

Hutun Baringtonia asiatica seed powder is extracted by maceration using technical methanol until all components are extracted. The obtained methanol extract is evaporated with a vacuum rotary evaporator until thick. Thick Hutun seed extract is then tested its toxicity to the larvae of Aedes aegypti as its bioindicator. The media of A. aegypti mosquito larvae is made by filling the container with water. The eggs of A. aegypti larvae are stored in a damp place until the eggs of the mosquito larvae hatch and ready for use in testing. Ten beakers are prepared for testing, where for each sample takes nine beakers and a beaker as a control. Concentrated extract is weighed as much as 0.02 g and diluted with 2 mL of ethanol. The solution is put in pipette as much as 5; 50; 500 µL. Each is put in a small bottle and the solvent is evaporated for 24 hours. Insert into the bottle 2 mL of water, 50 mL of dimethylsulfoxide, 10 mosquito larvae of A. aegypti. Then the Hutun seed extract solution is added with water until the volume is 5 mL in a concentration of 10; 100; 500: 1000 ppm. For control, into a small bottle put 2 mL of water, 50 mL of dimethylsulfoxide, 10 mosquito larvae A. aegypti then add water until the volume is 5 mL. The observations are made after 24 hours of the death of the mosquito larvae. Data analysis is performed to find the death concentration LC50.

2.3 Research Model