Toxicity of methanolic extract from cocoa pod husk (Theobroma cacao L) using brine shrimp lethality test (BSLT)

Indonesia is one of the countries that produces a large amount of Theobroma cacao L. in the world. The extract of water, ethyl acetate, chloroform, and n -hexane of cocoa pod husk contains several chemical constituents that exhibit potential biological activities. This study aimed to determine the toxicity of cocoa ( Theobroma cacao L.) pod husk extract. Cocoa pod husk (6 kg) was first macerated with methanol and then extracted using n -hexane to obtain n -hexane extract (TCH) weighing 68.81 g (40.91%). The methanol extract was further extracted with ethyl acetate to obtain 17.98 g (10.69%) ethyl acetate extract. The methanol extract thus obtained was 76.38 g (45.41%). The methanol extract was further extracted using acetone to give 11.91 g (15.59%) acetone extract (TCA) and 45.33 g (59.35%) methanol extract (TCM). Theobroma cacao Acetone extract (TCA) was further isolated by gravity chromatography column and 11 fractions (TCA 1-TCA 11) were obtained. TCA 3 was re-chromatographed using an eluent system of n -hexane: ethyl acetate (8.5:1.5). The toxicity of the TCA extract and its combined fractions (TCA 1-TCA 11) on Artemia salina Leach larvae was assessed. TCA fraction 3 was found to be toxic with an LC 50 value of 136.3 ppm.