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

Abstract

Abstract 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 LC50 value of 136.3 ppm.