Holothuria atra is one of the many species of sea cucumber that is commonly found in Indonesia and considered as a potential source of bioactive compounds. This study was conducted to analyze the proximate contents and obtain the best solvent to extract the bioactive compounds from H. atra according to the result of phytochemical screening and antibacterial activity. A total of 14 samples were collected with an average of total length value of 22.89 ± 6.28 cm, the width of 4.00 ± 0.81 cm, wet weight of 114.89 ± 54.86 g and gutted weight of 70.22 ± 35.58 g. The bioactive compounds of H. atra’s flesh were extracted using n-Hexane, ethyl acetate and methanol. The result of statistical analysis indicated that the type of solvents in bioactive extraction, extract concentration, and the type pathogen impacted the antibacterial activity. The crude extract from methanol contained alkaloid, flavonoid, glycosides, saponin, steroid and triterpenoid with 3 spots in TLC; ethyl acetate crude extract had alkaloid, flavonoid, glycosides, phenyl hydroquinone, saponin, steroid and triterpenoid with 10 spots in TLC; while n-hexane only contained glycosides, steroid and triterpenoid with 7 spots in TLC.

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