AbstrakPenelitian ini bertujuan untuk mengkaji rendemen, kandungan gizi dan uji in vitro ekstrak ikan buntal ikan buntal pisang dari perairan Kabupaten Cirebon. Metode yang digunakan pada penelitian ini meliputi analisis proksimat, analisis fitokimia dan uji in vitro ekstrak dari ketiga bagian ikan buntal pisang (daging, jeroan dan kulit). Ikan buntal pisang yang digunakan berukuran (10-13) cm. Rendemen yang didapatkan yaitu daging 37,80%, tulang 45,71%, jeroan 7,25% dan kulit 9,23%. Ikan buntal pisang memiliki potensi terbesar kandungan proteinnya, berkisar 18,54%-25,31%. Seluruh ekstrak memiliki kandungan alkaloid dan memiliki sedangkan ekstrak daging pada uji molisch. Hasil uji fitokimia menunjukkan bahwa daging ikan buntal pisang memiliki kandungan tetrodotoksin tetapi jumlahnya belum diketahui. Nilai LC 50 pada ekstrak daging sebesar 104.3498 μg/mL, ekstrak jeroan sebesar 105.4564 μg/mL, dan ekstrak kulit sebesar 104.9637 μg/mL. Hasil tersebut menggambarkan bahwa ekstrak daging, jeroan dan kulit pada ikan buntal pisang tidak toksik, walaupun terdapat indikasi adanya tetrodotoksin pada dagingnya.Kata kunci: ekstrak, fitokimia, ikan buntal pisang, kandungan gizi Abstrak This aims of study was to assess yield, nutrient content and in vitro assay of extract on green rough-backed puffer from Cirebon district. The method used in this study was the proximate analysis, phytochemicals, and BSLT test of extracts on three parts of puffer fish (meat, viscera and skin). Puffer fish used measure around 10-13 cm. The yield obtained was meat 37.80%, bone 45.71%, viscera 7.25% and skin 9.23%. Protein content showed greatest potential, protein in the fish meat was 18.54% while the skin was 25.31%. Phytochemical test showed the entire extract has a positive value for alkaloids and have a negative value on the other reagents, except meat extract on molisch test. Phytochemical test results indicated that the meat of green rough-backed puffer contain tetrodotoxin, but the amount was unknown. LC 50 values in meat extract was 104.3498μg/mL, extract the viscera of 105.4564μg/mL, and the skin extract was 104.9637μg/mL. These results showed that the extract of meat, viscera and skin of the puffer fish is not toxic, although there are indications of tetrodotoxin in the flesh.