ABSTRAK Penelitian ini bertujuan untuk mengetahui perubahan profil hematologi (eritrosit, hemoglobin, MCV, MCH, MCHC, hematokrit, leukosit, limfosit, mixed cells, dan trombosit) dan profil biokimia (ALT, AST, alkali fosfatase, total bilirubin, total protein, albumin, kolestrol, glukosa, kreatinin, dan serum urea) tikus putih (Rattus novergicus) jantan strain wistar yang diberikan ekstrak n-heksan kulit akar biduri (Calotropis gigantea). Dua puluh delapan ekor tikus jantan dibagi secara acak ke dalam 3 kelompok perlakuan dan 1 kelompok kontrol yang masing-masing kelompok terdiri atas 7 ekor tikus. Kelompok kontrol (K) hanya diberikan pelarut (Na-CMC 1 %), kelompok I (D1) diberikan ekstrak n-heksan kulit akar biduri (Calotropis gigantea) 125 mg/kg BB, kelompok II (D2) diberikan ekstrak n-heksan kulit akar biduri (Calotropis gigantea) 250 mg/kg BB, dan kelompok III (D3) diberikan ekstrak n-heksan kulit akar biduri (Calotropis gigantea) 500 mg/kg BB. Pemberian ekstrak dilakukan secara oral selama 28 hari berturut-turut. Hasil penelitian menunjukkan bahwa pemberian ekstrak n-heksan kulit akar biduri (Calotropis gigantea) selama 28 hari tidak memiliki perbedaan yang signifikan antara kelompok perlakuan dengan kelompok kontrol terhadap hampir semua profil hematologi kecuali pada parameter trombosit dan mixed-cells. Pada profil biokimia terjadi perbedaan yang signifikan terhadap hampir semua parameter biokimia (ALT, AST, ALP, albumin, total bilirubin, kolestrol, glukosa, dan serum urea) tikus putih (Rattus norvegicus) jantan strain wistar, kecuali pada parameter total protein dan kreatinin. Kata Kunci: Calotropis gigantea, uji toksisitas subakut, profil hematologi, profil biokimia ABSTRACT This study aims to determine any changes in hematological profiles (erythrocytes, hemoglobin, MCV, MCH, MCHC, hematocrit, leukocytes, lymphocytes, mixed cells, and platelets) and biochemical profiles (ALT, AST, alkalie phosphatase, total bilirubin, total protein, albumin, cholesterol, glucose, creatinine, and serum urea) in white rats (Rattus novergicus) of male wistar strain that were given n-hexane root bark extract of Calotropis gigantea. Twenty-eight male rats were randomly divided into 3 treatment groups and 1 control group, each group consisting of 7 rats. The control group (K) was only given a solvent (Na-CMC 1%), group I (D1) was given n-hexane root bark extract of Calotropis gigantea 125 mg/kg body weight, group II (D2) was given n-hexane root bark extract of Calotropis gigantea 250 mg/kg body weight, and group III (D3) was given n-hexane root bark extract of Calotropis gigantea 500 mg/kg body weight. The extract was administered orally for 28 consecutive days. The results showed that the administration of n-hexane root bark extract of Calotropis gigantea for 28 days did not have a significant difference between the treatment group and the control group for almost all hematological profiles except for platelet and mixed-cell parameters. In the biochemical profiles there were significant differences in almost all biochemical parameters (ALT, AST, ALP, albumin, total bilirubin, cholesterol, glucose, and serum urea) of wistar strain male rats (Rattus norvegicus), except in the parameters of total protein and creatinine. Keywords: Calotropis gigantea, subacute toxicity test, hematology profile, biochemical profile