Abstract: Objective To assess the value of Halena SAS-3 full automatic hemoglobin agarose electrophoresis in the diagnosis of thalassemia. Methods Hemoglobin A and A2 was detected by agarose electrophoresis in 200 blood samples collected from 170 healthy subjects, 2 patients with beta-thalassemia, 12 with acute lymphoblastic leukemia, 12 acute myelocytic leukemia and with 4 non-Hodgkin lymphoma. Results The Mean±SD of hemoglobin A in the healthy subjects and patients with beta-thalassemia, acute lymphoblastic leukemia, acute myelocytic leukemia and non-Hodgkin lymphoma was 97.55±0.51, 97.01±0.329, 97.42±0.57, 97.44±0.55, and that of A2 was2.44±0.51, 2.99±0.32, 2.84±0.28, 2.55±0.55, respectively. The values of electrophoresed hemoglobin F of the 2 patients with beta-thalassemia were higher than those of the normal adults. The within-run coefficient of variation (CV) and between-run CV was 0.17 and 7.76 for hemoglobin A, and 0.16 and 7.14 for hemoglobin A2, respectively. Conclusions Halena SAS-3 full automatic agarose electrophoresis system possesses strong resolving power in detecting trace abnormal hemoglobin and accurately identifies unusual evaluation of hemoglobin A2 and F to facilitate the diagnosis of thalassemia.