Fatty liver-hemorrhagic syndrome (FLHS) is a common nutritional disease in commercial layers and breeders. The most important clinical sign of FLHS is a sudden drop in egg production and increased mortality which causes significant economic loss in the poultry industry. However, the current diagnostic method for FLHS is based on the gross findings at necropsy which is not helpful to reduce the economic loss because of lateness of diagnosis. Therefore, we need early diagnosis and diagnostic methods before chickens were affected by FLHS. In this study we tried to evaluate the effectiveness of clinical pathology including blood chemistry as an early diagnostic method for FLHS in commercial chickens. Profiles of blood biochemistry were compared between two flocks selected in the same commercial layer farm based on the presence of FLHS clinical sings. A flock with clinical signs of FLHS was designated as FLHS and other flock without clinical signs of FLHS as Non-FLHS. Several parameters of blood biochemistry were selected and compared between FLHS and Non-FLHS to evaluate the possibility of early diagnosis. Average concentrations of serum cholesterol, serum calcium, aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and creatine kinase (CK) were 139.4 ± 87.2 (mg/dL), 24.5 ± 5.4 (mg/dL), 153.6 ± 23.1 (IU/L), 1238.3 ± 475.2 (IU/L) and 1107.3 ± 422.8 (IU/L) in Non-FLHS flock, respectively, and 210.2 ± 173.2 (mg/dL), 25.2 ± 4.1 (mg/dL), 174.3 ± 53.5 (IU/L), 1694.9 ± 691.3 (IU/L) and 1104.9 ± 472.9 (IU/L) in FLHS flock, respectively. The activities of serum cholesterol, AST and LDH except CK, were significantly higher in FLHS than those in Non-FLHS flock (p<0.05). Some birds of FLHS flock showed 2~17 times greater than in Non-FLHS flock. For the definitive diagnosis of FLHS in the flocks tested for blood chemistry, we analyzed fat content and histological lesion score in the liver sampled from both FLHS and Non-FLHS flock. Average liver fat contents based on dry weight were 16.1 ± 0.4 (%) in Non-FLHS flock and were 21.6 ± 16.0 (%) in FLHS flock. These result confirmed that FLHS flock was definitely affected by FLHS. The above results suggest that selected parameters of blood biochemistry, particularly AST, could be useful to diagnose FLHS before significant liver damage occurred in commercial layers.