To evaluate the biocompatibility of biomedical materials for hepatic tissue engineering, another new method was introduced to observe hepatocytes functions. In this experiment, hepatocytes were seeded onto four kinds of membranes of PLLA, PLGA (90:10), PLGA (75:25), and Chitosan cross-linked with collagen. The culture mediums were collected at 21 day after seeding, and then albumin (Alb), Urea (UN), glucose (Glu), total protein (TP), and triglyceride (TG) in the supernatant were detected by automatic biochemical analyzer. Results showed that hepatocytes on film of Chitosan cross-linked with collagen exhibited the highest level of TP and TG. These results were highly corresponding with the results of morphological observation. This data indicated that analyzing TP and TG in culture medium by automatic biochemical analyzer might be applied to evaluate hepatocytes biocompatibility on materials as a convenient and feasible method.