Clinical laboratory examinations frequently include biochemical analysis of the liver. The presence of alanine aminotransferase (ALT) and aspartate transaminase (AST) in serum can be used to identify liver damage. In this study, a centrifugal microfluidic-based clinical biochemical detection system was developed for the detection of liver function markers. Using the centrifugal microfluidic chip and centrifugal force on the chip, separation of blood cells and serum was performed. The extraction and mixing of quantitative serum and diluent were completed under the chip design of microchannels and microchambers. The lyophilized reagent beads in the chip interacted with the combined solution. The Fuzzy PID algorithm regulates the power of the heating film to deliver the ideal reaction temperature. In accordance with Beer–Lambert, the rate of change in the absorbance of the reaction solution at 340 nm of the light source was measured and a standard curve for the relationship between concentration and rate of change in absorbance was constructed. The system is portable, quick, and simple to use because it uses a centrifugal microfluidic chip instead of the conventional detection and analysis approach. In the future, it is anticipated that the system will have several applications in the detection of highly integrated on-chip point-of-care devices.