Background Serum creatinine (SCr) is a useful diagnostic marker for the assessment of renal function. Accurate quantitation of SCr is clinically important in calculation of glomerular filtration rate (GFR). Method To confirm whether there are differences in SCr between enzymatic kits of different manufacturers, the analytical performance of the matched and open test system in the measurement of SCr was evaluated. The analytical performance evaluation was conducted according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Precision, accuracy, linearity, dilution, lower limit of measurement and analytical interference were studied between the two test systems. Results The performance of SCr from the open test system was in compliance with the matched test system with good precision, accuracy, and linearity. In presence of most common interferents, both test systems could lead to accurate creatinine results except for the existence of specified drugs. For dobutamine, the open test system showed better anti-interference performance than the matched system. Conclusion This study provides referable opinions for clinical laboratory selection on the test system and a framework for future analogous studies based on different test systems.