Purpose: CA 19-9 is the most widely used tumor marker for the diagnosis of digestive system tumor, especially pancreatic and biliary tract cancer. This study was conducted to improve the result value near reference range by comparing the reagents of CA 19-9 one step method and two step method. In addition, it was intended to establish a standard for selecting reagents. Material and Methods: 120 patients who visited the National Cancer Center in 2023 were selected as subjects for this study. The reagents used in the study were CA 19-9 IRMA kits (Shinjin, Korea) and three types of reagents were compared. Two step method reagent that is currently being used (A), one step method reagent (B) and two step method reagent improved by request (C) were compared and regression analysis was performed on their data. And we also performed recovery test, linearity test and hook effect test for each reagent. Result: There were 46 cases of reagent B in which the concentration value was lower than the result measured in reagent A that was previously used, and 77 cases of reagent C. As a result of regression analysis of reagents A, B, and C, the coefficients of determination of reagents A and B, reagents A and C, and reagents B and C were 0.653, 0.577, and 0.875. In the recovery rate test and the linearity test, the results of all reagents were good, and in the hook effect test, reagent B showed a hook effect at a low value. Conclusion: The improved reagent C appears to have been improved based on the concentration value of reagent B, which the manufacturer judged to be more stable at low concentrations. The hook effect in reagent B can be a fatal reason for disqualification when selecting reagents in general patient samples which high-concentration samples appear frequently. The first improved reagent C will be able to be used once it is confirmed that it has more stability for various concentration values.