Background: It is not yet clear whether the trace blood gas analyzer can be used for biochemical detection of newborns. This study aimed to evaluate the reliability of the method for the detection of bilirubin in infants.Method: Based on the CLSI EP15-A2 document, the analytical performance of the blood gas analyzer method for bilirubin detection in neonates was validated. The resulting data of 363 simultaneous bilirubin detection with blood gas analyzer (optical method) and biochemical analyzer (enzymatic method) were reviewed. According to the CLSI EP9-A3 document, the relevance and consistency of the measurement results were evaluated by Pearson correlation analysis, Passing-Bablok regression, and Bland-Altman deviation analysis. Results:The precision and accuracy of the Werfen GEM 4000 blood gas analyzer for the detection of different levels of bilirubin samples adhered to the manufacturer's statement and industry quality standards.The bilirubin detection values of the 2 methods showed a good correlation, and both of them were significantly correlated (P<0.001). Passing-Bablok regression results showed that the regression equation of the bilirubin detection value of the 2 methods is y = −21.00 + 1.17x, with the slope as 1.17 [95% confidence interval (CI): 1.15 to 1.19], and the intercept was −21.00 (95% CI: −23.62 to −18.71), the data of the 2 sets were not consistent in each concentration range. The Bland-Altman plot demonstrated that the bilirubin detection value of 16/363 cases (4.4%) for the 2 methods exceeded the 95% limits of agreement (95% LoA); of which the maximum bias was −30.34 (95% CI: −38.48 to −22.26) and there were 5/76 cases (6.6%) outside the 95% LoA in the >300 μmol/L group. Conclusions:The method for detecting neonatal total bilirubin by trace blood gas analyzer basically meets the clinical requirements and can be used for the preliminary screening of neonatal jaundice. However, for severe hyperbilirubinemia that requires close monitoring of dynamics, a precise enzymatic quantification is required.