Objective To explore the correlation between vancomycin serum trough concentrations,24-hour area under the curve (AUC24h),and neonatal acute kidney injury (AKI),and determine the optimal exposure thresholds for rational therapy in neonates.Methods A retrospective analysis was conducted the neonates who received intravenous vancomycin treatment from June 2019 to June 2023.The clinical data were collected,including serum trough concentrations,demographic data,clinical information,and relevant laboratory tests.AUC24h was calculated using Bayesian methods.Spearman correlation coefficient was used to assess the relationship between trough vancomycin concentrations and AUC24h.Receiver Operating Characteristic (ROC) curves and calibration curve were used to evaluate the predictive performance of trough concentrations and AUC24h for AKI.The optimal cutoff value was selected using the Youden index.Multivariable Cox regression analysis was used to examine the correlation between trough concentration cutoff value,AUC24h cutoff value,and AKI.Survival curves of patients were plotted.Log-rank test was used to compare differences in the incidence of AKI between groups.Results A total of 135 cases were included.Overall,8.9% (12/135) of the patients experienced nephrotoxicity.Nephrotoxic patients showed higher trough vancomycin concentrations (16.10 mg/L vs.12.40 mg/L;P>0.05) and AUC24h (647.53 mg·h/L vs.503.25 mg·h/L;P<0.01) compared to those without AKI.Trough concentrations showed weak correlation with AUC24h (Rho=0.304).ROC curve analysis revealed a higher area under the curve of AUC24h than trough concentrations (0.763 vs.0.608),with Brier scores<0.1.The cutoff value of trough concentration and AUC24h was 24.90 mg/L and 575.78 mg·h/L respectively for AKI.Cox regression analysis demonstrated correlation between trough concentration and AUC24h and AKI risk.Log-Rank tests indicated significant difference in AKI rates for both dichotomized trough concentrations and AUC24h (P<0.01).Conclusions Vancomycin serum trough concentrations and AUC24h,calculated using Bayesian methods,can predict vancomycin-induced nephrotoxicity in neonates.Cutoff value of trough concentration>24.90 mg/L and AUC24h>575.78 mg·h/L can be considered as risk predictors in clinical prescription.