Background: Recent American guideline published in 2020 recommend using 24-h area under the curve (AUC)/minimum inhibitory concentration instead of vancomycin serum concentration (Ctrough) for vancomycin therapeutic drug monitoring (TDM). However, Ctrough-based TDM is widely used in clinical practice. Thus, this retrospective study aimed to compare Ctrough-based and AUC-based TDM. Methods: We evaluated patients' TDM data with at least one vancomycin trough measurement. Patients younger than 18 years, admitted to an intensive care unit, or on renal replacement therapy were excluded. The variables of Ctrough-based and AUC-based TDM were simulated using Mw-Pharm++ (Mediware, Czech Republic) with vancomycin two-compartment model. The therapeutic range was 400-600 mg*h/L and 15-20 mg/L for AUC and Ctrough, respectively. We evaluated the correlation between Ctrough and AUC, the attainment rate of AUC target range, and changes in vancomycin dose and Ctrough when AUC-based TDM is applied. Results: One hundred and four patients were enrolled. Ctrough and AUC correlated moderately (R=0.707, P <0.001). Among 31 patients with Ctrough of 15-20 mg/L, the AUC of only 18 patients was within the target range (18/31, 58.1%). In addition, most patients with Ctrough of 10-15 mg/L had the AUC within the target range (57/66, 86.4%). The respective vancomycin dose and Ctrough were expected to be significantly lower in AUC-based TDM simulation than those in Ctrough-based TDM simulation Conclusions: Ctrough of 15-20 mg/L for vancomycin monitoring is not appropriate for attaining AUC target range. Targeting either AUC or lower Ctrough is recommended for vancomycin TDM.