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Purpose: Recent vancomycin dosing and monitoring guidelines recommend monitoring vancomycin area under the 24-hour time-concentration curve instead of traditional trough-only monitoring. This study aimed to compare the total costs of vancomycin dosing and monitoring between trough-guided and AUC-guided approaches in a quaternary hospital from Brazil., Methods: In this retrospective cohort study, patients were divided into 2 groups according to the monitoring method. Patients with previous renal impairment were excluded. Vancomycin AUC was estimated by using 2 steady-state serum concentrations and first-order kinetics equations. The primary outcome was total cost of vancomycin therapy and monitoring from the hospital perspective, which included costs of cumulative doses, laboratory fees, materials used in blood collection, nursing time for collection, and pharmacist time for result interpretation., Findings: A total of 68 patients were included in the AUC/MIC-guided monitoring group, and 76 patients were included in the trough-guided monitoring group. There were no significant differences between groups regarding baseline serum creatinine level, duration of vancomycin therapy, and cumulative vancomycin dose. The median (interquartile range) total vancomycin drug and monitoring cost was $298.32 ($153.81-$429.85) for the AUC/MIC-guided group compared with $285.59 ($198.81-$435.57) for the trough-guided group (P = 0.9658)., Implications: Vancomycin AUC estimation using 2 steady-state serum concentrations and first-order kinetics equations is a feasible alternative for limited-resource institutions that intend to transition from a trough approach to AUC/MIC-guided monitoring., (Copyright © 2022 Elsevier Ltd. All rights reserved.)