Background: Surgical Antimicrobial Prophylaxis is administration of a short course of antimicrobial agent prior to surgery. It has long been recommended for routine use in preventing occurrence of surgical site infections. The choice of drug, dosage, route, duration and timing of antimicrobial prophylaxis has been specified in the clinical practice guideline issued by American Society of Health-System Pharmacists. In the present study, we aim to seek the practice of antibiotic prophylaxis at a tertiary care centre in central Nepal and report the adherence to American and National Antimicrobial Treatment Guideline where applicable. Methods: This hospital-based cross-sectional study was conducted between January to June 2023 among 232 patients undergoing surgery at Chitwan Medical College Teaching Hospital. Data collection tool prepared with slight modification from previous studies recorded the demographic details, surgical data, surgical antimicrobial prophylaxis usage data and assessment of Prophylaxis data. Data analysis was done using SPSS-20. Results: We observed compliance to selection of drug, route of administration, dosage of antimicrobial prophylaxis with American guideline. All (100%) prophylaxis was administered parenterally for more than 72 hours in over 40% cases. Choice of drug was appropriate in over 84% of cases and dosage adequacy was observed in over 61% cases. Surgical antimicrobial prophylaxis was administered most frequently 30 minutes before surgery (52.59%). Conclusions: All the surgical patients were found to receive surgical antimicrobial prophylaxis. Ceftriaxone was the most commonly administered drug followed by combination of piperacillin and tazobactam. The duration of treatment post-surgery was found to be unnecessarily broader than recommended guideline.