Background: Revised international clinical guidelines for the antibiotic vancomycin have changed the advice pharmacists need to provide to medical and nursing colleagues. Objectives: (1) To determine the self-reported confidence of hospital pharmacists to provide contemporary advice on vancomycin and (2) to evaluate hospital pharmacists’ knowledge to provide contemporary advice on vancomycin following a pilot continuing professional development (CPD) module. Methods: The study was a prospective two-phase design in an Australian teaching hospital. Phase one: a survey of pharmacist self-reported confidence to eight questions on providing contemporary advice on vancomycin. Responses were recorded using a Likert scales. Phase two: The provision of a pilot online CPD module on vancomycin containing knowledge-based assessment based on a clinical vignette. Likert scales recorded self-reported confidence were reported as median and interquartile range (IQR). Knowledge assessment was reported using descriptive statistics. The main outcome measure were the self-reported confidence, and knowledge of pharmacists regarding provision of contemporary advice on clinical vancomycin use. Results: Response rates for surveys; confidence n = 35 (72.9 %) and knowledge n = 31 (58.5 %). Phase one: confidence was highest regarding vancomycin dosing and monitoring with 71.4–81.6 % of respondents agreeing or strongly agreeing that they were confident in these domains. Respondents agreeing or strongly agreeing were least confident regarding intravenous administration and infusion related reactions, 57.1 and 45.7 % respectively. Respondents who provided advice on vancomycin >10 times in the prior 12 months reported significantly higher confidence in; therapeutic range 1 (IQR 1–2) versus 2 (IQR 1–3) p = 0.02; amending dosage based on therapeutic drug monitoring results 2 (IQR 1–3) versus 3 (IQR 2–3) p = <0.001, and providing general advice to prescribers on vancomycin 2 (IQR 1–3) versus 2 (IQR 2–4) p = <0.009. Knowledge questions were answered correctly post CPD by >75 % of pharmacists. Conclusion: Pharmacists’ self-reported confidence to managing vancomycin was variable but generally high. Knowledge scores were consistently high after pharmacists completed a pilot CPD module on vancomycin. These data provides impetus for a randomised controlled study across multiple sites to determine the extent to which pharmacist knowledge on vancomycin can be attributed to completion of an online CPD.