This study aims to analyse the influence of human resource quality, workload, and work environment on staff performance at the Blood Transfusion Unit of Ampana District General Hospital. The Blood Transfusion Unit plays a vital role in ensuring the availability of safe, high-quality, and timely blood supplies; therefore, staff performance is a crucial factor in supporting the quality of healthcare services. This research employed a quantitative approach using a survey method. The population comprised all staff of the Blood Transfusion Unit, and a census sampling technique was applied. Data were collected via questionnaires that had been tested for validity and reliability. Data analysis was conducted using descriptive and inferential statistics, specifically multiple linear regression, to examine both partial and simultaneous effects among the variables. The results indicate that human resource quality and the work environment have a positive and significant effect on staff performance, whilst workload has a negative and significant effect on staff performance. Simultaneously, the three independent variables significantly influence staff performance. The study concludes that improving staff competence and professionalism, managing workload proportionately, and creating a conducive work environment are key factors in enhancing staff performance and the overall quality of blood transfusion services.