Customer satisfaction is an essential request of the quality management system requirements by the International Organisation for Standardization for the medical laboratories. The current study aimed to evaluate the users' (doctors and head nurses) satisfaction with chemical laboratory services at the King Abdulaziz University Hospital (KAUH). A cross-sectional study was carried out for six months among the medical staff team who use the clinical chemistry laboratory of KAUH. The current survey covered the tests in all four laboratory areas, including routine chemistry, hormones, special chemistry, and therapeutic drug monitoring. A well-designed and validated questionnaire was used and distributed through official emails. The collected data were statistically analyzed and represented in tables. The response rates among the consultants, residents and specialists were 53.3%, 85.7%, and 75%, respectively. The clinical chemistry laboratory provides all tests needed by most of the participants (66.7%). The areas of hormones and TDM had got fewer satisfaction rates compared with the other laboratory areas. All the participants agreed that turnaround time (TAT) was acceptable (> 60–80%). The laboratory technologist responses were highly satisfied for most of the participants (≥ 70%). The survey outcome concluded that most of the participants were satisfied with KAUH clinical chemistry laboratory. A better understanding of the satisfaction rates of the clinical chemistry laboratory customer at KAUH, as well as the areas of weakness in hormone and TDM areas, will be useful in constructing an action plan for further improvement.