The article’s aim is to focus on the application of Six Sigma to measure defects and identify improvement opportunities as part of knowledge management in the sterilization department in a Colombian private hospital. The methodology was established in three stages: Recognize, Define and Measure as an exploratory study using data from all instrument sterilized (N = n = 12.846). Quality Management System’s information was considered to recognize the context because work team had not hired in the organization directly. Sigma level was calculated in three sterilization processes: autoclave, ethylene oxide and hydrogen peroxide using data from 2017 and 2018. Outcomes of this study provides information about behavior of sterilization processes to establish a baseline based on the historical data. Furthermore, employees require improve their skills and competences in statistics and data processing. Future works are related to reducing waste evidenced in excessive time of processing or storing, instrumental availability and knowledge management using lesson-learned in the processes as a result of Analyze and Control Stages of DMAIC cycle implementation.