The literature supports that organizations that create working conditions with positive practice environments improve nurses' job satisfaction and reduce turnover in hospital settings (Havens & Johnston, 2004; International Council of Nurses [ICN], 2007). Positive practice environments promote quality nursing care and better patient outcomes. However, in South America, there are no instruments to measure the nursing practice environment in a hospital setting. The purpose of this research was to examine the validity and reliability of a Spanish translation of the Practice Environment Scale of the Nursing Work Index (PES-NWI; Lake, 2002) in the Colombian clinical context. An instrument validation study using expert panel review, content validity indexing techniques, and a factor analysis of survey responses from 144 nurses was used. The scale-level content validity evaluation achieved high levels of acceptability, with values above 0.90. Construct validity results suggest that the instrument consists of 5 factors as prescribed by original instrument. The overall internal consistency value was a Cronbach's alpha level of 0.89. This is the first instrument to measure the work environment of nursing practice in South America, now validated in the Colombian context.