The aim of this study is to evaluating the antibacterial activity of Laurus nobilis leaves extract in hospital environment isolates. Maceration and Soxhlet apparatus were used to prepare aqueous and methanolic extracts. The total phenolic content and high-performance liquid chromatography (HPLC) were conducted to determine the active compounds in the extracts. The results showed that the methanolic and aqueous extracts contain four flavonoids derivatives (kaempferol, luteolin, quercetin and Rutin) were identified on the basis of matching retention time with the standards. The total phenolic contents were 56.81 and 81.56 mg/g in 50 mg/ml, in aqueous and methanolic extracts respectively. The antibacterial activity of Laurus nobilis leaves extracts showed that the methanolic extract was more effective than aqueous extract in concentration 64mg/ml. Moreover, the result of the minimum inhibitory concentration (MIC) showed that the methanolic extract on P. aeruginosa isolates was 32 mg\ml, while the MIC values of aqueous extract were 64 and128 mg\ml.