A series of 60 specimens were collected from patients suffering from different infections. Forty-five isolates of P. aeruginosea were isolated. Isolation of P.aeruginosa was done independence of utilization selective I culture techniques and differential media along with their ability to produce I pycyanin pigment. The results revealed that (10) isolates produce noticeable amount of pyocyanin, which used in separation of phenazine antibiotic. Isolation and identification of phenazine antibiotic and compared with standard antibiotic was carried out by column chromatography, thin layer cliromatograph and high performance liquid chromatography. j" Results of this study showed that isolates obtained from urine sample were ip more efficient in production of phenazine in contrast to those isolate. |- obtained from ears, which gave low production of phenazine. Accordingly we choose three isolate designated as PM3, PM8 and PM20 as best isolates production and PM8 to complete our study as the highest phenazine I producing isolate. |" A crude extract of phenazine from P.aeruginosa was identified and its v chemical, physical and biological properties were studied. siS; , ffect of different parameters on production of antibiotic from PM8 isolate was studied. Production parameters and nutritional requirement for . roduction was studied; including incubation time, temperature and pH. Results indicated that maximum production was found to be after (4) days incubation at 35°C and pH 7.2. The best source of carbon was found to ggrycerol (1.0)