Palicourea croceiodes (Rubiaceae) is a plant claimed in traditional medicine in the eastern part of Nigeria to cure various ailments such as urinary tract infections, vaginal infections which cause infertility in women,, sexually transmitted diseases and other related ailments. The GC-MS analysis of the ethyl acetate leaf extract revealed nineteen compound with prevailing compounds as 11, 14,-Eicosadienoic acid methyl ester,Hexadecenoic acid,2,6,10,15,19,23,-Hexamethyl-2,6,10,14,18,22-tetracosahexaee (squalene) and 10- octadecenoic acid methyl ester. Antimicrobial assay of the extract against some pathogenic bacteria and fungi including Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Klebsiella aerogenes, E.coli, Proteus mirabilis, Enterobacter aerogenes, Candida albican and Aspergillus niger using the agar well diffusion method. The extract was also found to exhibit selective activities against some of the organisms with Minimum Inhibitory Concentration (MIC) of between 0.25mg/ml and 0.0625mg and a Minimum Fungicidal Concentration of (MFC) of 0.125mg/ml and 0.0625mg/ml. The study justified the use of the plant as an antibiotic in ethno-medical applications