The objective of this research was study the phytochemical composition of Adiantum capillus-veneris and to evaluate the isolates for possible in vitro antifungal and antibacterial activities. The compound obtained were screened by GC-MS method. While agar-well diffusion method was employed to measure antimicrobial activity against five bacteria and fourteen fungi and yeast. Thirtyone bioactive phytochemical compounds were identified in the methanolic extract of Adiantum capillus-veneris. The identification of phytochemical compounds is based on the peak area, retention time molecular weight, molecular formula, MS Fragment- ions and Pharmacological actions. GC-MS analysis of Adiantum capillus-veneris revealed the existence of the ?-D-Glucopyranoside, O-?-D-glucopyranosyl-(1.fwdarw.3)-?-D-fruc, d-Mannose, 5,7-Dodecadiyn -1,12-diol, 3-Trifluoroacetoxypentadecane, 3-Trifluoroacetoxypentadecane, Pterin-6-carboxylic acid, Imidazole-4-carboxylic acid,2-fluoro-1-methoxymethyl-,ethyl ester, D-Carvone, Pyrrolizin-1,7-dione-6-carboxylic acid, methyl (ester), D-Glucose,6-O-?-D-galactopyranosyl, Estragole, Phenol,2-methyl-5-(1-methylethyl), 3-Allyl-6-methoxyphenol, Ppropiolic acid, 3-(1-hydroxy-2-isopropyl-5-methylcyclohexyl), 7-epi-trans-sesquisabinene hydrate, Tetraacetyl-d-xylonic nitrile, y-Sitosterol, Ergosta-5,22-dien-3-ol, acetate, (3?,22E), Curan-17-oic acid,2,16-didehydro-20-hydroxy-19-oxo,methyl ester, 9,10-Secocholesta -5,7,10(19)-triene-1,3-diol,25-[(trimethylsilyl)oxy], Cis-Vaccenic acid, L-Ascorbic acid, 6-octadecanoate, L-Ascorbic acid, 6-octadecanoate, Deoxyspergualin, Tributyl acetylcitrate, 10,13-Dioxatricyclo[7.3.1.0(4,9)]tridecan-5-ol-2-carboxylic acid, 18,19-Secoyohimban-19-oic acid, 16,17,20,21-tetradehydro-16, 9-Octadecenamide,(Z), Olean-12-ene-3,15,16,21,22,28,-hexol,(3?,15?,16?,21?,22?), (22S)-21-Acetoxy-6?,11?-dihydroxy-16?,17?propylmethylenedioxy, Ethyl iso-allocholate, Olean-12-ene-3,15,16,21,22,28-hexol,(3?,15?,16?,21?,22?) and Olean -13(18)-ene. The FTIR analysis of Adiantum capillus-veneris leaves proved the presence of Alkenes, Aliphatic fluoro compounds, Alcohols, Ethers, Carboxlic acids, Esters, Nitro Compounds, Hydrogen bonded Alcohols and Phenols. Adiantum capillus-veneris was highly active against Aspergillus terreus (7.09±0.32). Methanolic extract of bioactive compounds of Adiantum capillus-veneris was assayed for in vitro antibacterial activity against Bacillus subtilis, Pseudomonas eurogenosa, Streptococcus faecalis, Salmonella typhi and Staphylococcus aureus by using the diffusion method in agar. The zone of inhibition were compared with different standard antibiotics. The diameters of inhibition zones ranged from 3.07±0.21 to 7.09±0.32 mm for all treatments. © 2016, International Journal of Pharmacognosy and Phytochemical Research. All rights reserved.