Ethanol extracts of Basella alba leaves (EB) and Tribulus terrestris seeds (ET), and methanol extract of Asparagus racemosus roots (MA) and Mucuna pruriens seeds (MM) were found effective to induce masculinization in Nile tilapia. The present study intends to evaluate the anti-aromatase activity of EB, ET, MA and MM, to identify the androgenic bioactive compounds in the extracts, and to determine their pharmacokinetics. The crude plant extracts were first fractioned through column and thin layer chromatography (TLC). Three days old Nile tilapia juveniles (mean weight 0.025 ± 0.009g; n = 50 fish/replicate, 3 replicates/treatment) were fed diets fortified with the obtained fractions for 30 days. F ish were then sacrificed and gonad aromatase mRNA expression, and 11-ketotestosterone (11-KT) and estradiol (E2) levels were measured. Fractions yielding the highest male percentage for each plant were subject to gas chromatography-mass spectrometry (GC-MS) analysis. The in silico docking and SwissADME study were conducted with the components showing higher peak percentage in chromatogram. EB, ET, MM and MA yielded 6 (EB1 - EB6), 8 (ET1- ET8), 14 (MM1-MM14) and 5 (MA1- MA5) fractions, respectively. Fish fed EB2, ET2, MA2 and MM13 fraction fortified diets showed significantly (p < 0.05) higher male percentage (92.32%–98.39%) compared to other treatment groups. EB2, ET2, MA2 and MM13 fed fish showed significantly higher 11-KT level compared to control male and lower E2 level compared to control female. Aromatase mRNA expression was significantly down-regulated by all these four fractions (-1.32 to -5.65 folds ) with respect to control female. GC-MS analysis revealed the presence of 1-Octadecene (OD) in EB2, Phenol, 2,4-bis(1,1-dimethylethyl) (PD) in ET2 and MA2, 9,12-Octadecadienoic acid (Z,Z)- (ODDA) in MM13. In silico molecular docking indicated that PD was more effective than ODDA and OD to inhibit aromatase (Figure 1). PD showed better pharmacokinetics and more drug-likeness compared to OD and ODDA in SwissADME analysis.