This study highlights the characterization and biological activities (in vitro) of root extracts of Indigofera hirsuta (Ih-E) and Afrormosia laxiflora (Al-E). The results showed that total phenolic and flavonoid contents in dry Ih-E were 54.38 mg and 116.03 mg/gram, respectively, while in Al-E were 170.46 mg and 80.23 mg/gram. Both extracts exhibited antioxidant activities since they decreased lipid peroxidation, but increased antioxidant activities and anti-inflammatory activities as shown from inhibition of RBC haemolysis and inhibition of albumin denaturation. Also, they have cytotoxic effect against normal BHK-21 and HepG2 where IC50 for Ih-E were 309 µg/ml and 95µg/ml, respectively (selectivity index 1.58), while for Al-E 57.50µg/ml and 144.50 µg/ml, respectively (selectivity index 0.40). Ih-E has antimicrobial activities towards Pseudomonas aeruginosa, Staphylococcus aureus, Vibrio species and E-coli, while Al-E showed no antimicrobial activities. Conclusion: Both extracts possessed antioxidant, anti-inflammatory and anticancer activities, thus, can be used as pharmacological tools.