Objective: To determine the antioxidant activity, total phenolic and flavonoid content of Petroleum ether extract (PE), Dichloromethane extract (DCM), Ethanol extract (ET) and aqueous extract (AQ) of henna seeds. Methods: Total antioxidant assay (phosphomolybenum method), DPPH radical scavenging assay, reducing power assay and lipid peroxidation inhibition assay were used to ascertain the potential of seeds as an antioxidant. Results: In all the assays carried out ET showed a greater potential to scavenge DPPH radical, reduce MO (VI) to MO (V) complex and Fe (III) to Fe (II) and to inhibit lipid peroxidation. The IC 50 of ET was far greater than that of the standard, ascorbic acid (AS) in the lipid peroxidation assay. The activity of AQ was lesser when compared with that of ET but greater than PE and DCM. The amount of phenolics and flavonoids were present in higher amounts in ET followed by AQ. Trace amounts of phenolics were detected in PE and DCM, but the amount of flavonoids were below the detection level. The study showed that the antioxidant activity and the concentrations of phenolics and flavonoids are proportionate to each other. Conclusions: Ethanolic extract of henna seeds are efficient antioxidants, which can be utilized for further isolation of active compounds and pharmaceutical applications. {\textcopyright} 2011 Hainan Medical College.