BACKGROUND: Antioxidants protect spermatozoa against lipid peroxidation during freezing. OBJECTIVE: The study is designed to elucidate the suitable extender to preserve infertile smoker's semen against ROS damage using natural Tea polyphenol (T. arjuna bark extract). MATERIALS AND METHODS: Forty-two infertile subjects with smoking habit and 28 fertile subjects without smoking habit were considered for the study. Four semen extenders including our naturally derived antioxidant component were prepared and used to preserve semen sample from the study subjects for a period of one month. Standard semen parameters, biochemical and sperm DNA damage marker with inhibition were measured before and after cryopreservation. RESULTS: The motility and morphology of sperm cells were maintained better in E4 extender, and DNA damage is reduced. CONCLUSION: Extender recipe with natural antioxidants (E3&E4) was found to be apt for infertile semen preservation. © CryoLetters.