Complimentary or natural antioxidant type of alternative medicine is developed worldwide to treat male infertility. The aim of this study is to the extraction of T. arjuna bark and activity against human sperm DNA damage in asthenoteratospermic smoker’s subjects—an in vitro study. All preliminary and antioxidant assays (DPPH, H2O2, and total antioxidant, reducing power activity) were done. T. arjuna bark metal analysis was done with AAS. On the other hand, patients were asked to fill a direct questionnaire about smoking history; 25 infertile smokers were identified as asthenoteratospermic; 34 fertile non-smokers (control) were assessed for semen parameters by CASA, seminal plasma Zinc analysis by AAS, DNA fragmentation by colorimetric method and semen genomic DNA damage inhibition by modified non-enzymatic salting out extraction method. Most of the antioxidants are highly present in the aqueous extract; meanwhile, the major content in this extract is zinc 16 µg/g (Ca = 0.5 µg/g; Se = 2.2 µg/g and Mg = 1.6 µg/g) along with FT-IR peaks which also confirmed the metal presence. The semen parameters in smokers that were noticed are low sperm count and morphological changes. Meanwhile, in the seminal plasma of smokers, zinc and DNA fragmentation results were positively correlated with sperm morphology (p < 0.001). Repaired DNA bands were noticed in the in vitro study of aqueous T. arjuna bark, in smokers’ semen. T. arjuna bark will act as cryo protector as well as great zinc supplementary to maintain sperm motility and morphology in smokers. © 2017, Springer-Verlag GmbH Germany.