In this study, the cytological and biochemical effects of zingiberene (ZBN) on human lymphocytes cultures were investigated. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays were used for viability and cytotoxic evaluations. Micronucleus (MN) and chromosomal aberration (CA) tests were used for genetic evaluations. Moreover, total antioxidant capacity (TAC) and total oxidative status (TOS) analyses were used for biochemical evaluations. Utilizing the MTT and LDH assays, the cytotoxicity of ZBN was determined on lymphocyte cultures and the short-term lymphocyte cultures were incubated with various doses of the ZBN; the results demonstrated that the growth of lymphocytes cells was inhibited in a dose-dependent manner. In addition, MNs and CAs in lymphocytes were not influenced by exposure to ZBN. Moreover, ZBN treatment caused increases in TAC levels in human lymphocytes without changing TOS levels. In conclusion, ZBN could be used as a suggested natural antioxidant for therapeutic, pharmaceutical and food applications.