The present study aimed to identify some metabolites products obtained from different jojoba callus tissue extracts using gas chromatography/mass spectrometry analysis (GC-MS). It is known that Jojoba, a medicinal and oil-yielding, has multi-purpose uses. In addition, it produces toxins, fatty acids, phenolic compounds and other secondary metabolites from callus identified using GC-MS. Despite the direct effect of 2,4-D on directing the explant towards callus induction, it has interaction effect with Kin and achieves hormonal balance inside the explant. There is a significant interaction effect between Kin and 2,4-D. The increased concentration of both Kin and 2,4-D raises the extent of nodal segments response towards proliferation, development, and callus induction. Our results showed that the interaction effect between Kin and 2,4-D is the best results for phytochemical active constituents formation achieved from callus cultured. Qualitative results revealed that increasing cytokinins Kin concentration leads to slightly double increase in active constituents from the explants for Triodecanoic and Methoxyacetic acids. While, the gradual increase of 2,4-D to 2.0 mg l -1 caused a single increase in active constituents for Triodecanoic, Methoxyacetic and Octadecanoic acid, respectively. On the other hand, Decanoic acid has no effect by adding both hormones (Kin and 2,4-D) and also Octadecanoic acid when kin is added. Also, there is a direct relation between the development of callus and reproduction of active phytochemical constituents, which increase qualitatively by increasing callus development and 2,4-D hormone concentration at 2 mg l -1 for 1-Hexadecanol, Nonadecatriene, Aristolene and Tricosene, respectively compared to free hormone treatment.