Podophyllotoxin (PTOX), one of the most important natural medicinal compounds, has anticancer properties. Its effective medicinal derivatives, such as etoposide and teniposide, have been approved by the FDA for cancer treatment. This compound is found as a specialized metabolite in the Linum album Kotschy ex Boiss., belonging to the Linaceae family. PTOX is the major aryltetralin lignans resulting from the shikimic acid/phenylpropanoid pathway and accumulates in the shoot and roots of L. album. PTOX play a necessary role in plant defense systems, protecting against abiotic and biotic stresses and helping their adaptation to adverse environmental conditions. Therefore, the content of specialized metabolites increases under stress. In this study, the content of PTOX under stressful conditions (potassium deficiency stress at two-time points of 12 and 48 hours) was examined using HPLC in a completely randomized design with three replications. The results of HPLC showed that the content of PTOX first decreased in 12 hours, while after 48 hours of treatment compared with the control plants, it showed a significant increase with the value of 135.8 in the shoot. In the root, the results were consistent with the results of the aerial parts, and the amount increased significantly after 48 hours. In general, the results show that L. album as a suitable natural source for PTOX has great potential to generate Large-scale products for commercial and pharmaceutical purposes.