Tulsi, often referred to as Tulasi, is a revered herb with both culinary and therapeutic attributes. It is a member of the Lamiaceae family and is indigenous to the Indian subcontinent. Ayurvedic medicine has relied on it for more than 3000 years as a crucial component. In the Ayurveda system, Tulsi is widely recognized as the "Elixir of Life" because of its therapeutic characteristics and its efficacy in treating a range of common health conditions. The objective of this research study is to develop and evaluate transdermal patches containing an aqueous extract of Ocimum sanctum Linn. leaves. The process involves extracting and purifying the leaves to obtain raw extracts, which are then used in the production of transdermal patches. The manuscript offers a thorough explanation of the chemicals and equipment used in patch manufacturing, followed by an examination of phytochemicals to determine their bioactive constituents. The formulation process entails the creation of the transdermal drug delivery system (TDDS) and the calibration of the extract using UV spectrophotometry. Physicochemical investigations are conducted to assess the quality and effectiveness of the patches. Furthermore, the research study includes stability testing in challenging environments and antibiotic susceptibility testing using the disc diffusion method. The patches are subjected to antimicrobial and antifungal testing, with a specific focus on microbial strains, growth medium, and testing standards. The research study also incorporates FTIR analysis to evaluate the compatibility of medications and excipients, as well as to examine the hygroscopic properties of the patches. The results demonstrate the effectiveness and long-lasting nature of the developed patches, highlighting their potential for use in transdermal medication delivery systems.