The aim of the present investigation is to evaluate the antimicrobial potential of A. sativum and Z. officinalis extracts mediated glycerosomes formulation by thin film of lipid hydration method. The water content present in the A. sativum and Z. officinale was 42.3 and 27.2 % respectively. The yield of A. sativum and Z. officinale extract using ethanol as solvent through the Soxhlation process was found to be 4.7 ± 1.05 and 12.7 ± 1.108 % respectively. The crude drug of A. sativum and Z. officinale showed foreign organic was found to be 0.2 and 0.21 % and volatile oil was found to be 1.8 and 2.4 % only. The ash study revealed that total, acid-insoluble, water-soluble, and sulfated ash values were for the crude drugs of A. sativum and Z. officinale. The ethanol extracts showed maximum extractive yield. The phytochemicals detected in A. sativum bulb ethanol extract are Alkaloids, Carbohydrates, Cardiac glycosides, Phenols, Sterols, Triterpenoids, Saponins, Flavonoids, and Fixed oils and in Z. officinale rhizome ethanol extract are Alkaloids, Carbohydrates, Cardiac glycosides, Anthraquinone glycosides, Proteins and Amino acids, Triterpenoids, Saponins, Flavonoid, and Fixed oils. The prepared glycerosome was spherical with a slightly irregular shape. The negative potential of glycerosomes demonstrates the greater stability of the vesicular dispersion. The ASEE glycerosome formulations (F1 and F2) exhibited better antibacterial activity and greater antifungal activity against the gram-negative bacteria in comparison with the ZOEE glycerosome formulations (F3 and F4). The present research work strongly concluded that the A. sativum and Z. officinale and their glycerosome formulations are good candidate drugs and formulations for the safe and effective treatment of microbial infections