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Thyme (Thymus vulgaris L.) is a perennial sub shrub belonging to family Lamiaceae. Thyme essential oils have been used for many thousands of years, especially in food preservation, pharmaceuticals, alternative medicine and natural therapies. In order to determine the effect of harvest time on quality and quantity of essential oil in thyme, this experiment was done in a randomized complete block design with three replications at research station of Islamic Azad University, Khorasgan (Isfahan) during 2010-2011. Plants were harvested in five stages: before blooming, beginning of blooming, 50% blooming, full blooming and fruit set. The essential oils obtained from the phenological stages of thyme were analyzed by using GC/MS. The results obtained in our study showed that the phenological stages had very significant effects (P < 0.01) on essential oil yield and percentage as well as thymol percentage and yield. The highest essential oil content of thyme (2.42%) was extracted at the beginning of blooming stage. Analysis and identification of components showed that thymol is the main compound in all samples. The highest thymol content of thyme (74.8%) was extracted at the full blooming stage. According to the results of this research, harvesting the thyme at 50% blooming stage have maximum essential oil quality and quantity in Isfahan province.