Due to the importance of medicinal plants and their role in the treatment of a wide range of diseases, this experiment aims to study the effect of growth regulators and the hormone melatonin on the mint plant and the extent of improving its production and quality to obtain the best quantities of active substances from the growing mint plant in vitro using the explant (developing apexes and nodes). The plant parts were planted after cutting them to a size of 1 cm and it was sterilized with ethyl alcohol at a concentration of 70% by spraying for one minute and sodium hypochlorite solution at a concentration of 10% for 20 minutes in agriculture tubes containing MS medium for tissue culture, various concentrations of growth regulators and melatonin are added, then incubated in a growth room at a temp. of 20-25 and brightness of about 1000 lux for 16 hr. of light and 8 hr. darkness. The results and data were recorded after 45 days, where the treatment NAA(1.0mg/l) BA(0.5mg/l) without melatonin gave the highest percentage of the active compounds(Cineole and β linens). While the treatment NAA(2.0mg/l) BA(1.0mg/l)and melatonin(5mg/l) gave the highest percentage of the active compounds (D-Limonene and α pinene).