A pot experiment was conducted to evaluate the extract biochemical variations among 25 spearmint population. The experiment was a randomized complete design (RCD) with 25 treatments (population) and three replications. For this purpose, three rhizomes with 5 cm length were selected and planted in each pot. All pots were kept out side. The plant samples of all treatments were harvested at full flowering stage and the most important extract biochemical factors such as chlorophyll a, b and total, carotenoide, flavone and flavonol, total flavonoide, total phenolic compound, antioxidant activity, and carbohydrate content were measured. Moreover, all characteristics of studied population were subjected to cluster analysis and correlation between factors was determined. There were significant differences among population in all studied factors. The highest and lowest chlorophyll contents (35.77 and 10.5 mg/g FW) were obtained in Fars- Khafr 2 and Mazandaran-Nour population, respectively. Among studied population, Isfahan 2, Mazandaran-Qaemshahr, Mazandaran-Nour and Yasouj were superior in extract biochemical valuable properties like antioxidant activity, total phenolic compounds, total flavonoides, and total carbohydrates. Also, some population of Fars province constituted more carotenoide and chlorophyll contents and were superior to others. Furthermore, a positive correlation was detected between antioxidant activity, phenolic compound, and total flavonoides.