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The academic formation of an individual with good study opportunities in Mexico begins approximately at 3 years of age and remains constant until graduation from high school. At this stage, there are learning periods, that include classes and homework, and evaluation periods, that covers exams and class presentations. Evaluation periods concentrate two or three daily exams during one week. The present project was designed to determine through cortisol concentrations if there are differences in the expression of stress between males and females in high school in a private school in the city of Puebla, Mexico. We recruited 14 female and 13 male adolescent volunteers between the ages of 17 and 18. We obtained two blood samples, the first one was taken during the class period and the second one was taken on the week of final evaluations of the semester. The blood was processed according to the instructions of an ELISA Bio-cortisol (MEXLAB) kit in order to determine serum cortisol concentrations, and according to a Bio-glucose (MEXLAB) kit to determine serum glucose concentrations. The results show that males have higher blood cortisol levels during the class and examination periods compared to females. We also show that cortisol levels in males are significantly higher during the examination period than during the class period; whereas females did not show differences in cortisol levels between periods. In the case of glucose levels, we observed differences related to sex but not differences related to the evaluated periods. These data suggest that school activities at the high school level generate more stress in males than in females.