This paper examines the relationship between well-being, age, menopausal status, hormone levels and hot flashes. Data from the first 4 years of longitudinal observation from the Melbourne Women's Midlife Health project war utilized. This study involved a population-based sample of 405 women interviewed annually. Blood was taken during the follicular phase (if still menstruating) for estradiol, sex hormone binding globulin, follicle stimulating hormone and testosterone. A validated well-being scale was used. Positive affect increased with age while negative affect decreased with age but only in the postmenopausal category. Positive affect was significantly lower in the 2 years postmenopausal group but this effect of menopausal status did not remain when hot flashes were included in the analysis. Negative affect was highest in the 1-2 years postmenopausal group. Although hot flashes adversely affected negative moods, a significant effect of menopausal status remained. No direct association between any of the hormone levels and positive or negative affect scores was evident. In conclusion, this study found that well-being was decreased in the first 2 years after the first menstrual period but began to improve spontaneously after this time.