Migraineurs have been identified to have chronically decreased serotonin levels while its concentrations markedly increase during ictal periods. Regarding the importance of adequate tryptophan intake in regulating serotonin homeostasis and subsequent effect on migraine attacks, we designed the current study. The migraine group (n = 514, diagnosed according to the ICHDIII criteria) was recruited from a tertiary headache clinic. The controls consisted of 582 sex-matched healthy volunteers who were randomly selected from general population. After collecting demographic and anthropometric data, a validated 168-item semi-quantitative food frequency questionnaire (FFQ) was used for dietary intake assessments. Multiple regression models were applied to explore the relationship between migraine and tryptophan intake. The mean (SD) of the age of participants in the controls and migraine group was 44.85 (13.84) and 36.20 (9.78) years, respectively. The multiple regression models were adjusted for age (year), sex, body mass index (BMI) (kg/m), total daily energy intake (kcal/day), dietary intakes of total carbohydrates (g/day), animal-based protein (g/day), plant-based protein (g/day), total fat (g/day), saturated fat (g/day), and cholesterol (mg/day). It was shown that there is a negative association between tryptophan intake and migraine risk ((OR in the 3rd quartile = 0.46; 95% CI = 0.25-0.85) (OR in the 4th quartile = 0.40; 95% CI = 0.16-0.98) compared with the first quartile; P for trend = 0.045). Therefore, our results showed that subjects who had a median intake of 0.84-1.06 g of tryptophan per day had reduced odds of developing migraine by approximately 54-60%, relative to those consumed ≤ 0.56 g/day.