Backgrounds: The neurotransmitter serotonin (5-HT) contributes to the regulation of food intake and appetite behavior, and its rate of synthesis depends upon brain tryptophan (TRP) availability in relation to the large neutral amino acids (LNAAs). Thus, the ratio of TRP to the sum of LNAA (TRP/LNAA) will be representative of the 5-HT brain levels. Differences between men and women in the obesity-linked alterations in whole blood amino acid levels and compartmentation have been described. The aim of this work was to study the effect of obesity on brain TRP availability in men and women and to determine whether there are sex-linked differences. Methods: The plasma levels of TRP and LNAAs in 42 men and 46 women (classified according to their body mass index (BMI) into lean, overweight and obese) have been determined by HPLC. Results: The TRP/LNAA ratio shows a significant drop which is even more pronounced in men than in women as BMI increases. Moreover, the comparison of the values between the 3 BMI groups revealed that the drop in the TRP/LNAA ratio appears with overweight in men but only with obesity in women. Conclusion: These sex differences could affect hypothetical behavioral differences of feeling hunger when facing voluntary food restriction for weight loss in individuals of different genders and with varying degrees of obesity.