An experiment was carried out in growing chickens to study the effect of supplementation of a semi-synthetic diet containing soya-bean meal as the sole protein source with DL-methionine, to improve its biological value, on the excretion of endogenous protein and amino acids measured in lower ileum and total tract using traditional methods. Thirty-two White Rock male broilers (10 days old) were randomly divided into eight groups each of four birds, of similar body weight (mean live weight: 142·8 (s.e. 0·68) g), and individually housed in metabolism cages. Following a paired-feeding design based on metabolic body weight (kgM0·75), each group of birds was given, for an experimental period of 20 days, each of four levels of protein (60, 120, 180 or 240 g/kg; 5 days each) in two groups of isoenergetic (14·5 kJ metabolizable energy per g dry matter) and semi-synthetic diets based on soya-bean meal, either not supplemented or supplemented with 2 g/kg DL-methionine (diets S and SM, respectively). After 3 days of each treatment excreta were collected for 48 h, frozen and stored at –20ºC. At the end of the fourth treatment three chickens of each group were killed and their lower ileal contents collected. The remaining chick of each treatment was fasted for 24 h and given a protein-free diet for 8 days and excreta were collected for the last 4 days. Then (day 39 of age), chickens were killed and lower ileum contents removed and stored at –20ºC. Samples of excreta and lower ileum contents were subjected to nitrogen (N) analysis by Kjeldahl procedure and amino acid (AA) analysis by high-performance liquid chromatography. Supplementation with DL-methionine of the soya-bean meal-based diets halved total tract endogenous AA losses. Regression analysis produced a higher estimation of ileal and faecal endogenous AA excretion than feeding a protein-free diet. Endogenous AA excretion determined in the lower ileum was higher than in excreta no matter which estimation procedure was utilized. In conclusion, supplementation of dietary protein with the first limiting AA to improve its protein quality, causes an important drop in endogenous AA losses, that may have an important effect on the N economy and energy requirements in poultry. The use of regression analysis on excreta data where graded amounts of protein are given to growing chickens, seems a suitable method for determining endogenous AA losses provided that good quality proteins are used.