In order to study the relationship between sex, age and glucose, and the concentrations of various fuel related blood substrates in children during prolonged fasting, we have selected data of fasting procedures in 13 control children aged 3-5 yr, fasted 24 h, and 58 control children aged 6-15 yr, fasted 40 h. Compared to the blood results after overnight fast, glucose is decreased, and lactate, pyruvate, ketones and non-esterified fatty acids (NEFA's) are all clearly increased at the end of fast. The concentrations of alanine and triglycerides remain unchanged. The relation with sex, age and glucose has only been analyzed in the older children group. A sex-dependency is indicated for the ketones. Ketones are negatively related with age. NEFA's pyruvate and alanine are not age-related, whereas glucose, lactate and triglycerides are moderately age-dependent. Ketones are negatively related with glucose, whereas pyruvate, NEFA's and triglycerides are not glucose-related. Lactate and alanine are weakly related to glucose. The data demonstrate diminished glucose homeostasis and increased ketogenesis in younger children compared to older ones during prolonged fasting.