Anterior pituitary hormone deficiency could be isolated with only 1 hormone deficiency or combined with 2 or more hormone deficiencies (combined pituitary hormone deficiency, CPHD) of adrenocorticotropic hormone (ACTH), growth hormone (GH), thyroid-stimulating hormone (TSH), prolactin (PRL) and the gonadotropin follicle-stimulating hormone (FSH) and luteinising hormone (LH). Imaging studies and the patient’s phenotype can guide the diagnostic suspicion, with the biochemical approach being an important tool for confirmation. This is a critical review of the usefulness of baseline and functional tests in the diagnosis of ACTH, GH, TSH, LH and FSH deficiency in children, taking into account the limitations of current methods for hormone quantification.