Steroid measurement with LC-MS/MS. Application examples in pediatrics.

The correct measurement of steroids is vital for the diagnosis of congenital adrenal hyperplasia (CAH), apparent mineralocorticoid excess, familial hyperaldosteronism type I, primary aldosteronism, Cushing's disease, adrenal insufficiency, etc. Steroid diagnostics also plays an important role in disorders of sexual differentiation and gonadal function. Steroid metabolism is involved in evaluations for precocious puberty, premature thelarche, and polycystic-ovary disease. Finally, the hypothalamo-pituitary-adrenal (HPA) axis is considered to be one of the major systems involved in fetal programming or in stress regulation. Most methods for the determination of steroid hormones are based on immunoassays, which are rapid and easy to perform. However, the reliability of several steroid immunoassays has been shown to be questionable because of the lack of specificity and of matrix effects. Immunological methods, especially direct assays, often overestimate true steroid values. Patient follow-up over time or between laboratories, as well as longitudinal studies, are therefore extremely difficult. This is of particular importance in pediatrics. Liquid chromatography triple quadrupole mass spectrometry (LC-MS/MS) is an increasingly common tool in the clinical laboratory and has the potential to overcome the limitations of immunoassays. LC-MS/MS affords the specificity, imprecision, and limits of quantification necessary for the reliable measurement of steroids, expanding diagnostic capabilities. In addition to the high throughput, the method requires minimal sample preparation and a small sample volume. All these features make it an attractive method to use in a clinical setting. Moreover, LC-MS/MS has the advantage that a spectrum of steroid hormones can be measured simultaneously. Steroid profiling is a very effective method for distinguishing almost all steroid-related disorders. It allows accurate diagnosis and is very useful in many clinical situations. Steroid profiles open up new vistas. The applicability for clinical samples and questions in pediatric endocrinology will be discussed.