A sensitive and rapid mass spectrometric method for the simultaneous measurement of eight steroid hormones and CALIPER pediatric reference intervals

ObjectivesTo develop an accurate assay and establish the normal reference intervals for serum cortisol, corticosterone, 11-deoxycortisol, androstenedione, 21-hydroxyprogesterone, testosterone, 17-hydroxyprogesterone, and progesterone. These steroids are commonly used as biomarkers for the diagnosis and monitoring of endocrine diseases such as congenital adrenal hyperplasia. Appropriate age- and gender-stratified reference intervals are essential in accurate interpretation of steroid hormone levels.Design and methodsThe samples analyzed in this study were collected from healthy, ethnically diverse children in the Greater Toronto Area as part of the CALIPER program. A total of 337 serum samples from children between the ages of 0 and 18 years were analyzed. The concentrations were measured by using an LC–MS/MS method. The data were analyzed for outliers and age- and gender-specific partitions were established prior to establishing the 2.5th and 97.5th percentiles for the reference intervals.ResultsReference intervals for all hormones required significant age-dependent stratification while testosterone and progesterone required additional sex-dependent stratification.ConclusionsWe report a sensitive, accurate and relatively fast LC–MS/MS method for the simultaneous measurement of eight steroid hormones. Detailed reference intervals partitioned based on both age and gender were also established for all eight steroid hormones.

► Sensitive LC–MS method for the simultaneous measurement of eight steroid hormones.
► Provides rapid and sensitive diagnosis and monitoring of endocrine diseases.
► Detailed reference intervals have been established for all eight steroid hormones.
► These reference intervals were acquired from a healthy pediatric population.