Steroid analysis in clinical routine diagnostics – discussing crucial questions

Abstract: Quantitative steroid analysis via liquid chromatography-tandem mass spectrometry (LC-MS/MS) is applicable to clinical routine diagnostics by now, substituting immunoassays due to its superior selectivity and comparable sensitivity. Multiplexed assays covering a multitude of analytes represent the gold standard in this regard. There are commercially available kits which are easily adapted to individual LC-MS/MS systems required. Prior to and even after their appearance, in-house method development represented the flexible alternative in terms of solving specific analytical problems or focusing on a narrower steroid profile while maximizing sensitivity and high throughput applicability. In this work, commercial assays and in-house methods are discussed in relation to a benchmark LC-MS/MS method. Thereby, prerequisites and results are compared. Furthermore, the effect of concomitant medication on steroid assays was tested and requirements regarding quality assurance in routine steroid analysis are discussed. Most of the different commercially available or in-house LC-MS/MS methods for steroid analysis show a good or reasonable agreement of results. However, the harmonization in the methodology of mass spectrometric assays has to be improved to further reduce their variability. Such a procedure would facilitate the performance of diagnostic tests that involve the measurement of steroid hormones by the tremendous improvement of diagnostic sensitivity and specificity.