A practical ID-LC-MS/MS method for the most commonly analyzed steroid hormones in clinical laboratories

Abstract

Background: Analysis of steroid hormones rapidly and reliably remains a challenge in clinical laboratories as this plays an important role in evaluation of many endocrine disorders. The aim of this study was to create a steroid profiling panel by using a liquid chromatography tandem mass spectrometry (LC-MS/MS) method which was composed of the most commonly analyzed steroid hormones in clinical laboratories. Materials and methods: Protein precipitation was performed for sample preparation. Ultra performance liquid chromatography (UPLC) system and an analytical column with C18 selectivity was chosen for chromatographic seperation. Atmospheric pressure chemical ionization (APCI) ion source was preferred for ionization, and tandem MS with triple quadrupole was used. MS scan was performed using the selected reaction monitoring mode in positive polarity. During the method validation process, test performance was evaluated for each steroid hormone, and 40 serum samples were used for method comparison with immunoassays available in our core laboratory. Results: An isotope dilution (ID)-LC-MS/MS method was developed, in which 13 steroids can be analyzed in the same run. Test performance was quite good for the 11 steroids (cortisol, DHEA, DHFAS, total testosterone, progesterone, androstenedione, 11-deoxycortisol, cortisone, corticosterone and dihydrotestosterone) while estradiol and aldosterone performance was suboptimal considering the precision and trueness. Conclusion: This ID-IC-MS/MS method would be useful in clinical laboratories, especially for the immunoassays having insufficient test performance and when checking for interferences in available immunoassays.