Simultaneous measurement of 19 steroid hormones in dried blood spots using ultra-performance liquid chromatography-tandem mass spectrometry

Abstract

A rapid and accurate ultra-performance liquid chromatography-tandem-mass spectrometry (UPLC-MS/MS) method was developed for the measurement of 19 steroid hormones in dried blood spots and to achieve the highly traceable analysis of steroid hormones in dried blood samples. In this method, a BEH C8 column and UPLC-MS/MS were used for the separation of the steroid hormones. The extraction process was simple and accurate. The distribution ranges of 19 steroid hormones in four healthy individuals (2 males and 2 females) were determined simultaneously by positive ionization mode (ESI⁺) and negative ionization mode (ESI⁻) analysis, which were recorded by multiple reaction monitoring (MRM) modes. The linearity of the standard curves of 19 steroid hormones in dried blood spots was good, and the linear correlation coefficients R² were all ≥0.997. Meanwhile, the matrix effect of the method ranged from 87.1% to 131.3%. Across the analytical range, the inter-assay coefficient of variation (CV) was <12.07% and the intra-assay CV was <18.16%. The spiked recovery was >67.33%. The distribution ranges of 19 steroid hormones in four healthy individuals were in agreement with those in previous reports. A UPLC-MS/MS method for the simultaneous measurement of 19 steroid hormones in dried blood spots was developed to achieve traceable analysis of steroid hormones in dried blood samples.