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

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 R-2 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 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.

Automated summary

A rapid and accurate UPLC-MS/MS method was developed for measuring 19 steroid hormones in dried blood spots and achieving traceable analysis. The method utilized a BEH C8 column and UPLC-MS/MS for separation, with a simple and accurate extraction process. The distribution ranges of the 19 steroid hormones in four healthy individuals were determined simultaneously using positive and negative ionization modes, and the standard curves showed good linearity with correlation coefficients of R-2=0.997. The developed method provides important insights into the distribution of steroid hormones and enables traceable analysis in dried blood samples.