Steroid profiling helps various pathologies to be rapidly
diagnosed. Results from analyses investigating steroidogenic
pathways may be used as a tool for uncovering pathology
causations and proposals of new therapeutic approaches. The
purpose of this study was to address still underutilized application
of the advanced GC-MS/MS platform for the multicomponent
quantification of endogenous steroids. We developed and
validated a GC-MS/MS method for the quantification of
58 unconjugated steroids and 42 polar conjugates of steroids
(after hydrolysis) in human blood. The present method was
validated not only for blood of men and non-pregnant women
but also for blood of pregnant women and for mixed umbilical
cord blood. The spectrum of analytes includes common
hormones operating via nuclear receptors as well as other
bioactive substances like immunomodulatory and neuroactive
steroids. Our present results are comparable with those from our
previously published GC-MS method as well as the results of
others. The present method was extended for corticoids and
17α-hydroxylated 5α/β-reduced pregnanes, which are useful for
the investigation of alternative “backdoor” pathway. When
comparing the analytical characteristics of the present and
previous method, the first exhibit by far higher selectivity, and
generally higher sensitivity and better precision particularly for
17α-hydroxysteroids.