The Journal of steroid biochemistry and molecular biology
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J. Steroid Biochem. Mol. Biol. · May 2015
Validated LC-MS/MS simultaneous assay of five sex steroid/neurosteroid-related sulfates in human serum.
Conventionally, the concentration of steroidal sulfates was estimated by indirect or immuno‑based assays before the use of liquid-chromatography tandem mass spectrometry (LC-MS/MS). In the present study, a validated LC-MS/MS method is described for the simultaneous quantification of dehydroepiandrosterone sulfate (DHEA-S), estrone sulfate (E1‑S), androsterone sulfate (ADT‑S), pregnenolone sulfate (Preg‑S) and allopregnanolone sulfate (Allopreg‑S). E1‑S binding to serum proteins was observed, especially for the high concentration quality control serum samples, leading to -10 to -15% bias using a polymer-based SPE. ⋯ Recovery of the five compounds in stripped serum is equivalent to that in unstripped serum. The average recovery difference is less than 5% between stripped and unstripped serum for each compound. All results of other test parameters such as matrix, hemolysis and lipemic effects as well as stabilities meet the acceptance criteria of EndoCeutics SOPs and FDA guidelines.