Abstract: Mass-spectrometry based methods of analyzing estrogens and other steroids from biological samples are disclosed herein. Further disclosed are methods of detecting the presence of illegal steroids in a mammal. Methods of detecting a disease state or condition or elevated risk of a disease state or condition in a mammal are also disclosed. Also disclosed are kits for use in a method for detecting and/or quantifying one or more steroids in a sample by mass spectrometry.

Abstract: Methods for enhancing detection by mass spectroscopy (MS) and/or chromatographic separability of carbonyl-containing compounds such as steroids are disclosed. Reaction of a carbonyl compound with a sulfonhydrazide compound provides a sulfonhydrazone with enhanced ionization efficiency during the electrospray ionization process. In a particularly disclosed embodiment, derivatization of catechol estrogens with p-toluenesulfonhydrazide enhances both detection by atmospheric pressure ionization-MS (API-MS), such as electron spray ionization-MS (ESI-MS) and separation by liquid chromatography (such as HPLC) under reverse phase conditions. In yet other embodiments, the sulfonhydrazone is further reacted with a sulfonyl halide under alkaline conditions to derivatize hydroxyl groups in the compound. Prior formation of the sulfonhydrazide derivative protects the carbonyl bond of the compound during subsequent alkaline reaction with the sulfonyl halide.