Microbore high-performance liquid chromatography electrospray ionisation mass spectrometry of steroid sulphates

With the increasing abuse of steroids in both athletic and equine sports, there is a growing need for more sensitive and specific methods of detection of these compounds. Steroid sulphates are products of steroid metabolism and may be used as markers of steroid administration as well as endogenous steroid production. Electrospray negative ion mass spectra of 23 steroid monosulphates, 5 steroid disulphates and 4 bile acid sulphates have been recorded. The most intense ions in the mass spectra are due to molecular anions or dianions and in most cases these are the only ions present. The daughter ion mass spectra of these compounds vary depending on the location of the sulphate group in the molecule. Alicyclic-steroid sulphates have daughter ion mass spectra containing a single ion at m/z 97 (HSO4-) while, for aromatic sulphates, m/z 97 is absent. Combined microbore HPLC-electrospray ionisation mass spectrometry was used for the separation and identification of the individual components of a mixture of seven steroid monosulphates and detection of these compounds by scanning and multiple reaction monitoring is reported.