Inventors:
Xia Xu - Frederick MD, US
Regina G. Ziegler - Bethesda MD, US
David J. Waterhouse - Frederick MD, US
Joseph E. Saavedra - Thurmont MD, US
Larry K. Keefer - Bethesda MD, US
Assignee:
The United States of America as represented by the Department of Health and Human Services - Washington DC
International Classification:
G01N 33/00
G01N 1/00
US Classification:
436 87, 436173, 436817, 4352871
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.