LC-MS/MS using the Citrine MS/MS SystemSteroids regulate essential biological functions including growth and reproduction. Altered steroid homeostasis can result in various diseases and syndromes. To assess the role of these important lipids, specific and sensitive analytical strategies are used to measure steroids at sub-nanomolar concentrations in biological samples. Here we will discuss an enhanced LCâ€“MS/MS method for targeted quantification of multiple, relevant steroid molecules. Technical advantages of the Citrine MS/MS system for improving divergent steroid analysis will also be presented.