Simultaneous Determination of Prednisone and Prednisolone in Serum by Turbulent Flow Liquid Chromatography-Tandem Mass Spectrometry

Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative therapeutic treatment for patients with high-risk hematologic malignancies and bone marrow failure syndromes. While allo-HCT can be highly effective, it is met with significant regimen-related toxicities and complications such as graft-versus-host disease (GVHD), poor immune reconstitution, and infections. Prednisone is the preferred treatment for patients with both acute and chronic GVHD. While effective, high-dose prednisone can cause many complications, including weight gain, skin fragility, muscle weakness, bone demineralization, hyperglycemia, insomnia, and psychosis. Optimizing prednisone (and prednisolone) dosing by measuring their concentrations and calculating their pharmacokinetic parameters will allow for personalized treatments for patients, producing more effective and safer treatments for GVHD. This chapter describes a method to measure both compounds simultaneously. Prednisone and prednisolone are extracted from serum by the addition of methanol containing deuterated internal standards. Chromatographic separation is achieved using a reversed-phase HPLC column followed by tandem mass spectrometry performed in the positive ion mode. This assay is fast, accurate, sensitive and allows for rapid drug measurements and timely dose modifications.