Method Description

The synthetic glucocorticoids are extracted from 0.5 mL of urine using an acetonitrile protein precipitation followed by methylene chloride liquid extraction of the solvent. Cortisol-9, 11, 12, 12-d, and triamcinolone-d1 acetonide-d6 are added to each sample before the liquid extraction and serve as the internal standards. Then, 17 mcL of the reconstituted sample extract is injected into a high-performance liquid chromatography system and analyzed by tandem mass spectrometry. The mass spectrometer has an electrospray interface and is operated in the multiple reaction monitoring positive mode. The calibration utilizes a 4-point standard curve over a concentration range of 0 to 25 mcg/dL.(McWhinney BC, Ward G, Hickman PE. Improved HPLC method for simultaneous analysis of cortisol, 11-deoxycortisol, prednisolone, methylprednisolone, and dexamethasone in serum and urine. Clin Chem. 1996;42:979-981; Savu S, Silvestro L, Haag A, Sorgel F. A confirmatory HPLC-MS/MS method for ten synthetic corticosteroids in bovine urines. J Mass Spectrom. 1996;31[12]:1351-1363; Djedovic NK, Rainbow SJ. Detection of synthetic glucocorticoids by liquid chromatography-tandem mass spectrometry in patients being investigated for Cushing's syndrome. Ann Clin Biochem. 2011;48[Pt 6]:542-549. doi:10.1258/acb.2011.010250)