Development of a rapid, sensitive and authentic LCMS method for the determination of Dexamethasone, Testosterone and Estrone (E1) in human plasma

A simple, rapid, fast, precise and accurate reverse-phase HPLC method was developed and validated for the simultaneous determination of Dexamethasone, Testosterone and Estrone (E1) in pre-mixture by using RP-HPLC having photon diode array detector, Evaporative Light Scattering Detector (ELSD) and coupled with dual electrospray ionization mass spectrometry Quadrupole time-of-flight (LCMS). The method has shown adequate separation for Dexamethasone, Testosterone and Estrone (E1). Separation was achieved on reverse phase Shodex C18, 3 μm, 4.6 × 100 mm, using isocratic elution mode with a mobile phase Acetonitrile and 0.1% formic acid in water. The flow rate was 1 mL/min and effluents were monitored at 240 and 280 nm, using photodiode array (PDA) detector, ELSD and Ion detection was performed by dual electrospray ionization source (dual ESI) in positive mode. The recovery of steroids was found to be in the range of 98-102%. The described method was linear. The precision, ruggedness and robustness values were also within the prescribed limits.