Simultaneous quantification of progestogens in plasma and serum by UHPLC-HRMS employing multiplexed targeted single ion monitoring

Progesterone is the predominant gestagen in most mammals studied so far. It plays a substantial role regulation of female reproductive cycle and providing support for pregnancy maintenance. Despite its known functions, gaps knowledge are present regarding reduced metabolites that potentially exert biological activity. Therefore, new UHPLC-HRMS method based on Q Exactive™ mass spectrometer was developed to detect quantify simultaneously progesterone, hormone precursor pregnenolone 10 progestogens (20α-DHP, 20β-DHP, 3α,5α-THP, 3α,5β-THP, 3β,5α-THP, 3β,5β-THP, 3α-DHP, 3β-DHP, 5α-DHP 5β-DHP) plasma serum samples. Purification achieved by an optimized solid phase extraction (SPE) analysis conducted positive electrospray ionization (ESI) mode with application multiplexed selected ion monitoring (msx-t-SIM). The validation included study sensitivity, selectivity, curve fitting, carry-over, accuracy, precision, recovery matrix effects. poor properties underivatized steroids, high sensitivity range pg/mL achieved.