Simultaneous quantitative determination of norgestrel and progesterone in human serum by high-performance liquid chromatography-tandem mass spectrometry with atmospheric pressure chemical ionization

Abstract

A selective, reliable and rapid method for the simultaneous determination of progesterone and norgestrel concentrations in human serum after taking oral contraceptive tablet has been developed using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) with atmospheric pressure chemical ionization (APCI) interface. The norethisterone was used as the internal standard. Selected transitions of m/z 313/245 for norgestrel, m/z 315/97 for progesterone and m/z 299/109 for norethisterone were monitored using multiple reaction monitoring (MRM) mode for quantitation. The assay was linear over the concentration range of 0.2-50 ng mL-1 for norgestrel and progesterone. The lower level of quantitation in human serum was obtained at 0.2 ng mL-1 for both norgestrel and progesterone using optimum tuning parameters. The intra-assay precision and inter-assay precision do not exceed 10 and 9%, respectively. The method has been applied to the determination of norgestrel and progesterone in serum of female volunteers.