A novel,fast,sensitive and robust method based on ultra-performance liquid chromatography coupled to atmospheric pressure electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS)has been developed to separate ...A novel,fast,sensitive and robust method based on ultra-performance liquid chromatography coupled to atmospheric pressure electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS)has been developed to separate two Tibolone stereoisomers i.e.,3α-Hydroxy Tibolone and 3β-Hydroxy Tibolone and to quantify 3α-Hydroxy Tibolone using p-toulenesulfonyl isocyanate(PTSI)as a derivatizing reagent in human plasma.3α-Hydroxy Tibolone-13CD3 was used as an internal standard(IS).The analyte and IS were extracted from human plasma by liquid-liquid extraction using ethyl acetate.Extracted samples were analyzed by UPLC-ESI-MS/MS.Chromatography was performed using binary gradient on UPLC analytical column.A linear calibration curve over the range of 0.100-35.000 ng/mL was obtained and lower limit of quantification(LLOQ)was 0.100 ng/mL demonstrating acceptable accuracy and precision.This method was successfully applied to a pharmacokinetic study in order to compare a test Tibolone 2.5 mg formulation vs.a reference 2.5 mg Tibolone tablet formulation in 50 post-menopausal/surgical menopause female human volunteers under fasting conditions.It is concluded that test formulation of Tibolone is bioequivalent to reference formulation of Tibolone.展开更多
文摘A novel,fast,sensitive and robust method based on ultra-performance liquid chromatography coupled to atmospheric pressure electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS)has been developed to separate two Tibolone stereoisomers i.e.,3α-Hydroxy Tibolone and 3β-Hydroxy Tibolone and to quantify 3α-Hydroxy Tibolone using p-toulenesulfonyl isocyanate(PTSI)as a derivatizing reagent in human plasma.3α-Hydroxy Tibolone-13CD3 was used as an internal standard(IS).The analyte and IS were extracted from human plasma by liquid-liquid extraction using ethyl acetate.Extracted samples were analyzed by UPLC-ESI-MS/MS.Chromatography was performed using binary gradient on UPLC analytical column.A linear calibration curve over the range of 0.100-35.000 ng/mL was obtained and lower limit of quantification(LLOQ)was 0.100 ng/mL demonstrating acceptable accuracy and precision.This method was successfully applied to a pharmacokinetic study in order to compare a test Tibolone 2.5 mg formulation vs.a reference 2.5 mg Tibolone tablet formulation in 50 post-menopausal/surgical menopause female human volunteers under fasting conditions.It is concluded that test formulation of Tibolone is bioequivalent to reference formulation of Tibolone.
