G4204a pumps

For the detection of plasma progesterone and its metabolites, the extract of plasma samples was analyzed using a high-performance liquid chromatography station (Agilent, Santa Clara, CA) equipped with G4204A pumps, a G1367E auto-sampler, a G1316A column oven and a triple quadrupole 6490 (Agilent, Santa Clara, CA). The separation of drug metabolites was achieved using an Eclipse Plus C18 RRHD analytical column (3.0 mm × 50 mm, 1.8 μM; Agilent, Santa Clara, CA, USA) at 40°C with an isocratic mobile phase consisting of 10% buffer A (0.1% formic acid in methanol: water, 60:40) and 90% buffer B (0.1% formic acid in acetonitrile: water, 60:40), at a flow rate of 0.2 ml/min. The injection volume was 10 μL and sample injection was performed using an auto-sampler. All progesterone metabolites were ionized using electrospray ionization in the positive ion mode (ESI). The temperature of the drying gas in the ionization source was maintained at 225°C. The gas flow was 12 l/min, the nebulizer pressure was 35 psi, and the capillary voltage was 4000 V (positive) and 3000 V (negative). The analytes were quantified using multiple reaction monitoring with mass transitions and the parameters for each compound. Methanol and water were of LC–MS grade and all reagents were obtained from Thermo Fisher Scientific.

A liquid chromatography-mass spectrometry (LC-MS) method was established to detect all 8 abiraterone metabolites and daidzein at the same time.⁵⁸ The extract of plasma samples were analyzed on a high-performance liquid chromatography station (Agilent, Santa Clara, CA) equipped with G4204A pumps, a G1367E auto-sampler, a G1316A column oven and a triple quadrupole 6490 (Agilent, Santa Clara, CA). Separation of drug metabolites was achieved using an Eclipse plus C18 RRHD analytical column 3.0 mm × 50 mm, 1.8 μM (Agilent, Santa Clara, CA) at 40°C with an isocratic mobile phase consisting of 70% buffer A (0.1% formic acid in methanol: water, 60:40) and 30% buffer B (0.1% formic acid in acetonitrile: water, 60:40), at a flow rate of 0.2 mL/min. The injection volume was 10 μL and sample injection was performed with the auto-sampler. Androgen was ionized using electrospray ionization in positive ion mode (ESI). The temperature of the drying gas in the ionization source was 225°C. The gas flow was 12 L/min, the nebulizer pressure was 35 psi, and the capillary voltage was 4000 V (positive) and 3000 V (negative). The analytes were quantified using multiple reaction monitoring with the mass transitions and parameters for each compound. Methanol and water were of LC-MS grade and all reagents were obtained from Thermo Fisher Scientific.