Blood was obtained from prostate cancer patients with consent under a protocol approved by the Cleveland Clinic Institutional Review Board. Samples were analyzed on a ultra high-performance liquid chromatography station (Shimadzu, Kyoto, Japan) with a DGU-20A3R degasser, 2 LC-30AD pumps, a SIL-30AC autosampler, a CTO-10A column oven and a CBM-20A system controller in tandem with a QTRAP 5500 mass spectrometer (AB Sciex, Framingham, MA). The mobile phase consisted of LC-MS grade (Fisher) methanol: acetonitrile: water (44:36:20). Separation of drug metabolites was achieved using a Zorbax Eclipse plus 150 × 2.1 mm, 3.5μm C18 column (Agilent, Santa Clara, CA) at a flow rate of 0.2 ml/min. Drug metabolites were ionized using electrospray ionization in positive ion mode. Multiple reaction monitoring was used to follow mass transitions for D4A (m/z: 348.2/156.3), abiraterone (m/z: 350.3/156.1), and d4-cortisol (m/z: 367.1/121.1). Standard curves were generated using human serum spiked with known concentrations of each metabolite to enable determination of unknown concentrations in patient samples.
Dgu 20a3r degasser
Manufactured by AB Sciex
Sourced in Japan
The DGU-20A3R degasser is a laboratory equipment product designed to remove dissolved gases from liquid samples. It serves as a core component in liquid chromatography systems, helping to maintain the integrity and consistency of the mobile phase.
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4 protocols using dgu 20a3r degasser
1
Quantifying Prostate Cancer Metabolites
2
Quantifying Prostate Cancer Metabolites
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Blood was obtained from prostate cancer patients with consent under a protocol approved by the Cleveland Clinic Institutional Review Board. Samples were analyzed on a ultra high-performance liquid chromatography station (Shimadzu, Kyoto, Japan) with a DGU-20A3R degasser, 2 LC-30AD pumps, a SIL-30AC autosampler, a CTO-10A column oven and a CBM-20A system controller in tandem with a QTRAP 5500 mass spectrometer (AB Sciex, Framingham, MA). The mobile phase consisted of LC-MS grade (Fisher) methanol: acetonitrile: water (44:36:20). Separation of drug metabolites was achieved using a Zorbax Eclipse plus 150 × 2.1 mm, 3.5μm C18 column (Agilent, Santa Clara, CA) at a flow rate of 0.2 ml/min. Drug metabolites were ionized using electrospray ionization in positive ion mode. Multiple reaction monitoring was used to follow mass transitions for D4A (m/z: 348.2/156.3), abiraterone (m/z: 350.3/156.1), and d4-cortisol (m/z: 367.1/121.1). Standard curves were generated using human serum spiked with known concentrations of each metabolite to enable determination of unknown concentrations in patient samples.
3
UHPLC-MS/MS Quantification of Drug Metabolites
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Samples were analyzed on a ultra high-performance liquid chromatography station (Shimadzu, Kyoto, Japan) with a DGU-20A3R degasser, 2 LC-30AD pumps, a SIL-30AC autosampler, a CTO-10A column oven and a CBM-20A system controller in tandem with a QTRAP 5500 mass spectrometer (AB Sciex, Framingham, MA). Drug metabolites were ionized using electrospray ionization in positive ion mode. Multiple reaction monitoring was used to follow mass transitions for Abi, IS, and the metabolites (Table S1 ). Due to the similarity in structure and mass transitions for the metabolites it was necessary to separate them with chromatography. Separation of drug metabolites was achieved using a mobile phase consisting of LC-MS grade (Fisher) methanol: acetonitrile: water:formic acid (39:26:34:1) at a flow rate of 0.2 ml/min., and C18 analytical column; Zorbax Eclipse plus 150 × 2.1 mm, 3.5μm (Agilent, Santa Clara, CA).
4
UHPLC-MS/MS Quantification of Drug Metabolites
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