Uplc ltq orbitrap xl

Plasma extract samples (7 μL) were injected into an Acquity UPLC-BEH-C18 column (2.1 × 50 mm, 1.7 μm; Waters, Milford, MA) that was coupled in-line with a UPLC-LTQ-Orbitrap XL (Thermo Fisher Scientific, USA). The injected samples were equilibrated with water containing 0.1% formic acid. Samples were eluted with an acetonitrile gradient containing 0.1% formic acid at a flow rate of 0.35 mL/min for 20 min. Metabolites were separated by UPLC (Waters, Milford, MA), analyzed, and assigned by LTQ-Orbitrap-XL (Thermo Fisher Scientific, USA). The mass spectrometer was operated in ESI-positive mode. The spray voltage was 5 kV. The flow rate nitrogen sheath gas and the auxiliary gas were 50 and 5 (arbitrary units). The capillary voltage (V), tube-lens voltage (V), and capillary temperature (°C) were kept constant at 35 V, 80 V, and 370°C, respectively. The Orbitrap data were collected in the range of m/z 50–1,000. For quality control, a mixture of four standard compounds (acetaminophen, sulfadimethoxine, terfenadine, and reserpine) was injected into every 10th sample. The MS/MS spectra of metabolites were obtained by a collision-energy ramp from 55–65 eV, and analyzed with Xcalibur 2.1 and MS Frontier software (Thermo Fisher Scientific, USA).

PBMC and Plasma extract samples (4 μL) were injected into an Acquity UPLC-BEH-C18 column (2.1 × 50 mm, 1.7 μm; Waters, Milford, MA) that was coupled in-line with a UPLC-LTQ-Orbitrap XL (Thermo Fisher Scientific, Waltham, MA). The injected samples were equilibrated with water containing 0.1 % formic acid. Samples were eluted with an acetonitrile gradient containing 0.1 % formic acid at a flow rate of 0.35 mL/min for 20-min. Metabolites were separated by UPLC, analyzed, and assigned by LTQ-Orbitrap-XL. The mass spectrometer (MS) was operated in ESI-positive mode. The spray voltage was 5 kV. The flow-rate nitrogen sheath gas and the auxiliary gas were 50 and 5 (arbitrary units). The capillary voltage (V), tube-lens voltage (V), and capillary temperature (°C) were kept constant at 35 V, 80 V, and 370 °C. Orbitrap data were collected in the range of m/z 50–1,000. MS/MS spectra of metabolites were obtained by a collision-energy ramp from 55–65 eV, and conducted with Xcalibur 2.1 and MS Frontier software (Thermo Fisher Scientific).

The plasma extract samples (7 µL) were injected into an Acquity UPLC-BEH-C18 column (2.1×50 mm, 1.7 µm; Waters, Milford, MA) that was coupled in-line with a UPLC-LTQ-Orbitrap XL (Thermo Fisher Scientific, USA). The injected samples were equilibrated with water containing 0.1% formic acid. Samples were eluted with an acetonitrile gradient containing 0.1% formic acid at a flow rate of 0.35 mL/min for 20 minutes. Metabolites were separated by UPLC (Waters, Milford, MA), analyzed, and assigned by LTQ-Orbitrap-XL (Thermo Fisher Scientific, USA). The mass spectrometer was operated in ESI-positive mode. The spray voltage was 5 kV. The flow-rate nitrogen sheath gas and the auxiliary gas were 50 and 5 (arbitrary units). The capillary voltage (V), tube-lens voltage (V), and capillary temperature (°C) were kept constant at 35 V, 80 V, and 370°C. The Orbitrap data were collected in the range of m/z 50–1,000. For quality control, a mixture of four standard compounds (acetaminophen, sulfadimethoxine, terfenadine, and reserpine) was injected every ten samples. The MS/MS spectra of metabolites were obtained by a collision-energy ramp from 55–65 eV, and conducted with Xcalibur 2.1 and MS Frontier software (Thermo Fisher Scientific, USA).