Sciex 5500 qtrap mass spectrometer system

Metabolite identification was conducted on an Acquity UPLC H-Class system coupled with a XevoG2-S QTof™ mass spectrometer (Waters, Milford, MA, USA). Quantitative analysis was carried out using a Shimadzu ultra-fast liquid chromatography (UFLC) system (Shimadzu Corporation, Kyoto, Japan) coupled to an Applied Biosystems Sciex 5500 QTRAP^® mass spectrometer system (AB SCIEX, Foster City, CA, USA). Sample preparation and mobile phase preparation were carried out using various equipment: a KQ-400DE type ultrasonic cleaning machine from Kunshan Ultrasonic Instrument Co., Ltd. (Kunshan, China), a Milli-Q purification system from Millipore (Bedford, MA, USA), a GZX-9070MBE type electric blast drying oven from Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory (Shanghai, China), and BT25S 1/100,000 electronic analytical balance and BS210S 1/10,000 electronic analytical balance from Sartorius Scientific Instrument (Beijing) Co., Ltd. (Beijing, China).

Quantitative analysis was performed using a Shimadzu ultra-fast liquid chromatography (UFLC) system (Shimadzu Corporation, Kyoto, Japan) coupled to an Applied Biosystems Sciex 5500 QTRAP^® mass spectrometer system (AB SCIEX, Foster City, CA, USA). Chromatographic separation was achieved using an ACQUITY UPLC™ HSS T3 column (100 mm × 2.1 mm, 1.8 μm, Waters). The mobile phase consisted of water with acetonitrile (A) and 0.1% formic acid (B), with a gradient elution: 90–70% (B) from 0 to 2 min; 70–35% (B) from 2 to 10 min; 35–15% (B) from 10 to 11 min; 15–90% (B) from 11 to 12 min; followed by elution with 90% B for 3 min. The column was operated at a flow rate of 0.3 mL/min, and the injection volume was 2.0 μL.
Mass spectrometry detection was conducted using the electrospray ionization (ESI) source in positive mode. Curtain gas (CUR), nebulizer gas (GS1), and auxiliary gas (GS2) were set at 35, 50, and 50 psi, respectively. The ion spray voltage (IS) was set to 5500 V, and the source temperature was maintained at 550 °C. Multiple reaction monitoring (MRM) mode was employed for quantitation, with a dwell time of 50 ms for each MRM transition. Applied Biosystems Analyst software (version 1.6) was used for controlling the UFLC-QTRAP-MS/MS system, as well as for data acquisition and processing.

Quantitative analysis was performed using a Shimadzu ultra-fast liquid chromatography (UFLC) system (Shimadzu Corporation, Kyoto, Japan) coupled to an Applied Biosystems Sciex 5500 QTRAP^® mass spectrometer system (AB SCIEX, Foster City, CA, USA). Chromatographic separation was achieved using an ACQUITY UPLC™ HSS T3 column (100 mm × 2.1 mm, 1.8 μm, Waters). The mobile phase consisted of water with acetonitrile (A) and 0.1% formic acid (B), with a gradient elution: 90–70% (B) from 0 to 2 min; 70–35% (B) from 2 to 10 min; 35–15% (B) from 10 to 11 min; 15–90% (B) from 11 to 12 min; followed by elution with 90% B for 3 min. The column was operated at a flow rate of 0.3 mL/min, and the injection volume was 2.0 μL.
Mass spectrometry detection was conducted using the electrospray ionization (ESI) source in positive mode. Curtain gas (CUR), nebulizer gas (GS1), and auxiliary gas (GS2) were set at 35, 50, and 50 psi, respectively. The ion spray voltage (IS) was set to 5500 V, and the source temperature was maintained at 550 °C. Multiple reaction monitoring (MRM) mode was employed for quantitation, with a dwell time of 50 ms for each MRM transition. Applied Biosystems Analyst software (version 1.6) was used for controlling the UFLC-QTRAP-MS/MS system, as well as for data acquisition and processing.