Acquity uplc xevo tq s system

LC-MS/MS analysis was performed using an Acquity UPLC/Xevo TQ-S system (Waters, Milford, MA, USA) equipped with a Capcell Core ADME column (100 mm × 2.1 mm i.d., 2.7 μm, Shiseido, Tokyo, Japan) at a temperature of 40 °C. The injection volume and flow rate were 5.0 μL and 0.3 mL/min, respectively. Separation was performed by gradient elution using methanol (mobile phase A) and 5 mM ammonium formate in water (mobile phase B). The initial mobile phase of 10/90 (A/B, v/v) was retained for 3 min, changed to 90/10 (A/B, v/v) over 5 min and then held for 2 min, and finally changed to 10/90 (A/B, v/v) over 0.1 min and then retained until 10 min. MS/MS was conducted using positive electrospray ionization (ESI) and multiple reaction monitoring (MRM). The ion source temperature and the desolvation temperature were 150 °C and 500 °C, respectively, and the capillary voltage was 1.0 kV. The MRM conditions are summarized in Table 1.