1290 infinity 2 liquid

Media samples (10 μL) were chromatographed on a ZORBAX SSHD Eclipse Plus C18 column (3×50 mm, 1.8 μm, 959757-302; Agilent, Santa Clara, CA) with a guard column (2.1×5 mm, 1.8 μm, 821725-901; Agilent) using 1290 Infinity II liquid chromatograph (LC; Agilent). Column temperature and the LC flow rate were set at 40°C and 0.4 ml/min. Initial chromatographic condition was maintained at 90% mobile phase A (water with 0.1% formic acid, v/v) and 10% mobile phase B (acetonitrile with 0.1% formic acid, v/v) for one min, then increased to 80% B by 3 min, then to 95% B by 4 min, and then returned to initial condition at 5 min until 8 min for sufficient equilibrium. Unmodified mobile phases A (water) and B (acetonitrile) were used for the analysis of Troglitazone.
MS/MS analyses were performed in positive ion mode (all test drugs except Troglitazone) or negative ion mode (for Troglitazone) with an electrospray ionization (ESI) source using 6470 triple-quadruple MS (Agilent). The capillary voltage was set at 3500 V. The nebulizer gas pressure and gas temperature were set at 35 psi and 350°C, respectively. The MS/MS parameters for each test compound are summarized in Supplemental Table 1.