*3.1. LC-MS*/*MS Analysis of DWP16001, Dapagliflozin, and Ipragliflozin*

To compare the pharmacokinetics and tissue distribution of DWP16001, dapagliflozin, ipragliflozin in mice, analyses of the three SGLT2 inhibitors using LC-MS/MS were applied. Figure 2 shows the selected precursor and product ions of DWP16001, dapagliflozin, ipragliflozin, and D4-DWP16001 (IS). The selected precursor and product ions of dapagliflozin and ipragliflozin were consistent with previously published findings [12,15].

Representative multiple reaction-monitoring (MRM) chromatograms of DWP16001, D4-DWP16001 (IS), dapagliflozin, and ipragliflozin (Figure 3) showed that all the analyte peaks obtained using the liquid-liquid extraction method using MTBE were well separated with no interfering peaks at their respective retention times.

**Figure 2.** Product ion spectra of (**A**) DWP16001, (**B**) D4-DWP16001 (IS), (**C**) dapagliflozin, and (**D**) ipragliflozin.

**Figure 3.** Representative multiple reaction-monitoring (MRM) chromatograms of (**A**) DWP16001, (**B**) D4-DWP16001 (IS), (**C**) dapagliflozin, and (**D**) ipragliflozin in mouse double-blank plasma, blank plasma spiked with DWP16001, dapagliflozin, ipragliflozin at the lower limit of quantification (LLOQ) (5 ng/mL), and plasma samples at 1 h following single oral administration of DWP16001, dapagliflozin, or ipragliflozin at a dose of 1 mg/kg.
