Security guard cartridges rp 1

To analyze BCP from the in vivo and in vitro studies, the LC–MS/MS manufactured by Shimadzu (LCMS-8050 set, Kyoto, Japan) was used. The LC–MS/MS system consisted of an auto-sampler (SIL-30AC), LC pump (LC-30AD-1 and -2), column oven (CTO-20AC), and a coupled electrospray ionization (ESI) detector (LCMS-8050). A Zorbax SB-C8 column (150 × 2.1 mm, i.d. 3.5 µm, Agilent, Santa Clara, CA, USA) with Security Guard Cartridges RP-1 (4 × 3.0 mm; Phenomenex, CA, USA) was used to separate each compound. The mobile phase was adjusted using 95% ACN in 0.1% formic acid. Flow rate, column oven temperature, and injection volume were 0.5 mL/min, 40 °C, and 5 L, respectively. The electrospray ionization (ESI) source was operated in the negative mode for BCP and in the positive mode for IS. All samples were observed in multiple reaction monitoring (MRM) modes. In the MRM mode, the dwell time was 100 ms per MRM channel. Gas flow, source temperature, and nebulizing gas flow were set at 10 L/min, 300 °C, and 3 L/min, respectively. The collision energy was 26 V for BCP and −14 V for IS. The analytical conditions are summarized in Table S3.

High-performance liquid chromatography (HPLC) analysis of pyrogallol was performed using an Agilent 1290 Infinity LC (Agilent Technology, Waldbronn, Germany). Samples were analyzed by ultraviolet-visible spectroscopy (UV-Vis) detection at a wavelength of 200 nm. A Luna C18 (150 × 3.00 mm i.d. 3 μm) (Phenomenex, CA, USA) column with Security Guard Cartridges RP-1 (4 × 3.0 mm; Phenomenex) was used for pyrogallol analysis. Analysis was performed with isocratic conditions and a mobile phase consisting of 0.1% formic acid in DW (solvent A) and ACN (solvent B). The flow rate and injection volume were 0.35 mL/min and 10 μL, respectively. The detailed HPLC conditions for pyrogallol are shown in Table 2.