Application of On-line SPE-HPLC System in Pharmacokinetic Study of Highly Active Anti-cancer Compound TEB-415

An on-line solid phase extraction-high performance liquid chromatography (SPE-HPLC) system was developed for the plasma pharmacokinetic study of highly active anti-cancer compound TEB-415 in mouse. A C18 column (Venusil MP, 100 mm × 4.6 mm, 5 μm) was used as the analytical column and a mixed solution of acetonitrile-5 mM monopotassium phosphate buffer (pH 3.5) at a flow rate of 1.0 mL min−1 as the mobile phase for the isocratic elution. The on-line solid phase extraction was carried out on an MF Ph-1 column (10 mm × 4 mm, 5 μm) with purified water as the washing solvent and 5 mM monopotassium phosphate buffer (pH 3.5) as the elution solvent. The detected wavelength was set at 262 nm. The pharmacokinetic parameters were calculated by WinNonlin 5.2 software. The linear range of TEB-415 was 100–20000 μg L−1 with a limit of qualification of 20 μg L−1. The extraction recovery was 91.99%–99.14% and the RSD of intra- and inter-day precision was less than 3.5%. The accuracy of short-term stability, freeze-thaw stability and long-term stability were 91.5%–101.5%. After oral medication, the mean peak time (Tmax) of TEB-415 in mice was 5.29 h and the mean maximum concentration (Cmax) was 3403 μg L−1. The area under the curve (AUC) of TEB-415 was 24600 μg L−1 h−1. The mean half-life of drug in mice was 3.84 h, and the mean retention time (MRT) was 6.56 h. These parameters above suggest that TEB-415 has an appropriate rate of absorption and elimination for the mice. Meanwhile, it had preferable bioavailability.

Graphical AbstractAn on-line solid phase extraction-high performance liquid chromatography (SPE-HPLC) system was developed for the determination of highly active anti-cancer compound TEB-415 in mouse. The chromatograms of blank mice plasma sample, blank mice plasma sample spiked with 400 μg L−1 TEB-415 and plasma sample of mice after administration of TEB-415 for 6 h showed that there was no endogenous interference around the target peak, suggesting a good specificity for the method.Figure optionsDownload full-size imageDownload as PowerPoint slide