Title:A Rapid and Selective UPLC-MS/MS Assay for Accurate Analysis of Apatinib in Rat Plasma and its Application to a Pharmacokinetic Study
VOLUME: 17 ISSUE: 5
Author(s):Jinglin Gao, Zhangying Feng, Huan Ren, Mengdi Yu, Haidong Wang and Mingxia Wang*
Affiliation:Department of Clinical Pharmacology, Fourth Hospital of Hebei Medical University, 12 Jiankang Rd, Shijiazhuang, 050011, Department of Clinical Pharmacology, Fourth Hospital of Hebei Medical University, 12 Jiankang Rd, Shijiazhuang, 050011, Department of Clinical Pharmacology, Fourth Hospital of Hebei Medical University, 12 Jiankang Rd, Shijiazhuang, 050011, Department of Clinical Pharmacology, Fourth Hospital of Hebei Medical University, 12 Jiankang Rd, Shijiazhuang, 050011, Department of Clinical Pharmacology, Fourth Hospital of Hebei Medical University, 12 Jiankang Rd, Shijiazhuang, 050011, Department of Clinical Pharmacology, Fourth Hospital of Hebei Medical University, 12 Jiankang Rd, Shijiazhuang, 050011
Keywords:Apatinib, UPLC-MS/MS, rat plasma, pharmacokinetics, protein precipitation, method development, validation.
Abstract:
Objective: Apatinib, a novel small-molecule Tyrosine Kinase Inhibitor (TKI), is under development
to treat advanced gastric cancer. For the pharmacokinetic evaluation and routine drug monitoring
of apatinib, a quantitative ultra-performance liquid chromatography coupled with tandem mass
spectrometry (UPLC-MS/MS) method in rat plasma was developed with tinidazole used as an internal
standard (IS).
Methods: Protein precipitation (PPT) was selected as a sample pre-treatment method to extract apatinib.
Then, chromatography was performed on a Kinetex C8 column (2.1×100 mm, 2.6 μm) using a constant
mobile phase including 0.2% formic acid and 10 mM ammonium acetate in water and methanol
(30:70, v/v) with a gradient flow rate from 0.2 mL/min to 0.4 mL/min. Chromatographic analysis was
performed in only 4.5 min. Mass spectrometric detection was carried on positive electrospray ionization
(ESI+) mode with Multiple-Reaction Monitoring (MRM).
Results: The standard calibration curve showed good linearity in 2-1000 ng/mL with the correlation
coefficient (R2) > 0.99. The Lower Limit of Quantitation (LLOQ) was 2 ng/mL. The precision, accuracy,
extraction recovery, matrix effect, stability and carryover were all within the acceptable range.
Conclusion: This method was simple, accurate, selective and successfully used for a pharmacokinetic
study following seven rats orally administrated a single of 60 mg/kg apatinib.
