Introduction: In our research, a rapid, selective and sensitive analytical method based on
ultra-performance liquid chromatography (UPLC) was developed and validated for the quantitative
analysis of icotinib (an oral and novel EGFR-TKI) in rat plasma. This method was applied for pharmacokinetics.
Sample preparation was completed through protein precipitation with acetonitrile, and
chromatographic separation was performed on an Acquity UPLC BEH C18 column (2.1 mm x 50 mm,
1.7 µm) with a water-acetonitrile-0.1% trifluoroacetic acid mobile phase (0.3 mL/min), utilizing ultraviolet
detection at 248 nm. Just 100 µL plasma sample and 3.5 minutes’ running time were required,
representing an obvious improvement compared with previously described methods. The assay was
linear over the range of 0.05-10 µg/mL for icotinib, with a lower limit of quantitation (LLOQ) of 0.05
μ g/mL for icotinib and an excellent linearity (r2=0.9992) was achieved.
Conclusion: Intra and inter-day precisions of icotinib were no more than 10% and its accuracies were
in the range of 98.3-104.0%. Our method was successfully applied for the determination of icotinib in
rat plasma for pharmacokinetic study.