Abstract
This study describes the development and validation of a highly sensitive HPLC method with fluorescence detection for the determination of crizotinib (CZT) in human plasma. Telmisartan (TLM) was used as internal standard (IS). After a simple protein precipitation using methanol, CZT and TLM (IS) were separated by isocratic mode on a mu-Bondapack CN column, (150 mm length x 3.9 mm i.d., 5 mu m particle diameter) using a mobile phase consisted of 1% acetic acid: acetonitrile (70: 30, v/v). CZT and TLM were eluted at 2.18 and 3.42 min, respectively. The detection of CZT and TLM (IS) was carried out at a wavelength of 264 nm for excitation and 382 nm for emission. Under the optimum chromatographic conditions, linear relationships with excellent correlation coefficient (r = 0.9999, n = 6) were found between the peak area ratio of CZT to that of TLM and the concentration of CZT in the concentration range of 2-512 ng/mL. The accuracy of the method was also validated and the % recovery values from spiked human plasma for the intra-day analysis were 98.37 - 103.01% with a mean value of 100.69% +/- 7.26% whereas the values for the inter-day analysis were 98.37 - 102.44% with a mean value of 100.26% +/- 7.06%. The method had higher throughput as it involved simple sample preparation procedure and short run-time (<5 min). The results demonstrated the great value of the proposed method in pharmacokinetic studies of CZT.