Determination of crizotinib in human and mouse plasma by liquid chromatography electrospray ionization–tandem mass spectrometry (LC-ESI–MS/MS)

Abstract

An LC-ESI-MS/MS method using high-throughput solid-phase extraction (SPE) was developed and validated to measure crizotinib in human and mouse plasma to support ongoing clinical and preclinical pharmacokinetic studies. Chromatographic separation of mouse or human plasma extracts was performed on a Supelco Discovery c18 column (50 × 2.1mm, 5.0 µ) with gradient elution using a combination of acidified aqueous and methanol (MeOH) mobile phases. The mass-to-charge transition monitored for detection and quantitation of crizotinib was m/z 450.2>260.2 while the stable label internal standard (ISTD) was monitored at m/z 457.2>267.3. The validation studies demonstrated that the assay is both precise and accurate with %CV < 9% and accuracies within 8% of nominal target concentration across all concentrations tested for both the human and mouse plasma matrices. Sample volumes required for analysis were 50 µL and 25 µL for human plasma and mouse plasma, respectively. Calibration curves were linear over a range of 5 – 5000 ng/mL for human plasma and 2 – 2000 ng/mL for mouse plasma. The use of a 96-well plate format enabled rapid extraction as well as compatability with automated workflows. The method was successfully applied to analyze crizotinib concentrations in plasma samples collected from children enrolled on a phase I pediatric study investigating the use of crizotinib for treatment of pediatric brain tumors.

On-Line Sequential Chemiluminescence Detection of the Chemotherapy Crizotinib With Luminol-Potassium Ferricyanide/Silver Nanoparticles System in Pharmaceutical Preparations and Human Serum

“…These methods include, spectrofluorimetry, [3] spectrophotometry [4,5] and liquid chromatography-tandem mass spectrometry. [6][7][8] The use of nanotechnology in analytical chemistry field facilitates the evaluation and detection of pharmaceutical drugs in very small levels, increase the suitability of the analytical methods and cut down the cost of analytical analysis.…”

Recent developments in the chromatographic bioanalysis of approved kinase inhibitor drugs in oncology

“…The reproducibility is mostly affected by the repeated addition and elution of liquids, and can be corrected by the use of a preferably stable isotope labeled internal standard. While the most frequently stated reason for SPE in TKI bioanalysis is the reduced matrix effect compared to PP [15,[92][93][94], most papers do not state which factors influenced their choice for preparative methods. SPE provides clean samples, and can relatively easily be automated to in-line systems, but in-line SPE could drastically reduce sample throughput of the equipment.…”

“…With these steps, for normal size SPE columns, usually up to 1 ml per step is used. Micro-SPE column plates require less solvents, but still use 200 µl for washing and elution [15,92]. In TKI bioanalysis, SPE is mostly used for less sensitive and specific detection like UV [93,[96][97][98][99][100][101], or in multi-drug assays when short analytical run-times are desired [15], because there is not much space in the chromatography to accommodate ion-suppressive matrix components.…”

Enhanced spectrofluorimetric determination of the multitargeted tyrosine kinase inhibitor, crizotinib, in human plasma via micelle-mediated approach

“…Accordingly, reliable and accurate analytical methodology for CRZ determination is crucial for further pharmacokinetic studies. Literature review revealed that CRZ was assayed in plasma by liquid chromatography coupled with either tandem mass spectrometry [6][7][8][9] or fluorescence detection [10]. However, these chromatographic methods suffered from complexity and being expensive.…”