Simultaneous and Rapid Determination of Six Tyrosine Kinase Inhibitors in Patients with Non-Small Cell Lung Cancer Using HPLC-MS/MS

Abstract

Objective. To develop a new method for quantitatively analyzing six tyrosine kinase inhibitors (gefitinib, erlotinib, icotinib, afatinib, osimertinib, and crizotinib) used in the treatment of non-small cell lung cancer (NSCLC) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods. The analytes were detected in the selected reaction monitoring mode on a triple quadrupole mass spectrometer with the positive ionization mode. Carbamazepine was utilized as the internal standard. The pretreatment of the plasma sample was completed based on protein precipitation with acetonitrile, and the analytes were separated on an Agilent Zorbax SB-C18 reversed-phase column (2.1 mm × 100 mm, 3.5 μm, Agilent, USA) using gradient elution. The mobile phase consisted of 0.1% formic acid in water (phase A) and 0.1% formic acid in acetonitrile (phase B). The flow rate was 0.3 mL/min, and the injection volume was 5 μL. The column temperature was set and maintained at 35°C. Results. The calibration curves were linear over the range from 5.0 to 1000.0 ng/mL for gefitinib, crizotinib, and osimertinib; from 50.0 to 4000.0 ng/mL for icotinib and erlotinib; and from 5.0 to 400.0 ng/mL for afatinib. Linear correlation coefficients were >0.990 for all regression curves. The intra- and interday accuracy and precision of the method were within ±15.0% and not more than 15.0%, respectively. The mean recovery of all the analytes ranged from 70.18% to 110.76%, the matrix effect was from 88.85% to 127.58%, and stability was within ±15.0%. Conclusion. This newly developed method was sensitive, simple, and robust and could be used in therapeutic drug monitoring of six tyrosine kinase inhibitors in NSCLC patients.

Pretreatment and analysis techniques development of TKIs in biological samples for pharmacokinetic studies and therapeutic drug monitoring

“…In addition to optimizing stationary phases, it is crucial to compare and optimize mobile phases when establishing an appropriate HPLC method. To enhance separation effect and peak shape, influencing factors such as pH (i.e., formic acid) [ 33 ], buffer solution (i.e., ammonium formate) [ 3 ], as well as elution mode (i.e., isocratic elution or gradient elution) [ 80 ] should be focused on according to the characteristics of samples and TKIs.…”

“…The parameter was optimized using a 3-level factorial design approach, and the best results were obtained under the following conditions: C 18 column, ammonium format buffer consisting of acetonitrile (28.19:71.81, V / V ), and buffer with pH 3. This optimized method was successfully applied to a pharmacokinetic study, demonstrating high recovery (99.91%), specificity (99.94%), precision (RSD < 0.543%), and robustness (RSD < 0.500%) ( Table S1 ) [ 2 , 3 , 5 , 8 , 12 , [15] , [16] , [17] , [26] , [27] , [28] , [29] , [30] , [31] , [32] , [33] , [34] , [35] , [37] , [38] , [39] , 41 , [48] , [49] , [50] , 52 , 54 , 55 , 61 , 63 , 64 , 76 , 80 , 83 , 84 , 86 , [88] , [89] , [90] , [91] , [92] , [93] , [94] , [95] , [97] , [98] , [99] , [100] , [101] , [102] , [103] , [104] , [105] , 109 , [111] , [112] , [113] , [114] ,…”

Determination of Gefitinib Using Routine and Greener Stability-Indicating HPTLC Methods: A Comparative Evaluation of Validation Parameters

“…A system-biology-based in silico analysis was applied for the identification of off-targets of GFT [18]. Various "liquid chromatography mass spectrometry (LC-MS)" methods have been used to measure GFT in mouse and human plasma samples [19][20][21][22][23]. An ultra-performance liquid chromatography (UPLC) technique was also used for the GFT analysis in human plasma samples [24].…”