An LC–MS/MS method for rapid and sensitive high‐throughput simultaneous determination of various protein kinase inhibitors in human plasma

Abstract

A reliable, high-throughput and sensitive LC-MS/MS procedure was developed and validated for the determination of five tyrosine kinase inhibitors in human plasma. Following their extraction from human plasma, samples were eluted on a RP Luna®-PFP 100 Å column using a mobile phase system composed of acetonitrile and 0.01 m ammonium formate in water (pH ~4.1) with a ratio of (50:50, v/v) flowing at 0.3 mL min . The mass spectrometer was operating with electrospray ionization in the positive ion multiple reaction monitoring mode. The proposed methodology resulted in linear calibration plots with correlation coefficients values of r = 0.9995-0.9999 from concentration ranges of 2.5-100 ng mL for imatinib, 5.0-100 ng mL for sorafenib, tofacitinib and afatinib, and 1.0-100 ng mL for cabozantinib. The procedure was validated in terms of its specificity, limit of detection (0.32-1.71 ng mL ), lower limit of quantification (0.97-5.07 ng mL ), intra- and inter assay accuracy (-3.83 to +2.40%) and precision (<3.37%), matrix effect and recovery and stability. Our results demonstrated that the proposed method is highly reliable for routine quantification of the investigated tyrosine kinase inhibitors in human plasma and can be efficiently applied in the rapid and sensitive analysis of their clinical samples.

Development and validation of a rapid and sensitive UPLC–MS/MS assay for the quantification of tofacitinib in human plasma

“…Acetonitrile with ammonium acetate buffer gave better sensitivity and sharper peaks for TOF and TOF‐13C3 15 N than methanol. It was observed that equal proportions of aqueous and organic media in the mobile phase resulted in early elution of TOF as observed previously (Abdelhameed et al, ; Kadi et al, ) on Gemini C 18 and Cosmosil C 18 columns with considerable matrix interference. Further, when the aqueous phase was kept at ≥60%, there was excessive retention of TOF leading to an analysis time of ~10 min.…”

“…However, the recovery obtained was <70% and selectivity was significantly affected with considerable matrix interference owing to repeated clogging of the column. Kadi et al () and Abdelhameed et al () reported a mean recovery of ≥98% for TOF from 1000 μL of plasma by LLE with diethyl ether in the presence of glycine buffer. Thus, LLE was tested with different solvents like diethyl ether, ethyl acetate, n ‐hexane, dichloromethane and MTBE.…”

“…For the extraction of TOF from various biological matrices all three conventional procedures were employed namely, protein precipitation (PPT) (Paniagua et al, ), liquid–liquid extraction (LLE) (Abdelhameed et al, ; Kadi et al, ; Kumar et al, ) and solid‐phase extraction (Dowty et al, ; Krishnaswami, Wang, et al, ; Krishnaswami, Boy, et al, ; Menon et al, ; Sharma et al, ). Initially, PPT was tried with acetonitrile and methanol in the presence of a base.…”

“…To obtain better selectivity and sensitivity as well as shorter analysis time, appropriate selection of chromatographic conditions like mobile phase and its pH, column and flow rate was essential. In the majority of the methods (Abdelhameed et al, ; Dowty et al, ; Kadi et al, ; Kumar et al, ; Paniagua et al, ; Sharma et al, ) the analysis time was ≥4 min. Initially, some reversed‐phase columns like Gemini C 18 (150 × 4.6 mm, 5.0 μm), Cosmosil C 18 (100 × 4.6 mm, 5.0 μm) and Symmetry C 18 (100 × 4.6 mm, 5.0 μm) were tested using formic acid–ammonium formate, acetic acid–ammonium acetate buffer and methanol–acetonitrile in different ratios.…”

Quantification of cobimetinib, cabozantinib, dabrafenib, niraparib, olaparib, vemurafenib, regorafenib and its metabolite regorafenib M2 in human plasma by UPLC–MS/MS

“…Numerous LC–MS/MS methods have been described for quantification of the individual compounds or a combination of cobimetinib (Cardoso et al, ; Deng et al, ; Huynh et al, ; Rousset et al, ), cabozantinib (Abdelhameed, Attwa, & Kadi, ), dabrafenib (Cardoso et al, ; Huynh et al, ; Merienne et al, ; Rousset et al, ), niraparib (van Andel et al, ), olaparib (Nijenhuis, Lucas, Rosing, Schellens, & Beijnen, ; Pressiat et al, ), vemurafenib (Cardoso et al, ; Huynh et al, ; Nijenhuis, Rosing, Schellens, & Beijnen, ; Rousset et al, ) and regorafenib (Cardoso et al, ; Huynh et al, ; Luethi et al, ; Merienne et al, ; van Erp et al, ) in human plasma. However, a quantification method combining all of the above mentioned oncolytics in a single run has not been published yet.…”