Bioanalytical assay for the quantification of the ALK inhibitor lorlatinib in mouse plasma using liquid chromatography-tandem mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2018-04, Vol.1083, p.204-208
• Main Authors: Spatari, Claudia, Li, Wenlong, Schinkel, Alfred H., Ragno, Gaetano, Schellens, Jan H.M., Beijnen, Jos H., Sparidans, Rolf W.
• Format: Article
• Language: English
• Subjects: ALK inhibitor; Animals; Chromatography, Liquid - methods; Drug Stability; Female; Lactams, Macrocyclic - blood; LC-MS/MS; Linear Models; Lorlatinib; Mice; Mouse plasma; Receptor Protein-Tyrosine Kinases - antagonists & inhibitors; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry - methods
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2018.03.014

A bio-analytical assay for the first third generation ALK inhibitor lorlatinib in mouse plasma was developed and validated. Ten-μl plasma samples were prepared by adding rucaparib as the internal standard and precipitation of the plasma proteins. For LC-MS/MS analysis, compounds were eluted at 0.5 mL/min and separated on a 3-μm particle-size, polar embedded octadecyl silica column by gradient elution using 0.1% of formic acid (in water) and methanol. Compounds were monitored with positive electrospray ionization using a triple quadrupole mass spectrometer in selected reaction monitoring mode. The assay was fully validated in the 2–2000 ng/mL calibration range. Within−day (8.0–11.6%) and between−day (10.0–15.0%) precisions and accuracies (99.0–113.3%) were within acceptable range. Plasma samples were deemed stable for 6 h at ambient temperature, during three freeze-thaw cycles and for 2 months at −30 °C. Finally, the new assay was applied successfully to pilot pharmacokinetic studies in male and female wild-type mice. •The first completely validated bio-analytical assay for lorlatinib is presented.•The LC-MS/MS method has successfully been validated in mouse plasma.•The assay was applied for a pharmacokinetic pilot study in wild type mice.