Application of newly developed and validated LC-MS/MS method for pharmacokinetic study of adagrasib and pembrolizumab simultaneously in rat plasma

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2024-07, Vol.1241, p.124171, Article 124171
• Main Authors: Harsha Sri, Kamma, Ravisankar, Panchumarthy, Kumar Konidala, Sathish, Srinivasa Babu, P.
• Format: Article
• Language: English
• Subjects: Adagrasib; Animals; Antibodies, Monoclonal, Humanized - blood; Antibodies, Monoclonal, Humanized - chemistry; Antibodies, Monoclonal, Humanized - pharmacokinetics; Bio-analytical method; LC-MS/MS; Limit of Detection; Linear Models; Liquid Chromatography-Mass Spectrometry; Liquid-Liquid Extraction; Male; Pembrolizumab; Pharmacokinetic study; Rats; Reproducibility of Results; Tandem Mass Spectrometry
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2024.124171

•Developed an LC-MS/MS method for the simultaneous determination of new FDA approved anticancer drugs in rat plasma.•The developed method was validated as per USFDA guidelines including stability parameters.•The developed LC-MS/MS method was efficiently employed in an in vivo pharmacokinetic study of anticancer drugs in rat plasma.•The developed LC-MS/MS can act as the therapeutic drug monitoring in combined therapy of selected drugs in non-small cell lung cancer. Non-small cell lung cancer (NSCLC) is a significant subtype of lung cancer, and poses a dangerous global threat. One of the current approaches of NSCLC treatment is a combination therapy of adagrasib and pembrolizumab. Accurate monitoring of these drug concentrations in biological fluids is critical for treatment efficacy. Since no method was reported for simultaneous estimation of these drugs, this study focuses on the development of a validated LC-MS/MS bioanalytical method for simultaneous quantification of Adagrasib and Pembrolizumab in rat plasma. The analytes were extracted from the biological matrix through liquid–liquid extraction techniques using acetonitrile as extraction solvent. The analytes were separated on a Waters X-bridge phenyl C18 column, with a mixture of acetonitrile: 0.1 % TFA in water (50: 50 v/v) as mobile phase at an isocratic flow rate of 1.0 mL/min with a runtime of about 5 min. Adagrasib (m/z 605.12 → 201.62), Pembrolizumab (m/z 146.32 → 85.15), and Sotorasib (m/z 561.59 → 218.92) were determined by recording the mass spectra through multiple reaction monitoring in positive mode. The method was validated according to USFDA guidelines. The results demonstrate satisfactory linearity with an r2 value of 0.9998 in the ranges of 40–800 and 10–200 ng/mL, accuracy with mean percentage recovery of 95.22–98.59 % and 96.98–98.57 %, precision indicated with %RSD ranged between 0.39–1.91 % and 0.85–9.03 % for Adagrasib and Pembrolizumab respectively, and other key parameters. The developed method can determine the pharmacokinetic parameters to indicate the efficacy and safety of the drugs, and also can quantify selected drugs simultaneously in biological samples.