The current technique was developed to estimate the amount of alectinib present in spiked rabbit plasma using liquid chromatographic mass spectrometry. The liquid-liquid extraction method was used, and chromatographic separation was carried out on a C18 (4.6mm id x 50mm) analytical column with a mobile phase consisting of acetonitrile and water with 0.1% formic acid at a volume ratio of 75:25. Alectinib's product m/z +483.2 (parent) 396.1 (product) and the internal standard m/z +447.5 (parent) 380.3 (product) were both obtained using positive ion mode. The calibration curve was linear from 0.5 to 600 ng/ml. The percentage extraction recovery (98.15% → 98.86%), demonstrated excellent matrix and analyte selectivity (% interference = 0), and satisfactory stability study results in all types (% nominal 94.94% → 99.63%). The intra and interday accuracy with % nominal 97 → 98.8%, precision % CV ≤ 2% in all quality control levels. The rabbit model's pharmacokinetic parameters were examined, and alectinib's area under the curve (AUC 0—∞) was 4269 ± 8.13 hr.ng/ml. The half-life of elimination (t1/2) is 8.52 ± 6.66 hours. The currently established approach was used in rabbit blood samples for pharmacokinetic investigations of commercial formulations since it was thought to be a novel, verified bioanalytical method based on experimental results.