The hyphenated methodology that combines spectroscopic and chromatographic techniques is gaining high interest in the pharmaceutical industry. Hence, the objective of present research work was to investigate robust and sensitive LC–MS/MS method for quantification of zanubrutinib in biological samples such as human plasma. The extraction of zanubrutinib from spiked plasma was performed by adopting liquid – liquid extraction with diethyl ether as extracting solvent and dacomitinib as internal standard. The extracted zanubrutinibstandard and internal standard were chromatographed on an Hypersil Gold C18 (50 mm×3.0 mm, 5 µm) columnwith pH 4.2,ammonium formate(5.0 mM) and acetonitrile in 75: 25 (v/v) as solvent A, pH 4.2 ammonium formate(5.0 mM) and methanol in 60: 40 (v/v) as solvent B. Equal volumes of solvent A and B was used as mobile phasepumped at 0.5 mL/minflow in isocratic mode. The analysis was completed within a total chromatographic run time of 4 min that facilitates less time and solvent consumption. The column separated analytes were recorded using mass detector with positive ion electrospray ionization source. The mass spectrum shows precursor-to-product ion transitions at m/z of 472/167 (m+1) for zanubrutiniband 470/321 (m+1) for dacomitinib. The method produces calibration curve linear in 1-500 ng/mL concentration range with sensitive detection limit of 0.30 ng/mL for Zanubrutinib. The mean plasma spiked extraction recoveries of both zanubrutinib and internal standard was very high with acceptable % RSD in all precision studies.The analytes were noticed to be stable in a variety of stability studies performed.The method was validated to be sensitive, accurate and was suitable for determination of zanubrutinibin human plasma and applicable for regular quality analysis studies.