Determination of vonoprazan and Metabolite Vonoprazan Carboxylic Acid(M1) by UPLC-MS/MS and its application to pharmacokinetic study in rats Plasma

OBJECTIVE To establish a ultra-high-performance liquid chromatography mass spectrometry (UPLC-MS/MS) method for the rapid simultaneous detection of vonoprazan and its metabolite vonoprazan carboxylic acid (M1) in rat plasma and their application to in vivo pharmacokinetic studies in rats.METHODS Vonoprazan and M1 were separated on an ACQUITY UPLC^® BEH C₁₈ column (100 mm×2.1 mm,1.7 μm) with column temperature at 40℃.Acetonitrile-water (containing 0.1% formic acid) was used as the mobile phase and gradient elution was applied with the flow rate at 0.4 mL·min^-1.Electrospray positive ionization source (ESI⁺) combined with multiple reaction monitoring mode was selected,with quantitative transition m/z 346.04→314.97 for vonoprazan,and m/z 347.08→205.06 for metabolite M1.The protein was removed by acetonitrile precipitation after adding internal standard and 2 μL supernatant was injected.Therefore,all plasma data were analyzed by DAS 3.2.7 software to obtain pharmacokinetic parameters.RESULTS The retention times of vonoprazan and M1 were 1.07 min and 1.25 min,respectively.The concentration range of 5-1 000 ng·mL^-1 and 10-2 000 ng·mL^-1 of vonoprazan and M1 were shown with good linear relationship.The precision and accuracy of vonoprazan and M1 were-4.41%-11.68%,and the extraction recovery rate were 78.85%-86.05%,the matrix effect were 98.54%-104.08%.Vonoprazan and M1 showed good stability under various experimental conditions.The stability results of vonoprazan and M1 showed that all RSD were<15.0%.After intragastric administration of 10 mg·kg^-1 vonoprazan,the area under the curve AUC_(0-t)of the drug and metabolite M1 in vivo were 1 972.51 and 13 232.42 μg·L^-1·h,respectively;the half-life t_1/2 were 2.97 and 2.13 h,respectively;the plasma clearance CLz were 5.13 and 0.76 L·h^-1·kg^-1,respectively.CONCLUSION The method has the advantages of short analysis time and simple operation,which meets the relevant requirements of biological sample analysis.Thus,the method can successfully applied to the concentration determination and pharmacokinetic study of vonoprazan and M1 in rats.