UPLC-MS/MS检测大鼠血浆中沃诺拉赞、代谢产物沃诺拉赞羧酸(M1)及其药动学研究

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

关键词:  沃诺拉赞  代谢产物  药动学  超高效液相色谱串联质谱法

Key words:  vonoprazan  metabolite  pharmacokinetics  UPLC-MS/MS

摘要:

目的 建立一种同时快速检测大鼠血浆中沃诺拉赞及其代谢产物沃诺拉赞羧酸(M1)的超高效液相色谱串联质谱方法(UPLC-MS/MS)，并应用该方法开展其在大鼠体内的药动学研究。方法 使用ACQUITY UPLC^® BEH C₁₈柱(100 mm×2.1 mm, 1.7 μm)对沃诺拉赞和M1进行分离，柱温为40 ℃；流动相为乙腈-水(含0.1%甲酸)，梯度洗脱，流速0.4 mL·min^–1；采用ESI⁺电喷雾离子源结合多反应监测模式进行检测，沃诺拉赞的定量离子对为m/z 346.04→314.97，代谢产物M1的离子对为m/z 347.08→205.06；大鼠血浆加入内标后经乙腈沉淀去除蛋白，离心后取2 μL进样。所有数据应用DAS 3.2.7软件进行分析得到药动学参数。结果 沃诺拉赞和M1的保留时间分别为1.07 min和1.25 min；标准曲线显示沃诺拉赞和M1分别在5~1 000 ng·mL^–1和10~2 000 ng·mL^–1内呈良好线性关系；沃诺拉赞和M1的精密度和准确度为–4.41%~11.68%，提取回收率为78.85%~86.05%，基质效应为98.54%~104.08%；沃诺拉赞和M1的稳定性结果RSD均< 15.0%。大鼠灌胃10 mg·kg^–1沃诺拉赞后，体内药物和代谢产物M1的曲线下面积AUC_(0~t)分别为1 972.51，13 232.42 μg·L^–1·h，半衰期t_1/2分别为2.97，2.13 h，血浆清除率CLz分别为5.13，0.76 L·h^–1·kg^–1。结论 该方法分析时间短、操作简便，方法学均符合生物样品分析相关要求，可以适用于大鼠体内沃诺拉赞及其代谢产物M1浓度检测和药动学研究。

Abstract:

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.