UPLC-MS/MS法测定人体血浆中伏诺拉生浓度及其2种片剂生物等效性研究  

Determination of vonoprazan in human plasma by UPLC-MS/MS and study on the bioequivalence of two types of tablets

作　　者：文琪 蔡娟 孙春艳[2] 唐维英 李莎 杨敏 吴强 WEN Qi;CAI Juan;SUN Chun-yan;TANG Wei-ying;LI Sha;YANG Min;WU Qiang(Bioanalytical Service Center of Sichuan Institute for Drug Control,NMPA Key Laboratory for Technical Research on Drug Products In Vitro and In Vivo Correlation,Chengdu 611731,China;Yangtze River Pharmaceutial Group,National Key Laboratory of Complex Drug Formulations for Overcoming Delivery Barriers,Taizhou 225300,China)

机构地区：[1]四川省药品检验研究院生物样本检测中心,药物制剂体内外相关性技术研究重点实验室工业和信息化部产业技术基础公共服务平台,成都611731 [2]扬子江药业集团克服递药屏障高端制剂全国重点实验室,泰州225300

出　　处：《药物分析杂志》2024年第12期2033-2040,共8页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立并验证一种简便、快速、灵敏的UPLC-MS/MS法,测定人体血浆中伏诺拉生浓度并用于其2种片剂生物等效性研究。方法:采用单次给药、两周期、两制剂、双交叉对照试验设计进行生物等效性研究,健康人体受试者分别在空腹和餐后条件下单剂量口服20 mg富马酸伏诺拉生片受试制剂或参比制剂后在不同时间点采集血浆样品。乙腈蛋白沉淀处理血浆样品后用UPLC-MS/MS法分析。采用Waters ACQUITY UPLC~?BEH C_(18)(50 mm×2.1 mm,1.7μm)色谱柱,柱温40℃,以0.1%甲酸水-0.1%甲酸乙腈为流动相梯度洗脱。ESI离子源,正离子模式,多反应离子监测(MRM),用于定量分析的离子对为m/z 346.1→315.4,内标化合物伏诺拉生-d_(4)为m/z 350.1→316.2。结果:线性范围为0.30~50.00 ng·mL^(-1)(r>0.9989),定量下限为0.30 ng·mL^(-1);批内和批间精密度RSD均<5.7%,准确度在-2.15%~0.82%范围内;提取回收率、选择性、基质效应、稳定性等均符合接受标准。该方法被成功应用于富马酸伏诺拉生片的生物等效性研究。受试制剂在餐后和空腹试验中C_(max)分别为(29.08±11.59)ng·mL^(-1)和(26.87±8.14)ng·mL^(-1),AUC_(0-t)分别为(258.90±87.71)h·ng·mL^(-1)和(223.08±43.27)h·ng·mL^(-1)。参比制剂在餐后和空腹试验中C_(max)分别为(28.73±10.25)ng·mL^(-1)和(26.93±8.09)ng·mL^(-1),AUC_(0-t)分别为(250.33±73.13)h·ng·mL^(-1)和(227.56±46.26)h·ng·mL^(-1)。餐后试验中,受试制剂和参比制剂的C_(max)、AUC_(0-t)、和AUC_(0-∞)几何均值比的90%置信区间分别为88.64%~112.28%、96.1%~108.2%、96.6%~108.7%;空腹试验中,受试制剂和参比制剂的的C_(max)、AUC_(0-t)和AUC_(0-∞)几何均值比的90%置信区间分别为94.0%~106.2%、94.7%~102.0%和95.2%~102.5%。结论:该方法具有简单、快速,灵敏度高等优点。富马酸伏诺拉生片受试制剂和参比制剂具有生物等效性。Objective:To develop a rapid,specific and sensitive UPLC-MS/MS method for the determination of vonoprazan in human plasma and its application in a bioequivalence study of two types of tablets.Methods:A single dose,two-cycle,two products,and self-cross controlled trial design on bioequivalence was used.Plasma samples were collected from healthy human volunteers at different time points after oral administration with the test or reference product of 20 mg fumarate vonoprazan tablets under both fasting and fed conditions,respectively.The plasma samples were treated by acetonitrile protein precipitation and then analyzed by UPLC-MS/MS.Chromatographic separation of vonoprazan was achieved using a Waters ACQUITY UPLC?BEH C_(18)(50 mm×2.1 mm,1.7μm)column at 40℃.The mobile phase consisted of water(containing 0.1%formic acid)for eluent A and acetonitrile(containing 0.1%formic acid)for eluent B under a gradient elution.An electrospray ionization(ESI)with multiple reaction monitoring(MRM)mode was used to monitor the precursor-product ion transitions of m/z 346.1→315.4 for vonoprazan and m/z 350.1→316.2 for vonoprazan-d_(4).Results:The rang of linearity was 0.30-50.00 ng·mL^(-1)(r>0.9989),and the LLOQ was 0.30 ng·mL^(-1).Intra-and inter-day precision values were within 5.7%,and intra-and inter-day accuracy values were ranged from-2.15%to 0.82%.Recovery,specificity,matrix effect and stability met the guiding principles.This method has been successfully applied to study the bioequivalence of vonoprazan fumarate tablets.The C_(max)of the test product in postprandial and fasting tests were(29.08±11.59)ng·mL^(-1)and(26.87±8.14)ng·mL^(-1),respectively,and the AUC_(0-t)was(258.90±87.71)h·ng·mL^(-1)and(223.08±43.27)h·ng·mL^(-1),respectively.The C_(max)of the reference product in postprandial and fasting tests were(28.73±10.25)ng·mL^(-1)and(26.93±8.09)ng·mL^(-1),respectively,and the AUC_(0-t)was(250.33±73.13)h·ng·mL^(-1)and(227.56±46.26)h·ng·mL^(-1),respectively.In the postprandial trial,the 90%CIs for t

关 键 词：超高效液相串联质谱 伏诺拉生 生物等效性 药代动力学 酸相关性疾病 

分 类 号：R917[医药卫生—药物分析学]