HPLC-MS/MS法测定人血浆中维格列汀浓度及其应用  

HPLC-MS/MS method for the quantification of vildagliptin in human plasma and its application

作　　者：王医成[1,2] 贺康 彭静波[1,2] 饶泰 陈尧[1,2] 郭莹[1,2] 谭志荣 WANG Yi-cheng;HE Kang;PENG Jing-bo;RAO Tai;CHEN Yao;CUO Ying;TAN Zhi-rong(Department of Clinical Pharmacology,Xiangya Hospital,Central South University,Changsha 410078,China;Institute of Clinical Pharmacology,Hunan Key Laboratory of Pharmacogenetics,Central South University,Changsha 410078,China)

机构地区：[1]中南大学湘雅医院临床药理研究所,长沙410078 [2]中南大学临床药理研究所,长沙410078

出　　处：《药物分析杂志》2024年第1期68-75,共8页Chinese Journal of Pharmaceutical Analysis

基　　金：湖南省自然科学基金资助项目(2020JJ9057、2022JJ80113、2022JJ80097)。

摘　　要：目的:建立HPLC-MS/MS法测定人EDTA抗凝血浆中维格列汀的浓度,并将其应用于药代动力学研究。方法:以稳定同位素标记^(13)C-^(15)N-维格列汀为内标,血浆样品用乙腈进行蛋白沉淀处理,采用Hypurity C_(18)(150 mm×2.1 mm, 5μm)色谱柱,以甲醇-5 mmol·L^(-1)甲酸铵水溶液为流动相,梯度洗脱,流速0.5 mL·min^(-1),柱温40℃,进样量2μL,采用电喷雾离子源(ESI源),多反应监测正离子模式进行检测。用于定量分析的维格列汀监测离子对m/z 304.3→154.2,内标监测离子对m/z 310.3→160.3。考察其专属性、标准曲线、定量限、精密度、回收率、基质效应、稳定性,并使用该方法对健康受试者的血浆维格列汀浓度进行测定。结果:血浆中维格列汀质量浓度在1.11~534.0 ng·mL^(-1)范围内线性关系良好。4个浓度水平的批内、批间精密度(RSD)均在0.9%~8.5%,准确度在99.8%~109.3%。低浓度血浆样品室温放置0.5、1、2 h的准确度分别为92.0%、87.6%、71.2%,冰上放置0.5、1、2 h的准确度分别为102.0%、94.5%、86.6%,该结果提示维格列汀在血浆中可能存在不稳定现象。提取回收率、基质效应以及其他稳定性结果等均符合生物样品分析的相关要求。8例健康受试者药代动力学研究结果:t_(1/2)为(1.49±0.37) h,t_(max)为(2.06±1.11) h,C_(max)为(290.94±100.36) ng·mL^(-1),AUC_(0-24 h)为(1 343.46±186.89) ng·h·mL^(-1),AUC_(0-∞)为(1 351.31±188.79) ng·h·mL^(-1)。结论:该方法操作简便,特异性好,灵敏度高,成功应用于8名健康受试者空腹口服给药50 mg维格列汀片后的药代动力学研究,可作为一种可靠的检测方法用于人体药动学研究和治疗药物监测。Objective: To develop a high performance liquid chromatography-mass spectrometry(HPLC-MS/MS) method for the determination of vildagliptin in human anticoagulant plasma with ethylenediamine tetra acetic acid and apply it to the study of pharmacokinetics. Methods:^(13)C-^(15)N-vildagliptin was used as internal standard(IS). After extraction from human plasma by protein precipitation with acetonitrile, all components were separated by a Hypurity C_(18) column(150 mm×2.1 mm, 5 μm), using a gradient elution procedure consisting of methanol and 5 mmol·L^(-1) ammonium formate at a flow rate of 0.5 mL·min^(-1),and the column temperature was 40 ℃. Injection volume was just 2 μL. Positive electrospray ionization was performed using multiple reaction monitoring(MRM) with transitions of m/z 304.3→154.2 for vildagliptin and m/z 310.3→160.3 for internal standard. Specificity, standard curve, lower limit of quantification, precision, recovery, matrix effect and stability were examined. Then this method was used to determine the plasma concentration of veragliptin in healthy subjects. Results:The calibration curve of vildagliptin in human plasma was linear over the concentration range of 1.11 to 534.0 ng·mL^(-1). The lower limit of quantitation was 1.11 ng·mL^(-1). The intra-and inter-day precisions at four quality control levels were within 0.9%-8.5%,and the accuracy was within 99.8%-109.3%. The data of short-term stability at room temperature displayed that the accuracy percentage of LQC samples was 92.0% for 0.5 h exposure, 87.6% for 1 h exposure, 71.2% for 2 h exposure. These of LQC samples chilled on ice was 102.0% for 0.5 h exposure, 94.5% for 1 h exposure, 86.6% for 2 h exposure. These results showed a phenomenon that there was a possible degradation of vildagliptin in plasma. The results of extraction recovery and matrix effect and other stability met the requirements of biological sample analysis. The pharmacokinetic study results of 8 healthy subjects showed that t_(1/2 )was(1.49±0.37) h, t_(max )was(2.06

关 键 词：高效液相色谱质谱联用法 维格列汀 二肽基肽酶Ⅳ(DDP-4) 稳定性 药代动力学 糖尿病 治疗药物监测(TDM) 

分 类 号：R917[医药卫生—药物分析学]