高效液相色谱串联质谱法测定人血浆中氢氯噻嗪和氨苯蝶啶的浓度  

Simultaneous determination of hydrochlorothiazide and triamterene in human plasma by HPLC-MS/MS

作　　者：齐文渊[1] 李博[1,2] 薛薇[1] 严蓓[1] 杨蕾 李可欣[1] 史爱欣[1] QI Wen-yuan;LI Bo;XUE Wei;YAN Bei;YANG Lei;LI Ke-xin;SHI Ai-xin(Clinical Trial Centre,Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing 100730,China;School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,Liaoning Province,China)

机构地区：[1]北京医院国家老年医学中心、中国医学科学院老年医学研究院临床试验研究中心,北京100730 [2]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《中国临床药理学杂志》2021年第16期2232-2235,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家重点研发计划基金资助项目(2020YFC2008303)。

摘　　要：目的建立HPLC-MS/MS法同时测定人血浆中氢氯噻嗪和氨苯蝶啶的浓度。方法血浆样品用液液萃取进行前处理,用HPLC-MS/MS测定。色谱柱:Waters Atlantis HILIC Silica色谱柱(2.1 mm×50 mm,3.5μm);流动相:5 mmol·L^(-1)乙酸铵水溶液(A)-乙腈(B)=15∶85(v∶v),等度洗脱;流速:0.4 mL·min^(-1);柱温:40℃;进样量:5μL。质谱方法用电喷雾离子化(ESI),正负离子切换模式,多重反应监测(MRM)进行化合物检测,其中氢氯噻嗪及其内标氢氟噻嗪在负离子模式下检测,监测离子对分别为m/z 296.0→m/z 269.0、m/z 330.0→m/z 239.0;氨苯蝶啶及内标d5-氨苯蝶啶在正离子模式下检测,监测离子对分别为m/z 254.0→m/z 237.0和m/z 259.0→m/z 242.0。结果血浆中氢氯噻嗪和氨苯蝶啶线性范围分别为0.5~200 ng·mL^(-1)和0.2~120 ng·mL^(-1),标准曲线的相关系数(r~2)均在0.99~1.00。日内、日间准确度在-15%~15%,精密度均小于15%,血浆基质对氢氯噻嗪和氨苯蝶啶测定无干扰。结论本研究建立的同时定量测定人血浆中氢氯噻嗪和氨苯蝶啶的HPLC-MS/MS方法,灵敏度高、特异性强、耗时短,能够快速测定血浆样品中氢氯噻嗪和氨苯蝶啶浓度,提供准确可靠的分析结果,可用于临床样品的分析。Objective To establish a method for simultaneous determination of hydrochlorothiazide and triamterene in human plasma by HPLC-MS/MS. Methods The plasma sample was extracted using liquid liquid extraction with acetic ether. The resulting supernatant liquid after being centrifuged was supernatant was transferred and evaporated, then the residue was analyzed on a Waters Atlantis HILIC silica column(2.1 mm×50 mm, 3.5 μm). Mobile phase was comprised of acetonitrile(B) and 5 mmol·L-1 ammonium acetate(v/v) aqueous solution(A) in 85% B isocratic elution, with a flow rate of 0.4 mL·min-1, column temperature at 40 ℃, and injected sample volume of 5 μL. The mass spectrometry method used electrospray ionization(ESI), positive and negative ion switching mode, and multiple reaction monitoring(MRM) to detect the compounds, in which hydrochlorothiazide and its internal standard hydroflumethiazide were detected in negative ion mode with the ion transition: m/z 296.0→m/z 269.0 and m/z 330.0→m/z 239.0, respectively;triamterene and its internal standard d5-triamterene were detected in positive ion mode with ion transition: m/z 254. 0→m/z 237. 0 and m/z 259. 0→m/z 242. 0,respectively. Results A good linearity of hydrochlorothiazide and triamterene was shown in the range of 0. 5-200 ng·m L-1 and 0. 2-120 ng·m L-1( r2= 0. 99-1. 00). Both of intra-batch RSD and inter-batch RSD were less than 15 %. The plasma matrix had no effect on the quantification of hydrochlorothiazide and triamterene. Conclusion A rapid,sensitive and accurate HPLC-MS/MS method was developed and it was proved to be effective for the determination of hydrochlorothiazide and triamterene in human plasma.

关 键 词：氢氯噻嗪 氨苯蝶啶 血浆 液液萃取 高效液相色谱串联质谱法 

分 类 号：R972.4[医药卫生—药品]