液相色谱串联质谱法检测血浆中血管紧张素Ⅰ方法的建立  

Quantitation of angiotensin I in plasma by liquid chromatography-mass spectrometry

作　　者：檀鑫[1] 李凤娟 王雪[1] 王绿娅[1] 王媛[1] 杜杰[1] TAN Xin;LI Fengjuan;WANG Xue;WANG Luya;WANG Yuan;DU Jie(Beijing Anzhen Hospital,Capital Medical University,Key Laboratory of Remodeling-related Cardiovascular Diseases,Ministry of Education,The Collaborative Innovation Center for Cardiovascular Disorders,Beijing Institute of Heart,Lung and Blood Vessel Disease,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心血管生物研究室,100029

出　　处：《心肺血管病杂志》2021年第6期624-627,638,共5页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金青年科学基金(81600392)。

摘　　要：目的:建立液相色谱串联质谱法检测人血浆中的血管紧张素I,并探讨不同抗凝剂对检测结果的影响。方法:采集2018年7月在北京安贞医院住院的20例心血管疾病患者的EDTA抗凝血浆、枸橼酸钠抗凝血浆和血清。利用TSQ quantum ultra三重四极杆质谱仪,在正离子电喷雾条件下,质谱检测模式为选择反应监控,液相系统为Ultimate 3000 UPLC,流动相是含甲酸0.1%的水相和甲醇,流速为0.3 mL/min, 5 min梯度,用蛋白沉淀法进行样本前处理,检测样本中的血管紧张素I水平。并评价该方法的线性范围、灵敏度、精密度、提取回收率和基质效应等性能。结果:该方法的定量检出下限是0.2μg/L,线性范围为0.2~50μg/L,批内和批间准确度和精密度变异系数均<15%。提取回收率在83.7%~106.9%,基质效应在82.4%~93.0%。不同抗凝剂对血管紧张素I检测结果是有影响的,其中EDTA抗凝血浆、枸橼酸钠抗凝血浆和血清中血管紧张素I浓度分别为(299.2±113.6)、(26.65±11.10)和(51.48±12.99) ng/L,差异有统计学意义(P<0.0001)。结论:建立了检测人血浆中的血管紧张素I的液相色谱串联质谱法,该方法色谱梯度时间短,前处理简单易操作。Objective: To establish a method to detect Angiotensin I by liquid chromatography-mass spectrometry(LC-MS), and evaluate the effect of different collection tubes on the Angiotensin I result. Methods: Samples of 20 cardiovascular disease inpatients from Beijing Anzhen Hospital in July 2018 were obtained in three different collection tubes: EDTA plasma, sodium citrate plasma, and serum. The detection was carried out by selected reaction monitoring using a TSQ Quantiva tandem quadrupole mass spectrometer under positive electrospray ionization conditions. Chromatography was performed on an Ultimate 3000 UPLC system with a constant flow rate of 0.3 mL/min under chromatographic conditions of water(0.1% formic acid) and methanol, min, giving a total chromatographic analysis time of 5 min. Samples were extracted by protein precipitation. The linear range, sensitivity, precision, recovery and matrix effect of the method were evaluated. Results: The method was found to be linear over the analytical range of 0.2-50μg/L giving the limit of detection of 0.2μg/L. The coefficients of variation of intra-and inter-assay accuracies and precisions were less than 15%. The extraction recovery ranged from 83.7% to 106.9% and the matrix effect varied from 82.4% to 93.0%. The result of Angiotension I were distinct in different collection tubes and the concentrations of angiotensin I in EDTA plasma, sodium citrate plasma and serum were(299.2 ± 113.6),(26.65 ± 11.10) and(51.48 ± 12.99) ng/L, respectively, with significant differences(P < 0.0001). Conclusions: We developed a liquid chromatography-tandem mass spectrometry method for the quantitation of Angiotensin I with shorter chromatographic analysis time and easier operation.

关 键 词：血管紧张素I 液相色谱-质谱联用技术 血浆 抗凝剂 

分 类 号：R54[医药卫生—心血管疾病]