用液相色谱-串联质谱技术研究禁用物质氢氯噻嗪在人尿中的代谢情况  

The Metabolic Profile and Detection Window of Hydrochlorthiazide in Human Urine

作　　者：马艳华[1] 闫宽 董颖[1] 

机构地区：[1]国家体育总局反兴奋剂中心,北京100029

出　　处：《中国运动医学杂志》2016年第1期67-70,73,共5页Chinese Journal of Sports Medicine

基　　金：国家体育总局反兴奋剂中心科研项目(2013ZR002)

摘　　要：目的：使用液相色谱-串联质谱法研究口服利尿剂氢氯噻嗪（Hydrochlorothiazide）后,尿样中药物原形、其代谢物氯噻嗪（Chlorothiazide）和降解产物4-氨基-6-氯-1,3苯二磺酰氯（4-Amino-6-chlorobenzene-1,3-disulphonamide）的代谢规律和检测窗口期。方法：选取男女各1名年龄为26岁和27岁的健康志愿者,进行氢氯噻嗪人体受试实验,收集服药前的空白尿和服药后3天内的所有尿样,以及之后14天的晨尿。尿样经同等体积的5%醋酸铅水溶液沉淀蛋白质后,上清液用多反应离子监测模式（MRM）直接进行检测。结果：氢氯噻嗪、氯噻嗪和4-氨基-6-氯-1,3苯二磺酰氯的检测限分别为2.5、1和5 ng/ml;定量限分别为10、3和15 ng/ml;3种药物在低、中和高3个浓度的日内和日间精密度均小于10%;准确度均介于88%至110%之间;平均基质效应在75%-120%范围内。对受试者的尿样进行分析后,发现氢氯噻嗪原形的检测窗口期为7天,代谢物氯噻嗪的检测窗口期为3天,降解产物ACB的检出时间为13天。结论：该检测方法对氢氯噻嗪原形、其代谢物氯噻嗪和降解产物ACB具有前处理简单、灵敏度高、准确可靠和重现性好等特点。与之前的文献相比,3种禁用物质的检测窗口期都有不同程度的延长。Objective To study the metabolic profile and detection window period of hydrochlorothiazide（HCT）and its metabolite（chlorothiazide,CT）and degradation product（4-amino-6-chloro-1,3-benzenedisulphonamide,ACB）in human urine for the purpose of doping control. Method Two volunteers including 1 male（aged 26）and 1 female（aged 27）were orally administered one tablet of hydrochlorothiazide containing 25 mg HCT after signing the statement of informed consent. Their urine samples were collected before,within 3 days and in the morning from 4th to 17 th day after the administration of hydrochlorothiazide.After precipitation of protein with 5% Pb Ac2 aqueous solution,the supernatant of the samples were injected into liquid chromatography-tandem mass spectrometry（LC-MS/MS）without sample pre-concentration. Results The limits of detection for HCT,CT and ACB were 2.5 ng/ml,1 ng/ml and 5 ng/ml,respectively. The limits of quantification of HCT,CT,and ACB were 10 ng/ml,3 ng/ml and 15 ng/ml,respectively. The intra-and interday precision,accuracy and matrix effects of the HCT,CT and ACB in low-moderate-and high-concentration were 〈10%,88%-110% and 75%-120%,respectively. The detection windows of HCT,CT and ACB were 7,3 and 13 days,respectively after the administration of hydrochlorothiazide. Conclusions This approach provided easy pre-processing of sample,high sensitivity,satisfactory accuracy and reproducibility for the detection of HCT,CT and ACB. The detection window period for HCT,CT and ACB in this study was somewhat prolonged as compared with the published literature.

关 键 词：液相色谱-串联质谱 人体受试 氢氯噻嗪 代谢曲线 兴奋剂控制 

分 类 号：R446.12[医药卫生—诊断学] R872[医药卫生—临床医学]