高效液相色谱-串联质谱法测定人脑脊液中一线抗结核药浓度  

Concentration Determination of First-Line Anti-Tuberculosis Drugs in Human Cerebrospinal Fluid by High Performance Liquid Chromatography-Mass Spectrometry/Mass Spectrometry

作　　者：西娜 赵冠人 陈明 张策 王雅薇 吴首蓉 XI Na;ZHAO Guanren;CHEN Ming;ZHANG Ce;WANG Yawei;WU Shourong(Dept.of Pharmacy,Eighth Medical Center of China PLA General Hospital,Beijing 100091,China)

机构地区：[1]中国人民解放军总医院第八医学中心药剂科

出　　处：《中国医院用药评价与分析》2020年第1期69-72,共4页Evaluation and Analysis of Drug-use in Hospitals of China

基　　金：解放军总医院第八医学中心面上课题(No.2017MS-012)

摘　　要：目的:建立同时测定人脑脊液中异烟肼、利福平、吡嗪酰胺及乙胺丁醇等4种抗结核药浓度的高效液相色谱-串联质谱(high performance liquid chromatography-mass spectrometry/mass spectrometry,HPLC-MS/MS)方法,实现对结核性脑膜炎患者脑脊液中抗结核药浓度的准确测定,为临床用药提供依据。方法:色谱柱为Agilent Poroshell 120SB C18柱(4. 6 mm×50 mm,2. 7μm),甲醇-0. 2%醋酸溶液为流动相(梯度洗脱),采用电喷雾离子源,以正离子多反应监测方式进行检测。以对乙酰氨基酚为内标,脑脊液经甲醇沉淀蛋白处理后检测。异烟肼m/z 138. 0→121. 0,利福平m/z 823. 2→791. 2,吡嗪酰胺m/z 124. 1→79. 0,乙胺丁醇m/z205. 2→116. 1,对乙酰氨基酚m/z 152. 0→110. 0。分析时间为12 min。结果:脑脊液中内源性物质对测定无干扰,基质效应小。异烟肼、利福平、吡嗪酰胺及乙胺丁醇的线性范围分别为0. 196~9. 8、0. 6~30. 0、1. 0~50. 0及0. 16~8. 0μg/ml,线性范围内线性关系良好。本方法对各药物日内、日间精密度的RSD均<10%,提取回收率高。结论:本方法稳定性好,特异性强,测定结果准确,适合临床脑脊液样本的高通量分析。OBJECTIVE: To establish the method for concentration determination of isoniazid, rifampicin,pyrazinamide and ethambutol in human cerebrospinal fluid by high performance liquid chromatography-mass spectrometry/mass spectrometry( HPLC-MS/MS),to achieve the accurate determination of anti-tuberculosis drugs in cerebrospinal fluid of patients with tuberculous meningitis,so as to provide evidence for clinical medication guidance.METHODS: The supernatant uid was separated on the Agilent Poroshell 120 SB C18 column( 4. 6 mm × 50 mm,2. 7 μm),methanol-0. 2% acetic acid solution was mobile phase( gradient elution). Electrospray ionization source was applied and operated in the positive multiple reaction monitoring( MRM) mode. Acetaminophen was used as internal standard. The proteins of cerebrospinal fluid was precipitated with methanol. The transition of m/z was138. 0→121. 0 for isoniazid,823. 2 → 791. 2 for rifampicin,124. 1 → 79. 0 for pyrazinamide,205. 2 → 116. 1 for ethambutol,and 152. 0 → 110. 0 for acetaminophen. Each analysis was completed within 12 minutes. RESULTS:Endogenous substances in cerebrospinal fluid had no interference with the determination,and the matrix effect was small. The linear ranges of isoniazid,rifampicin,pyrazinamide and ethambutol were 0. 196-9. 8,0. 6-30. 0,1. 0-50. 0 and 0. 16-8. 0 μg/ml,respectively,and the linear relationships were good. The intra-day and inter-day precision RSD of this method for each drug were < 10%,and the extraction recovery rate was relatively high. CONCLUSIONS: The method is stable,specific and accurate,and proved to be suitable for the concentration determination of isoniazid,rifampicin,pyrazinamide and ethambutol in human cerebrospinal fluid from clinical samples.

关 键 词：结核性脑膜炎 人脑脊液 抗结核药物浓度 测定方法 

分 类 号：R927.2[医药卫生—药学]