中国防痨杂志2024，Vol.46Issue(5)：549-556,8.DOI:10.19982/j.issn.1000-6621.20240059
高效液相色谱-质谱联用法测定血浆中异烟肼及其代谢物浓度的研究
Study on the determination of isoniazid and its metabolites concentration in plasma by high-performance liquid chromatography-mass spectrometry
摘要
Abstract
Objective:To establish a method for simultaneous determination of isoniazid and its metabolites—acetylhydrazine and hydrazine in the plasma of pulmonary tuberculosis patients using high-performance liquid chromatography-mass spectrometry(HPLC-MS/MS).Methods:Plasma samples were collected from 104 confirmed pulmonary tuberculosis patients(all receiving standardized anti-tuberculosis treatment,taking isoniazid at a dose of 300-500 mg once a day,and venous blood was collected 2 hours after medication)from Beijing Chest Hospital,Capital Medical University.After protein precipitation in methanol,plasma samples were subjected to derivatization using isoniazid-D4 as the internal standard and p-methylbenzaldehyde as the derivatization reagent.The analyte was separated using an InfinityLab Poroshell 120 HILIC-Z column(100 mm×2.1 mm,2.7 μm).The binary elution system(A phase was an aqueous solution containing 0.1％formic acid and 5 mmol ammonium formate;B phase was acetonitrile)was used for gradient elution,with a column temperature of 30 ℃ and a flow rate of 0.4 ml/min);electro spray ionization positive ion mode detection and multi-reaction monitoring mode scanning was adopted,and the operation time was 8 min.Results:This determination method was not affected by endogenous substances in plasma.Isoniazid,acetylhydrazine,and hydrazine had good linearity in the range of 50-6000,25-3000,and 1-120 ng/ml,respectively(determination coefficients(R2)＞0.99);intra-day and inter-day precision＜15％;absolute accuracy value＜15％;the extraction recovery rates were all above 85％;the matrix effect ranges from 85％to 115％;the stability was good.The method was applied to detect the concentrations of isoniazid,acetylhydrazine,and hydrazine in 104 patients with pulmonary tuberculosis,which were(2402.33± 1248.57)ng/ml,(1902.51±596.82)ng/ml,and(26.50±17.13)ng/ml,respectively.Conclusion:The HPLC-MS/MS method established in this study was easy to operate,rapid,specific,and sensitive.It could be applied to monitor the therapeutic drug concentration of isoniazid and its metabolites(acetylhydrazine and hydrazine)in patients with pulmonary tuberculosis.关键词
结核,肺/异烟肼/液相色谱-质谱法/药物监测Key words
Tuberculosis,pulmonary/Isoniazid/Liquid chromatography-mass spectrometry/Drug monitoring分类
医药卫生引用本文复制引用
葛菲,朱慧,程凯,陆宇,徐建..高效液相色谱-质谱联用法测定血浆中异烟肼及其代谢物浓度的研究[J].中国防痨杂志,2024,46(5):549-556,8.基金项目
高层次公共卫生技术人才建设项目(G2024-2-006) Talent Development Plan for High-level Public Health Technical Personnel Project(G2024-2-006) （G2024-2-006）
