UPLC-MS/MS同时测定结核患者体内异烟肼、利福平的浓度
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摘要
目的建立超高效液相色谱-串联质谱法(UPLC-MS/MS)检测结核患者体内异烟肼、利福平的浓度。方法血液经过蛋白沉淀,以对乙酰氨基酚为内标采用Thermo Syncronis C_(18)柱(2.1 mm×100 mm,1.7μm)色谱分离,以甲醇和醋酸水溶液(0.1%)梯度洗脱进行UPLC-MS/MS检测。结果异烟肼在0.05～5μg·mL~(-1)内,利福平在0.01～10μg·mL~(-1)内响应值与质量浓度呈良好线性关系,相关系数均大于0.999 0,加标回收率分别为100.5%～102.5%和92.0%～96.4%,检出限分别为0.05和0.01μg·mL~(-1),相对标准偏差(RSD)分别为0.5%～4.3%和1.1%～3.9%。结论采用内标法建立了可同时定量检测INH和RFP的检测方法,该方法特异性好、灵敏度高、重现性和准确度可靠,可用于检测结核患者血液中INH和RFP的浓度。
OBJECTIVE To establish an UPLC-MS/MS method for the determination of isoniazid and rifampin in tuberculosis patients. METHODS The blood protein was precipitated by methanol after adding acetaminophen as internal standard,chromatography separation was performed on Thermo Syncronis C_(18) column( 2. 1 mm × 100 mm,1. 7 μm),gradient elution was conducted using methanol and acetic acid aqueous solution( 0. 1%) as mobile phase,and MS/MS was used for detection. RESULTS The correlation coefficients were greater than 0. 999 0 for isoniazid in the range of 0. 05-5 μg·mL~(-1) and rifampin in the range of 0. 01~(-1)0 μg·mL~(-1),the recoveries were 100. 5%~(-1)02. 5% and 92. 0%-96. 4%,the limits of detection were 0. 05 and 0. 01 μg·mL~(-1),and the relative standard deviations( RSDs) were 0. 5%-4. 3% and 1. 1%-3. 9%,respectively. CONCLUSION The established internal standard method can simultaneously detect rifampin and isoniazid with good specificity,high sensitivity,reproducibility and accuracy,which can be used to determine the blood concentrations of rifampin and isoniazid in tuberculosis patients.
OBJECTIVE To establish an UPLC-MS/MS method for the determination of isoniazid and rifampin in tuberculosis patients. METHODS The blood protein was precipitated by methanol after adding acetaminophen as internal standard,chromatography separation was performed on Thermo Syncronis C_(18) column( 2. 1 mm × 100 mm,1. 7 μm),gradient elution was conducted using methanol and acetic acid aqueous solution( 0. 1%) as mobile phase,and MS/MS was used for detection. RESULTS The correlation coefficients were greater than 0. 999 0 for isoniazid in the range of 0. 05-5 μg·mL~(-1) and rifampin in the range of 0. 01~(-1)0 μg·mL~(-1),the recoveries were 100. 5%~(-1)02. 5% and 92. 0%-96. 4%,the limits of detection were 0. 05 and 0. 01 μg·mL~(-1),and the relative standard deviations( RSDs) were 0. 5%-4. 3% and 1. 1%-3. 9%,respectively. CONCLUSION The established internal standard method can simultaneously detect rifampin and isoniazid with good specificity,high sensitivity,reproducibility and accuracy,which can be used to determine the blood concentrations of rifampin and isoniazid in tuberculosis patients.
引文
[1]World Health Organization.Global tuberculosis report 2017[EB/OL].WHO,2017,[2017-10-10].https://www.who.int/tb/publications/global_report/zh/
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[2]CHENEVIER P,MASSIAS L,GUEYLARD D,et al.Determination of ethambutol in plasma by high-performance liquid chromatography after precolumn derivatization[J].J Chromatogr B,1 9 9 8,7 0 8:Doi.1 0.1 0 1 6/s0 3 7 8-4 3 4 7(9 7)0 0 6 4 1-5.
[3]GUO T,YAN M,SONG H T,et al.HPLC Determination of active components in Yi'an Biqin Lifu Yiyan tablests[J].Chin J Pharm Anal(药物分析杂志),2007,27(4):528-531.
[4]QI Y,YANG Q Y,WU S.Ion pair RP-HPLC Determination of ethambutol hydrochloride in the combined preparation of antituberculotic[J].Chin J Pharm Anal(药物分析杂志),2009,29(4):645-647.
[5]LIU X Q,WU S P,LI X,et al.HPLC-ELSD Determination of content and dissolution of ethambutolhydrochloride tablets[J].Chin J Pharm Anal(药物分析杂志),2008,28(5):804-806.
[6]WU L,SU M X,DI B,et al.Concentration determination of ethambutol in human plasma by LC-MS/MS[J].Chin J New Drugs Clin Rem(中国新药与临床杂志),2009,28(11):852-855.
[7]ZHAO G R,PENG M L,SHEN J,et al.Simultaneous determination of isoniazid,rifampicin,ethambutol,pyrazinamide and levofloxacin in human plasma by HPLC-MS/MS[J].Chin J Drug Appl Monitor(中国药物应用与监测),2015,12(1):16-19.
[8]AN J,DONG Z J,ZHANG X D,et al.Simultaneous determination of six antituberculosis drugs in human plasma by UPLC-MS/MS[J].Chin J Pharm Anal(药物分析杂志),2016,36(2):208-216.