超高效液相色谱—质谱联用法同时检测结核患者体内6种抗结核药的浓度

发布时间：2018-08-10 19:05

【摘要】：结核病是一种古老的传染病。近年来,在世界范围内结核发病率呈上升趋势,给人类健康带来了巨大的威胁。目前我们常用的一线抗结核药物有很多种,由于各种药物对结核杆菌的杀伤力不同,所以在临床上通常联合使用几种药物,这样才能更有效地杀死结核杆菌。但各种药物的联合应用会带来许多不良反应和结核菌的耐药性,这使得临床医生很难把握药物的使用剂量。如何应用恰当的剂量达到最好的治疗效果,将不良反应和结核菌的耐药性降至最低,对药师和临床医师都是一个挑战。目前临床上通常应用HPLC-ELSD法和反相高效液相色谱法来测定抗结核药的血药浓度,对临床医生调整抗菌药物剂量有一定帮助。但此类方法最多只能同时检测到四种抗结核药物,而且成本较高,分析时间长,敏感性不够,其临床应用价值有限。为了解决这一难题,本研究采用UPLC-MS/MS法对患者抗结核药物血药浓度的进行测定,此方法成本较低、分析时间短,敏感性高,并可同时检测人血浆中乙胺丁醇(EMB)、异烟肼(INH)、吡嗪酰胺(PZA)、利福平、利福喷丁、利福布汀6种抗结核药物的血药浓度,有望为临床医生调整抗结核药物剂量提供更加可靠的依据。目的:评价UPLC-MS/MS法对于检测人类血浆中抗结核药血药浓度的价值,为临床医生调整抗结核药物剂量提供可靠的依据。方法:利用蛋白沉淀法将被分析物与生物基质分离,Waters ACQUITY UPLC HSS T3(1.8μm,2.1×100 mm)色谱柱进行色谱分离,采用乙腈-15 m M甲酸铵-0.05%甲酸梯度洗脱,流速为0.2 m L/min,柱温:40℃,进样量:5μL。并采用ESI源正离子模式,多反应监测进行测定。结果:乙胺丁醇、异烟肼、吡嗪酰胺、利福平、利福喷丁、利福布汀分别在0.05~5μg/m L、0.1-10μg/m L、0.2-20μg/m L、0.05-10μg/m L、0.05-10μg/m L、0.05-5μg/m L范围内线性关系良好,相关系数均大于0.991。除吡嗪酰胺在低浓度点的日间精密度小于15.4%,其它药物低、中、高3个浓度的日内和日间精密度RSD均小于15%,准确度为88.1%~109.1%,准确度良好。结论:UPLC-MS/MS法操作简单、快速,灵敏度高,线性范围宽,可同时对6种抗结核药物进行检测,一步完成,减少了分析成本,可为临床医生调整抗结核药物剂量提供重要依据,对结核病患者的合理用药具有重要意义。
[Abstract]:Tuberculosis is an ancient infectious disease. In recent years, the incidence of tuberculosis in the world is on the rise, which poses a great threat to human health. At present, there are many kinds of first-line antituberculotic drugs. Because of the different killing power of various drugs to tuberculosis bacilli, we usually use several drugs together in clinic, so we can kill tuberculosis bacillus more effectively. However, combined use of various drugs will lead to many adverse reactions and drug resistance of tuberculous bacteria, which makes it difficult for clinicians to grasp the dosage of drugs used. It is a challenge for pharmacists and clinicians to use the appropriate dose to achieve the best therapeutic effect and to minimize adverse reactions and drug resistance of tuberculous bacteria. At present, HPLC-ELSD method and RP-HPLC are usually used to determine the concentration of antituberculotic drugs in blood, which is helpful for clinicians to adjust the dosage of antimicrobial agents. However, this method can only detect four antituberculous drugs at the same time, and the cost is high, the analysis time is long, the sensitivity is not enough, and its clinical application value is limited. In order to solve this problem, UPLC-MS/MS method was used to determine the blood concentration of antituberculotic drugs in patients. This method has the advantages of low cost, short analysis time and high sensitivity. The plasma concentrations of ethambutanol (EMB), isoniazid (INH), pyrazinamide (PZA), rifampicin rifambutin and rifambutin in human plasma may provide a more reliable basis for clinicians to adjust the dosage of antituberculosis drugs. Objective: to evaluate the value of UPLC-MS/MS method in detecting the concentration of antituberculous drugs in human plasma and to provide reliable basis for clinicians to adjust the dosage of antituberculous drugs. Methods: Waters ACQUITY UPLC HSS T3 (1.8 渭 m ~ (2.1 脳 100mm) column was separated by protein precipitation method. Acetonitrile -15 mm ammonium formate was eluted with gradient of 0.05% formic acid, flow rate was 0.2 mL / min, column temperature was 40 鈩，

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