【摘要】 目的 采用高效液相色谱一质谱/质谱联用(HPLC-MS/MS)技术检测人血浆中贝达喹啉浓度,并对其应用效果进行评价。方法 人血浆样品经乙腈沉淀蛋白等处理后检测,液相色谱采用Agilent ZORBAX SB-C18色谱柱(2.1mm×100mm, 3.5μm)为分析柱,以甲醇-5mmol/L甲酸铵水溶液(含0.1%甲酸)(85:15, V/V)为流动相,质谱使用电喷雾离子源(ESI),以正离子多反应监测(MRM)方式进行检测,分析时间为3 min。结果 本测定方法不受血浆中内源性物质干扰,贝达喹啉母离子和定量子离子的质荷比(m/z)为555. 2→58. 3,标准曲线线性范围为0.1～5.0μg/ml,方法的日内、日间精密度(相对标准偏差,RSD)均小于10%,准确率为97.8%～107%。用建立的方法检测了12例服用贝达喹啉的结核病患者,服药剂量为每日400 mg时,贝达喹啉峰浓度为(3. 17±1. 14)μg/ml,谷浓度为(1. 12±0. 64)μg/ml;服药剂量为200 mg/次,每周3次时,贝达喹啉峰浓度为(2.01±0.87)μg/ml,谷浓度为(0.65±0.30)μg/ml,48份样本血药浓度均在检测范围内(0.1～5.0μg/ml)。结论 本研究建立了抗结核新药贝达喹啉血浆中药物浓度的HPLC-MS/MS联用方法,该方法适用于临床结核病患者血浆样品中贝达喹啉浓度的快速分析。更多还原

【Abstract】 Objective To establish an high performance liquid chromatography-mass spectrometry/mass spectrometry(HPLC-MS/MS) method for the determination of bedaquiline in human plasma and to evaluate the application of the method. Methods The protein of plasma samples were precipitated with acetonitrile. HPLCMS/MS was performed using an Agilent ZORBAX SB-C18 column(2.1 mm×100 mm,3. 5 μm) as the analytical column. The mobile phase consisted of methanol-5 mmol/L ammonium formate(containing 0.1% formic acid solution)(85 : 15,v/v). Electrospray ionization source(ESI) was applied and operated in the positive multiple reaction monitoring(MRM) mode. One analysis was completed within 3 min. Results Chromatograms showed no endogenous interfering peaks with blank samples. The mass-to-charge ratio(m/z) of the parental ion and the quantitative ion of bedaquiline was 555. 2→58. 3. The linear calibration curve was obtained over the concentration on range of 0. 1-5. 0 μg/ml. The inter-and intra-day precision(RSD) were less than 10%. The accuracy were 97. 8%-107. 0%. 48 plasm samples of 12 tuberculosis patients were detected using the established method. When the dosage was 400 mg daily,the peak concentration of bedaquiline was(3. 17±1.14) μg/ml, and the trough concentration was(1. 12±0. 64) μg/ml; when the dosage was 200 mg per time,3 times a week, the peak concentration of bedaquiline was(2. 01 ± 0. 87) μg/ml, and the trough concentration was(0.65 ± 0.30) μg/ml. The concentrations of bedaquiline in the 48 samples were in the range of calibration curve(0.1-5. 0 μg/ml). Conclusion A HPLC-MS/MS method was established for the quick analysis and determination of bedaquiline(a new anti-tuberculosis drug) in plasma samples of tuberculosis patients.更多还原