Shou Dan, Dong Yu, Shen LiFeng, Wu RenJie, Zhang Yang, Zhang Chun, Zhu Yan
Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou 310007, China
Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou 310007, China.
J Chromatogr Sci. 2014 Jul;52(6):501-7. doi: 10.1093/chromsci/bmt069. Epub 2013 Jun 5.
Tobramycin and vancomycin are the most commonly used antibiotics for the treatment of osteomyelitis. A sensitive and rapid method was developed for the analysis of tobramycin and vancomycin in human drainage tissue fluid. The procedure involved a simple liquid-liquid extraction of tobramycin, vancomycin and atenolol (internal standard) and separation by ultra-high performance liquid chromatography on an Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 µm) with a mobile phase consisting of 0.1% (v/v) formic acid water solution and 0.1% (v/v) formic acid acetonitrile solution at a flow rate of 0.3 mL/min. Detection was performed by positive ion electrospray ionization in multiple reaction monitoring mode (m/z 468 → 163 transitions for tobramycin; m/z 725 → 144 for vancomycin; m/z 267 → 74 for the internal standard). The retention times of tobramycin, vancomycin, and the internal standard were 0.68, 3.62 and 3.03 min, respectively. The total analysis time was less than 10 min. Excellent linear relationships (correlation coefficient > 0.99) were demonstrated between the area under the peak ratios of tobramycin and vancomycin to the internal standard in the drainage tissue fluid, and the concentration ranges were 1.25-100.00 mg/L and 0.50-150.00 mg/L for tobramycin and vancomycin, respectively. The intra-day and inter-day accuracy and precision (coefficient of variation) acceptance criteria for each quality control was ≤ 7.8% and the mean accuracy values were < 5.0% for tobramycin and < 4.0% for vancomycin. All experiments suggested the high-throughput potential of the proposed method. The method was successfully applied to investigate local delivery of tobramycin and vancomycin in four calcaneal osteomyelitis patients who had accepted drug-loaded artificial bone implantation.
妥布霉素和万古霉素是治疗骨髓炎最常用的抗生素。开发了一种灵敏快速的方法用于分析人体引流组织液中的妥布霉素和万古霉素。该方法包括对妥布霉素、万古霉素和阿替洛尔（内标）进行简单的液液萃取，并在Acquity UPLC BEH C18柱（2.1×100 mm，1.7 µm）上通过超高效液相色谱分离，流动相由0.1%（v/v）甲酸水溶液和0.1%（v/v）甲酸乙腈溶液组成，流速为0.3 mL/min。采用正离子电喷雾电离在多反应监测模式下进行检测（妥布霉素的m/z 468→163跃迁；万古霉素的m/z 725→144；内标的m/z 267→74）。妥布霉素、万古霉素和内标的保留时间分别为0.68、3.62和3.03分钟。总分析时间少于10分钟。引流组织液中妥布霉素和万古霉素与内标峰面积比的峰面积与浓度范围之间呈现出良好的线性关系（相关系数>0.99），妥布霉素和万古霉素的浓度范围分别为1.25 - 100.00 mg/L和0.50 - 150.00 mg/L。每个质量控制的日内和日间准确度和精密度（变异系数）接受标准≤7.8%，妥布霉素的平均准确度值<5.0%，万古霉素的平均准确度值<4.0%。所有实验表明该方法具有高通量潜力。该方法成功应用于研究4例接受载药人工骨植入的跟骨骨髓炎患者中妥布霉素和万古霉素的局部递送情况。
