LC-MS/MS定量检测人血浆中多黏菌素E及其临床应用

    Quantitative Determination of Polymyxin E in Human Plasma by LC-MS/MS and Its Clinical Application

    • 摘要:
      目的 建立一种简便、低成本、快速的高效液相色谱串联质谱法(LC-MS/MS)以检测人血浆中多黏菌素E的浓度。
      方法 以100 mmol·L−1硫酸锌水溶液和乙腈为沉淀剂,多黏菌素B1为内标。采用Thermo Accucore aQ(50 mm×2.1 mm,2.6 μm)色谱柱进行分离,流动相采用甲醇-含0.1%甲酸的水,梯度洗脱方式,柱温为50 ℃。在电喷雾正离子模式下进行质谱检测,并使用多反应监测模式进行定量。定量离子对分别为m/z 585.512→535.256(多黏菌素E1),m/z 578.462→528.489(多黏菌素E2)和m/z 602.675→100.971(内标多黏菌素B1)。
      结果 建立了一种基于LC-MS/MS测定多黏菌素E的方法。多黏菌素E1和E2分别在0.058~17.407 μg·mL−1和0.100~30.000 μg·mL−1线性关系良好。日内和日间的精密度、基质效应等验证均符合要求。
      结论 本方法前处理步骤简单快捷、经济成本低,且经过验证,同时适用于临床使用多黏菌素E甲磺酸盐和硫酸多黏菌素E患者体内多黏菌素E血药浓度监测和药动学等相关研究。

       

      Abstract:
      OBJECTIVE To establish a simple, low-cost and rapid high performance liquid chromatography tandem mass spectrometry(LC-MS/MS) method for the detection of polymyxin E concentration in plasma.
      METHODS Zinc sulfate solution and acetonitrile were used as precipitating agent, polymyxin B1 was used as internal standard. A Thermo Accucore aQ(50 mm×2.1 mm, 2.6 μm) column was used for separation. The mobile phase consisted of methanol-water containing 0.1% formic acid. Gradient elution mode was used, and the column temperature was set at 50 °C. The mass spectrometry was performed in the electrospray positive ion mode, and the multi-reaction monitoring mode was used for quantitative determination. The ion transitions of polymyxin E1, polymyxin E2, polymyxin B1 as internal standard were m/z 585.512→535.256, m/z 578.462→528.489 and m/z 602.675→100.971.
      RESULTS A method based on LC-MS/MS was developed for the determination of polymyxin E. The linear relationship between concentration of polymyxin E1 and polymyxin E2 was found to be well within the ranges of 0.058–17.407 µg·mL−1 and 0.100–30.000 µg·mL−1, respectively. The intra day and inter day precision and the verification of matrix effects were both met the established criteria.
      CONCLUSION The pre-treatment procedure of this method is simple and economical, and has been validated. It is also suitable for monitoring the plasma concentration of polymyxin E and pharmacokinetic studies in patients with polymyxin E mesylate and polymyxin E sulfate.

       

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