LC-MS/MS定量检测人血浆多黏菌素E及其临床应用

Developing a LC-MS/MS Method to Quantify Colistin in Human Plasma and Its Application

关键词:  多黏菌素E  高效液相色谱串联质谱法  治疗药物监测

Key words:  colistin  LC-MS/MS  Therapeutic drug monitoring

摘要:

目的 多黏菌素E存在较大的个体差异,为了优化其临床疗效并减少药物暴露相关毒性,需要一种简单可靠的定量检测方法来对多黏菌素E进行药物浓度监测。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,色谱柱进行分离,流动相采用含0.5%甲酸的水-甲醇,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 建立了一种基于高效液相色谱-质谱联用技术(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 ug·mL-1和0.100-30.000 ug·mL-1线性关系良好。批内和批间的精密度、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、相关治疗窗和药动学等研究。

Abstract:

ABSTRACT: OBJECTIVE To optimize the clinical efficacy of colistin and reduce the toxicity associated with drug exposure, a simple and reliable quantitative detection method is needed to quantify the concentration of colistin. METHODS Zinc sulfate solution and acetonitrile were used as precipitating agent, polymyxin B1 was used as internal standard. The mobile phase was separated using methanol-water containing 0.5% formic acid. 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 colistin A, colistin B and IS 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 high performance liquid chromatography-mass spectrometry (LC-MS/MS) was developed for the determination of colistin. The linear relationship between concentration of colistin A and colistin B 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 precision within and between batches and the verification of matrix effects were both met the established criteria. CONCLUSION The pre-treatment procedure of this method is simple and economical. It is suitable for the study of plasma concentration and pharmacokinetics of colistin mesylate and colistin sulfate in patients.