A fast and sensitive LC-MS/MS method for the quantification of fosfomycin in human urine and plasma using one sample preparation method and HILIC chromatography
A fast and sensitive LC-MS/MS method for the quantification of fosfomycin in human urine and plasma using one sample preparation method and HILIC chromatography
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DOI：
10.1016/j.jchromb.2017.07.036
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发表时间：
2017-09-01
影响因子：
3
通讯作者：
Koch, Birgit C. P.
中科院分区：
文献类型：
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作者：
Wijma, Rixt A.;Bahmany, Sorna;Koch, Birgit C. P.
磷霉素是一种古老的抗生素，由于抗生素耐药性的出现和多重耐药感染的发生率不断上升，因此越来越多地使用磷霉素。令人惊讶的是，人们对其药代动力学(PK)和药效学(PD)知之甚少。尿液和血浆中磷霉素的定量检测有助于深入了解磷霉素的PK/PD特性，这对于优化治疗方案和预防耐药的出现至关重要。因此，需要一种分析方法来定量测定尿液和血浆中的磷霉素。建立了一种快速灵敏的串联质谱法，并结合HILIC色谱柱，建立了尿样和血浆中磷霉素的通用样品前处理方法，并根据FDA指南进行了验证。通用制样方法只需要上百亩L样品，加入内标磷霉素-13C(3)苯甲胺，超滤步骤即可。该方法适用于包括临床相关浓度范围的0.75-375 mg/L(两种基质的R-2均为0.9998)，该浓度范围是基于临床环境中可能的病原体的敏感性。对所有质控水平的尿液和血浆的验证结果，精密度分别为2.1%和3.2%，日内精密度为1.5%和1.7%，日间精密度分别为5.0%和3.8%。无基质影响，尿液和血浆总回收率高(102.5%和99.4%)。制备的样品在4℃和15℃下至少稳定72小时，并且在-80℃下储存的样品至少稳定6个月。确定了选择性和灵敏度，没有观察到残留。该方法已成功地应用于健康志愿者和患者的两个药代动力学研究。
Fosfomycin is an old antibiotic that is increasingly prescribed because of emergence of the antibiotic resistance and the growing incidence of multi-drug resistant infections. Surprisingly, little is known about its pharmacokinetics (PK) and the pharmacodynamics (PD). Quantification of fosfomycin in both urine and plasma provides insight into the PK/PD characteristics of fosfomycin, which is crucial for the optimization of the therapy and the prevention of the emergence of resistance. An analytical method is therefore needed for the quantification of fosfomycin in both urine and plasma. A fast and sensitive tandem mass spectrometry method in combination with HILIC chromatography for the quantification of fosfomycin with a universal sample preparation method for urine and plasma was developed and validated according to FDA guidelines. The universal sample preparation method only requires 100 mu L of a sample, the addition of the internal standard fosfomycin-13C(3) benzylamine and an ultrafiltration step. The method is applicable for the concentration range of 0.75-375 mg/L (R-2 of 0.9998 in both matrices) encompassing the clinically relevant concentration range based on the susceptibility of possible (uro)pathogens in the clinical setting. The validation results for urine and plasma for all QC levels, were < 2.1% and < 3.2% for accuracy, < 1.5% and < 1.7% for within day precision and < 5.0% and < 3.8% for between day precision, respectively. No matrix effects were encountered and the total recovery in urine and plasma was high (102.5% and 99.4%). Prepared samples were stable at 4 degrees C and 15 degrees C for at least 72 h and stored samples at -80 degrees C were stable for at least 6 months. Selectivity and sensitivity were confirmed and no carry-over was observed. The method was successfully applied in two pharmacokinetic studies in healthy volunteers and patients respectively.