多黏菌素E血药浓度LC-MS/MS测定方法的建立及其在MDRGN感染患儿中的临床应用

章梦; 聂晶; 王君燕; 缪静

doi:10.13748/j.cnki.issn1007-7693.20252289

多黏菌素E血药浓度LC-MS/MS测定方法的建立及其在MDRGN感染患儿中的临床应用

Development and Validation of an LC-MS/MS Assay for the Quantification of Polymyxin E in Human Serum and its Clinical Application in Children with MDRGN Infections

摘要

摘要:
目的建立一种液相色谱串联质谱(LC-MS/MS)方法，用多黏菌素B1作为内标，测定多药耐药革兰氏阴性菌(multidrug-resistant Gram-negative，MDRGN)感染患儿血清中多黏菌素E1和多黏菌素E2的含量，并对MDRGN患儿多黏菌素E药动学进行研究。
方法采用Waters ACQUITY UPLC^® BEH C₁₈(2.1 mm×50 mm，1.7 μm)色谱柱对多黏菌素E的主要成分进行色谱分离，建立时间程序为5.0 min的LC-MS/MS，以含2%甲酸的乙腈为沉淀剂进行蛋白沉淀的样品前处理，用正离子扫描以及多反应监测模式进行分析物测定。
结果血清中多黏菌素E1在0.11~4.14 μg·mL⁻¹内线性关系良好(r²>0.9971)，多黏菌素E2在0.15~5.60 μg·mL⁻¹内线性关系良好(r²>0.9902)。批内和批间精密度和准确度的RSD≤15%。测定1例使用多黏菌素E MDRGN患儿的体内暴露量， MDRGN患儿的多黏菌素E血药浓度药动学表明，首次负荷剂量给药(4 mg·kg⁻¹)时，静脉滴注开始后0.5 h，多黏菌素甲磺酸钠(colistimethate sodium，CMS)的浓度达到峰值；静脉滴注开始后4 h，CMS在体内水解吸收，多黏菌素E达到峰浓度。第2剂开始改为维持剂量(2.5 mg·kg⁻¹ CMS，q12 h)，连续治疗10 d，患儿在第3天开始，多黏菌素E的谷浓度趋于稳态。根据两点法估算AUC_{0-24 h}为(41.40±1.25)mg·h·L⁻¹。
结论本研究建立了一种稳健的测定多黏菌素E总含量的方法。该方法快速、简单、经济，适合应用于临床MDRGN患儿多黏菌素E的药物浓度监测研究。该方法是临床医生精确监测感染MDRGN患儿中多黏菌素E治疗的有价值的工具。

Abstract:
OBJECTIVE To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS) method for quantifying serum concentrations of polymyxin E1 and polymyxin E2 in pediatric patients with multidrug-resistant Gram-negative(MDRGN) infections, using polymyxin B1 as an internal standard, and to characterize the pharmacokinetics of polymyxin E in this population.
METHODS The Waters ACQUITY UPLC^® BEH C₁₈(2.1 mm×50 mm, 1.7 μm) column was used to separate the main components of polymyxin E, and an LC-MS/MS with a time program of 5.0 min was established. Acetonitrile(containing 2% formic acid) was used as precipitator for sample pretreatment of protein precipitation. Positive ion scanning and multiple reaction monitoring mode were used for analyte determination.
RESULTS A linear response was observed for polymyxin E1 in serum concentrations ranging from 0.11 to 4.14 μg·mL⁻¹ (r²>0.9971) and for polymyxin E2 from 0.15 to 5.60 μg·mL⁻¹ (r²>0.9902). The intra- and inter-assay precision and accuracy(RSD) were all within 15%. Application of this validated method to characterize the pharmacokinetics of polymyxin E in pediatric patients with MDRGN infections revealed that following an initial loading dose(4 mg·kg⁻¹), the prodrug colistimethate sodium(CMS) rapidly attained its maximum plasma concentration(C_max) at 0.5 h after the initiation of intravenous infusion. The active metabolite, polymyxin E reached its C_max at 4 h post-dose, consistent with the known hydrolysis and conversion process of CMS in vivo. The maintenance dose(2.5 mg·kg⁻¹ CMS, q12 h) was initiated starting from the second dose and continued for 10 d. The trough concentration of polymyxin E in the pediatric patient began to approach steady state from day 3 onward. The area under the concentration-time curve from 0 to 24 hours(AUC_0–24h), estimated using a limited sampling strategy, was (41.40±1.25)mg·h·L⁻¹.
CONCLUSION In this study, a quantitative assay for polymyxin E was established employing a Waters ACQUITY UPLC I-Class/Xevo TQD IVD LC-MS/MS system. The developed method is rapid, straightforward, and cost-effective, making it well-suited for therapeutic drug monitoring of polymyxin E in pediatric patients with MDRGN infections. This approach serves as a valuable tool for clinicians to precisely monitor polymyxin E therapy in children with MDRGN infections.

HTML全文

参考文献(23)

施引文献

资源附件(0)