An ID-HPLC-MS/MS based candidate reference measurement procedure for the quantification of imipenem, meropenem and ertapenem in human plasma

IF 2.5 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2026-02-01 Epub Date: 2025-12-27 DOI:10.1016/j.jchromb.2025.124908

Jing Lin , Qingqing Pan , Weihua Wang , Shuangshuang Chen , Guangliang Wu , Min Shen , Huoyan Ji

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引用次数: 0

Abstract

Background

Carbapenems require precise plasma levels for optimal therapy, yet no LC-MS/MS method offered SI-traceability or reference-grade performance. In this study, a candidate reference measurement procedure (RMP) using isotope dilution high-performance liquid chromatography-tandem mass spectrometry (ID-HPLC-MS/MS) was developed and validated for accurate quantification of imipenem (IPM), meropenem (MEM) and ertapenem (ETP) in human plasma.

Methods

To ensure traceability to SI units, the absolute content of the IPM, MEM and ETP primary reference materials (PRMs) were determined by quantitative nuclear magnetic resonance (qNMR) spectroscopy. Samples were prepared by protein precipitation followed by dilution. Method validation was performed according to guidances from the Clinical and Laboratory Standards Institute (CLSI), including selectivity/specificity, matrix effect, carryover, linearity, limit of quantification (LOQ) and limit of detection (LOD), trueness and precision, and stability. Measurement uncertainty was evaluated according to the Guide to the Expression of Uncertainty in Measurement (GUM). Moreover, the candidate RMP was compared between two independent laboratories.

Results

The method showed high selectivity and stability. Linear ranges were 0.208 to 52.087 μg/mL (IPM, r = 0.9997), 0.412 to 103.427 μg/mL (MEM, r = 0.9999) and 0.083 to 20.791 μg/mL (ETP, r = 0.9999). Mean recoveries were 100.05 to 104.80 %, 92.34 to 94.77 % and 95.55 to 100.04 %, respectively. The method demonstrated high sensitivity, with limits of LOQ of 0.103, 0.103 and 0.021 μg/mL and corresponding LOD of 0.0077, 0.0018 and 0.0006 μg/mL for IPM, MEM and ETP, respectively. No obvious matrix effect and carryover were observed. The expanded uncertainties (k = 2) were ≤ 6.12 %.

Conclusion

The qNMR-based candidate RMP provides SI-traceable, accurate simultaneous quantification of IPM, MEM and ETP, enabling standardization of clinical assays and reliable patient-sample measurement.

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基于ID-HPLC-MS/MS的人血浆亚胺培南、美罗培南和厄他培南候选参比测定方法

背景：碳青霉烯类需要精确的血浆水平以获得最佳治疗，但LC-MS/MS方法无法提供si可追溯性或参考级性能。本研究建立了同位素稀释高效液相色谱-串联质谱（ID-HPLC-MS/MS）候选参比测量方法（RMP），并对其进行了验证，用于准确定量人血浆中亚胺培南（IPM）、美罗培南（MEM）和厄他培南（ETP）。方法：采用定量核磁共振（qNMR）法测定IPM、MEM和ETP主要标准物质（PRMs）的绝对含量，确保其可溯源至SI单位。样品是通过蛋白质沉淀和稀释制备的。根据临床与实验室标准协会（CLSI）的指南进行方法验证，包括选择性/特异性、基质效应、结转、线性、定量限（LOQ）和检出限（LOD）、真实度和精密度以及稳定性。根据《测量不确定度表达指南》（GUM）评定测量不确定度。此外，候选RMP在两个独立实验室之间进行了比较。结果：该方法选择性高，稳定性好。线性范围分别为0.208 ~ 52.087 μg/mL （IPM, r = 0.9997）、0.412 ~ 103.427 μg/mL （MEM, r = 0.9999）和0.083 ~ 20.791 μg/mL （ETP, r = 0.9999）。平均加样回收率分别为100.05 ~ 104.80%、92.34 ~ 94.77%和95.55 ~ 100.04%。方法灵敏度高，IPM、MEM和ETP的检出限分别为0.103、0.103和0.021 μg/mL，检出限分别为0.0077、0.0018和0.0006 μg/mL。未见明显基质效应和结转现象。扩展不确定度(k = 2)≤6.12%。结论：基于qnmr的候选RMP提供si可追溯，准确的同时定量IPM， MEM和ETP，使临床分析标准化和可靠的患者-样品测量成为可能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.