Objective To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method capable of simultaneously determining the concentrations of imipenem, meropenem, posaconazole and polymyxin E (colistin) in human plasma, and to preliminarily evaluate its applicability in the detection of clinical plasma samples.

Methods Plasma samples were pretreated using a protein precipitation procedure. Chromatographic separation was achieved on a reversed-phase C₁₈ column, and quantitative analysis was performed in multiple reaction monitoring (MRM) mode. According to relevant guidelines, the method was systematically evaluated in terms of selectivity, linearity, lower limit of quantification (LLOQ), precision, accuracy, extraction recovery, matrix effect, and stability. In addition, de-identified residual clinical plasma samples were analyzed to preliminarily assess the applicability of the method.

Results Imipenem, meropenem, posaconazole, colistin E₁, and colistin E₂ exhibited good linearity within their respective concentration ranges, with correlation coefficientsr not lower than 0.992 0. Except for the stability of the low-concentration imipenem sample after storage at room temperature for 3 h, which slightly exceeded the acceptable limit, the remaining stability results and the LLOQ, precision, accuracy, extraction recovery, and matrix effect met the requirements for bioanalytical method validation. Analysis of residual clinical plasma samples showed concentration ranges of 0.74-18.62 μg/mL for imipenem, 0.86-52.40 μg/mL for meropenem, 0.68-2.45 μg/mL for posaconazole, 0.56-6.82 μg/mL for colistin E₁, and 0.73-8.96 μg/mL for colistin E₂, all of which fell within the corresponding linear ranges.

Conclusion The established LC-MS/MS method can simultaneously quantify the concentrations of imipenem, meropenem, posaconazole, and polymyxin E in human plasma. The method demonstrated satisfactory analytical performance in accordance with the requirements for bioanalytical method validation and showed good feasibility in a limited number of authentic clinical plasma samples. Therefore, it provides a reliable analytical foundation for future studies on therapeutic drug monitoring (TDM) of these anti-infective agents.

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