Department of Analytical Chemistry, Faculty of Chemistry, Universitat de València, 46100 Burjassot, Spain.
Department of Physical and Analytical Chemistry, ESTCE, Universitat Jaume I, 12071 Castelló, Spain.
J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Jun 1;1225:123777. doi: 10.1016/j.jchromb.2023.123777. Epub 2023 Jun 3.
Combined prescription of the antimicrobial drugs linezolid and meropenem is a common strategy to treat multidrug-resistant nosocomial infections. We propose an innovative method to determine these two drugs in plasma and urine, based on micellar liquid chromatography. Both biological fluids were diluted in mobile phase, filtered and directly injected, without any extraction step. Using a C18 column and a mobile phase of 0.1 M sodium dodecyl sulfate - 10 % methanol, phosphate buffered at pH 3, running under isocratic mode, both antibiotics were eluted without overlapping in<15 min. Detection was by absorbance: 255 nm for linezolid and 310 nm for meropenem. The influence of sodium dodecyl sulfate and methanol concentration on retention factor was established for both drugs using an interpretative approach assisted by chemometrics. The procedure was successfully validated following the guidelines of 2018 Bioanalytical Method Validation Guidance for Industry in terms of: linearity (determination coefficients over 0.99990), calibration range (1 - 50 mg/L), instrumental and method sensitivity, trueness (bias of -10.8 to + 2.4%), precision (relative standard deviation of < 10.2%), dilution integrity, carry-over effect, robustness and stability. It should be emphasized that the method uses low volumes of toxic and volatile solvents and can be achieved in a short period. The procedure was found useful for routine analysis, as it was cost-affordable, more eco-friendly and safer than hydroorganic HPLC, easy-to-handle and highly sample-throughput. Finally, it was applied to incurred samples of patients taking this medication.
联合使用抗菌药物利奈唑胺和美罗培南是治疗多药耐药性医院感染的常用策略。我们提出了一种基于胶束液相色谱法测定这两种药物在血浆和尿液中浓度的创新方法。两种生物体液均在流动相中稀释、过滤并直接进样，无需任何提取步骤。使用 C18 柱和 0.1 M 十二烷基硫酸钠-10 %甲醇的磷酸缓冲液（pH 3）作为流动相，在等度模式下运行，两种抗生素均能在 15 分钟内洗脱且无重叠。检测波长分别为利奈唑胺的 255nm 和美罗培南的 310nm。通过化学计量学辅助的解释性方法，建立了两种药物的保留因子与十二烷基硫酸钠和甲醇浓度的关系。该方法按照 2018 年行业生物分析方法验证指南的要求成功进行了验证，包括线性（决定系数均大于 0.99990）、校准范围（1 - 50mg/L）、仪器和方法灵敏度、准确度（偏差为-10.8%至+2.4%）、精密度（相对标准偏差小于 10.2%）、稀释完整性、交叉污染效应、稳健性和稳定性。值得强调的是，该方法使用了低体积的有毒和挥发性溶剂，且能够在短时间内完成。该方法已被证明可用于常规分析，因为它比水有机 HPLC 更具成本效益、更环保、更安全、更易于操作且具有更高的样品通量。最后，我们将该方法应用于正在服用该药物的患者的实际样本中。
