Tropical Diseases Research Program, School of Veterinary Medicine, 86-3000, National University, Heredia, Costa Rica.
Centro Nacional de Ciencia y Tecnología de Alimentos, 11501-2060, Sede Rodrigo Facio, Universidad de Costa Rica, Costa Rica.
J Microbiol Methods. 2022 Mar;194:106436. doi: 10.1016/j.mimet.2022.106436. Epub 2022 Feb 25.
Herein we described a versatile liquid chromatographic method for detection and quantification of the total levels of two antimicrobials [i.e., streptomycin (STM) and doxycycline (DOX)], in mice plasma and selected tissues, with the aid of a single quadrupole as a detection method. The method included a few sample preparation steps, including freeze-drying and in situ triphasic solvent-assisted defatting, precipitation, and extraction, allowing easy and fast tissue sample processing and avoiding analyte loss. Using a murine model, we demonstrated that mass spectrometry detects simultaneously and with high specificity two of the most widespread antimicrobials used against Brucellosis. An accurate [recoveries varied from 75.23 (bone marrow) to 101.33% (liver)] and sensitive (LoD in the ng g range) method to assess STM and DOX in murine tissue, including subtherapeutic and therapeutic doses of the antimicrobials, was achieved. This validated method can be successfully used to monitor the depletion of STM and DOX in several mice tissues and plasma during metabolism after administration.
本文描述了一种通用的液相色谱法，用于检测和定量检测小鼠血浆和选定组织中两种抗生素（即链霉素（STM）和强力霉素（DOX））的总水平，该方法借助单四极杆作为检测方法。该方法包括几个样品制备步骤，包括冷冻干燥和原位三相溶剂辅助脱脂、沉淀和提取，使组织样品的处理变得简单快捷，并避免了分析物的损失。使用小鼠模型，我们证明质谱法可以同时高特异性地检测两种最广泛用于治疗布鲁氏菌病的抗生素。该方法具有较高的准确度（回收率在骨髓为 75.23%到肝脏为 101.33%之间）和灵敏度（LOD 在 ng·g 范围内），可用于评估小鼠组织中的 STM 和 DOX，包括抗生素的亚治疗和治疗剂量。该经过验证的方法可成功用于监测给药后代谢过程中 STM 和 DOX 在几种小鼠组织和血浆中的消耗情况。
