Recently, an opportunity to perform a broad ruggedness assessment of our liquid chromatography-tandem mass spectrometry (LC-MS/MS) system presented itself during the analytical planning phase of a large-scale human fecal microbiome study. The specific aim of this project was to study the microbial-mediated metabolism of a targeted set of bile acids/salts by mixed bacterial communities cultured from the feces of 12 healthy volunteers when grown in a custom growth medium and following exposure to different clinically-relevant antibiotics. The magnitude of this study offered a rare opportunity to significantly stress procedures and LC-MS/MS system components comprised in our bile acid/salt targeted metabolomics method. With this second specific aim in mind, we modified the sample analysis plan to include a series of figure-of-merit (FoM)-based tests that are commonly used in regulated bioanalytical labs to assess LC and MS system ruggedness for a specific assay – these FoM-based testing parameters were monitored continuously over the course of sample analysis and the results are presented in this report. In total, the assessment included 1206 sequential injections (180 calibration standards, 136 blank-internal standard samples, and 890 diluted medium samples) that took place over 8-days. Completion of the 8-days of non-stop sample analysis revealed no critical hardware or software failures, and the analysis of the FoM-based tests indicated no observable degradation of system performance over the number of samples and time tested. The FoM-based test metrics presented may be used as a template to assess the ruggedness of any LC-MS/MS-based targeted metabolomics workflow.

最近，在大规模人类粪便微生物组研究的分析计划阶段，出现了对我们的液相色谱-串联质谱分析（LC-MS / MS）系统进行广泛耐用性评估的机会。该项目的具体目标是研究在自定义生长培养基中生长并暴露于不同临床相关因素后，由12名健康志愿者粪便培养的混合细菌群落对目标胆汁酸/盐的微生物介导的代谢。抗生素。这项研究的规模为难于强调我们的胆汁酸/盐靶向代谢组学方法中的程序和LC-MS / MS系统组件提供了难得的机会。有了第二个具体目标，我们修改了样品分析计划，使其包括一系列基于品质因数（FoM）的测试，这些测试通常在受监管的生物分析实验室中用于评估特定测定法的LC和MS系统的耐用性–监控了这些基于FoM的测试参数在样品分析过程中连续不断地进行，结果在本报告中介绍。总共评估了8天内进行的1206次连续进样（180次校准标样，136个空白内标样品和890个稀释培养基样品）。完成为期8天的不间断样本分析后，没有发现严重的硬件或软件故障，并且基于FoM的测试分析表明，在测试的样本数量和时间范围内，系统性能没有明显下降。