Chhonker Yashpal S, Kanvinde Shrey, Ahmad Rizwan, Singh Amar B, Oupický David, Murry Daryl J
Clinical Pharmacology Laboratory, Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Center for Drug Delivery and Nanomedicine, Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Metabolites. 2021 Feb 12;11(2):106. doi: 10.3390/metabo11020106.
Eicosanoids are key mediators and regulators of inflammation and oxidative stress that are often used as biomarkers for severity and therapeutic responses in various diseases. We here report a highly sensitive LC-MS/MS method for the simultaneous quantification of at least 66 key eicosanoids in a widely used murine model of colitis. Chromatographic separation was achieved with Shim-Pack XR-ODSIII, 150 × 2.00 mm, 2.2 µm. The mobile phase was operated in gradient conditions and consisted of acetonitrile and 0.1% acetic acid in water with a total flow of 0.37 mL/min. This method is sensitive, with a limit of quantification ranging from 0.01 to 1 ng/mL for the various analytes, has a large dynamic range (200 ng/mL), and a total run time of 25 min. The inter- and intraday accuracy (85-115%), precision (≥85%), and recovery (40-90%) met the acceptance criteria per the US Food and Drug Administration guidelines. This method was successfully applied to evaluate eicosanoid metabolites in mice subjected to colitis versus untreated, healthy control mice. In summary, we developed a highly sensitive and fast LC-MS/MS method that can be used to identify biomarkers for inflammation and potentially help in prognosis of the disease in inflammatory bowel disease (IBD) patients, including the response to therapy.
类二十烷酸是炎症和氧化应激的关键介质和调节因子，常被用作各种疾病严重程度和治疗反应的生物标志物。我们在此报告一种高灵敏度的液相色谱-串联质谱（LC-MS/MS）方法，用于在一种广泛使用的小鼠结肠炎模型中同时定量至少66种关键类二十烷酸。使用Shim-Pack XR-ODSIII（150×2.00 mm，2.2 µm）实现色谱分离。流动相在梯度条件下运行，由乙腈和0.1%乙酸水溶液组成，总流速为0.37 mL/min。该方法灵敏度高，各种分析物的定量限为0.01至1 ng/mL，动态范围大（200 ng/mL），总运行时间为25分钟。根据美国食品药品监督管理局的指南，日间和日内准确度（85-115%）、精密度（≥85%）和回收率（40-90%）均符合验收标准。该方法成功应用于评估患结肠炎小鼠与未治疗的健康对照小鼠体内类二十烷酸代谢物。总之，我们开发了一种高灵敏度且快速的LC-MS/MS方法，可用于识别炎症生物标志物，并可能有助于炎症性肠病（IBD）患者疾病的预后评估，包括对治疗的反应。
