A simple, sensitive, and rapid liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method was developed for the simultaneous determination of arginine and its pathway-related metabolites (ornithine, proline, citrulline, glutamate, agmatine, spermidine, and spermine) in cellular extracts. Cells were lysed and cellular proteins precipitated by the addition of acetonitrile followed by ultra-sonication. Supernatants were analyzed using a Chromolith High Resolution RP-18 endcapped column (100 × 4.6 mm, 1.15 μm, 150 Å), with mobile phases of 0.1% formic acid solution and 0.1% formic acid in acetonitrile. Detection was carried out in multiple reaction monitoring (MRM) mode. Calibration curves showed linearity (r² > 0.99) for all metabolites over the calibration ranges used. The intra- and inter-day precision was less than 13.5%, and the accuracy was between 91.3 and 114.7%. The method developed in this study was successfully applied to measure arginine and its pathway-related metabolites, which are related to nitric oxide synthase/arginase pathways in mouse bone marrow-derived dendritic cells (BMDCs). The ability to simultaneously measure arginine and its pathway-related metabolites is valuable for better understanding local and systemic inflammatory processes.

开发了一种简单，灵敏，快速的液相色谱（LC）-串联质谱（MS / MS）方法，用于同时测定精氨酸及其途径相关的代谢产物（鸟氨酸，脯氨酸，瓜氨酸，谷氨酸，胍丁胺，亚精胺和精胺）在细胞提取物中。裂解细胞，并通过添加乙腈，然后进行超声波处理，沉淀出细胞蛋白。使用Chromolith高分辨率RP-18封端色谱柱（100×4.6 mm，1.15μm，150Å）分析上清液，流动相为0.1％甲酸溶液和0.1％甲酸的乙腈溶液。检测是在多反应监测（MRM）模式下进行的。校正曲线显示线性度（r ²> 0.99）超过所用校准范围内的所有代谢物。日内和日间精度均低于13.5％，且精度在91.3至114.7％之间。在这项研究中开发的方法已成功地用于测量精氨酸及其途径相关的代谢产物，这些代谢产物与小鼠骨髓源性树突状细胞（BMDC）中的一氧化氮合酶/精氨酸酶途径有关。同时测量精氨酸及其途径相关代谢产物的能力对于更好地了解局部和全身性炎症过程非常有价值。