College of Pharmaceutical Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan.
Department of Pathology and Laboratory Medicine, Institute of Biomedical Science, Tokushima University Graduate School, Tokushima 770-8503, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Nov 1;1210:123432. doi: 10.1016/j.jchromb.2022.123432. Epub 2022 Aug 27.
An analytical platform is required to characterize the short-chain fatty acids (SCFAs) in a mouse model of pathological immune conditions. Therefore, liquid chromatography tandem mass spectrometry combined with 2-picolylamine derivatization and a comprehensive study of SCFAs distribution based on serum, saliva, feces, liver, and brain from a mouse model of Sjögren's syndrome (SS) is performed. The design of experiments is used to achieve efficient 2-picolylamine derivatization, and optimize the reaction conditions. Twelve SCFAs are derivatized, and separated on a reversed-phase C column. All SCFAs show high linearity (r > 0.995) and intra/inter-day accuracy values from 71.6% to 115.6% (precision < 13.7%). This method was used to determine SCFAs concentrations in the serum, saliva, feces, liver, and brain of an SS model mice, and isobutyric acid, valeric acid, isovaleric acid, and 2-methylbutyric acid in liver from SS were significantly different compared with control group. Moreover, the preliminary evaluation of propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid in saliva is conducted based on the respective SS stages and are correlated with these histological scores. This analytical platform for the widely SCFAs profiling in several tissues can be a clue for studying unclear immune pathophysiology.
需要建立一种分析平台来对病理免疫条件下的小鼠模型中的短链脂肪酸(SCFAs)进行特征分析。因此,我们使用液相色谱串联质谱法结合 2-吡啶甲胺衍生化方法,对干燥综合征(SS)小鼠模型的血清、唾液、粪便、肝脏和大脑中的 SCFA 分布进行了全面研究。实验设计用于实现高效的 2-吡啶甲胺衍生化,并优化反应条件。衍生化了 12 种 SCFAs,并在反相 C 柱上进行分离。所有 SCFAs 均表现出高线性(r>0.995)和日内/日间精密度值为 71.6%至 115.6%(<13.7%)。该方法用于测定 SS 模型小鼠血清、唾液、粪便、肝脏和大脑中的 SCFA 浓度,与对照组相比,SS 模型小鼠肝脏中的异丁酸、戊酸、异戊酸和 2-甲基丁酸显著不同。此外,还根据各自的 SS 阶段对唾液中的丙酸、丁酸、异丁酸、戊酸和异戊酸进行了初步评估,并与这些组织学评分相关。该用于广泛分析多种组织中 SCFAs 的分析平台可以为研究不明确的免疫病理生理学提供线索。