Nagatomo Ryosuke, Okada Yasuki, Ichimura Mayuko, Tsuneyama Koichi, Inoue Koichi
College of Pharmaceutical Sciences, Ritsumeikan University.
Graduate School of Biomedical Sciences, Tokushima University.
Anal Sci. 2018 Sep 10;34(9):1031-1036. doi: 10.2116/analsci.18SCP10. Epub 2018 Aug 3.
We present a sensitive and selective method for the simultaneous determination of short-chain fatty acids (SCFAs), such as acetic acid (AA), propionic acid, butyric acid (BA), isobutyric acid, valeric acid, isovaleric acid, hydroangelic acid, caproic acid, 4-methylvaleric acid and succinic acid (SA) in feces samples using a ultra-high performance liquid-chromatography tandem mass spectrometry (UHPLC-MS/MS) with simple derivatization of 2-picolylamine. The main SFCAs were derivatized in the same condition, and showed the specific product ion (m/z 109) in the electrospray positive mode regarding to 2-picolylamine. The derivatized SA showed a different pattern of the product ion (m/z 191). The derivatized analytes showed LOD < 75 nM, LOQ < 100 nM and r in the calibration curve > 0.991. The QuEChERS was used for sample preparation of feces samples. In the recovery test, the recovery values appeared from 89.7 to 100.2% (RSD: 2.1 to 9.2%, n = 6). This developed method was applied to evaluate obese diabetes model mice. In the result, the branched-chain SCFAs levels in feces from model mice of spontaneous obese type II diabetes were on a declining trend compared with normal. The AA levels from model mice with high-calorie/fat diet are owed a declining trend for 3 to 9 months. The BA levels showed that normal mice were increasing, and model mice had decreased tendency for breeding months. High-calorie/fat diet showed that the SA levels increased.
我们提出了一种灵敏且具选择性的方法,用于同时测定粪便样本中的短链脂肪酸(SCFAs),如乙酸(AA)、丙酸、丁酸(BA)、异丁酸、戊酸、异戊酸、羟基戊酸、己酸、4-甲基戊酸和琥珀酸(SA)。该方法采用超高效液相色谱串联质谱法(UHPLC-MS/MS),并对2-吡啶甲胺进行简单衍生化。主要的短链脂肪酸在相同条件下进行衍生化,在电喷雾正模式下,针对2-吡啶甲胺显示出特定的产物离子(m/z 109)。衍生化的SA显示出不同的产物离子模式(m/z 191)。衍生化后的分析物检测限(LOD)< 75 nM,定量限(LOQ)< 100 nM,校准曲线中的相关系数(r)> 0.991。采用QuEChERS法对粪便样本进行制备。在回收率测试中,回收率值在89.7%至100.2%之间(相对标准偏差:2.1%至9.2%,n = 6)。该开发的方法应用于评估肥胖糖尿病模型小鼠。结果显示,与正常小鼠相比,自发性肥胖II型糖尿病模型小鼠粪便中的支链短链脂肪酸水平呈下降趋势。高热量/高脂肪饮食喂养的模型小鼠在3至9个月内AA水平呈下降趋势。BA水平显示正常小鼠升高,而模型小鼠在繁殖期呈下降趋势。高热量/高脂肪饮食显示SA水平升高。