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2-吡啶甲胺衍生化超高效液相色谱串联质谱法在粪便样本中短链脂肪酸测定中的应用

Application of 2-Picolylamine Derivatized Ultra-high Performance Liquid Chromatography Tandem Mass Spectrometry for the Determination of Short-chain Fatty Acids in Feces Samples.

作者信息

Nagatomo Ryosuke, Okada Yasuki, Ichimura Mayuko, Tsuneyama Koichi, Inoue Koichi

机构信息

College of Pharmaceutical Sciences, Ritsumeikan University.

Graduate School of Biomedical Sciences, Tokushima University.

出版信息

Anal Sci. 2018 Sep 10;34(9):1031-1036. doi: 10.2116/analsci.18SCP10. Epub 2018 Aug 3.

Abstract

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.

摘要

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。

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