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首页> 外文期刊>Talanta: The International Journal of Pure and Applied Analytical Chemistry >A sensitive GC/MS detection method for analyzing microbial metabolites short chain fatty acids in fecal and serum samples
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A sensitive GC/MS detection method for analyzing microbial metabolites short chain fatty acids in fecal and serum samples

机译：一种敏感的GC / MS检测方法，用于分析粪便和血清样品中的微生物代谢物短链脂肪酸

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摘要

Gut microbiota and their major metabolites, short-chain fatty acids (SCFAs), are recognized as important players in gut homeostasis and metabolic disease occurance. A convenient and sensitive detection method is needed to profile SCFAs in limited and complex biological samples. The gas chromatography/mass spectrometry (GC/MS) is the most common method for SCFAs profiling in biological samples. Trimethylsilyl (TMS) derivatization reagents such as N, O-bis(trimethyl-silyl)-trifluoroacetamide (BSTFA) are commonly used in GC/MS analysis to improve sensitivity and accuracy, but they were barely used in SCFA analysis due to their sensitivity to moisture and the volatility of SCFAs. Here, we developed a rapid, convenient and reliable method for SCFAs profiling in small amounts of fecal and serum samples by GC/MS using BSTFA in combination with sodium sulfate dehydration pretreatment. SCFAs were extracted with anhydrous ether from acidified fecal water extract or serum samples, followed by dehydration with sodium sulfate and BSTFA derivatization at a reduced temperature. Select ion monitoring mode was used for highly sensitive quantification of SCFAs by GC/MS. The derivation with BSTFA at 37 degrees C or 22 degrees C showed an excellent linearity (R-2 0.999), good recoveries (81.27-128.42%), high repeatability (RSD 2%) and low limit of detections (LODs) of different SCFAs ranging from 0.064 to 0.067 mu M. All major SCFAs including acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid were identified and quantified accurately in fecal and serum samples. In conclusions, a reliable, convenient and sensitive method was developed for the measurement of SCFA and other volatile compounds in small biological samples using sodium sulfate dehydration pretreatment and BSTFA derivatization-based GC/MS analyses.
机译：Gut Microbiota及其主要代谢物，短链脂肪酸（SCFA）被认为是肠道稳态和代谢疾病的重要参与者。需要一种方便敏感的检测方法来在有限且复杂的生物样品中剖面到SCFA。气相色谱/质谱（GC / MS）是生物样品中SCFA分析中最常见的方法。三甲基甲硅烷基（TMS）衍生化试剂如N，O-BIS（三甲基甲硅烷基） - 三氟乙酰胺（BSTFA）通常用于GC / MS分析中以提高敏感性和精度，但由于它们对SCFA分析而言，它们几乎没有用于SCFA分析水分和SCFA的波动性。在这里，我们通过GC / MS使用BSTFA与硫酸钠脱水预处理结合使用GC / MS，开发了一种快速，方便，可靠的SCFA分析，少量粪便和血清样品。通过从酸化的粪便水提取物或血清样品中用无水乙醚萃取SCFA，然后用硫酸钠和BSTFA衍生物在降低的温度下脱水。选择离子监控模式用于GC / MS的SCFA的高度敏感量化。 BSTFA在37摄氏度或22℃下的衍生显示出优异的线性度（R-2＆ 0.999），良好的回收率（81.27-128.42％），高可重复性（RSD <2％）和检测下限（LOD）不同的SCFA范围从0.064至0.067 mu m.所有主要的SCFA，包括乙酸，丙酸，异丁酸，丁酸，异戊酸和戊酸，并在粪便和血清样品中精确地鉴定和定量。在结论中，使用硫酸钠脱水预处理和基于BSTFA衍生化的GC / MS分析，开发了可靠，方便和敏感的方法用于测量小生物样品中的小生物样品中的SCFA和其他挥发性化合物。

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