University of Pittsburgh, United States of America Department of Medicine, Division of Gastroenterology, Hepatology, and Nutrition, 200 Lothrop Street, Room W1111/2, Biomedical Science Tower, Pittsburgh, PA, 15213, USA.
Stellenbosch University, South Africa Department of Internal Medicine, Room 3055, Clinical Building, Faculty of Medicine & Health Sciences, Francie Van Zjil Drive, Tygerberg, 7505, USA.
Anal Biochem. 2021 Jan 1;612:114016. doi: 10.1016/j.ab.2020.114016. Epub 2020 Nov 12.
A one-vial extraction method for the quantitation of short-chain fatty acids (SCFAs) in human stool was developed. Samples were extracted with an acidified aqueous internal standard solution, sodium sulfate, and diethyl ether, followed by analysis with GC-FID. Accuracy, in terms of relative recovery, was typically between 90 and 110% for most analytes; without internal standard, the accuracy was about 5-34%; the linear dynamic range (LDR) was 0.05-50 μmol per gram; the limit of detection (LOD) was less than or equal to 0.05 μmol per gram; and the (lower) limit of quantitation (LOQ) was 1 μmol per gram. The method is suitable for quantitating acetic acid, propanoic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, isohexanoic acid, hexanoic acid, and heptanoic acid. It is not suitable for the quantitation of formic acid. Application to human biological research was tested by the measurement of SCFA in heathy humans. This confirmed that the method performed adequately, and even better than expected, with values up to 150 μmol per gram.
建立了一种用于定量检测人粪便中短链脂肪酸（SCFA）的单瓶提取方法。样品用酸化的水内标溶液、硫酸钠和乙醚提取，然后用 GC-FID 分析。大多数分析物的相对回收率的准确度通常在 90%至 110%之间；没有内标时，准确度约为 5-34%；线性动态范围（LDR）为 0.05-50 μmol/克；检测限（LOD）小于或等于 0.05 μmol/克；定量下限（LOQ）为 1 μmol/克。该方法适用于定量检测乙酸、丙酸、异丁酸、丁酸、异戊酸、戊酸、异己酸、己酸和庚酸。它不适用于甲酸的定量。通过测量健康人体中的 SCFA 对人体生物学研究的应用进行了测试。这证实了该方法表现良好，甚至优于预期，其值高达 150 μmol/克。
