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A simplified method for the quantitation of short-chain fatty acids in human stool

Author:
B. Loye Eberhart, Annette S. Wilson, Stephen J.D. O'Keefe, Matsepo C. Ramaboli, Lucky T. Nesengani
Source:
Analytical biochemistry 2021 v.612 pp. 114016
ISSN:
0003-2697
Subject:
accuracy, butyric acid, chemical species, detection limit, ethyl ether, extraction, feces, formic acid, hexanoic acid, humans, isobutyric acid, measurement, propionic acid, research, sampling, sodium sulfate, valeric acid
Abstract:
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.
Agid:
7177988