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

Eberhart, B. Loye; Wilson, Annette; O'Keefe, Stephen J.D.; Ramaboli, Matsepo C.; Nesengani, Lucky T.

doi:10.1016/j.ab.2020.114016

Cited by 18 publications

(10 citation statements)

References 28 publications

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“…Although this method does not assess the composition of the microbiota, it determines the content of its metabolite products, which also allows for an indirect assessment of the intestinal microbiota [ 15 ]. In addition, information is also obtained about the type of fermentation taking place in the gut that results in the production of SCFAs [ 16 ]. SCFA levels vary depending on the composition of the intestinal microflora and food intake [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Alterations in fecal short chain fatty acids (SCFAs) and branched short-chain fatty acids (BCFAs) in men with benign prostatic hyperplasia (BPH) and metabolic syndrome (MetS)

Ratajczak

Mizerski

Rył

et al. 2021

Aging

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Gut microbiome-derived short-chain fatty acids (SCFAs) emerge in the process of fermentation of polysaccharides that resist digestion (dietary fiber, resistant starch). SCFAs have a very high immunomodulatory potential and ensure local homeostasis of the intestinal epithelium, which helps maintain the intestinal barrier. We analyzed the association between stool SCFAs levels acetic acid (C 2:0), propionic acid (C 3:0), isobutyric acid (C 4:0i), butyric acid (C 4:0n), isovaleric acid (C 5:0i) valeric acid (C 5:0n), isocaproic acid (C 6:0i), and caproic acid (C 6:0n)) in aging man with benign prostatic hyperplasia (BPH) and healthy controls. The study involved 183 men (with BPH, n = 103; healthy controls, n = 80). We assessed the content of SCFAs in the stool samples of the study participants using gas chromatography. The levels of branched SCFAs (branched-chain fatty acids, BCFAs): isobutyric acid (C4:0i) (p = 0.008) and isovaleric acid (C5:0i) (p < 0.001) were significantly higher in patients with BPH than in the control group. In healthy participants isocaproic acid (C6:0i) predominated (p = 0.038). We also analyzed the relationship between stool SCFA levels and serum diagnostic parameters for MetS. We noticed a relationship between C3:0 and serum lipid parameters (mainly triglycerides) in both healthy individuals and patients with BPH with regard to MetS. Moreover we noticed relationship between C4:0i, C5:0i and C6:0i and MetS in both groups. Our research results suggest that metabolites of the intestinal microflora (SCFAs) may indicate the proper function of the intestines in aging men, and increased BCFAs levels are associated with the presence of BPH.

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Section: Introductionmentioning

confidence: 99%

Alterations in fecal short chain fatty acids (SCFAs) and branched short-chain fatty acids (BCFAs) in men with benign prostatic hyperplasia (BPH) and metabolic syndrome (MetS)

Ratajczak

Mizerski

Rył

et al. 2021

Aging

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“…By comparative analysis of the retention time and the mass spectrometric fragment ions of iso C5:0 and anteiso C5:0 standards, it was noticed that both iso C5:0 and anteiso C5:0 were consistent in retention time and similar in mass spectrometric fragment ions, which could not be separated effectively by characteristic ions. In prior research, the large majority of the literature has effectively quantified only anteiso C5:0 and C5:0 in biological samples [ 27 , 32 , 33 ]. Coelution of iso C5:0 and anteiso C5:0 after acidification has been reported in other literature [ 34 ], and in that research the HP-inno Wax column with the stationary phase of nitro-p-phenylene terephthalic acid-modified polyethylene glycol was also used for chromatographic separation and mass spectrometric quantification, where the problem of co-eluting peaks was remarkably consistent with the results of this study.…”

Section: Resultsmentioning

confidence: 99%

Quantification of Free Short-Chain Fatty Acids in Raw Cow Milk by Gas Chromatography-Mass Spectrometry

Wang

Chen

et al. 2023

Foods

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Free short-chain fatty acids (FSCFAs) are a momentous contributor to the flavor of the raw cow milk. Hence, the purpose of this research was to build an approach for the quantification of 10 FSCFAs in raw cow milk. Raw cow milk samples are acidified by hydrochloric acid ethanol (0.5%) solution pretreatment and then processed on the gas chromatography-mass spectrometry. With the exception of iso C5:0 and anteiso C5:0 co-flux, the remaining eight FSCFAs were effectively separated by chromatography. The methodological validation data revealed that the linear relationship satisfied the assay requirements (coefficient of determination >0.999), the limits of quantification were 0.167 to 1.250 μg mL−1, the recoveries ranged from 85.62% to 126.42%, the coefficients of variation were 1.40~12.15%, and no SCFAs in the triglyceride form were potential degradation, and the precision ranging from 0.56% to 9.09%. Our easy, fast, and robust method successfully determined three FSCFAs in raw cow milk without derivatization. Some characteristic features of FSCFAs have been discovered in raw cow milk such as its higher percentages of C4:0 and C6:0. Our research has provided a very valuable method for the future quality and safety control of raw milk and nutritional studies.

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“…Therefore, lyophilzation is recommended to reduce this variance between and among participants. Overall, the developed method provides a shorter run time, improved linear regression, less variance, improved sensitivity and lower LOD and LOQ values compared to previous methods [18,19]. The selected population for the study included a total of 29 male and female adults (IR-group n = 20; R-group n = 9).…”

Section: Sample Preparationmentioning

confidence: 99%

An Improved Validated Method for the Determination of Short-Chain Fatty Acids in Human Fecal Samples by Gas Chromatography with Flame Ionization Detection (GC-FID)

Smith,

Polite,

Christy

et al. 2023

Metabolites

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Short-chain fatty acids (SCFAs) are metabolites produced by the gut microbiota through the fermentation of non-digestible carbohydrates. Recent studies suggest that the gut microbiota composition, diet and metabolic status play an important role in the production of SCFAs. The primary objective of this study was to develop a simplified method for SCFA analysis in human fecal samples by gas chromatography with flame ionization detection (GC-FID). The secondary objective was to apply the method to fecal samples collected from a clinical trial. The developed GC-FID method showed excellent linearity (R2 > 0.99994), with a limit of detection (LOD) ranging from 0.02 to 0.23 µg/mL and a limit of quantification (LOQ) ranging from 0.08 to 0.78 µg/mL. Recovery for the method ranged between 54.24 ± 1.17% and 140.94 ± 2.10%. Intra- and inter-day repeatability ranged from 0.56 to 1.03 and from 0.10 to 4.76% RSD, respectively. Nine SCFAs were identified and quantified (acetic, propionic, iso-butyric, butyric, iso-valeric, valeric, 4-methyl valeric, hexanoic and heptanoic acids) in freeze-dried fecal samples. The clinical trial compared participants with prediabetes mellitus and insulin resistance (IR-group, n = 20) to metabolically healthy participants (reference group, R-group, n = 9) following a 4-week intervention of a daily red raspberry smoothie (RRB, 1 cup fresh-weight equivalent) with or without fructo-oligosaccharide (RRB + FOS, 1 cup RRB + 8 g FOS). The statistical analysis (Student’s t-test, ANCOVA) was performed on PC-SAS 9.4 (SAS Institute). Acetic acid was higher in the R-group compared to the IR-group at baseline/week 0 (p = 0.14). No significant changes in fecal SCFA content were observed after 4 weeks of either RRB or RRB + FOS.

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

Cited by 18 publications

References 28 publications

Alterations in fecal short chain fatty acids (SCFAs) and branched short-chain fatty acids (BCFAs) in men with benign prostatic hyperplasia (BPH) and metabolic syndrome (MetS)

Alterations in fecal short chain fatty acids (SCFAs) and branched short-chain fatty acids (BCFAs) in men with benign prostatic hyperplasia (BPH) and metabolic syndrome (MetS)

Quantification of Free Short-Chain Fatty Acids in Raw Cow Milk by Gas Chromatography-Mass Spectrometry

An Improved Validated Method for the Determination of Short-Chain Fatty Acids in Human Fecal Samples by Gas Chromatography with Flame Ionization Detection (GC-FID)

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