Quantitative determination of C2-C8 volatile fatty acids in human serum by vacuum distillation and gas chromatography

Tangerman, A.; Schaik, A. Van; Meuwese-Arends, Maria T.; Tongeren, J. H. M. Van

doi:10.1016/0009-8981(83)90281-4

Cited by 27 publications

(11 citation statements)

References 16 publications

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“…Vacuum transfer has the added advantage that it concentrates the sample and thus reduces the volumes of blood required. Initially, as an internal standard, we used isobutyric acid (27), but found that its concentration fell off rapidly on standing in orthophosphoric acid solution, due probably to micellar separation (27,32 (21,(35)(36)(37)(38)(39). In normal fasting animals, for instance, circulating blood acetate is mainly endogenous (liver) in origin in contrast to the fed state, as in the present study, where the gut is the major source (38,39).…”

Section: Discussionmentioning

confidence: 54%

Carbohydrate fermentation in the human colon and its relation to acetate concentrations in venous blood.

Pomare¹,

Branch²,

Cummings³

1985

J. Clin. Invest.

283

118

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There is now substantial evidence that some dietary polysaccharides, notably dietary fiber, escape absorption in the small bowel and are then broken down in the large intestine of man. The main end products of this colonic digestive process, which is anerobic, are short chain fatty acids (SCFA), and acetic, propionic, and butyric acids. Although these acids are known to be absorbed from the colon, their subsequent fate and significance is unknown. We have measured venous blood SCFA levels in healthy subjects after a 16-h fast, and then following oral doses of either 50 mmol SCFA,5,10, or liter. Pectin fermentation was much slower, with blood acetate levels starting to rise after 6 h and remaining elevated at about twice fasting levels for the subsequent 18 h. However, areas under the blood acetate curves were closely related (r = 0.97; n = 5), whatever the source of acetate. These studies show that the large intestine makes an important contribution to blood acetate levels in man and that fermentation may influence metabolic processes well beyond the wall of this organ.

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Section: Discussionmentioning

confidence: 54%

Carbohydrate fermentation in the human colon and its relation to acetate concentrations in venous blood.

Pomare¹,

Branch²,

Cummings³

1985

J. Clin. Invest.

283

118

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“…For vacuum distillation of fecal samples, we adapted the procedure as described earlier for serum (21). In short, to 3.0 ml of fecal super natant or fecal homogenate 0.3 ml of the IS solution (150 mmol/liter of 2-ethylbutyric acid in 100% formic acid) was added, resulting in a 9% formic acid solution (pH 2-2.5).…”

Section: Vacuum Distillation Of Fecal Samplesmentioning

confidence: 99%

A Gas Chromatographic Analysis of Fecal Short-Chain Fatty Acids, Using the Direct Injection Method

Tangerman

Nagengast

1996

Analytical Biochemistry

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A simple, reproducible, and rapid gas chromato graphic method for short-chain fatty acid determina tion in human feces was developed. It involves direct injection of fecal supernatants into the gas chromato graph, without any pretreatment. Contamination of the gas chromatographic column with nonvolatile fe cal material was prevented by the use of a glass liner in the injector. This liner, which acted as a precolumn, was stoppered with a glass wool plug at the lower end of the liner. Injection was performed against the glass wall of the liner, ensuring an immediate contact of the injected sample with the hot glass wall. More than 100 injections of fecal supernatants could be carried out before the liner had to be replaced by a new one. Peak tailing and ghosting was prevented by the use of for mic acid in the fecal samples. The method gave sharp peaks with baseline separation for all the fatty acids.© 1996 Academic Press, Inc.Short-chain fatty acids (SCFA)2 are produced largely as a result of the breakdown of dietary carbohydrate in the gut by anaerobic bacterial fermentation (1). Acetic, propionic, and ¿7-butyric acid are quantitatively the most important ones. The SCFA present in minor amounts in the human colon (i-butyric, ^-valeric, i-valeric, and i3-caproic acid (2)) primarily originate from protein catabolism and in particular from degradation of certain amino acids (3).About 80-90% of the SCFA are absorbed; the rest are excreted in the feces (4, 5). Despite their presence in the colon in high millimolar concentrations, little information exists regarding the role of SCFA in health and disease (6-8). This is partly due to the fact that 1 To whom correspondence should be addressed. Fax: 0031 243540103.2 Abbreviations used; SCFA, short-chain fatty acid; GC, gas chro matography; IS, internal standard.

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“…As expected, this fragment was found to contain all original C and H atoms of acetate and it was therefore suitable for the studies of exchange of the H atoms. Isolation of acetate from biological fluids may be done by extraction [22,23], steam distillation [24] or vacuum distillation [12,13,24]. In the latter method only volatile compounds are obtained, which makes the sample suitable for g.c.…”

Section: Discussionmentioning

confidence: 99%

“…Due to its high volatility, acetic acid was isolated by another method based on vacuum distillation in an allglass equipment at 13-33 Pa [12,13]. Aliquots (50 10) of bile were acidified to pH 2 with 25 1dl of phosphoric acid (3.5 mol/l), and [1-14C]acetic acid (about 0.05 ,tg) was added in 50,1u of water to allow corrections for incomplete recovery.…”

Section: Isolation and Derivatization Of Carboxylic Acidsmentioning

confidence: 99%

Compartmentation of acetyl CoA studied by analysis of tricarboxylic acid cycle acids and 3-hydroxybutyrate in bile of rats given [2,2,2-2H3]ethanol

Norsten

Cronholm

1990

Biochemical Journal

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Acetate, 3-hydroxybutyrate, pyruvate, lactate, citrate, 2-oxoglutarate, succinate, fumarate and malate were analysed in rat bile by gas chromatography and gas chromatography/mass spectrometry of their O-melthyloxime-t-butyldimethylsilyl derivatives. The concentration of acetate increased to about 1.8 mmol/l after administration of [2,2,2-2H3]ethanol. Acetate was formed from ethanol to an extent of about 82% and retained all of the 2H at C-2, whereas 15% of the 2H had been lost in the tricarboxylic acid cycle intermediates and 24% in 3-hydroxybutyrate. Thus the exchange of 2H for 1H takes place after formation of acetyl CoA. For citrate and 3-hydroxybutyrate, 41% and 11% respectively was formed from [2,2,2-2H3]ethanol. These results indicate that different pools of acetyl CoA are used for the synthesis of ketone bodies and citrate, with the latter being derived from ethanol to a much larger extent. Smaller fractions of 2-oxoglutarate (16%) and succinate (5%) were derived from [2,2,2--2H3]ethanol, indicating significant contributions from amino acids.

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Quantitative determination of C2-C8 volatile fatty acids in human serum by vacuum distillation and gas chromatography

Cited by 27 publications

References 16 publications

Carbohydrate fermentation in the human colon and its relation to acetate concentrations in venous blood.

Carbohydrate fermentation in the human colon and its relation to acetate concentrations in venous blood.

A Gas Chromatographic Analysis of Fecal Short-Chain Fatty Acids, Using the Direct Injection Method

Compartmentation of acetyl CoA studied by analysis of tricarboxylic acid cycle acids and 3-hydroxybutyrate in bile of rats given [2,2,2-2H3]ethanol

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