Separation and estimation of saturated C2–C8 fatty acids by buffered partition columns

Moyle, Vivien; Baldwin, Ernest; Scarisbrick, R.

doi:10.1042/bj0430308

Cited by 96 publications

(36 citation statements)

References 3 publications

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“…Although an external indicator (12) and fractional titration (11,16) have been used for the detection of fatty acids as they are eluted from a chromaCan. J. Chem.…”

Section: Preparation Of Silica Colz~rnnsmentioning

confidence: 99%

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C₂–C₈FATTY ACIDS

Fairbairn¹,

Harpur²

1951

Can. J. Chem.

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Thirty-l;\-e milliliters of an aqueous solutioil roilt:linii~g 5 to 70 nlicromoles of \.olatilc C! to Cs acids was dislilled to constant volu~llc without reflusing. The tlis~illa~c xvas titrated, made alkaline, and exxporated LO ciryllcss a t 110°C. The free acicls were regenerated by the addition of 0.03 1111. of 10 N sulphi~ric acid.absorbecl O I I lilter paper, and estracted clc~atltitati\.ely ivith 3 ml. ol chloroforml,~~tanol (09: 1). Acids present ill the estract were separated by chromatography or1 a colnposite silica-gel col~~nlll employiilg tivo ir~ter~lal indicators, and after elution were tlcterrninetl by Litration, the tlctails lor which are given. The ~~r e p n r a t i o~l ot a reproducible silica is descril~ed. Introduction,. 1 he use of silica gel for the quantitative separation of snzall a m o u~l t s of iatty acids, mnging from acetic t o butyric (2, 12, 17). valeric to clecanoic (lSj, and ~~lldecanoic to stearic (10), has been reportecl from several laboratories. In general, different solvents (both deve1ol)ing aiicl stationar!.), separate col~umns, and separate samples of acids have been rccl~lirccl for the analysis of each group of acids. T h e only methocl reported which attelnpts to bridge the gap between two groups of acids is not presented in suf5cient cletail (15). Difficulties have been encountered in the preparation of rcproclucible silicas ( G ) , ancl there is rlo method available for the quantitative tralisier of milligram quantities of thc lon.er f a t t l~ acids from the aqueous s o l u t i o~~s in n.hich the)-usuallj, occur to the s~nall v o l~~m e s of organic solvents r e q~~i r e d lol-t h c chromatographic anall-sis ( 3 ) .Our investigation of the fatty acids excreted b). nematode parasites made the use oi a single small salnple imperative, clespite the fact that these acicls varied in coustitut-ion from acetic to hexanoic and higher. Moyle, Baldnlin, ancl Scarisbrick (11) successfully separated the acids in Lhe acetic to octanoic range by the use of 1-hree separate buffered-silica colunlns, together with a fractional titration of the eluate. Unfortunately, their methocl is laborious and ~lnsuitable for nul-nerous analyses.As already reported (4) it has beell possible to modify the original methods of Ramsey and Patterson (17) and of Elsden (2) in such a way t h a t acetic, propionic, butyric, valeric, hesanoic, and octalloic acids may be quantitatively 1

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“…Although an external indicator (12) and fractional titration (11,16) have been used for the detection of fatty acids as they are eluted from a chromaCan. J. Chem.…”

Section: Preparation Of Silica Colz~rnnsmentioning

confidence: 99%

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C₂–C₈FATTY ACIDS

Fairbairn¹,

Harpur²

1951

Can. J. Chem.

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“…The acids were analysed quantitatively, after a second distillation with mercuric salts (Friedemann, 1938) by evaporating the alkaline extract to dryness, and extracting with a chloroform + butanol mixture (Elsden, 1946). A portion of this solution was titrated with ethanolic NaOH to determine the total acid present, and further samples were analysed on silica columns as described by Moyle, Baldwin & Scarisbrick (1948). Gases were determined by cultivating the organism in a completely filled flask connected by a capillary to a mercury filled gas burette.…”

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Some Studies on the Occurrence and Properties of a Large Gram-Negative Coccus from the Rumen

Hobson¹,

Mann²,

Oxford³

1958

Journal of General Microbiology

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SUMMARY: A large Gram-negative coccus was isolated in a lactate-containing medium from the rumen of young calves. This organism is anaerobic and ferments only lactate, glucose, fructose, maltose, sucrose, mannitol and sorbitol. The coccus is found in large numbers (c. lOg/ml.) in the rumen contents of young calves, but is present only in small numbers or is absent from the rumens of older cattle and sheep. An attempt made to establish the organism in large numbers in a sheep rumen was unsuccessful. The possibility of the coccus being a normal inhabitant of the throat of adult animals was also examined. Five serological types were obtained. The distribution of these types in the same animal over a period, and in different animals, appears to be random. The distribution of a small lactate-fermenting coccus ( VeiZZoneZZa gazogenes) is also described.During the last few years a large number of bacteria have been isolated from the rumens of cattle and sheep, and some of these have been the subjects of extensive study. The present paper deals with the properties and occurrence of a large Gram-negative coccus which has been found to be common in the rumens of young animals, but less so in older animals. The organism was first isolated by chance in media used for cultivating lactobacilli and when its properties became known it was sought specifically in rumen contents. A preliminary communication about this organism has already been made (Hobson, Mann & Oxford, 1958). METHODS Media.Tomato extract agar was prepared by the method of Rogosa et al. (1953). The semi-solid medium contained Bacto casitone, 6 g. ; Bacto yeast extract, 2 g. ; L-cystine, 0.3 g.; thioglycollic (mercaptoacetic) acid, 0-12 ml. ; NaCl, 2 g. ; agar, 0.3 g. ; sodium lactate solution (70 %, w/w), 5.7 ml. ; resazurin, 0.4 ml. of 0.1 yo (w/v) solution; distilled water, 400 ml. Sterilized by autoclaving at 121' for 15 min. The pH value of the medium was 6-9-7-0. Increasing the agar concentration to 2 % gave solid thioglycollate medium. Johns's medium was prepared as described by Johns (1951). This was made into Chocolate Johns's Agar (CJA) medium by addition of 5 yo (v/v) The semi-solid medium (9 ml.) was incubated in tubes (6 in. x Q in.) in air, other media under 5 % (vlv) CO, + 95 yo (v/v) H,, and all at 38'. Sodium lactate solution was a 70% (wlw) solution (Hopkin and Williams Ltd.,

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“…The method selected in the present work for this type of estimation is that due to James & Martin (1952), and it has proved eminently satisfactory, being speedier than that of Moyle, Baldwin & Scarisbrick (1948) and giving reproducible results which the method of Reid & Lederer (1952) did not give in our hands.…”

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confidence: 86%

The determination of the volatile fatty acids formed during the fermentation of grass/water mixtures

Barnett

Duncan

1953

J. Agric. Sci.

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1. The use of the technique of gas-partition chromatography in the investigation of the formation of volatile acids in minced grass/water slurries is discussed.2. Slurries kept anaerobically at 17, 37 and 62° for 13 weeks have been examined, and the amounts of the different volatile acids formed have been estimated. It has been noted that butyric acid forms most readily in the slurry at 37°, while in aerated slurries at 17°, the formation of volatile fatty acids is more speedy than under anaerobic conditions.3. Only acetic, propionic and butyric acids have been noted so far, and the highest concentrations of the acids observed in any slurry were 5, 1·25 and 4% respectively.

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Separation and estimation of saturated C2–C8 fatty acids by buffered partition columns

Cited by 96 publications

References 3 publications

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C₂–C₈FATTY ACIDS

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C₂–C₈FATTY ACIDS

Some Studies on the Occurrence and Properties of a Large Gram-Negative Coccus from the Rumen

The determination of the volatile fatty acids formed during the fermentation of grass/water mixtures

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Separation and estimation of saturated C2–C8 fatty acids by buffered partition columns

Cited by 96 publications

References 3 publications

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C2–C8FATTY ACIDS

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C2–C8FATTY ACIDS

Some Studies on the Occurrence and Properties of a Large Gram-Negative Coccus from the Rumen

The determination of the volatile fatty acids formed during the fermentation of grass/water mixtures

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THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C₂–C₈FATTY ACIDS

THE DISTILLATION, EXTRACTION, AND CHROMATOGRAPHIC SEPARATION OF THE C₂–C₈FATTY ACIDS