Determination of fatty acids as phenacyl esters in rat adipose tissue and blood vessel walls by high-performance liquid chromatography

Hanis, T.; Smrž, M.; Klír, P; Macek, Karel; Klima, Josef; Baśe, J; Deyl, Z.

doi:10.1016/s0021-9673(01)81467-9

Cited by 35 publications

(16 citation statements)

References 33 publications

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“…Using the conditions described here, we injected a standard sample that contained 0.1 ng of each of four fatty acids (C8:0, C12:0, C17:0 and C22:0), which allowed the detection of all components. The limit of detection (assuming a signal to noise ratio of 3) was then found to be about 0.1 ng of fatty acid per injection, which was more than 10 times less than previous values reported for fatty acid phenacyl esters [6,13,17] and fatty acid naphthacyl esters [15].…”

Section: Fatty Acid Naphthacyl Ester Absorbance Spectrum and Sensitivitymentioning

confidence: 56%

“…2-bromo-2'-acetonaphthone has already been used to prepare naphthacyl derivatives [13,15], but 2-bromoacetophenone has often been preferred to prepare fatty acid phenacyl esters, because of shorter retention times [6,7,13,16,17]. In our study, naphthacyl ester separation resulted in an improved resolution of the fatty acids (see the section describing the separation profile).…”

Section: Fatty Acid Naphthacyl Ester Preparationmentioning

confidence: 67%

“…The more commonly used derivatization with 2-bromoacetophenone enhanced the sensitivity enough to allow the detection of nanograms of fatty acids in complex biological samples [6,7,13,16,17]. However, compared with GC methods, HPLC methods still suffer from a lower resolving power.…”

Section: Introductionmentioning

confidence: 99%

“…Two main new improvements are presented allowing a shorter resolution (50 min) than previous HPLC analyses [16,17], for all components of a complex mixture of fatty acids, from C7:0 to C22:6 n -3. Firstly, a little-known chromophore, 2-bromo-2'-acetonaphthone [13,15], was employed for the synthesis of fatty acid naphthacyl esters.…”

Section: Introductionmentioning

confidence: 99%

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High performance liquid chromatography of fatty acids as naphthacyl derivatives

Rioux¹,

Catheline²,

Bouriel³

et al. 1999

Analusis

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IntroductionStudying fatty acid metabolism in living cells requires not only analytical methods leading to the separation and quantification of each fatty acid, but also preparative methods allowing the recovery of each eluted fatty acid. Numerous fatty acid separation systems based on chromatographic methods (gas chromatography, high performance liquid chromatography) have been developed in the past years [1]. However, none of them seem to be both resolutive and preparative enough to investigate the metabolic bioconversion of fatty acids with physiological significance (biosynthesis, elongation, desaturation, retroconversion and β-oxidation).Gas chromatography (GC) has been the method of choice for separation and quantification of fatty acids for a long time, usually after their conversion to methyl esters [2,3]. Indeed the use of a capillary column coupled to a flame ionization detector (FID) provides high resolution for fatty acid analysis in complex mixtures, including separation of some positional and geometrical isomers. However, there are several inherent limits to GC methods. First of all, short-chain fatty acid methyl esters are volatile and may be lost on refluxing the esterification medium [1]. Secondly, problems originate from possible thermal degradation and structural modification of unsaturated fatty acids during their conversion to methyl esters [4,5]. Thirdly, the main limit to GC is that FID detection destroys the fatty acids and does not allow the recovery of any separated material for further analysis.A great advantage of high performance liquid chromatography (HPLC) methods over GC methods is that the resolved fatty acids are not destroyed during their detection, which enables further analyses to be performed. Recently, several HPLC systems for the separation of fatty acids have been reported, especially with biological applications [6,7]. Abstract. A procedure for the separation of fatty acids, after their derivatization as fatty acid naphthacyl esters, by reverse-phase high performance liquid chromatography is described. A reproducible resolution (50 min), shorter than former HPLC analyses, of a standard mixture of fatty acid naphthacyl esters (25 fatty acids; C7:0 -C22:6 n -3), was achieved by a ternary elution gradient of methanol-acetonitrile-water. Compared with gas chromatography, HPLC analysis of fatty acid naphthacyl esters showed similar percentages of molar distribution of long-chain fatty acids (≥14 carbons). The separation was monotered by UV absorbance at 246 nm, which was highly sensitive (the detection limit was about 0.1 ng), did not destroy the fatty acid naphthacyl esters and thus allowed individual recovery for further analysis (specific radioactivity determination or positional isomer identification). This preparative HPLC method provides, therefore, a useful process for the study of fatty acid metabolism in biological systems.Key words. Fatty acids -fatty acid naphthacyl esters -high performance liquid chromatography -fatty acid metabolism.

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Section: Fatty Acid Naphthacyl Ester Absorbance Spectrum and Sensitivitymentioning

confidence: 56%

Section: Fatty Acid Naphthacyl Ester Preparationmentioning

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

High performance liquid chromatography of fatty acids as naphthacyl derivatives

Rioux¹,

Catheline²,

Bouriel³

et al. 1999

Analusis

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“…This necessitates the use of advanced and fast analysis techniques. Gas and liquid chromatography have found to be very suited for the analysis of fatty acids [3][4][5]. Both techniques are widely applied but there is still a need to speed up the analysis because of the innumerable applications in life.…”

Section: Introductionmentioning

confidence: 99%

Application of micellar electrokinetic chromatography and indirect UV detection for the analysis of fatty acids

Erim

Kraak

1995

Journal of Chromatography A

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Application of micellar electrokinetic chromatography and indirect UV detection for the analysis of fatty acids Erim, F.B.; Xu, X.; Kraak, J.C. General rightsIt is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. AbstractA micellar electrokinetic chromatographic system using indirect UV detection was developed for the analysis of saturated fatty acids. Sodium dodecyl benzenesulfonate (SDBS) was used as micellar surfactant as well as chromophore for the indirect detection. The effects of the addition of acetonitrile and Brij on the migration and solubility of the fatty acids was investigated. With a mixture of 10 mM SDBS + 50% acetonitrile + 30 mM Brij, a complete separation of the C8-C2o fatty acids was obtained. The applicability of the developed system for the analysis of fatty acids in butter is demonstrated.

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Insoluble eggshell matrix proteins – their peptide mapping and partial characterization by capillary electrophoresis and high‐performance liquid chromatography

et al. 2003

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Avian eggshell matrix proteins were studied by two analytical approaches. Peptide mapping was done by trypsin and pepsin followed by collagenase cleavage; analyses were carried out by capillary electrophoresis and reversed-phase high-performance liquid chromatography (HPLC). Comparison of peptide maps obtained by both methods revealed a complex mixture of peptides in the insoluble layers of the eggshell; it was concluded that there are at least three different insoluble protein/peptide layers in the avian eggshell (cuticle, palisade, and mammillary layer). Partial characterization of peptides in each layer was made by HPLC-mass spectrometry analysis. There is an evidence that the eggshell insoluble proteins contain species susceptible to collagenase cleavage, however, the sequences split by this enzyme probably are not those typical for the main triple-helical core of collagenous proteins. It is proposed that the action of collagenase upon eggshell proteins is caused by the side effect of collagenase described previously with synthetic peptides. Some of the proteins present are probably glycosylated. Fatty acid content in the insoluble eggshell layers (after decalcification) was in the range of 2-4% (which reflected both lipid and lipoproteins bound fatty acids). Porphyrin pigments are dominant in the cuticle layer.

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Determination of fatty acids as phenacyl esters in rat adipose tissue and blood vessel walls by high-performance liquid chromatography

Cited by 35 publications

References 33 publications

High performance liquid chromatography of fatty acids as naphthacyl derivatives

High performance liquid chromatography of fatty acids as naphthacyl derivatives

Application of micellar electrokinetic chromatography and indirect UV detection for the analysis of fatty acids

Insoluble eggshell matrix proteins – their peptide mapping and partial characterization by capillary electrophoresis and high‐performance liquid chromatography

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