L'analyse des acides gras en électrophorèse capillaire

Mofaddel, Nadine; Desbènes-Monvernay, A.

doi:10.1051/analusis:1999270120

Analusis

1999

DOI: 10.1051/analusis:1999270120

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L'analyse des acides gras en électrophorèse capillaire

Nadine Mofaddel¹,

A. Desbènes-Monvernay²

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Cited by 8 publications

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References 30 publications

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“…Moreover, ∼8% of organic compounds of pharmaceutical and biomedical interest possess a carboxyl group . Although it would seem that their analysis by gas chromatography (GC), high-performance liquid chromatography (HPLC), or capillary electrophoresis would be straightforward, − extracts from biological fluids are of such great complexity and difference in concentration there is a problem with resolving and detecting all the components. One of the ways to deal with this problem is through mass spectrometry (MS).…”

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confidence: 99%

Enhancement of the LC/MS Analysis of Fatty Acids through Derivatization and Stable Isotope Coding

2007

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This paper focuses on the development of an enhanced LC/ESI-MS method for the identification and quantification of fatty acids through derivatization. Fatty acids were derivatized with 2-bromo-1-methylpyridinium iodide and 3-carbinol-1-methylpyridinium iodide, forming 3-acyloxymethyl-1-methylpyridinium iodide (AMMP). This process attaches a quaternary amine to analytes and enabled ESI-MS in the positive mode of ionization with common LC mobile phases. Moreover, detection sensitivity was generally 2500-fold higher than in the negative mode of ionization used with underivatized fatty acids. The limits of detection were roughly 1.0-4.0 nM (or 10 pg/injection) for standard fatty acids from C10 to C24 and spanned approximately 2 orders of magnitude in linearity. AMMP derivatives had unique tandem mass spectra characterized by common ions at m/z 107.0, 124.0, and 178.0. Individual fatty acids also had unique fingerprint regions that allowed identification of their carbon skeleton number, number of double bonds, and double bond position. The derivatization method also allowed coding of analytes as a means of recognizing derivatives and enhancing quantification. 2H-Coding was achieved through derivatization with deuterated 3-carbinol-1-methyl-d3-pyridinium iodide. The 2H-coded derivatization reagent, 3-acyloxymethyl-1-methyl-d3-pyridinium iodide, was used in two ways. One was to differentially label equal fractions of a sample such that after being recombined and analyzed by ESI-MS all fatty acids appeared as doublet clusters of ions separated by roughly 3 amu. This greatly facilitated identification of fatty acids in complex mixtures. Another use of stable isotope coding was in comparative quantification. Control and experimental samples were differentially labeled with nondeuterated and deuterated isotopomers of CPM, respectively. After mixing the two samples, they were analyzed by ESI-MS. The abundance of a fatty acid in an experimental sample relative to the control was established by the isotope ratio of the isotopomeric fatty acids. Absolute quantification was achieved by adding differentially labeled fatty acid standards to experimental samples containing unknown quantities of fatty acids. Utility of the method was examined in the analysis of human serum samples.

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Enhancement of the LC/MS Analysis of Fatty Acids through Derivatization and Stable Isotope Coding

2007

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Separation of Nile Blue‐labelled fatty acids by CE with absorbance detection using a red light‐emitting diode

et al. 2007

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The separation of fatty acids derivatised with Nile Blue (NB) by CE with detection using a red light-emitting diode (LED) was examined. NB was selected as the derivatisation agent due to its high molar absorption coefficient of 76,000 M(-1) cm(-1) at 633 nm, making it well suited for sensitive absorbance detection using a red 635 nm LED. NB-labelled fatty acids were separated by both MEKC using SDS micelles, i-PrOH and n-BuOH and by NACE in a number of solvents including MeOH, EtOH and ACN. The sensitivity of NACE was superior to MEKC, with detection limits of 5x10(-7)-7x10(-7) M obtained for each acid, approximately 20 times lower than the MEKC method. The NACE detection limits are approximately 100 times lower than previous reports on the separation of fatty acids by CE using indirect absorbance detection, ten times lower than using indirect fluorescence detection and are inferior only to those obtained using precapillary derivatisation and direct fluorescence detection. The efficiency of the NACE method was also superior to MEKC and allowed the separation of unsaturated fatty acids to be examined, although it was not possible to baseline-resolve linoleic (C18:2) and linolenic (C18:3) acids in a reasonable time. The method was used to analyse the fatty acid profile of two edible oils, namely sunflower and sesame oils, after alkali hydrolysis, where it was possible to identify both the saturated and unsaturated fatty acids in each sample.

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Identification of drying oils in mixtures of natural binding media used for artistic and historic works by capillary electrophoresis

Harrison

Kaml

Rainer³

et al. 2005

J of Separation Science

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To characterise the technologies and to find the appropriate treatments, the identification of the binding media of artistic and historic objects is of high importance for the restorer and conservator. Often applied together with other binders, in the present paper, the applicability of a CZE method was ascertained for the identification of the constituents of drying oils in mixtures with animal glues and/or plant gums. The drying oils are identified after alkaline hydrolysis due to their content of long-chain saturated and unsaturated fatty acids (palmitic, stearic, oleic, linoleic, linolenic) and shorterchain dicarboxylic acids (pimelic, suberic, azelaic, sebacic). The dicarboxylic acids occur as products of the unsaturated acids by oxidative degradation during the drying process. The possible interferences in CE caused by the other binders are amino acids and/or monosaccharides. Such interferences could be expected as indirect UV detection has to be used, which is highly unspecific. It was found that none of the constituents of the animal glues or plant gums migrate in the time window of the analytes, thus not obstructing the identification of the analytes in any of the oil mixtures with one binder, or with a combination of both. No interference is observed even in excess of the glues. The method was applied for the identification of a drying oil in a paint layer from a ceremonial mask originating from Papua New Guinea. There is evidence that the oil is most probably from candlenut tree (Aleurites moluccana).

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L'analyse des acides gras en électrophorèse capillaire

Cited by 8 publications

References 30 publications

Enhancement of the LC/MS Analysis of Fatty Acids through Derivatization and Stable Isotope Coding

Enhancement of the LC/MS Analysis of Fatty Acids through Derivatization and Stable Isotope Coding

Separation of Nile Blue‐labelled fatty acids by CE with absorbance detection using a red light‐emitting diode

Identification of drying oils in mixtures of natural binding media used for artistic and historic works by capillary electrophoresis

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