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


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Breadmore, MC, Henderson, RD, Fakhari, AR, Macka, M and Haddad, PR 2007 , 'Separation of Nile Blue-labelled fatty acids by CE with absorbance detection using a red light-emitting diode' , Electrophoresis, vol. 28, no. 8 , pp. 1252-1258 , doi: 10.1002/elps.200600580.

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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 M21cm21 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 561027–761027 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

Item Type: Article
Authors/Creators:Breadmore, MC and Henderson, RD and Fakhari, AR and Macka, M and Haddad, PR
Keywords: Carboxylic acids / Fatty acids / Light-emitting diode / NACE / Precolumn derivatisation
Journal or Publication Title: Electrophoresis
Publisher: Wiley-VCH
ISSN: 0173-0835
DOI / ID Number: 10.1002/elps.200600580
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