10 research outputs found
Simple methodology for the quantitative analysis of fatty acids in human red blood cells
In the last years, there has been an increasing
interest in evaluating possible relations between fatty acid
(FA) patterns and the risk for chronic diseases. Due to the
long life span (120 days) of red blood cells (RBCs), their
FA profile reflects a longer term dietary intake and was
recently suggested to be used as an appropriate biomarker
to investigate correlations between FA metabolism and diseases.
Therefore, the aim of this work was to develop and
validate a simple and fast methodology for the quantification
of a broad range of FAs in RBCs using gas chromatography
with flame ionization detector, as a more common
and affordable equipment suitable for biomedical and
nutritional studies including a large number of samples. For
this purpose, different sample preparation protocols were
tested and compared, including a classic two-step method
(Folch method) with modifications and different one-step methods, in which lipid extraction and derivatization were
performed simultaneously. For the one-step methods, different
methylation periods and the inclusion of a saponification
reaction were evaluated. Differences in absolute FA
concentrations were observed among the tested methods,
in particular for some metabolically relevant FAs such as
trans elaidic acid and eicosapentaenoic acid. The one-step
method with saponification and 60 min of methylation time
was selected since it allowed the identification of a higher
number of FAs, and was further submitted to in-house validation.
The proposed methodology provides a simple, fast
and accurate tool to quantitatively analyse FAs in human
RBCs, useful for clinical and nutritional studies.This work received financial support from the
European Union (FEDER funds through COMPETE) and National
Funds (FCT, Fundação para a Ciência e Tecnologia) through project
PTDC/SAU-ENB/116929/2010 and EXPL/EMS-SIS/2215/2013.
ROR acknowledges PhD scholarship SFRH/BD/97658/2013 attributed
by FCT (Fundação para a Ciência e Tecnologia).info:eu-repo/semantics/publishedVersio