Shelby Bleile – 3/1/13
A Review of “Identification of the Individual Molecular Species of Ceramides
Derived from Human Erythrocytes using HPLC/MS and HPLC/MS/MS”
Korzhenevskii D, Kuptsov V, Mityanina V, Selishcheva A, Saveliev S, Kalashnikova T.
Identification of the individual molecular species of ceramides derived from human erythrocytes
using HPLC/MS and HPLC/MS/MS. Journal Of Analytical Chemistry [serial online]. December 15,
2011;66(13):1270-1275. Available from: Academic Search Premier, Ipswich, MA. Accessed
March 1, 2013.
This study was concerned with ceramides that are present in human
erythrocytes. They are important waxy lipids present in membranes and plasma that are
believes to function as signal molecules and play a role in apoptosis of erythrocytes.
Future research point to ceramides as biochemical markers because they might be
indicators of different diseases. Before that research can occur, all ceramides molecular
species must be identified and studied. Therefore, the purpose of this study was to
obtain a complete profile of ceramides that are present in human erythrocytes, identified
and quantified by HPLC/MS and HPLC/MS/MS.
They first obtained blood and using centrifugation, obtained and washed the
erythrocytes and then performed several complicated extraction using methanol and
chloroform to obtain just the crude lipids. They used a packed silica column
chromatography to obtained just the ceramides from the crude lipids. They then used
the HPLC/MS to identify and quantify individual lipid molecular species. This showed
the different types of ceramides present, and also showed that they complex lipid
extraction that they developed was very efficient. After analysis of the results from the
HPLC/MS, a previously undefined ceramide was identified, Cer 24:2/S18.
This study had a very clear purpose, which was to identify ceramides in
erythrocytes, which they did and discovered a new fraction. They had a very
Shelby Bleile – 3/1/13
complicated lipid extraction that worked very efficiently. However, they only obtained
blood from one healthy volunteer, if they were to repeat this study, they should obtain
more volunteers and compare results to see if ceramides differ between healthy
individuals. Normally with research, preparation of the sample normally has to occur
and there was a great deal of preparation, to obtain just the ceramides fraction, so that
they could analyze the results properly. The choice of an HPLC/MS was appropriate in
that ceramides are organic, not volatile and they are water based. So other instruments
such, as the GC or IC would not be efficient in what they were attempted to study. The
MS results allowed the researchers to quantify the data.
This study could be improved by obtained blood samples from multiple
volunteers to be able to compare and contrast results. Especially due to the fact that a
new ceramides fraction was identified, maybe this is not present in all individuals. They
also could have drawn more blood from the individual to obtain a larger crude lipid mass
and see how they could increase their purity and rate of extraction.
Lastly, this is a great example of how instrumental analysis and biology are so
closely intertwined and how helpful it can be. The researchers did a great job of
developing a complex lipid extraction method and thereby identifying a new ceramide.
Further studies should include more blood samples, so that future research into
ceramides usefulness and biochemical markers can be accurate.