Validation of a lipidomic analysis procedure by nuclear magnetic resonance

Identifier:  TFG:1616

Handle: https://hdl.handle.net/20.500.11797/TFG1616

Authors:  Ozcariz García, Enrique

Abstract:
Currently lipidomics is in full swing. However, there are still certain limitations associated with any lipidomic study, among which are: an adequate extraction method (the butanol: methanol method being the most appropriate) or a correct analysis of the results. Mass spectrometry is the most widely used tool, but others such as nuclear magnetic resonance are gaining prominence in this field given the advantages that it presents as the non-need for derivatization of the sample or its high reproducibility and sensitivity. In this work, we intend to validate a lipidomic analysis protocol using nuclear magnetic resonance. To do this, it was studied which extraction procedure is the most precise and the lipid stability after being stored the samples at different temperatures during different periods of time. It was observed that the most precise extraction procedure was the manual one since it was the one that presented the lowest coefficients of variation. However, it was shown that the recovery of glycerophospholipids and sphingolipids was superior when extractions were carried out with the robot. As regards the lipid stability, the expected stability to the established storage conditions was not observed, but neither was a lipid degradation process detected. Factors such as experimental error and the intrinsic variability of lipids have been able to contribute to obtaining such variable results. The results obtained show the need to optimize the extraction protocol with the robot since the manual is not compatible with a high performance technique such as nuclear magnetic resonance. Likewise, it is necessary to study additional factors (such as the freezing and thawing cycles to which the samples are subjected) for a complete validation of the protocol presented in this paper.