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Studijní obor:Analytická chemie
Abstrakt:
Lipidomics is a subgroup of metabolomics, which is the last step of the omics cascade
closest to the organism function, and therefore the analysis of metabolites (including lipids) can
provide an important information about ongoing processes in the human body, such as diseases
or disorders reflected in dysregulations of lipid concentrations. The main goal of this Ph.D.
thesis is the development of high-throughput methods for the quantitation of lipids in biological
samples using the following mass spectrometry (MS) based strategies: lipid species separation
and lipid class separation, which have own requirements for the lipid quantitation mainly based
on the selection and the number of internal standards (IS). The lipid species separation is applied
for the analysis of oxylipins using a reversed-phase ultrahigh-performance liquid
chromatography (RP-UHPLC) coupled to tandem mass spectrometry with prior solid phase
extraction from biological samples. The lipid class separation is used for the comprehensive
quantitation of multiple lipid classes including monoacylglycerols, diacylglycerols,
triacylglycerols, cholesteryl-esters, ceramides, sphingomyelins, phosphatidylcholines, and
lysophosphatidylcholines. The lipid class separation approach is achieved by hydrophilic
interaction ultrahigh-performance liquid chromatography (HILIC-UHPLC) or ultrahighperformance
supercritical fluid chromatography (UHPSFC) coupled to mass spectrometry.
Advantages and limitations of both approaches for the lipidomic quantitation are discussed
including the method validation, the use of quality control samples, and the data normalization
using NIST SRM 1950 Reference Material human plasma. Finally, all developed analytical
methods have been applied for sets of clinical samples of cancer patients and healthy controls
followed by multivariate data analysis to distinguish both groups.