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Evaluation of Metabolism and Excretion of Endogenous Substances and Xenobiotics
Uher, Martin ; Hroch, Miloš (advisor) ; Peš, Ondřej (referee) ; Štěrbová, Petra (referee)
Despite the significant progress in analytical techniques, the development and validation of methods for determining endogenous compounds is still a challenging task due to the natural presence of the analyte in the investigated biological matrix. In contrast to xenobiotics, which do not normally occur in biological material, endogenous substances represent a fundamental problem leading to the impossibility of obtaining an analyte-free sample, the so-called blank, which is used in the development and validation of a new method for the preparation of reference samples. Several generally accepted procedures to solve this problem are described, such as the use of surrogate matrices, analyte-purified matrices, or surrogate analytes; however, the used procedures do not always meet the requirements for developing a reliable analytical method according to the given guidelines and regulations. This study aimed to propose an alternative approach in the preparation of validation reference samples while preserving the original biological matrix and solving the problem with the natural presence of analyzed compounds in these samples and, subsequently, apply the validated method as part of metabolomic studies focused on the research of selected liver diseases. The presented procedure is based on the standard...
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