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Optimisation of preanalytical tissue processing workflow for mass spectrometry based quantification of animal and human pancreatic tissue lipidomes
Authors: Parchem Karol Rafał | Manzi Malena | Jirásko Robert | Vaňková Zuzana | Holčapek Michal | Mohelníková-Duchoňová Beatrice | Loveček Martin | Melichar Bohuslav | Kuda Ondřej
Year: 2023
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Optimisation of preanalytical tissue processing workflow for mass spectrometry based quantification of animal and human pancreatic tissue lipidomes Many serious disorders, such as fatty liver disease, type 2 diabetes, chronic kidney disease, or various cancer types, are associated with lipid metabolism dysregulation [1]. Consequently, qualitative and qualitative changes in systematic and/or tissue-specific lipidomes can be observed. Therefore, the search for new lipid biomarkers of clinical significance appears to be of great value because of the possibility of their application in the detection of various pathological conditions. Of particular note are some cancer types, including pancreatic ductal adenocarcinoma (PDAC), which does not show specific symptoms in the early stages, making the diagnosis by commonly used screening tests especially difficult [2]. Among the biological samples, biofluids, including plasma, serum, and urine, are commonly used in cancer research [3], while tissue biopsies are more difficult to obtain and tend to be challenging to analyse. On the other hand, tissue lipidomics can provide additional information regarding potential biomarkers and contribute to a deeper understanding of the mechanism of pathology, e.g., when using animal disease models. Compared to biofluids, tissues are highly metabolically active [4]. Pancreatic tissue is one of the most difficult because of high activity of enzymes including triacylglycerol lipase, phospholipase A2, cholesterol esterase, or galactolipase. Therefore, the aim of this study was to develop a preanalytical tissue processing workflow to minimize lipolytic enzyme activity. During the optimization, the effects of sample freeze-drying, tissue cutting and weighting temperature as well as sample disintegration solvent were taken into account. The most preserved conditions, with no processing time-dependent increase in the concentrations of hydrolysis products, required the employment of liquid nitrogen and the processing of deep-frozen tissue until the sample was immersed in the extraction solvent. Finally, the optimised protocol was applied to co tissue lipidomics, pancreatic tissue, biomarkers, quantitative analysis

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