| Summary: | 碩士 === 長庚大學 === 醫學生物技術暨檢驗學系 === 100 === Metabolomics is getting more and more popular in biomedical study. The metabolomic data reflect the regulation of transcription and translation processes, the regulation of protein–protein interactions, and the allosteric regulation of enzymes and their interactions with metabolites. Metabolomics tools are gaining tremendous popularity in different fields of life sciences; from the discovery of biomarkers, diagnostic and biochemical characterization of diseases to drug analysis. Since the chemical properties of metabolites are diverse, a wide range of different sample preparation procedures are used for different research objectives. Eighty percentage (80%) methanol is the most commonly used chemical extraction method to extract intracellular metabolites and generate metabolite profiles. However, different extraction protocols of intracellular metabolites will obtain different metabolomic results leading to different biological interpretation. In this study, we used three different methods including neutral, alkaline, and acidic 80% methanol for intracellular metabolites of HepG2 cells extraction. After extraction and LC - TOFMS analysis of metabolites, we did not only detect different numbers of compounds depending on the extraction method used, but we also obtained distinct metabolite levels for the compounds commonly detected by all methods. Our results showed that different solvent-based extraction methods can yield significantly different metabolite profiles, which maybe subjected to different biological interpretations These different extraction methods also were used for seven redox related metabolites, ATP, ADP, AMP, NAD+, NADH, NADP+, and NADPH evaluation and quantification. The different solvent-based extraction methods also yielded significantly different quantities of these metabolites. Thus, development of different extraction protocols for sample preparation and combine these results from these methods will for metabolomics should get more correct biological interpretation.
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