PR002745 (Project)

Description:Zebrafish (Danio rerio) has served as a crucial experimental model in biomedical and environmental research. Compared to the more commonly used embryonic and juvenile stages, adult zebrafish possess a complete inter-organ metabolic network and has advantages for chronic disease model development. However, there are very few comprehensive research on metabolomics methods for adult zebrafish. This study implemented an integrated approach combining hydrophilic interaction liquid chromatography (HILIC) and reversed-phase liquid chromatography (RPLC) to maximize metabolite detection, and optimized the key parameters in sample pretreatment, including tissue homogenization, extraction solvent, extraction solvent addition method, re-dissolution solvent. Eventually, an untargeted metabolomics analysis method utilizing ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) techonology was established suitable for the whole tissue of analysis. This method demonstrated satisfactory precision, accuracy, linearity, and sample stability in methodology validation tests. Through metabolite databases retrieval using custom scripts, a total of 620 metabolites were annotated, highlighting the complementarity of RPLC and HILIC, as well as the method’s extensive coverage. Its applicability was verified through a 20-day observation period experiment, screening out 110 differential variables associated with zebrafish growth. Of these, 23 were known compounds, primarily related to glycerophospholipid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and phenylalanine metabolism. This study clarified the experimental conditions for metabolomics studies based on adult zebrafish, offering novel methodological approaches for relating disease or environmental exposure research.