High-Throughput Untargeted Serum Metabolomics Analysis of Hyperuricemia Patients by UPLC-Q-TOF/MS

Abstract

Hyperuricemia (HUA) as a metabolic disease is closely associated with metabolic disorders. The etiology and pathogenesis of HUA are not fully understood, so there is no radical cure so far. Metabolomics, a specialized study of endogenous small molecule substances, has become a powerful tool for metabolic pathway analysis of selected differential metabolites, which is helpful for initially revealing possible development mechanisms of various human diseases. Twenty HUA patients and 20 healthy individuals participated in the experiment, and ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was employed to investigate serum samples to find differential metabolites. The statistical techniques used were principal component analysis and orthogonal partial least-squares discriminant analysis. The differences in metabolomics results of samples after pretreatment with different solvents were compared, 38, 20, 26, 28, 33, 50, and 40 potential differential metabolites were found, respectively, in HUA patient samples, and each group involved different metabolic pathways. Repetitive metabolites were removed, 138 differential metabolites in HUA serum were integrated for analysis, and the human body was affected by 7 metabolic pathways of glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, linoleic acid metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and α-linolenic acid metabolism. In this work, the metabolomics approach based on UPLC-Q-TOF/MS was employed to investigate serum metabolic changes in HUA patients, 138 potential differential metabolites related to HUA were identified, which provided associations of lipids, amino acids, fatty acids, organic acids, and nucleosides profiles of HUA individuals. Metabolic pathways involved in glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, linoleic acid metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and a-linolenic acid metabolism shed light on the understanding of the etiology and pathogenesis process of HUA.

Anti-Gout Effects of the Medicinal Fungus Phellinus igniarius in Hyperuricaemia and Acute Gouty Arthritis Rat Models

“…Metabolomics is an emerging field of research that has been used to evaluate metabolic perturbations associated with different stages of diseases, identify disease biomarkers, explore potential targets and predict drug safety and efficacy ( Shan et al, 2020 ; Zhao et al, 2020 ; Song et al, 2021 ). Non-targeted metabolomics has been conducted extensively to explore biomarkers related to effective hyperuricaemia treatments ( Qin et al, 2021 ; Zhang et al, 2021 ). Therefore, metabolomics based on UHPLC-QE-MS, which is suitable for simultaneous and systemic analysis of multiple metabolite fingerprinting, was employed to generate metabolite profiles of urine to characterize the metabolic changes related to hyperuricaemia and uncover the underlying metabolic mechanism of the therapeutic effects of WPP and CPP on gout.…”

Investigation of pathogenesis of hyperuricemia based on untargeted and targeted metabolomics

“…In this research-based on the previous research method 20 , MeOH/MeCN (1:9, v/v) was used to extract serum samples of HUA patients. The extraction effect was not only better than that of pure solvent but also more complete non-targeted metabolic information could be obtained.…”

“…If a serum precipitation solvent suitable for HUA metabolomics analysis must be recommended, then MeOH/MeCN (1:9, v/v) can be used for analysis 20 . Frozen serum samples were thawed at 4 °C.…”

“…All supernatant was evaporated to dryness. Afterward, the residues were reconstituted in 100 µL of 80% MeOH aqueous, vortexed for 5 min, and incubated for 10 min on ice 20 ; then, they were centrifuged at 12,000 r/min for 10 min at 4 °C. The supernatant was analyzed by UPLC-Q-TOF/MS.…”

“…In recent years, studies on HUA based on metabolomics have been widely reported at home and abroad 16 – 19 . Qin et al 20 divided 20 HUA serum samples and 20 healthy serum samples into seven equal samples, which were pre-treated by different solvent systems, and then analyzed on the ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) analysis platform under the same chromatographic and mass spectrometry conditions. Differences in each group at each analysis stage and final pathway analysis results were subsequently compared.…”