Urinary metabolomics study of patients with gout using gas chromatography-mass spectrometry
Description
The aim of the study was to detect urinary metabolic changes in gout patients which may contribute to understanding the pathological mechanism of gout. A gas chromatography-mass spectrometry (GC-MS) method was employed to explore the distinctive metabolic patterns in patients with gout.
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Morning and mid-stream urine were collected from all the participants. 50 µL of the urine sample was mixed with 80 µL of urease solution, and was hydrolyzed in a 37°C water bath. Cold acetonitrile was added to precipitate the protein. The supernatant was lyophilized under vacuum. Before analysis, the lyophilized metabolites were dissolved in methoxyamine pyridine solution (20 mg/mL), and then reacted for 90 min in a 37°C water bath. Next, MSTFA was added for trimethylsilylation for 60 min. After derivatization, the supernatant was transferred to a vial insert for analysis. GC–MS analysis was performed on an Agilent 7890/5975C GC–MS system with a 30m *0.25mm *0.25μm DB5-MS fused silica capillary column. MSD ChemStation was employed for peak integration of all samples. Total peak area normalization was performed on the data.