Data was acquired in MRM mode, and the MRM transitions and retention times otpimized on synthetic standards were used as identifiers for the quantified metabolites.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - alternating - hilic"],"chromatography_protocol":["The urinary metabolites were separated and quantified by using an LC-MS/MS system consisting of an Agilent 1290 UPLC connected to an Agilent 6495D triple quadrupole (Agilent, CA, USA). The separation was achieved by injecting 3 µL of a sample to a BEH amide column (30×2.1 mm, 1.7 µm, Waters, MA, USA). The mobile phase consisted of 10 mM ammonium fomate of pH 3.0 (adjusted with formic acid) in (A) water and in (B) acetonitrile/water 90/10 (v/v). The mobile phase was delivered on column by a flow rate of 0.6 mL/min with the following gradient: 0.0 min (99% B), 1.5 min (99% B), 3.0 min (30% B), 4.0 min (30% B), 4.1 min (99% B), 7.0 min (99% B). Column and autosampler were thermostated at 40 °C and 4 °C, respectively.
"],"publication":["Rapid and targeted HILIC-MS/MS quantification of urinary metabolites reveals metabolic alterations in COVID-19 patients."],"submitter_affiliation":["Umea University","Swedish Metabolomics Centre"],"submitter_name":["Annika Johansson","Ondrej Hodek"],"organism_part":["urine"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["Prior to the analysis the urine samples were thawed at room temperature, vortex mixed, and 10 µL of urine was diluted 150× with 80% acetonitrile containing the internal standards (1 µM creatine-D3, 1 µM 1-methyl-histamine-D3, 1 µM 1-methyl-histidine-D3, 1 µM 3-methyl-histidine-D3). For quantification of the most abundant metabolites – creatinine, urea, uric acid – the 150× diluted urine was further diluted 40-fold with 80% acetonitrile containing internal standards of 10 µM creatinine-D3, 10 µM urea-15N2, and 10 µM uric acid-15N2.
"],"organism":["Homo sapiens"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS13986"],"author":["Annika Johansson. Umeå University. annika.johansson01@umu.se.","Ondrej Hodek. Swedish University of Agricultural Sciences. ondrej.hodek@slu.se."],"data_transformation_protocol":["The LC-MS data was processed by using the Agilent MassHunter Qualitative 12.0.430.0 and QQQ quantitative analysis version B.12.0.893.1 (Agilent Technologies Inc., Santa Clara, CA, USA) and Excel (Microsoft, Redmond, Washington, USA) software.
"],"study_factor":["Disease","Sample type"],"submitter_email":["ondrej.hodek@slu.se","annika.johansson01@umu.se"],"sample_collection_protocol":["The human urine samples used for validation were obtained from healthy adults with age between 20 and 73 years (n = 13). The samples were frozen at -80 °C within 4 hours after sampling. Urine samples from patients with mild (n = 10) and severe (n = 10) COVID-19 were obtained from the prospective, observational cohort study CoVUm (clinicaltrials.gov identifier: NCT04368013) and used for further clinical validation of the method.
"],"omics_type":["Metabolomics"],"study_design":["ultra-performance liquid chromatography-mass spectrometry","COVID-19","human urinary metabolite","targeted analysis"],"curator_keywords":["ultra-performance liquid chromatography-mass spectrometry","COVID-19","human urinary metabolite","targeted analysis"],"mass_spectrometry_protocol":["Analytes were ionized in an electrospray ion source operated both in positive and negative modes. The source and gas parameters were set as follows: ion spray voltage -3.5 kV in negative and +4.0 kV in positive mode, gas temperature 150 °C, drying gas flow 11 L/min, nebulizer pressure 20 psi, sheath gas temperature 325 °C, sheath gas flow 12 L/min, fragmentor 166 V.
"],"metabolite_name":["guanidineacetic acid","3-methyl-L-histidine","creatinine","1-methylhistamine","1-methyl-4-imidazoleacetic acid","creatine","1-methyl-L-histidine","1-methyl-5-imidazoleacetic acid","uric acid","urea"],"additional_accession":[]},"is_claimable":false,"name":"Rapid and targeted HILIC-MS/MS quantification of urinary metabolites reveals metabolic alterations in COVID-19 patients","description":"Urinary metabolites and their concentrations serve as biomarkers for identification of metabolic pathways that relate to specific diseases; therefore, fast and accurate quantification of the metabolites in urine is essential in health assessment and diagnosis. As many urinary metabolites are of polar nature, hydrophilic interaction liquid chromatography (HILIC) has been used over the last several years because it offers faster and more reproducible analyses compared to traditional techniques such as reversed-phase chromatography or capillary electrophoresis. In our study, we developed a HILIC method by using a 3-cm analytical column in connection with tandem mass spectrometry detection for quantification of 10 urinary metabolites including creatinine as the reference for normalization. As all tested metabolites contain ionizable functional groups, pH of the mobile phase was optimized to achieve baseline separation of 2 isomeric pairs (1-methyl-4-imidazoleacetic acid/1-methyl-5-imidazoleacetic acid and 1-methylhistidine/3-methylhistidine) and to obtain overall better separation efficiency resulting in a 7-min analysis. The developed method was validated in terms of sensitivity, carry-over, linearity, matrix effects, accuracy, and precision. The metabolite concentrations in healthy subjects determined by the developed method correspond well with the normal reference values found in the literature. Moreover, the method was tested on a small cohort of COVID-19 patients, where it enabled identification of differences in metabolite levels. Thus, the developed method has potential to be used routinely in a diagnostic field for high-throughput analysis of urine samples.
","dates":{"publication":"2026-06-08","submission":"2026-03-06"},"accession":"MTBLS13986","cross_references":{"MetaboLights":["MTBLC50599","MTBLC27596","MTBLC70966","MTBLC132918","MTBLC16919","MTBLC29009","MTBLC16344","MTBLC16737","MTBLC17775","MTBLC16199"],"ChEBI":["CHEBI:50599","CHEBI:27596","CHEBI:70966","CHEBI:132918","CHEBI:16919","CHEBI:29009","CHEBI:16344","CHEBI:16737","CHEBI:17775","CHEBI:16199"]}}