{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T13:14:48Z","timestamp":1792588488905,"version":"4.5.0"},"reference-count":40,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,4,11]],"date-time":"2021-04-11T00:00:00Z","timestamp":1618099200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Bladder cancer (BC) is a common malignancy of the urinary system and a leading cause of death worldwide. In this work, untargeted metabolomic profiling of biological fluids is presented as a non-invasive tool for bladder cancer biomarker discovery as a first step towards developing superior methods for detection, treatment, and prevention well as to further our current understanding of this disease. In this study, urine samples from 24 healthy volunteers and 24 BC patients were subjected to metabolomic profiling using high throughput solid-phase microextraction (SPME) in thin-film format and reversed-phase high-performance liquid chromatography coupled with a Q Exactive Focus Orbitrap mass spectrometer. The chemometric analysis enabled the selection of metabolites contributing to the observed separation of BC patients from the control group. Relevant differences were demonstrated for phenylalanine metabolism compounds, i.e., benzoic acid, hippuric acid, and 4-hydroxycinnamic acid. Furthermore, compounds involved in the metabolism of histidine, beta-alanine, and glycerophospholipids were also identified. Thin-film SPME can be efficiently used as an alternative approach to other traditional urine sample preparation methods, demonstrating the SPME technique as a simple and efficient tool for urinary metabolomics research. Moreover, this study\u2019s results may support a better understanding of bladder cancer development and progression mechanisms.<\/jats:p>","DOI":"10.3390\/molecules26082194","type":"journal-article","created":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T05:52:00Z","timestamp":1618206720000},"page":"2194","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":37,"title":["Metabolic Evaluation of Urine from Patients Diagnosed with High Grade (HG) Bladder Cancer by SPME-LC-MS Method"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7254-0864","authenticated-orcid":false,"given":"Kamil","family":"\u0141uczykowski","sequence":"first","affiliation":[{"name":"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3418-9692","authenticated-orcid":false,"given":"Natalia","family":"Warmuzi\u0144ska","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylwia","family":"Operacz","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iga","family":"Stryjak","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4429-6854","authenticated-orcid":false,"given":"Joanna","family":"Bogusiewicz","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Jacyna","sequence":"additional","affiliation":[{"name":"Department of Biopharmacy and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gda\u0144sk, 80-416 Gda\u0144sk, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0763-4589","authenticated-orcid":false,"given":"Renata","family":"Wawrzyniak","sequence":"additional","affiliation":[{"name":"Department of Biopharmacy and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gda\u0144sk, 80-416 Gda\u0144sk, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3701-0849","authenticated-orcid":false,"given":"Wiktoria","family":"Struck-Lewicka","sequence":"additional","affiliation":[{"name":"Department of Biopharmacy and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gda\u0144sk, 80-416 Gda\u0144sk, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Micha\u0142 J.","family":"Markuszewski","sequence":"additional","affiliation":[{"name":"Department of Biopharmacy and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gda\u0144sk, 80-416 Gda\u0144sk, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3971-9816","authenticated-orcid":false,"given":"Barbara","family":"Bojko","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.mpmed.2011.09.008","article-title":"Bladder cancer","volume":"40","author":"Turo","year":"2012","journal-title":"Medicine (Baltimore)"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/S0140-6736(09)60491-8","article-title":"Bladder cancer","volume":"374","author":"Kaufman","year":"2009","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1021\/pr500966h","article-title":"Metabonomic profiling of bladder cancer","volume":"14","author":"Chan","year":"2015","journal-title":"J. Proteome Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fonc.2018.00494","article-title":"Metabolomics of Non-muscle Invasive Bladder Cancer: Biomarkers for Early Detection of Bladder Cancer","volume":"8","author":"Cheng","year":"2018","journal-title":"Front. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bouatra, S., Aziat, F., Mandal, R., Guo, A.C., Wilson, M.R., Knox, C., Bjorndahl, T.C., Krishnamurthy, R., Saleem, F., and Liu, P. (2013). The Human Urine Metabolome. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0073076"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1002\/ijc.30016","article-title":"Biomarkers in bladder cancer: A metabolomic approach using in vitro and ex vivo model systems","volume":"139","author":"Rodrigues","year":"2016","journal-title":"Int. J. Cancer"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.trac.2012.08.011","article-title":"Preparation of urine samples prior to targeted or untargeted metabolomics mass-spectrometry analysis","volume":"41","year":"2012","journal-title":"TrAC\u2014Trends Anal. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.trac.2014.07.005","article-title":"Solid-phase microextraction in metabolomics","volume":"61","author":"Bojko","year":"2014","journal-title":"TrAC\u2019Trends Anal. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2605","DOI":"10.4155\/bio.12.250","article-title":"In vivo solid-phase microextraction for tissue bioanalysis","volume":"4","author":"Cudjoe","year":"2012","journal-title":"Bioanalysis"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1021\/acs.analchem.7b04502","article-title":"Advances in Solid Phase Microextraction and Perspective on Future Directions","volume":"90","author":"Gionfriddo","year":"2018","journal-title":"Anal. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-36481-2","article-title":"The use of solid phase microextraction for metabolomic analysis of non-small cell lung carcinoma cell line (A549) after administration of combretastatin","volume":"9","author":"Jaroch","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.14814\/phy2.13981","article-title":"Cigarette smoking promotes bladder cancer via increased platelet-activating factor","volume":"7","author":"Kispert","year":"2019","journal-title":"Physiol. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-27538-3","article-title":"Bladder cancer recurrence surveillance by urine metabolomics analysis","volume":"8","author":"Loras","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/j.aca.2018.01.055","article-title":"Targeted metabolomics in bladder cancer: From analytical methods development and validation towards application to clinical samples","volume":"1037","author":"Mpanga","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1007\/s11306-012-0433-5","article-title":"Holistic metabonomic profiling of urine affords potential early diagnosis for bladder and kidney cancers","volume":"9","author":"Huang","year":"2013","journal-title":"Metabolomics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1021\/pr901173v","article-title":"Noninvasive Urinary Metabonomic Diagnosis of Human Bladder Cancer","volume":"9","author":"Pasikanti","year":"2010","journal-title":"J. Proteome Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3865","DOI":"10.1021\/pr4000448","article-title":"Urinary Metabotyping of Bladder Cancer Using Two-Dimensional Gas Chromatography Time-of-Flight Mass Spectrometry","volume":"12","author":"Pasikanti","year":"2013","journal-title":"J. Proteome Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1007\/978-1-4939-8757-3_7","article-title":"High-Throughput Solid-Phase Microextraction\u2013Liquid Chromatography\u2013Mass Spectrometry for Microbial Untargeted Metabolomics","volume":"1859","author":"Mousavi","year":"2019","journal-title":"Microb. Metabolomics Methods Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.aca.2013.11.056","article-title":"Introduction of solid-phase microextraction as a high-throughput sample preparation tool in laboratory analysis of prohibited substances","volume":"809","author":"Bojko","year":"2014","journal-title":"Anal. Chim. Acta"},{"key":"ref_20","unstructured":"Pubchem (2019, October 15). U.S. National Library of Medicine National Center for Biotechnology Information, Available online: https:\/\/pubchem.ncbi.nlm.nih.gov\/."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.cca.2018.05.022","article-title":"Association between hippuric acid and left ventricular hypertrophy in maintenance hemodialysis patients","volume":"484","author":"Yu","year":"2018","journal-title":"Clin. Chim. Acta"},{"key":"ref_22","first-page":"1","article-title":"Benzoic Acid Used as Food and Feed Additives Can Regulate Gut Functions","volume":"2019","author":"Mao","year":"2019","journal-title":"BioMed Res. Int."},{"key":"ref_23","first-page":"1","article-title":"The early diagnosis and monitoring of metabolomics","volume":"4","author":"Wang","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1007\/s00432-009-0751-9","article-title":"Fragile histidine triad protein: Structure, function, and its association with tumorogenesis","volume":"136","author":"Hassan","year":"2010","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.urolonc.2009.01.016","article-title":"Prevention of urinary bladder cancer in the FHIT knock-out mouse","volume":"28","author":"Lenoir","year":"2010","journal-title":"Urol. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3113","DOI":"10.12659\/MSM.906721","article-title":"The Clinical Relevance of Fragile Histidine Triad Protein (FHIT) in Patients with Bladder Cancer","volume":"24","author":"Liu","year":"2018","journal-title":"Med. Sci. Monit."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1177\/147323001204000212","article-title":"The Significance of Fragile Histidine Triad Protein as a Molecular Prognostic Marker of Bladder Urothelial Carcinoma","volume":"40","author":"Zhang","year":"2012","journal-title":"J. Int. Med. Res."},{"key":"ref_28","first-page":"1","article-title":"Biomarkers for bladder cancer management: Present and future","volume":"2","author":"Ye","year":"2014","journal-title":"Am. J. Clin. Exp. Urol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1007\/s12291-010-0018-x","article-title":"Antioxidant effect of carnosine pretreatment on cisplatin-induced renal oxidative stress in rats","volume":"25","author":"Noori","year":"2010","journal-title":"Indian J. Clin. Biochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1007\/s00726-012-1271-5","article-title":"Carnosine and cancer: A perspective","volume":"43","author":"Gaunitz","year":"2012","journal-title":"Amino Acids"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1475-2867-11-8","article-title":"Levels of certain tumor markers as differential factors between bilharzial and non-biharzial bladder cancer among Egyptian patients","volume":"11","author":"Metwally","year":"2011","journal-title":"Cancer Cell Int."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"390","DOI":"10.2174\/187152609788922555","article-title":"The Implication of Platelet Activating Factor in Cancer Growth and Metastasis: Potent Beneficial Role of PAF-Inhibitors and Antioxidants","volume":"9","author":"Tsoupras","year":"2009","journal-title":"Infect. Disord.\u2014Drug Targets"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.cbi.2018.06.001","article-title":"Antiproliferative effect of p-Coumaric acid targets UPR activation by downregulating Grp78 in colon cancer","volume":"291","author":"Sharma","year":"2018","journal-title":"Chem. Biol. Interact."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.ejphar.2015.09.044","article-title":"The effects of caffeic, coumaric and ferulic acids on proliferation, superoxide production, adhesion and migration of human tumor cells in vitro","volume":"766","author":"Bouzaiene","year":"2015","journal-title":"Eur. J. Pharmacol."},{"key":"ref_35","first-page":"1","article-title":"Anticancer Properties of Phenolic Acids in Colon Cancer\u2014A Review","volume":"6","author":"Rosa","year":"2016","journal-title":"J. Nutr. Food Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1002\/ptr.4718","article-title":"Antiangiogenic Effects of P-Coumaric Acid in Human Endothelial Cells","volume":"27","author":"Kong","year":"2013","journal-title":"Phytother. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1007\/s13105-010-0061-3","article-title":"In vitro study of the antioxidative properties of the glucose derivatives against oxidation of plasma components","volume":"67","author":"Kolodziejczyk","year":"2011","journal-title":"J. Physiol. Biochem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1603","DOI":"10.1016\/j.freeradbiomed.2010.09.006","article-title":"Oxidative stress, inflammation, and cancer: How are they linked?","volume":"49","author":"Reuter","year":"2010","journal-title":"Free Radic. Biol. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6018","DOI":"10.1021\/ac2010185","article-title":"Optimization of the Coating Procedure for a High-Throughput 96-Blade Solid Phase Microextraction System Coupled with LC \u00c0 MS\/MS for Analysis of Complex Samples","volume":"83","author":"Mirnaghi","year":"2011","journal-title":"Anal. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1944","DOI":"10.1021\/ac102614v","article-title":"Systematic Evaluation of Solid-Phase Microextraction Coatings for Untargeted Metabolomic Profiling of Biological Fluids by Liquid Chromatography\u2014Mass Spectrometry","volume":"83","author":"Vuckovic","year":"2011","journal-title":"Anal. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/8\/2194\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:59:25Z","timestamp":1760363965000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/8\/2194"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,11]]},"references-count":40,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["molecules26082194"],"URL":"https:\/\/doi.org\/10.3390\/molecules26082194","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,11]]}}}