{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T03:51:21Z","timestamp":1761709881821,"version":"build-2065373602"},"reference-count":42,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2021,7,29]],"date-time":"2021-07-29T00:00:00Z","timestamp":1627516800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003447","name":"State Scholarships Foundation","doi-asserted-by":"publisher","award":["MIS-5000432"],"award-info":[{"award-number":["MIS-5000432"]}],"id":[{"id":"10.13039\/501100003447","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Cocaine toxicity has been a subject of study because cocaine is one of the most common and potent drugs of abuse. In the current study the effect of cocaine on human liver cancer cell line (HepG2) was assessed. Cocaine toxicity (IC50) on HepG2 cells was experimentally calculated using an XTT assay at 2.428 mM. The metabolic profile of HepG2 cells was further evaluated to investigate the cytotoxic activity of cocaine at 2 mM at three different time points. Cell medium and intracellular material samples were analyzed with a validated HILIC-MS\/MS method for targeted metabolomics on an ACQUITY Amide column in gradient mode with detection on a triple quadrupole mass spectrometer in multiple reaction monitoring. About 106 hydrophilic metabolites from different metabolic pathways were monitored. Multivariate analysis clearly separated the studied groups (cocaine-treated and control samples) and revealed potential biomarkers in the extracellular and intracellular samples. A predominant effect of cocaine administration on alanine, aspartate, and glutamate metabolic pathway was observed. Moreover, taurine and hypotaurine metabolism were found to be affected in cocaine-treated cells. Targeted metabolomics managed to reveal metabolic changes upon cocaine administration, however deciphering the exact cocaine cytotoxic mechanism is still challenging.<\/jats:p>","DOI":"10.3390\/molecules26154610","type":"journal-article","created":{"date-parts":[[2021,7,29]],"date-time":"2021-07-29T21:21:21Z","timestamp":1627593681000},"page":"4610","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Evaluation of Cocaine Effect on Endogenous Metabolites of HepG2 Cells Using Targeted Metabolomics"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1284-2013","authenticated-orcid":false,"given":"Adamantios","family":"Krokos","sequence":"first","affiliation":[{"name":"Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3642-7367","authenticated-orcid":false,"given":"Olga","family":"Deda","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Medicine and Toxicology, Department of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"},{"name":"Biomic_AUTh, Center for Interdisciplinary Research and Innovation (CIRI-AUTH), Balkan Center, B1.4, 10th km Thessaloniki-Thermi Rd, P.O. Box 8318, 57001 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5900-1011","authenticated-orcid":false,"given":"Christina","family":"Virgiliou","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"},{"name":"Biomic_AUTh, Center for Interdisciplinary Research and Innovation (CIRI-AUTH), Balkan Center, B1.4, 10th km Thessaloniki-Thermi Rd, P.O. Box 8318, 57001 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1893-937X","authenticated-orcid":false,"given":"Helen","family":"Gika","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Medicine and Toxicology, Department of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"},{"name":"Biomic_AUTh, Center for Interdisciplinary Research and Innovation (CIRI-AUTH), Balkan Center, B1.4, 10th km Thessaloniki-Thermi Rd, P.O. Box 8318, 57001 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolaos","family":"Raikos","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Medicine and Toxicology, Department of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eleni","family":"Aggelidou","sequence":"additional","affiliation":[{"name":"Laboratory of Physiology and Pharmacology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2067-8842","authenticated-orcid":false,"given":"Aristeidis","family":"Kritis","sequence":"additional","affiliation":[{"name":"Laboratory of Physiology and Pharmacology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2015-108X","authenticated-orcid":false,"given":"Georgios","family":"Theodoridis","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"},{"name":"Biomic_AUTh, Center for Interdisciplinary Research and Innovation (CIRI-AUTH), Balkan Center, B1.4, 10th km Thessaloniki-Thermi Rd, P.O. Box 8318, 57001 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"163","DOI":"10.4155\/fmc.11.181","article-title":"Accelerating cocaine metabolism as an approach to the treatment of cocaine abuse and toxicity","volume":"4","author":"Schindler","year":"2012","journal-title":"Future Med. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1093\/jat\/30.8.501","article-title":"Major and minor metabolites of cocaine in human plasma following controlled subcutaneous cocaine administration","volume":"30","author":"Kolbrich","year":"2006","journal-title":"J. Anal. Toxicol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"8408479","DOI":"10.1155\/2016\/8408479","article-title":"Cardiovascular and Hepatic Toxicity of Cocaine: Potential Beneficial Effects of Modulators of Oxidative Stress","volume":"2016","author":"Graziani","year":"2016","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1002\/hep.1840110418","article-title":"Cocaine-induced liver cell injury: Comparison of morphological features in man and in experimental models","volume":"11","author":"Kanel","year":"1990","journal-title":"Hepatology"},{"key":"ref_5","first-page":"494","article-title":"Metabolomics of cocaine: Implications in toxicity","volume":"25","year":"2015","journal-title":"Toxicol. Mech. Methods"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5601","DOI":"10.2174\/092986712803988938","article-title":"Contribution of Oxidative Metabolism to Cocaine-Induced Liver and Kidney Damage","volume":"19","author":"Valente","year":"2012","journal-title":"Curr. Med. Chem."},{"doi-asserted-by":"crossref","unstructured":"Dwyer, T.M. (2018). Chemical Signaling in the Nervous System. Fundamental Neuroscience for Basic and Clinical Applications, Elsevier. [5th ed.].","key":"ref_7","DOI":"10.1016\/B978-0-323-39632-5.00004-9"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cbi.2018.01.017","article-title":"In vitro model to study cocaine and its contaminants","volume":"285","author":"Steinmetz","year":"2018","journal-title":"Chem. Interact."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.dmpk.2015.10.002","article-title":"Application of metabolomics to toxicology of drugs of abuse: A mini review of metabolomics approach to acute and chronic toxicity studies","volume":"31","author":"Zaitsu","year":"2016","journal-title":"Drug Metab. Pharmacokinet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6299","DOI":"10.1007\/s00216-013-7046-3","article-title":"Determination of cocaine and metabolites in hair by column-switching LC-MS-MS analysis","volume":"405","author":"Alves","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.trac.2009.12.005","article-title":"Analytical procedures for determination of cocaine and its metabolites in biological samples","volume":"29","author":"Janicka","year":"2010","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1007\/s00216-015-9248-3","article-title":"QuEChERS sample preparation prior to LC-MS\/MS determination of opiates, amphetamines, and cocaine metabolites in whole blood","volume":"408","author":"Dulaurent","year":"2016","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"977","DOI":"10.4155\/bio.09.72","article-title":"Bioanalytical methods for the determination of cocaine and metabolites in human biological samples","volume":"1","author":"Barroso","year":"2009","journal-title":"Bioanalysis"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.forsciint.2016.12.024","article-title":"Determination of cocaine and its derivatives in hair samples by liquid phase microextraction (LPME) and gas chromatography\u2013mass spectrometry (GC\u2013MS)","volume":"274","author":"Pego","year":"2017","journal-title":"Forensic Sci. Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.forsciint.2018.07.004","article-title":"Determination of drugs of abuse and pharmaceuticals in skeletal tissue by UHPLC\u2013MS\/MS","volume":"290","author":"Orfanidis","year":"2018","journal-title":"Forensic Sci. Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.jchromb.2016.11.011","article-title":"A GC\u2013MS method for the detection and quantitation of ten major drugs of abuse in human hair samples","volume":"1047","author":"Orfanidis","year":"2017","journal-title":"J. Chromatogr. B"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.jflm.2014.01.003","article-title":"Determination of cocaine and its major metabolite benzoylecgonine in several matrices obtained from deceased individuals with presumed drug consumption prior to death","volume":"23","author":"Alvear","year":"2014","journal-title":"J. Forensic Leg. Med."},{"doi-asserted-by":"crossref","unstructured":"Marcos, A., Moreno, M., Orihuel, J., Ucha, M., De Paz, A.M., Higuera-Matas, A., Capell\u00e1n, R., Crego, A.L., Mart\u00ednez-Larra\u00f1aga, M.-R., and Ambrosio, E. (2020). The effects of combined intravenous cocaine and ethanol self-administration on the behavioral and amino acid profile of young adult rats. PLoS ONE, 15.","key":"ref_18","DOI":"10.1371\/journal.pone.0227044"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/j.molmed.2017.12.005","article-title":"Using Metabolomics to Investigate Biomarkers of Drug Addiction","volume":"24","author":"Ghanbari","year":"2018","journal-title":"Trends Mol. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.trac.2008.01.008","article-title":"LC-MS-based methodology for global metabolite profiling in metabonomics\/metabolomics","volume":"27","author":"Theodoridis","year":"2008","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.aca.2011.09.042","article-title":"Liquid chromatography\u2013mass spectrometry based global metabolite profiling: A review","volume":"711","author":"Theodoridis","year":"2012","journal-title":"Anal. Chim. Acta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1002\/mas.20306","article-title":"Mass spectrometry-based holistic analytical approaches for metabolite profiling in systems biology studies","volume":"30","author":"Theodoridis","year":"2011","journal-title":"Mass Spectrom. Rev."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"70","DOI":"10.2174\/1874143600802010070","article-title":"Mechanisms of Acute Cocaine Toxicity","volume":"2","author":"Heard","year":"2008","journal-title":"Open Pharmacol. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.1007\/s00216-013-7234-1","article-title":"Metabolic profiling of urine and blood plasma in rat models of drug addiction on the basis of morphine, methamphetamine, and cocaine-induced conditioned place preference","volume":"406","author":"Zaitsu","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1007\/s00213-009-1625-1","article-title":"Alterations in tryptophan and purine metabolism in cocaine addiction: A metabolomic study","volume":"206","author":"Patkar","year":"2009","journal-title":"Psychopharmacology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1016\/j.neuroscience.2012.05.019","article-title":"1H NMR-based metabonomics in brain nucleus accumbens and striatum following repeated cocaine treatment in rats","volume":"218","author":"Li","year":"2012","journal-title":"Neuroscience"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2318","DOI":"10.1194\/jlr.M027656","article-title":"Lipidomic profiling reveals protective function of fatty acid oxidation in cocaine-induced hepatotoxicity","volume":"53","author":"Shi","year":"2012","journal-title":"J. Lipid Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2215","DOI":"10.1002\/elps.201500208","article-title":"Development and validation of a HILIC-MS\/MS multitargeted method for metabolomics applications","volume":"36","author":"Virgiliou","year":"2015","journal-title":"Electrophoresis"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.ejphar.2003.09.056","article-title":"Cocaine induces apoptosis in cerebral vascular muscle cells: Potential roles in strokes and brain damage","volume":"482","author":"Su","year":"2003","journal-title":"Eur. J. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/1056-8719(94)00065-C","article-title":"Response of the human hepatic tissue cultures HEP-G2 and WRL-68 to cocaine","volume":"33","author":"Falk","year":"1995","journal-title":"J. Pharmacol. Toxicol. Methods"},{"doi-asserted-by":"crossref","unstructured":"Deda, O., Virgiliou, C., Orfanidis, A., and Gika, H.G. (2019). Study of Fecal and Urinary Metabolite Perturbations Induced by Chronic Ethanol Treatment in Mice by UHPLC-MS\/MS Targeted Profiling. Metabolites, 9.","key":"ref_31","DOI":"10.3390\/metabo9100232"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4071","DOI":"10.1021\/acs.jproteome.0c00440","article-title":"Metabolic Phenotyping Study of Mouse Brains Following Acute or Chronic Exposures to Ethanol","volume":"19","author":"Deda","year":"2020","journal-title":"J. Proteome Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1177\/026119299001700323","article-title":"Hepatotoxicity of Opiates and Cocaine on Different Hepatic Cellular Systems","volume":"17","author":"Jover","year":"1990","journal-title":"Altern. Lab. Anim."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.brainres.2006.03.061","article-title":"Mitochondrial dysfunction and caspase activation in rat cortical neurons treated with cocaine or amphetamine","volume":"1089","author":"Rego","year":"2006","journal-title":"Brain Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2710","DOI":"10.1038\/s41598-018-21182-7","article-title":"Identification of biochemical and cytotoxic markers in cocaine treated PC12 cells","volume":"8","author":"Badisa","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1007\/s11064-009-0053-2","article-title":"Cocaine Induces Alterations in Mitochondrial Membrane Potential and Dual Cell Cycle Arrest in Rat C6 Astroglioma Cells","volume":"35","author":"Badisa","year":"2009","journal-title":"Neurochem. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"E2101","DOI":"10.1073\/pnas.1602305113","article-title":"Benzoic acid is not the only important product of accelerated metabolism of cocaine","volume":"113","author":"Curry","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1093\/nar\/27.1.29","article-title":"KEGG: Kyoto Encyclopedia of Genes and Genomes","volume":"27","author":"Ogata","year":"1999","journal-title":"Nucleic Acids Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1007\/s00216-012-6638-7","article-title":"Neuronal metabolomics by ion mobility mass spectrometry: Cocaine effects on glucose and selected biogenic amine metabolites in the frontal cortex, striatum, and thalamus of the rat","volume":"405","author":"Kaplan","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.jpba.2017.03.061","article-title":"Investigation on the combined effect of cocaine and ethanol administration through a liquid chromatography\u2013mass spectrometry metabolomics approach","volume":"140","author":"Marcos","year":"2017","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.brainresbull.2008.12.014","article-title":"Cocaine challenge enhances release of neuroprotective amino acid taurine in the striatum of chronic cocaine treated rats: A microdialysis study","volume":"79","author":"Agovic","year":"2009","journal-title":"Brain Res. Bull."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1002\/cbf.3212","article-title":"The expression of tumor suppressor gene Cyldis upregulated by histone deacetylace inhibitors in human hepatocellular carcinoma cell lines","volume":"34","author":"Kotantaki","year":"2016","journal-title":"Cell Biochem. Funct."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/15\/4610\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:37:01Z","timestamp":1760164621000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/15\/4610"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,29]]},"references-count":42,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["molecules26154610"],"URL":"https:\/\/doi.org\/10.3390\/molecules26154610","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2021,7,29]]}}}