{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T20:07:24Z","timestamp":1788898044657,"version":"build-2803163510"},"reference-count":47,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T00:00:00Z","timestamp":1782432000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Medical University-Sofia","award":["BG-RRP-2.004-0004-C01"],"award-info":[{"award-number":["BG-RRP-2.004-0004-C01"]}]},{"award":["BG-RRP-2.004-0004-C01"],"award-info":[{"award-number":["BG-RRP-2.004-0004-C01"]}],"id":[{"id":"https:\/\/ror.org\/01n9zy652","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Dopamine dysregulation is connected to several neurological disorders, including Parkinson\u2019s disease, Huntington\u2019s disease, and addiction. A new, precise, accurate, and specific reversed-phase high-performance liquid chromatographic method was developed for dopamine determination in different biological media (human\/mouse plasma and mouse brain homogenate). The chromatographic assay was performed using Avantor ACE\u00ae RP-18 (250 \u00d7 4.6 mm, 5 \u00b5m) column equipped with a suitable ODS pre-column. The temperature was ambient, and the mobile phase was composed of 10 mM potassium dihydrogen phosphate buffer (pH = 3) with 0.25 g\/L sodium octanesulfonate, methanol, and acetonitrile at a volume-to-volume ratio of 75:20:5. Isocratic elution mode, flow rate 1.0 mL\/min, and ultraviolet detection (280 nm) were applied. The procedure was validated for linearity, and all calibration curves were linear over the selected range with determination coefficients greater than 0.998. Intraday repeatability, expressed as the coefficient of variation, did not exceed 4.88% for the plasma and 3.32% for the mouse brain homogenate samples across all tested concentration levels. The proposed chromatographic method was evaluated in terms of greenness, whiteness, and blueness using three ecological metrics (the Analytical Greenness software, White Analytical Chemistry model, and Blue Applicability Grade Index). The optimized procedure was proven to be suitable for implementation in the routine analytical practice.<\/jats:p>","DOI":"10.3390\/molecules31132255","type":"journal-article","created":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T02:16:44Z","timestamp":1782699404000},"page":"2255","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Bioanalytical HPLC-UV Determination of Dopamine in Plasma and Mouse Brain Homogenate with Greenness, Whiteness, and Blueness Assessment"],"prefix":"10.3390","volume":"31","author":[{"given":"Miglena","family":"Smerikarova","sequence":"first","affiliation":[{"name":"Department of Chemistry, Faculty of Pharmacy, Medical University-Sofia, Dunav Str. no. 2, 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stanislav","family":"Bozhanov","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Pharmacy, Medical University-Sofia, Dunav Str. no. 2, 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1640-5099","authenticated-orcid":false,"given":"Jana","family":"Tchekalarova","sequence":"additional","affiliation":[{"name":"Institute of Neurobiology, Bulgarian Academy of Sciences, 1431 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petja","family":"Ivanova","sequence":"additional","affiliation":[{"name":"Institute of Neurobiology, Bulgarian Academy of Sciences, 1431 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Violina T.","family":"Angelova","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Pharmacy, Medical University-Sofia, Dunav Str. no. 2, 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vania","family":"Maslarska","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Pharmacy, Medical University-Sofia, Dunav Str. no. 2, 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1124\/pharmrev.122.000618","article-title":"Dopamine, immunity, and disease","volume":"75","author":"Channer","year":"2023","journal-title":"Pharmacol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1007\/s10571-018-0632-3","article-title":"Dopamine: Functions, signaling, and association with neurological diseases","volume":"39","author":"Klein","year":"2019","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.conb.2019.01.001","article-title":"Mechanisms and regulation of dopamine release","volume":"57","author":"Liu","year":"2019","journal-title":"Curr. Opin. Neurobiol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Zhou, Z.D., Yi, L.X., Wang, D.Q., Lim, T.M., and Tan, E.K. (2023). Role of dopamine in the pathophysiology of Parkinson\u2019s disease. Transl. Neurodegener., 12.","DOI":"10.1186\/s40035-023-00378-6"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Lee, H., Elkamhawy, A., Rakhalskaya, P., Lu, Q., Nada, H., Quan, G., and Lee, K. (2024). Small molecules in Parkinson\u2019s disease therapy: From dopamine pathways to new emerging targets. Pharmaceuticals, 17.","DOI":"10.3390\/ph17121688"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2094","DOI":"10.1093\/brain\/awn112","article-title":"Parkinson\u2019s disease and dopaminergic therapy\u2014Differential effects on movement, reward and cognition","volume":"131","author":"Rowe","year":"2008","journal-title":"Brain"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Abrantes Dias, A.S., Amaral Pinto, J.C., Magalh\u00e3es, M., Mendes, V.M., and Manadas, B. (2020). Analytical methods to monitor dopamine metabolism in plasma: Moving forward with improved diagnosis and treatment of neurological disorders. J. Pharm. Biomed. Anal., 187.","DOI":"10.1016\/j.jpba.2020.113323"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.aca.2012.12.030","article-title":"Liquid chromatographic methods for the quantification of catecholamines and their metabolites in several biological samples\u2014A review","volume":"768","author":"Bicker","year":"2013","journal-title":"Anal. Chim. Acta"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Chen, F., Fang, B., and Wang, S. (2021). A Fast and Validated HPLC method for simultaneous determination of dopamine, dobutamine, phentolamine, furosemide, and aminophylline in infusion samples and injection formulations. J. Anal. Methods Chem., 2021.","DOI":"10.1155\/2021\/8821126"},{"key":"ref_10","first-page":"116","article-title":"A validated HPLC-UV method for determination of dopamine HCl in injectable solutions","volume":"3","author":"Aydin","year":"2020","journal-title":"Eurasian J. Biol. Chem. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1080\/22297928.2022.2121662","article-title":"Characterization and HPLC method validation for determination of dopamine hydrochloride in prepared nano particles and pharmacokinetic application","volume":"12","author":"Neha","year":"2022","journal-title":"Anal. Chem. Lett."},{"key":"ref_12","first-page":"1286","article-title":"Assessment and quantification of Dopamine in rat urine by a validated HPLC-Photodiode Array method (HPLC-PDA)","volume":"13","author":"Bhalchim","year":"2022","journal-title":"J. Pharm. Negat. Results"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.jpba.2014.05.039","article-title":"A rapid and simple method for the determination of 3,4-dihydroxyphenylacetic acid, norepinephrine, dopamine, and serotonin in mouse brain homogenate by HPLC with fluorimetric detection","volume":"98","author":"Fico","year":"2014","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.jpba.2014.01.041","article-title":"Preparation of a stir bar coated with molecularly imprinted polymer and its application in analysis of dopamine in urine","volume":"94","author":"Lei","year":"2014","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.jpha.2011.04.003","article-title":"A rapid method for the determination of dopamine in porcine muscle by pre-column derivatization and HPLC with fluorescence detection","volume":"1","author":"Zhao","year":"2011","journal-title":"J. Pharm. Anal."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1949","DOI":"10.4155\/bio.11.170","article-title":"Quantitative determination of free and total dopamine in human plasma by LC-MS\/MS: The importance of sample preparation","volume":"3","author":"Hendriks","year":"2011","journal-title":"Bioanalysis"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.jpba.2015.09.003","article-title":"Quantitative determination of dopamine in human plasma by a highly sensitive LC-MS\/MS assay: Application in preterm neonates","volume":"117","author":"Zhang","year":"2016","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1248\/bpb.b12-00689","article-title":"Simultaneous measurement of serotonin, dopamine and their metabolites in mouse brain extracts by high-performance liquid chromatography with mass spectrometry following derivatization with ethyl chloroformate","volume":"36","author":"Park","year":"2013","journal-title":"Biol. Pharm. Bull."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3382","DOI":"10.1016\/j.chroma.2011.02.006","article-title":"Monitoring of dopamine and its metabolites in brain microdialysates: Method combining freeze-drying with liquid chromatography-tandem mass spectrometry","volume":"1218","author":"Rambousek","year":"2011","journal-title":"J. Chromatogr. A"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Guiard, B.P., and Gotti, G. (2024). The high-precision liquid chromatography with electrochemical detection (HPLC-ECD) for monoamines neurotransmitters and their metabolites: A review. Molecules, 29.","DOI":"10.3390\/molecules29020496"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.jpba.2018.02.025","article-title":"Development of a diagnostic method for Parkinson\u2019s disease by reverse-phase high-performance liquid chromatography coupled with integrated pulsed amperometric detection","volume":"153","author":"Oh","year":"2018","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.vascn.2015.06.002","article-title":"Determination of dopamine concentrations in brain extracellular fluid using microdialysis with short sampling intervals, analyzed by ultra-high performance liquid chromatography tandem mass spectrometry","volume":"74","author":"Gottas","year":"2015","journal-title":"J. Pharmacol. Toxicol. Methods"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zorina, M., Dotsenko, V.V., Nesterenko, P.N., Temerdashev, A., Dmitrieva, E., Feng, Y.Q., and Atapattu, S.N. (2023). Phthalylglycyl chloride as a derivatization agent for UHPLC-MS\/MS determination of adrenaline, dopamine and octopamine in urine. Molecules, 28.","DOI":"10.3390\/molecules28072900"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.aca.2013.11.058","article-title":"Development of a simple and rapid solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry method for the analysis of dopamine, serotonin and norepinephrine in human urine","volume":"810","author":"Naccarato","year":"2014","journal-title":"Anal. Chim. Acta"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ogawa, Y., Tsugita, S., Torii, Y., Iwamoto, H., Sato, T., Kasahara, J., Takeuchi, M., Kuwabara, T., Iiyama, M., and Takayanagi, T. (2024). Microdialysis-integrated HPLC system with dual-electrode detection using track-etched membrane electrodes for in vivo monitoring of dopamine dynamics. J. Chromatogr. B Anal. Technol. Biomed. Life Sci., 1247.","DOI":"10.1016\/j.jchromb.2024.124318"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6088","DOI":"10.1021\/acs.analchem.5b00633","article-title":"In vivo monitoring of dopamine by microdialysis with 1 min temporal resolution using online capillary liquid chromatography with electrochemical detection","volume":"87","author":"Gu","year":"2015","journal-title":"Anal. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2704","DOI":"10.1039\/c3ay40222c","article-title":"Quantitation of dopamine, serotonin and adenosine content in a tissue punch from a brain slice using capillary electrophoresis with fast-scan cyclic voltammetry detection","volume":"5","author":"Fang","year":"2013","journal-title":"Anal. Methods"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1016\/j.electacta.2017.05.183","article-title":"Electrochemical analysis of dopamine: Perspectives of specific in vivo detection","volume":"245","author":"Ferapontova","year":"2017","journal-title":"Electrochim. Acta"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Falahi, S., Kubiak, A., Voronkina, A., Ehrlich, H., Joseph, Y., and Rahimi, P. (2024). Simultaneous electrochemical detection of dopamine and tryptophan using 3D goethite-spongin composites. Biomimetics, 9.","DOI":"10.3390\/biomimetics9060357"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Chen, J., Ran, X., Ding, X., and Zhang, D. (2023). Dual electrochemical quantitative analysis of dopamine based on flow injection amperometry with double glassy carbon electrodes. Electrochem. Commun., 151.","DOI":"10.1016\/j.elecom.2023.107511"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Al-Salahi, N.O.A., Hashem, E.Y., and Abdel-kader, D.A. (2022). Molecular and biomolecular spectroscopy spectrophotometric methods for determination of dopamine hydrochloride in bulk and in injectable forms. Spectrochim. Acta Part A Mol. Biomol. Spectrosc., 278.","DOI":"10.1016\/j.saa.2022.121278"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.microc.2018.10.004","article-title":"A simple dopamine detection method based on fluorescence analysis and dopamine polymerization","volume":"145","author":"Wei","year":"2019","journal-title":"Microchem. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"993","DOI":"10.3897\/pharmacia.70.e110336","article-title":"Determination of dopamine in blood serum and urine samples by differential pulse voltammetry at an iodine-coated platinum electrode","volume":"70","author":"Hourani","year":"2023","journal-title":"Pharmacia"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"74","DOI":"10.58502\/DTT.23.0034","article-title":"Recent advances in analytical approaches to determine neurotransmitters in biological samples","volume":"3","author":"Cho","year":"2024","journal-title":"Drug Targets Ther."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Orzel, J., Herian, M., and Swit, P. (2025). Neurotransmitter analysis using liquid chromatography-electrochemical detection in the 21st century: Critical review from an analytical chemistry perspective. J. Pharm. Biomed. Anal., 266.","DOI":"10.1016\/j.jpba.2025.117120"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"10076","DOI":"10.1021\/acs.analchem.0c01887","article-title":"AGREE-Analytical GREEnness Metric Approach and Software","volume":"92","author":"Wojnowski","year":"2020","journal-title":"Anal. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7598","DOI":"10.1039\/D3GC02347H","article-title":"Blue applicability grade index (BAGI) and software: A new tool for the evaluation of method practicality","volume":"25","author":"Manousi","year":"2023","journal-title":"Green Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5546","DOI":"10.1039\/D4GC05298F","article-title":"Red Analytical Performance Index (RAPI) and Software: The Missing Tool for Assessing Methods in Terms of Analytical Performance","volume":"27","author":"Nowak","year":"2025","journal-title":"Green. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10343","DOI":"10.1021\/acs.analchem.9b01872","article-title":"What color is your method? Adaptation of the RGB additive color model to analytical method evaluation","volume":"91","author":"Nowak","year":"2019","journal-title":"Anal. Chem."},{"key":"ref_40","unstructured":"(2026, May 07). International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, ICH, Guideline on Analytical Procedure Development Q14. Current Version, June 2024. Available online: https:\/\/www.ema.europa.eu\/en\/ich-q14-analytical-procedure-development-scientific-guideline."},{"key":"ref_41","unstructured":"(2026, May 07). International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, ICH Harmonized Tripartite Guideline, Bioanalytical Method Validation: Text and Methodology M10. Current Step 2b version. Available online: https:\/\/www.ema.europa.eu\/en\/ich-m10-bioanalytical-method-validation-scientific-guideline."},{"key":"ref_42","unstructured":"(2026, May 07). Bioanalytical Method Validation. Guidance for Industry. US Department of Health and Human Services Food and Drug Administration, Available online: https:\/\/www.fda.gov\/files\/drugs\/published\/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf."},{"key":"ref_43","unstructured":"(2026, May 07). FDA Reviewer Guidance, Validation of Chromatographic Methods. Center for Drug Evaluation and Research (CDER). Available online: https:\/\/www.gmp-compliance.org\/files\/guidemgr\/1-6-13.pdf."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Franco, R., Reyes-Resina, I., and Navarro, G. (2021). Dopamine in Health and Disease: Much More Than a Neurotransmitter. Biomedicines, 9.","DOI":"10.3390\/biomedicines9020109"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Chang, K.V., Wu, W.T., Chen, Y.H., Chen, L.R., Lin, Y.L., and Han, D.S. (2025). Plasma levels of adrenaline, noradrenaline, dopamine, serotonin, and cortisol in sarcopenia: Comparison with non-sarcopenic individuals and the impact of resistance exercise and nutrition interventions. J. Int. Soc. Sports Nutr., 22.","DOI":"10.1080\/15502783.2025.2586555"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1016\/j.jfo.2020.08.032","article-title":"Plasma dopamine and catecholamine levels in patients with central serous chorioretinopathy","volume":"44","author":"Yusufoglu","year":"2021","journal-title":"J. Fran\u00e7ais d\u2019Ophtalmologie"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Tchekalarova, J., Ivanova, P., Angelova, V.T., Bogdanova, N., Bozhanov, S., Smerikarova, M., and Maslarska, V. (2026). N-benzylpyrrolidine compounds with MAO-B inhibitory activity in an experimental model of Parkinson\u2019s disease. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27052496"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/13\/2255\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T04:16:36Z","timestamp":1782965796000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/13\/2255"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,26]]},"references-count":47,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["molecules31132255"],"URL":"https:\/\/doi.org\/10.3390\/molecules31132255","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,26]]}}}