{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T22:13:09Z","timestamp":1791497589640,"version":"4.3.4"},"reference-count":36,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,1,22]],"date-time":"2022-01-22T00:00:00Z","timestamp":1642809600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006595","name":"Unitatea Executiva Pentru Finantarea Invatamantului Superior a Cercetarii Dezvoltarii si Inovarii","doi-asserted-by":"publisher","award":["PN-III-P4-ID-PCCF-2016-0050"],"award-info":[{"award-number":["PN-III-P4-ID-PCCF-2016-0050"]}],"id":[{"id":"10.13039\/501100006595","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>The use of the Liebermann\u2013Burchard reaction in this study has been explored in the development of a simple, reliable, and robust quantitative electrochemical method to assay cholesterol, and hence provide a good alternative to colorimetric methods. The optimization of batch mode operation for electrochemical oxidation of cholesterol in the Liebermann\u2013Burchard reagents included the applied potential and acidic volume. Tested using chronoamperometry, the developed method showed a high sensitivity (14.959 \u03bcA mM\u22121) and low detection limit (19.78 nM) over a 0.025\u20133 mM concentration range, with remarkable linearity (R2 = 0.999), proving an analytical performance either higher or comparable to most of the cholesterol sensors discussed in literature. The influence of possible interfering bioactive agents, namely, glucose, uric acid, ascorbic acid, KCl and NaCl, has been evaluated with no or negligible effects on the measurement of cholesterol. Our study was directed at finding a new approach to chemical processing arising from the use of external potential as an additional level of control for chemical reactions and the transfer of electrons between surfaces and molecules. Finally, the optimized method was successfully applied for the determination of cholesterol content in real blood samples.<\/jats:p>","DOI":"10.3390\/s22030828","type":"journal-article","created":{"date-parts":[[2022,1,23]],"date-time":"2022-01-23T20:36:27Z","timestamp":1642970187000},"page":"828","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Innovative Non-Enzymatic Electrochemical Quantification of Cholesterol"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5910-6342","authenticated-orcid":false,"given":"Oana Elena","family":"Carp","sequence":"first","affiliation":[{"name":"Centre of Advanced Research in Bionanoconjugates and Biopolymers, \u201cPetru Poni\u201d Institute of Macromolecular Chemistry, Grigore Ghica Voda Alley 41A, 700487 Iasi, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4021-982X","authenticated-orcid":false,"given":"Mariana","family":"Pinteala","sequence":"additional","affiliation":[{"name":"Centre of Advanced Research in Bionanoconjugates and Biopolymers, \u201cPetru Poni\u201d Institute of Macromolecular Chemistry, Grigore Ghica Voda Alley 41A, 700487 Iasi, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9827-6528","authenticated-orcid":false,"given":"Adina","family":"Arvinte","sequence":"additional","affiliation":[{"name":"Centre of Advanced Research in Bionanoconjugates and Biopolymers, \u201cPetru Poni\u201d Institute of Macromolecular Chemistry, Grigore Ghica Voda Alley 41A, 700487 Iasi, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1166\/mat.2014.1161","article-title":"Spectroscopic Studies of Cholesterol: Fourier Transform Infra-Red and Vibrational Frequency Analysis","volume":"3","author":"Gupta","year":"2014","journal-title":"Mater. Focus"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"794","DOI":"10.1093\/clinchem\/20.7.794","article-title":"Mechanisms of the Liebermann-Burchard and Zak color reactions for cholesterol","volume":"20","author":"Burke","year":"1974","journal-title":"Clin. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/S0165-022X(98)00036-0","article-title":"Fluorometric method for the enzymatic determination of cholesterol","volume":"38","author":"Amundson","year":"1999","journal-title":"J. Biochem. Biophys. Methods"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1179","DOI":"10.1007\/s11051-012-1179-8","article-title":"Fluorescent cholesterol sensing using enzyme-modified CdSe\/ZnS quantum dots","volume":"14","author":"Kim","year":"2012","journal-title":"J. Nanopart. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.foodchem.2014.09.109","article-title":"Cholesterol determination in foods: Comparison between high performance and ultra-high performance liquid chromatography","volume":"193","author":"Albuquerque","year":"2016","journal-title":"Food Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1896","DOI":"10.1093\/clinchem\/43.10.1896","article-title":"Determination of total cholesterol in serum by liquid chromatography\u2013isotope dilution mass spectrometry","volume":"43","author":"Kock","year":"1997","journal-title":"Clin. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1735","DOI":"10.1007\/s11746-015-2736-y","article-title":"Does the Saponification-GC Method Underestimate Total Cholesterol Content in Samples Having Considerable Cholesterol Esters?","volume":"92","author":"Hansen","year":"2015","journal-title":"J. Am. Oil Chem. Soc."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1006\/abio.1995.1110","article-title":"Determination of Cholesterol Oxidation Products in Human Plasma by Isotope Dilution-Mass Spectrometry","volume":"225","author":"Dzeletovic","year":"1995","journal-title":"Anal. Biochem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2646","DOI":"10.14233\/ajchem.2014.15780","article-title":"Determination of Total Cholesterol in Serum by Gas Chromatography-Mass Spectrometry","volume":"26","author":"Lian","year":"2014","journal-title":"Asian J. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1016\/j.jpba.2011.03.017","article-title":"A validated liquid chromatography\u2013tandem mass spectrometry method for the quantitative determination of 4\u03b2-hydroxycholesterol in human plasma","volume":"55","author":"Bronsema","year":"2011","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1002\/elan.201900669","article-title":"An Overview on Electrochemical Determination of Cholesterol","volume":"32","author":"Amiri","year":"2020","journal-title":"Electroanalysis"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1080\/21691401.2017.1337028","article-title":"Recent approaches to ameliorate selectivity and sensitivity of enzyme based cholesterol biosensors: A review","volume":"46","author":"Gahlaut","year":"2018","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.steroids.2018.12.003","article-title":"Cholesterol biosensors: A review","volume":"143","author":"Narwal","year":"2019","journal-title":"Steroids"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"113538","DOI":"10.1016\/j.jpba.2020.113538","article-title":"Non-enzymatic electrochemical approaches to cholesterol determination","volume":"191","author":"Derina","year":"2020","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1016\/j.bios.2007.10.018","article-title":"Recent advances in cholesterol biosensor","volume":"23","author":"Arya","year":"2008","journal-title":"Biosens. Bioelectron."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2356","DOI":"10.1016\/j.bios.2010.03.036","article-title":"A nonenzymatic cholesterol sensor constructed by using porous tubular silver nanoparticles","volume":"25","author":"Li","year":"2010","journal-title":"Biosens. Bioelectron."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.jelechem.2018.02.011","article-title":"Electrochemically deposited dendritic poly (methyl orange) nanofilm on metal-carbon-polymer nanocomposite: A novel non-enzymatic electrochemical biosensor for cholesterol","volume":"814","author":"Bairagi","year":"2018","journal-title":"J. Electroanal. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.jcis.2019.10.007","article-title":"Simultaneous determination of cholesterol, ascorbic acid and uric acid as three essential biological compounds at a carbon paste electrode modified with copper oxide decorated reduced graphene oxide nanocomposite and ionic liquid","volume":"560","author":"Arotiba","year":"2020","journal-title":"J. Colloid Interface Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6301","DOI":"10.1039\/C5TB00908A","article-title":"Electrodeposition of flower-like nickel oxide on CVD-grown graphene to develop an electrochemical non-enzymatic biosensor","volume":"3","author":"Rengaraj","year":"2015","journal-title":"J. Mater. Chem. B"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.snb.2012.04.070","article-title":"Nonenzymatic cholesterol sensor based on spontaneous deposition of platinum nanoparticles on layer-by-layer assembled CNT thin film","volume":"171\u2013172","author":"Yang","year":"2012","journal-title":"Sens. Actuators B Chem."},{"key":"ref_21","unstructured":"G\u00f6r\u00f6g, S. (2012). Quantitative Analysis of Steroids, Elsevier."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.jelechem.2005.09.003","article-title":"Direct electrochemical acetoxylation of cholesterol at the allylic position","volume":"585","author":"Kowalski","year":"2005","journal-title":"J. Electroanal. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1016\/j.carres.2010.03.018","article-title":"A selective electrochemical method of glycosylation of 3\u03b2-hydroxy-\u03945-steroids","volume":"345","author":"Morzycki","year":"2010","journal-title":"Carbohydr. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6412","DOI":"10.1016\/j.electacta.2009.06.005","article-title":"Electrochemical oxidation of cholesterol in acetonitrile leads to the formation of cholesta-4,6-dien-3-one","volume":"54","author":"Hosokawa","year":"2009","journal-title":"Electrochim. Acta"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1007\/s11745-006-3013-5","article-title":"The Liebermann\u2013Burchard Reaction: Sulfonation, Desaturation, and Rearrangment of Cholesterol in Acid","volume":"42","author":"Xiong","year":"2007","journal-title":"Lipids"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"917","DOI":"10.1021\/jf00088a020","article-title":"Overestimation of the cholesterol content of eggs","volume":"37","author":"Beyer","year":"1989","journal-title":"J. Agric. Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/0021-9673(92)85689-Q","article-title":"Chromatographic separation of cholesterol in foods","volume":"624","author":"Fenton","year":"1992","journal-title":"J. Chromatogr. A"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/S0009-9120(67)80002-X","article-title":"The estimation of cholesterol in serum:A classification and critical review of methods","volume":"1","author":"Tonks","year":"1967","journal-title":"Clin. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1139\/y59-099","article-title":"A rapid method of total lipid extraction and purification","volume":"37","author":"Bligh","year":"1959","journal-title":"Can. J. Biochem. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9045938","DOI":"10.1155\/2019\/9045938","article-title":"Validation of a Simple and Robust Liebermann\u2013Burchard Colorimetric Method for the Assay of Cholesterol in Selected Milk Products in Ghana","volume":"2019","author":"Adu","year":"2019","journal-title":"Int. J. Food Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1021\/ac60258a057","article-title":"Equilibrium between sulfuric and acetylsulfuric acids in acetic acid-acetic anhydride","volume":"40","author":"Tanghe","year":"1968","journal-title":"Anal. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/0026-265X(91)90033-L","article-title":"A review of reactions of some sterols in strongly acidic media","volume":"43","author":"Zuman","year":"1991","journal-title":"Microchem. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1747","DOI":"10.1248\/cpb.28.1747","article-title":"Study on the correspondence of color change with polyenyl cation formation of cholesterol in strong acids","volume":"28","author":"Niiya","year":"1980","journal-title":"Chem. Pharm. Bull."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.chroma.2007.08.020","article-title":"Determination of total cholesterol in serum by high-performance liquid chromatography with electrochemical detection","volume":"1166","author":"Hojo","year":"2007","journal-title":"J. Chromatogr. A"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"127200","DOI":"10.1016\/j.snb.2019.127200","article-title":"Development of non-enzymatic cholesterol bio-sensor based on TiO2 nanotubes decorated with Cu2O nanoparticles","volume":"302","author":"Khaliq","year":"2020","journal-title":"Sens. Actuators B Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1016\/j.steroids.2011.09.003","article-title":"Amperometric detection of cholesterol using an indirect electrochemical oxidation method","volume":"76","author":"Chiang","year":"2011","journal-title":"Steroids"}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/22\/3\/828\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:05:53Z","timestamp":1760133953000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/22\/3\/828"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,22]]},"references-count":36,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["s22030828"],"URL":"https:\/\/doi.org\/10.3390\/s22030828","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,22]]}}}