{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T07:22:24Z","timestamp":1770448944446,"version":"3.49.0"},"reference-count":27,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2017,4,7]],"date-time":"2017-04-07T00:00:00Z","timestamp":1491523200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>An enzymatic method for quantitative detection of the reduced form of nicotinamide-adenine dinucleotide (NADH) using surface-enhanced Raman scattering was developed. Under the action of NADH oxidase and horseradish peroxidase, NADH can generate hydrogen peroxide (H2O2) in a 1:1 molar ratio, and the H2O2 can oxidize a chromogen into pigment with a 1:1 molar ratio. Therefore, the concentration of NADH can be determined by detecting the generated pigment. In our experiments, eight chromogens were studied, and o-tolidine (OT) was selected because of the unique Raman peaks displayed by its corresponding pigment. The optimal OT concentration was 2 \u00d7 10\u22123 M, and this gave the best linear relationship and the widest linear range between the logarithmic H2O2 concentration and the logarithmic integrated SERS intensity of the peak centered at 1448 cm\u22121. Under this condition, the limit of detection for NADH was as low as 4 \u00d7 10\u22127 M. Two NADH samples with concentrations of 2 \u00d7 10\u22124 and 2 \u00d7 10\u22125 M were used to validate the linear relationship, and the logarithmic deviations were less than 3%.<\/jats:p>","DOI":"10.3390\/s17040788","type":"journal-article","created":{"date-parts":[[2017,4,7]],"date-time":"2017-04-07T10:49:53Z","timestamp":1491562193000},"page":"788","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Quantitative Detection of NADH Using a Novel Enzyme-Assisted Method Based on Surface-Enhanced Raman Scattering"],"prefix":"10.3390","volume":"17","author":[{"given":"Haiyan","family":"Teng","sequence":"first","affiliation":[{"name":"Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mingyang","family":"Lv","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luo","family":"Liu","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2301-3336","authenticated-orcid":false,"given":"Xin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongmei","family":"Zhao","sequence":"additional","affiliation":[{"name":"Engineering Research Center for Semiconductor Integrated Technology, Institute of Semiconductors, Chinese Academy of Sciences, Beijing 100083, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhenglong","family":"Wu","sequence":"additional","affiliation":[{"name":"Analytical and Testing Center, Beijing Normal University, Beijing 100875, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haijun","family":"Xu","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2017,4,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1042\/BJ20061638","article-title":"The power to reduce: Pyridine nucleotides-small molecules with a multitude of functions","volume":"402","author":"Pollak","year":"2007","journal-title":"Biochem. J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/0304-4157(95)00003-A","article-title":"The mitochondrial permeability transition","volume":"1241","author":"Zoratti","year":"1995","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"12366","DOI":"10.1021\/ja204014s","article-title":"Reversible redox of NADH and NAD+ at a hybrid lipid bilayer membrane using ubiquinone","volume":"133","author":"Ma","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"4941","DOI":"10.1021\/bi00365a033","article-title":"Raman spectroscopy of oxidized and reduced nicotinamide adenine dinucleotides","volume":"25","author":"Yue","year":"1986","journal-title":"Biochemistry"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1006\/mben.2002.0227","article-title":"Metabolic engineering of escherichia coli: Increase of NADH availability by over expressing an NAD+\u2014Dependent formate dehydrogenase","volume":"4","author":"Bennett","year":"2002","journal-title":"Metab. Eng."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3244","DOI":"10.1021\/ac034888w","article-title":"Chemical reversibility and stable low-potential NADH detection with nonconventional conducting polymer nanotubule modified glassy carbon electrodes","volume":"76","author":"Valentini","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2309","DOI":"10.1039\/c1lc20303g","article-title":"Artificial photosynthesis on a chip: Microfluidic cofactor regeneration and photoenzymatic synthesis under visible light","volume":"11","author":"Lee","year":"2011","journal-title":"Lab Chip"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3775","DOI":"10.1016\/j.tetlet.2010.05.044","article-title":"Fluorescein derivative-based, selective and sensitive chemosensor for NADH","volume":"51","author":"Jung","year":"2010","journal-title":"Tetrahedron Lett."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"11532","DOI":"10.1021\/jp810931u","article-title":"Monitoring cellular metabolism with fluorescence lifetime of reduced nicotinamide adenine dinucleotide","volume":"113","author":"Ghukasyan","year":"2009","journal-title":"J. Phys. Chem. C"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1572","DOI":"10.1002\/anie.200503314","article-title":"Elementary Arithmetic Operations by Enzymes: A Model for Metabolic Pathway Based Computing","volume":"45","author":"Baron","year":"2006","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/j.snb.2006.04.075","article-title":"Low potential detection of NADH with Prussian Blue bulk modified screen-printed electrodes and recombinant NADH oxidase from thermus thermophilus","volume":"121","author":"Radoi","year":"2007","journal-title":"Sens. Actuators B"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1002\/elan.200703867","article-title":"Electroaddressable selective functionalization of electrode arrays: Catalytic NADH detection using aryl diazonium modified gold electrodes","volume":"19","author":"Harper","year":"2007","journal-title":"Electroanalysis"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/S1355-0306(02)71831-5","article-title":"On the potential of Raman microscopy for the forensic analysis of coloured textile fibres","volume":"42","author":"Jochem","year":"2002","journal-title":"Sci. Justice"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kasera, S., Herrmann, L.O., del Barrio, J., Baumberg, J.J., and Scherman, O.A. (2014). Quantitative multiplexing with nano-self-assemblies in SERS. Sci. Rep., 4.","DOI":"10.1038\/srep06785"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5888","DOI":"10.1021\/ac200707t","article-title":"Highly sensitive detection of proteins and bacteria in aqueous solution using surface-enhanced Raman scatteringand optical fibers","volume":"83","author":"Yang","year":"2011","journal-title":"Anal. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.bios.2015.02.023","article-title":"Approach for determination of ATP: ADP molar ratio in mixed solution by surface-enhanced Raman scattering","volume":"69","author":"Fang","year":"2015","journal-title":"Biosens. Bioelectron."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1016\/j.snb.2014.01.002","article-title":"A sensitive and label-free electrochemical impedance biosensor for protein detection based on terminal protection of small molecule-linked DNA","volume":"194","author":"Wang","year":"2014","journal-title":"Sens. Actuators B"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.snb.2015.10.003","article-title":"Alizarin dye based ultrasensitive plasmonic SERS probe for trace level cadmium detection in drinking water","volume":"224","author":"Dasary","year":"2016","journal-title":"Sens. Actuators B"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2431","DOI":"10.1002\/adma.201305106","article-title":"Highly sensitive, uniform and reproducible surface-enhanced Raman spectroscopy from hollow Au-Ag Alloy nanourchins","volume":"26","author":"Liu","year":"2014","journal-title":"Adv. Mater."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.bios.2014.05.013","article-title":"Nanofabricated SERS-active substrates for single-molecule to virus detection in vitro: A review","volume":"61","author":"Luo","year":"2014","journal-title":"Biosens. Bioelectron."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5980","DOI":"10.1063\/1.1833570","article-title":"Theoretical study of coated spherical metallic nanoparticles for single molecule surface-enhanced spectroscopy","volume":"85","author":"Xu","year":"2004","journal-title":"Appl. Phys. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1143","DOI":"10.1088\/0953-8984\/4\/5\/001","article-title":"Surface-enhanced Raman scattering","volume":"4","author":"Otto","year":"1992","journal-title":"J. Phys. Condens. Matter"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.1021\/nl070625w","article-title":"Electromigrated nanoscale gaps for surface-enhanced Raman spectroscopy","volume":"7","author":"Ward","year":"2007","journal-title":"Nano Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1039\/a800255j","article-title":"3,3\u2032,5,5\u2032-Tetramethylbenzidine as electrochemical substrate for horseradish peroxidase based enzyme immunoassays. A comparative study","volume":"123","author":"Volpe","year":"1998","journal-title":"Analyst"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"011905","DOI":"10.1063\/1.4889850","article-title":"Recyclable surface-enhanced Raman scattering template based on nanoporous gold film\/Si nanowire arrays","volume":"105","author":"Zhang","year":"2014","journal-title":"Appl. Phys. Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5252","DOI":"10.1021\/jp9530459","article-title":"Resonance Raman, surface-enhanced resonance Raman, infrared, and ab initio vibrational spectroscopic study of tetraazaannulenes","volume":"100","author":"Bell","year":"1996","journal-title":"J. Phys. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1021\/ac1024039","article-title":"Quantitative detection of human tumor necrosis factor \u03b1 by a resonance Raman enzyme-linked immunosorbent assay","volume":"83","author":"Laing","year":"2011","journal-title":"Anal. Chem."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/17\/4\/788\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:32:08Z","timestamp":1760207528000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/17\/4\/788"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,4,7]]},"references-count":27,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2017,4]]}},"alternative-id":["s17040788"],"URL":"https:\/\/doi.org\/10.3390\/s17040788","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,4,7]]}}}