DNA sensor based on carbon black/Methylene Blue arylamine analogue composite for doxorubicin detection

Dmitry Stoikov; Kamila Karaguzina; Dominika Kappo; Pavel Padnya; Ivan Stoikov

doi:10.15826/chimtech.9234

DNA sensor based on carbon black/Methylene Blue arylamine analogue composite for doxorubicin detection

Dmitry Stoikov, Kamila Karaguzina, Dominika Kappo, Pavel Padnya, Ivan Stoikov

Abstract

In modern analytical chemistry, various methods are being actively developed to simplify and reduce the cost of electrochemical sensor fabrication, thereby increasing the accessibility of these technologies for a wide range of users, including medical institutions, industrial enterprises, and research laboratories. In this study, we present a DNA sensor based on a screen‑printed carbon electrode modified with a carbon black suspension in propylene carbonate and diffusion‑free 3,7‑bis(4‑aminophenylamino)phenothiazin‑5‑ium chloride for doxorubicin detection. The sensor operates via drop‑based analysis using a printed graphite electrode, which ensures compactness and enhanced practical usability. The addition of carbon black to the SPСE surface significantly enhances the signal of diffusion‑free phenothiazine in buffer solution, thereby improving both sensitivity and the quality of the analytical signal during biospecific analyte–DNA interactions. The specificity of this interaction was confirmed by substituting sodium polystyrene sulfonate for DNA. The sensor demonstrated high sensitivity in detecting doxorubicin both in buffer solutions and in model biological fluid samples containing high levels of interfering components. The detectable doxorubicin concentration ranges from 1 pM to 1 μM, with a limit of detection of 0.8 pM. The developed DNA sensor offers several advantages that render it promising for clinical applications, including minimal reagent requirements, high sensitivity, and ease of use.

Keywords

DNA sensor; biosensor; doxorubicin; phenothiazine; carbon black; amperometry

Full Text:

PDF

References

Singh R, Gupta R, Bansal D, Bhateria R, Sharma M. A review on recent Trends and future Developments in electrochemical Sensing. ACS Omega. 2024;9(7):7336-7356. doi:10.1021/acsomega.3c08060

Electrochemical sensors market size and share analysis – growth trends and forecasts (2025-2032). Report by Coherent Market Insights [Internet]. 2025[cited 2025];165. English. Available from: https://www.coherentmarketinsights.com/industry-reports/electrochemical-sensors-market, Accessed on 27 October 2025.

Mollarasouli F, Zor E, Ozcelikay G, Ozkan SA. Magnetic nanoparticles in developing electrochemical sensors for pharmaceutical and biomedical applications. Talanta. 2021;226:122108. doi:10.1016/j.talanta.2021.122108

Liu D, Wang J, Wu L, Huang Y, Zhang Y, Zhu M, Wang Y, Zhu Z, Yang C. Trends in miniaturized biosensors for point-of-care testing. TrAC Trends Anal Chem. 2020;122:115701. doi:10.1016/j.trac.2019.115701

Feuz L, Jönsson P, Jonsson MP, Höök F. Improving the limit of detection of nanoscale Sensors by directed Binding to High-sensitivity Areas. ACS Nano. 2010;4(4):2167-2177. doi:10.1021/nn901457f

Liu W, Zhang Z, Geng X, Tan R, Xu S, Sun L. Electrochemical sensors for plant signaling molecules. Biosens Bioelectron. 2025;267:116757. doi:10.1016/j.bios.2024.116757

Ye C, Lukas H, Wang M, Lee Y, Gao W. Nucleic acid-based wearable and implantable electrochemical sensors. Chem Soc Rev. 2024;53(15):7960-7982. doi:10.1039/D4CS00001C

Moulahoum H, Ghorbanizamani F. The LOD paradox: when lower isn't always better in biosensor research and development. Biosens Bioelectron. 2024;264:116670. doi:10.1016/j.bios.2024.116670

Karimian P, Johnston E, Kasprzak A, Liu JW, Stockwell S, Vanhercke T, Hartley CJ. Use of a dual biosensor for identification of novel secretion signal peptides and efficient screening of precision fermentation production strains. Future Foods. 2025;11:100640. doi:10.1016/j.fufo.2025.100640

Shah RS, Singh RK, Kumar R. Development of 2D Materials-based surface Plasmon resonance Biosensor for the detection of healthy and pathological Tissues. Plasmonics. 2024;20(7):4811-4820. doi:10.1007/s11468-024-02608-7

Hsu CY, Rizaev JA, Pallathadka H, Mansouri S, Bokov DO, Sharma S, Rathore G, Rajput P, Mustafa YF, Abosaoda MK. A review of new emerging biosensors based on bacteria-imprinted polymers towards pathogenic bacteria: promising new tools for selective detection. Microchem J. 2024;207:111918. doi:10.1016/j.microc.2024.111918

Jia Y, Chen S, Wang Q, Li J. Recent progress in biosensor regeneration techniques. Nanoscale. 2024;16(6):2834-2846. doi:10.1039/d3nr05456j

Kwak H, Lee E, Karki R. DNA sensors in metabolic and cardiovascular diseases: molecular mechanisms and therapeutic prospects. Immunol Rev. 2025;329(1):13382. doi:10.1111/imr.13382

Sun W, Qin P, Gao H, Li G, Jiao K. Electrochemical DNA biosensor based on chitosan/nano-V2O5/MWCNTs composite film modified carbon ionic liquid electrode and its application to the LAMP product of yersinia enterocolitica gene sequence. Biosens Bioelectron. 2010;25(6):1264-1270. doi:10.1016/j.bios.2009.10.011

Low KF, Karimah A, Yean CY. A thermostabilized magnetogenosensing assay for DNA sequence-specific detection and quantification of vibrio cholerae. Biosens Bioelectron. 2013;47:38-44. doi:10.1016/j.bios.2013.03.004

Verma R, Sood S, Singh R, Sumana G, Bala M, Sharma VK, Samantaray JC, Pandey RM, Malhotra BD. Coupling electrochemical response of a DNA biosensor with PCR for neisseria gonorrhoeae detection. Diagn Microbiol Infect Dis. 2014;78(1):16-23. doi:10.1016/j.diagmicrobio.2013.09.010

Manzano M, Viezzi S, Mazerat S, Marks RS, Vidic J. Rapid and label-free electrochemical DNA biosensor for detecting hepatitis A virus. Biosens Bioelectron. 2018;100:89-95. doi:10.1016/j.bios.2017.08.043

Bacchu MS, Ali MR, Das S, Akter S, Sakamoto H, Suye SI, Rahman MM, Campbell K, Khan MZH. A DNA functionalized advanced electrochemical biosensor for identification of the foodborne pathogen salmonella enterica serovar typhi in real samples. Anal Chim Acta. 2022;1192:339332. doi:10.1016/j.aca.2021.339332

Zhang C, Xu S, Zhang X, Huang D, Li R, Zhao S, Wang B. Electrochemical detection of specific DNA sequences related to bladder cancer on CdTe quantum dots modified glassy carbon electrode. J Electroanal Chem. 2014;735:115-122. doi:10.1016/j.jelechem.2014.09.014

Benvidi A, Dehghani Firouzabadi A, Dehghan Tezerjani M, Moshtaghiun SM, Mazloum-Ardakani M, Ansarin A. A highly sensitive and selective electrochemical DNA biosensor to diagnose breast cancer. J Electroanal Chem. 2015;750:57-64. doi:10.1016/j.jelechem.2015.05.002

Wheeler OPG, Unterholzner L. DNA sensing in cancer: Pro-tumour and anti-tumour functions of cGAS–STING signalling. Essays Biochem. 2023;67(6):905-917. doi:10.1042/EBC20220241

Gao Y, Xiong M, Gong C, Wang B, Bai L, Zhang XB. Smart DNA sensors‐based molecular identification for cancer subtyping. Smart Mol. 2023;1(3):20230020. doi:10.1002/smo.20230020

Chiorcea-Paquim AM, Oliveira-Brett AM. DNA electrochemical Biosensors for in Situ probing of pharmaceutical Drug oxidative DNA Damage. Sensors. 2021;21(4):1125. doi:10.3390/s21041125

Ulianas A, Heng LY, Ahmad M, Lau HY, Ishak Z, Ling TL. A regenerable screen-printed DNA biosensor based on acrylic microsphere–gold nanoparticle composite for genetically modified soybean determination. Sens Actuators B Chem. 2014;190:694-701. doi:10.1016/j.snb.2013.09.040

Bisht A, Avinash D, Sahu KK, Patel P, Das Gupta G, Kurmi BD. A comprehensive review on doxorubicin: mechanisms, toxicity, clinical trials, combination therapies and nanoformulations in breast cancer. Drug Deliv Transl Res. 2025;15(1):102-103. doi:10.1007/s13346-024-01648-0

Linders AN, Dias IB, López Fernández T, Tocchetti CG, Bomer N, Van der Meer P. A review of the pathophysiological mechanisms of doxorubicin-induced cardiotoxicity and aging. NPJ Aging. 2024;10(1):9. doi:10.1038/s41514-024-00135-7

Luo S, Wu Q, Wang L, Qu H, Zheng L. Direct detection of doxorubicin in whole blood using a hydrogel-protected electrochemical aptamer-based biosensor. Talanta. 2025;285:112854. doi:10.1016/j.talanta.2024.127289

Sharifi J, Fayazfar H. Highly sensitive determination of doxorubicin hydrochloride antitumor agent via a carbon nanotube/gold nanoparticle based nanocomposite biosensor. Bioelectrochem. 2021;139:107741. doi:10.1016/j.bioelechem.2021.107741

Taei M, Salavati H, Hasanpour F, Shafiei A. Biosensor based on ds-DNA-decorated Fe2O3/SnO2-chitosan Modified multiwalled Carbon nanotubes for biodetection of Doxorubicin. IEEE Sens J. 2016;16(1):24-31. doi:10.1109/JSEN.2015.2474262

Taei M, Hasanpour F, Salavati H, Mohammadian S. Fast and sensitive determination of doxorubicin using multi-walled carbon nanotubes as a sensor and CoFe2O4 magnetic nanoparticles as a mediator. Microchim Acta. 2016;183(1):49-56. doi:10.1007/s00604-015-1588-3

Guo Y, Chen Y, Zhao Q, Shuang S, Dong C. Electrochemical sensor for ultrasensitive Determination of doxorubicin and methotrexate Based on Cyclodextrin‐graphene Hybrid Nanosheets. Electroanal. 2011;23(10):2400-2407. doi:10.1002/elan.201100259

Bergamini MF, Santos DP, Zanoni MVB. Determination of isoniazid in human urine using screen-printed carbon electrode modified with poly-l-histidine. Bioelectrochem. 2010;77(2):133-138. doi:10.1016/j.bioelechem.2009.07.010

Porfireva A, Evtugyn G. Electrochemical DNA sensor Based on the copolymer of proflavine and azure B for doxorubicin Determination. Nanomater. 2020;10(5):924. doi:10.3390/nano10050924

Soleymani J, Hasanzadeh M, Eskandani M, Khoubnasabjafari M, Shadjou N, Jouyban A. Electrochemical sensing of doxorubicin in unprocessed whole blood, cell lysate, and human plasma samples using thin film of poly-arginine modified glassy carbon electrode. Mater Sci Eng C. 2017;77:790-802. doi:10.1016/j.msec.2017.03.257

Kulikova T, Porfireva A, Evtugyn G, Hianik T. Electrochemical DNA sensors with layered Polyaniline—DNA coating for detection of specific DNA Interactions. Sensors. 2019;19(3):469. doi:10.3390/s19030469

Stoikov DI, Porfir’eva AV, Shurpik DN, Stoikov II, Evtyugin GA. Electrochemical DNA sensors on the basis of electropolymerized thionine and azure B with addition of pillar[5]arene as an electron transfer mediator. Russ Chem Bull. 2019;68(2):431-437. doi:10.1007/s11172-019-2404-8

Stoikov D, Ivanov A, Shurpik D, Stoikov I, Evtugyn G. Flow-through Electrochemical biosensor with a replaceable Enzyme reactor and Screen-printed Electrode for the determination of uric Acid and Tyrosine. Anal Lett. 2022;55(8):1281-1295. doi:10.1080/00032719.2021.2000621

Ivanov A, Stoikov D, Shafigullina I, Shurpik D, Stoikov I, Evtugyn G. Flow-through Acetylcholinesterase sensor with replaceable Enzyme Reactor. Biosensors. 2022;12(9):676. doi:10.3390/bios12090676

Padnya PL, Khadieva AI, Stoikov II. Current achievements and perspectives in synthesis and applications of 3,7-disubstituted phenothiazines as methylene Blue analogues. Dyes Pigments. 2023;208:110806. doi:10.1016/j.dyepig.2022.110806

Kuzin YI, Padnya PL, Stoikov II, Gorbatchuk VV, Stoikov DI, Khadieva AI, Evtugyn GA. Electrochemical behavior of the monomeric and polymeric forms of N-phenyl-3-(phenylimino)-3H-phenothiazin-7-amine. Electrochim Acta. 2020;345:136195. doi:10.1016/j.electacta.2020.136195

Stoikov D, Ivanov A, Shafigullina I, Gavrikova M, Padnya P, Shiabiev I, Stoikov I, Evtugyn G. Flow-through Amperometric biosensor System based on functionalized Aryl derivative of phenothiazine and PAMAM-Calix-dendrimers for the determination of uric Acid. Biosensors. 2024;14(3):120. doi:10.3390/bios14030120

Stoikov D, Shafigullina I, Shurpik D, Stoikov I, Evtugyn G. A Flow-through Biosensor system Based on Pillar[3]Arene[2]quinone and ferrocene for determination of hydrogen Peroxide and uric Acid. Chemosensors. 2024;12(6):98. doi:10.3390/chemosensors12060098

Khadieva A, Mostovaya O, Padnya P, Kalinin V, Grishaev D, Tumakov D, Stoikov I. Arylamine analogs of methylene Blue: substituent Effect on aggregation Behavior and DNA Binding. Int J Mol Sci. 2021;22(11):5847. doi:10.3390/ijms22115847

Kuzin YI, Khadieva AI, Padnya PL, Khannanov AA, Kutyreva MP, Stoikov II, Evtugyn GA. Electrochemistry of new derivatives of phenothiazine: electrode kinetics and electropolymerization conditions. Electrochim Acta. 2021;375:137985. doi:10.1016/j.electacta.2021.137985

Parham H. Determination of isosorbide dinitrate in arterial plasma, synthetic serum and pharmaceutical formulations by linear sweep voltammetry on a gold electrode. Talanta. 2001;55(2):255-262. doi:10.1016/s0039-9140(01)00416-7

Mazzaracchio V, Tomei MR, Cacciotti I, Chiodoni A, Novara C, Castellino M, Scordo G, Amine A, Moscone D, Arduini F. Inside the different types of carbon black as nanomodifiers for screen-printed electrodes. Electrochim Acta. 2019;317:673-683. doi:10.1016/j.electacta.2019.05.117

Wang C, Wang T, Li Z, Xu X, Zhang X, Li D. An electrochemical Enzyme biosensor for ammonium Detection in aquaculture Using Screen-printed Electrode modified by gold Nanoparticle/polymethylene Blue. Biosensors. 2021;11(9):335. doi:10.3390/bios11090335

Vongsetskul T, Taylor DJF, Zhang J, Li PX, Thomas RK, Penfold J. Interaction of a cationic Gemini surfactant with DNA and with sodium Poly(styrene sulphonate) at the Air/water Interface: A neutron Reflectometry Study. Langmuir. 2009;25(7):4027-4035. doi:10.1021/la802816s

Li M, Zhang Y, Wu B, Qiu R, Zhao C, Chen B, Shang H. Optimizing dose selection for doxorubicin-induced cardiotoxicity in mice: A comprehensive analysis of single and multiple-dose regimens. Eur J Pharmacol. 2025;1003:177883. doi:10.1016/j.ejphar.2025.177883

Zhang C, Zhou X, Yan F, Lin J. N-doped Graphene quantum Dots confined within silica Nanochannels for enhanced Electrochemical detection of Doxorubicin. Molecules. 2023;28(18):6443. doi:10.3390/molecules28186443

Hassani Moghadam F, Taher MA, Karimi-Maleh H. Doxorubicin anticancer Drug monitoring by ds-DNA-based Electrochemical biosensor in clinical Samples. Micromachines. 2021;12(7):808. doi:10.3390/mi12070808

Peng A, Xu H, Luo C, Ding H. Application of a disposable Doxorubicin sensor for direct Determination of clinical Drug concentration in patient Blood. Int J Electrochem Sci. 2016;11(7):6266-6278. doi:10.20964/2016.07.38

Ghanbari MH, Norouzi Z. A new nanostructure consisting of nitrogen-doped carbon nanoonions for an electrochemical sensor to the determination of doxorubicin. Microchem J. 2020;157:105098. doi:10.1016/j.microc.2020.105098

Sun S, Xu X, Niu A, Sun Z, Zhai Y, Li S, Xuan C, Zhou Y, Yang X, Zhou T, Tian Q. Novel electrochemical Sensor based on acetylene Black for the determination of doxorubicin in serum Samples. Int J Electrochem Sci. 2022;17(11):221187. doi:10.20964/2022.11.82

DOI: https://doi.org/10.15826/chimtech.9234

Copyright (c) 2025 Dmitry Stoikov, Kamila Karaguzina, Dominika Kappo, Pavel Padnya, Ivan Stoikov

This work is licensed under a Creative Commons Attribution 4.0 International License.

Chimica Techno Acta, 2014–2025
eISSN 2411-1414
Copyright Notice

Username
Password
Remember me