Nanozyme-based Aptasensor for colorimetric detection of vitamin D3 in milk

Articles in Press

Document Type : Research Paper

Authors

1 Department of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran

2 Immunogenetics Research Center, Mazandaran University of Medical Sciences, Sari, Iran

3 The Health of Plant and Livestock Products Research Center, Mazandaran University of Medical Sciences, Sari, Iran

10.22038/nmj.2025.83282.2083

Abstract

Objective(s): The growing demand for food production necessitates the development of advanced detection methods to ensure food quality and prevent adulteration. This study presents a colorimetric aptasensor specifically designed to detect vitamin D3 in milk, utilizing the unique properties of nanozymes in combination with aptamers.
Materials and Methods: The method utilizes the peroxidase-like activity of bare iron oxide magnetic nanoparticles, which interact electrostatically with aptamers, leading to a color change in the TMB-H2O2 solution through a Fenton-like redox reaction.
Results: The results demonstrated that both the choice of buffer and the concentration of vitamin D3 significantly impact the catalytic activity of the nanozymes. In addition, using iron oxide nanozymes offers several advantages, including enhanced stability, straightforward interactions, tunable activity, and effective background color removal through magnetic separation.
Conclusions: This innovative approach enhances the reliability of vitamin D3 detection in dairy products and holds broader implications for food safety and quality assurance. The findings highlight the potential of integrating nanotechnology and biosensing techniques to address critical challenges in food monitoring and safety. By tackling these issues, this research contributes to the development of effective strategies for ensuring food integrity and safeguarding consumer health in an era where food adulteration continues to be a pressing concern.

Keywords

References

  1. Murphy SC, Martin NH, Barbano DM, Wiedmann M. Influence of raw milk quality on processed dairy products: how do raw milk quality test results relate to product quality and yield? J Dairy Sci. 2016;99:10128-10149.
  2. Han J, Wang J. Dairy cow nutrition and milk quality. Agriculture. 2023;13:702.
  3. Swar S, Abbas R, Asrar R, Yousuf S, Mehmood A, Shehzad B, Farhan H, Aleem M, Marcelino L, Mohsin M. Milk adulteration and emerging health issues in humans and animals (a review). Contemp Vet J. 2021;1:1-8.
  4. Polzonetti V, Pucciarelli S, Vincenzetti S, Polidori P. Dietary intake of vitamin D from dairy products reduces the risk of osteoporosis. Nutrients. 2020;12:1743.
  5. Mandrioli M, Boselli E, Fiori F, Rodriguez-Estrada MT. Vitamin D3 in high-quality cow milk: an Italian case study. Foods. 2020;9:548.
  6. Trenerry VC, Plozza T, Caridi D, Murphy S. The determination of vitamin D3 in bovine milk by liquid chromatography mass spectrometry. Food Chem. 2011;125:1314-1319.
  7. Rahman A, Rahman M, Hossain M, Jahan M, Jahan N, Bari L. A simple and alternative UV spectrometric method for the estimation of vitamin D3. Microbial Bioact. 2019;2:E098-E105.
  8. Kasalová E, Aufartová J, Krčmová LK, Solichová D, Solich P. Recent trends in the analysis of vitamin D and its metabolites in milk—a review. Food Chem. 2015;171:177-190.
  9. Gupta R, Kaul S, Singh V, Kumar S, Singhal NK. Graphene oxide and fluorescent aptamer-based novel biosensor for detection of 25-hydroxyvitamin D3. Sci Rep. 2021;11:23456.
  10. Wu H, Cheng H, Tian J, Wang X. The determination of vitamin D in fortified milk and other foods by high performance liquid chromatography (HPLC). Se Pu. 1997;15:43-45.
  11. Kamankesh M, Shahdoostkhany M, Mohammadi A, Mollahosseini A. Fast and sensitive low density solvent-based dispersive liquid–liquid microextraction method combined with high-performance liquid chromatography for determining cholecalciferol (vitamin D3) in milk and yogurt drink samples. Anal Methods. 2018;10:975-982.
  12. Sazali NH, Alshishani A, Saad B, Chew KY, Chong MM, Miskam M. Salting-out assisted liquid–liquid extraction coupled with high-performance liquid chromatography for the determination of vitamin D3 in milk samples. R Soc Open Sci. 2019;6:190952.
  13. Citartan M, Gopinath SC, Tominaga J, Tan SC, Tang TH. Assays for aptamer-based platforms. Biosens Bioelectron. 2012;34:1-11.
  14. Lan L, Yao Y, Ping J, Ying Y. Recent progress in nanomaterial-based optical aptamer assay for the detection of food chemical contaminants. ACS Appl Mater Interfaces. 2017;9:23287-23301.
  15. Wang L, Wang R, Wei H, Li Y. Selection of aptamers against pathogenic bacteria and their diagnostics application. World J Microbiol Biotechnol. 2018;34:1-11.
  16. Han K, Liang Z, Zhou N. Design strategies for aptamer-based biosensors. Sensors. 2010;10:4541-4557.
  17. Xu R, Ouyang L, Chen H, Zhang G, Zhe J. Recent advances in biomolecular detection based on aptamers and nanoparticles. Biosensors. 2023;13:474.
  18. Davydova A, Vorobyeva M. Aptamer-based biosensors for the colorimetric detection of blood biomarkers: paving the way to clinical laboratory testing. Biomedicines. 2022;10:1606.
  19. Eilers A, Witt S, Walter J. Aptamer-modified nanoparticles in medical applications. In: Urmann K, Walter JG, eds. Aptamers in biotechnology. Advances in Biochemical Engineering/Biotechnology. Cham: Springer; 2020;161-193.
  20. Melikishvili S, Piovarci I, Hianik T. Advances in colorimetric assay based on AuNPs modified by proteins and nucleic acid aptamers. Chemosensors. 2021;9:281.
  21. Rezaei M. A review of recent advances in iron oxide nanoparticles as a magnetic agent in cancer diagnosis and treatment. Intern Med Today. 2022;28:280-299.
  22. Montiel Schneider MG, Martín MJ, Otarola J, et al. Biomedical applications of iron oxide nanoparticles: current insights progress and perspectives. Pharmaceutics. 2022;14:204.
  23. Farinha P, Coelho JMP, Reis CP, Gaspar MM. A comprehensive updated review on magnetic nanoparticles in diagnostics. Nanomaterials (Basel). 2021;11:3432.
  24. Park JY, Jeong HY, Kim MI, Park TJ. Colorimetric detection system for Salmonella typhimurium based on peroxidase‐like activity of magnetic nanoparticles with DNA aptamers. J Nanomat. 2015;2015:1-9.
  25. Li S, Zhao X, Yu X, Wan Y, Yin M, Zhang W, Cao B, Wang H. Fe3O4 nanozymes with aptamer-tuned catalysis for selective colorimetric analysis of ATP in blood. Anal Chem. 2019;91:14737-14742.
  26. Kim YS, Jurng J. A simple colorimetric assay for the detection of metal ions based on the peroxidase-like activity of magnetic nanoparticles. Sens Actuators B Chem. 2013;176:253-257.
  27. Wu L, Zhou S, Wang G, Yun Y, Liu G, Zhang W. Nanozyme applications: a glimpse of insight in food safety. Front Bioeng Biotechnol. 2021;9:727886.
  28. Liu X, Huang D, Lai C, Qin L, Zeng G, Xu P, Li B, Yi H, Zhang M. Peroxidase‐like activity of smart nanomaterials and their advanced application in colorimetric glucose biosensors. Small. 2019;15:1900133.
  29. Mukherjee A, Ashrafi AM, Svec P, Richtera L, Přibyl J, Brtnický M, Kynicky J, Adam V. The effect of synthesis procedure on hydrogen peroxidase-like catalytic activity of iron oxide magnetic particles. Appl Sci. 2020;10:6756.
  30. Maharjan A, Dikshit PK, Gupta A, Kim BS. Catalytic activity of magnetic iron oxide nanoparticles for hydrogen peroxide decomposition: optimization and characterization. J Chem Technol Biotechnol. 2020;95:2495-2508.
  31. Jo H, Ban C. Aptamer–nanoparticle complexes as powerful diagnostic and therapeutic tools. Exp Mol Med. 2016;48:e230.
  32. Wang L, Zhou H, Hu H, Wang Q, Chen X. Regulation mechanism of ssDNA aptamer in nanozymes and application of nanozyme-based aptasensors in food safety. Foods. 2022;11:544.
  33. Zeng C, Lu N, Wen Y, Liu G, Zhang R, Zhang J, Wang F, Liu X, Li Q, Tang Z. Engineering nanozymes using DNA for catalytic regulation. ACS Appl Mater Interfaces. 2018;11:1790-1799.
  34. Song Y, Qiao J, Liu W, Qi L. Norfloxacin detection based on the peroxidase-like activity enhancement of gold nanoclusters. Anal Bioanal Chem. 2021;413:979-985.
  35. Tian J. Aptamer-based colorimetric detection of various targets based on catalytic Au NPs/Graphene nanohybrids. Sens Bio-Sens Res. 2019;22:100258.
  36. Zhao L, Wang J, Su D, Zhang Y, Lu H, Yan X, Bai J, Gao Y, Lu G. The DNA controllable peroxidase mimetic activity of MoS2 nanosheets for constructing a robust colorimetric biosensor. Nanoscale. 2020;12:19420-19428.
  37. Anusha T, Bhavani KS, Hassan RYA, Brahman PK. Ferrocene tagged primary antibody generates electrochemical signal: An electrochemical immunosensing platform for the monitoring of vitamin D deficiency in clinical samples. Int J Biol Macromol. 2023;239:124269.
  38. Anusha T, Bhavani KS, Kumar JVS, Brahman PK, Hassan RYA. Disposable impedimetric nano-immunochips for the early and rapid diagnosis of Vitamin-D deficiency. Biosens Bioelectron. 2022;10:100124.
  39. Anusha T, Bhavani KS, Shanmukha Kumar JV, Brahman PK, Hassan RYA. Fabrication of electrochemical immunosensor based on GCN-β-CD/Au nanocomposite for the monitoring of vitamin D deficiency. Bioelectrochemistry. 2022;143:107935.
  40. Anusha T, Bhavani KS, Kumar JVS, Brahman PK. Synthesis and characterization of novel lanthanum nanoparticles-graphene quantum dots coupled with zeolitic imidazolate framework and its electrochemical sensing application towards vitamin D3 deficiency. Colloids Surf A Physicochem Eng Asp. 2021;611:125854.
  41. Anusha T, Bhavani KS, Kumar JVS, Bonanni A, Brahman PK. Fabrication of handmade paper sensor based on silver-cobalt doped copolymer-ionic liquid composite for monitoring of vitamin D3 level in real samples. Microchem J. 2021;161:105789.
  42. Cai T, Chen M, Yang J, Tang C, Lu X, Wei Z, Jiang H, Hou Y, Zhao J, Yu P. An AuNPs-based electrochemical aptasensor for the detection of 25-hydroxy vitamin D3. Anal Sci. 2024;40(4):599-607.
  43. Wadhwa S, John AT, Nagabooshanam S, Mathur A, Narang J. Graphene quantum dot-gold hybrid nanoparticles integrated aptasensor for ultra-sensitive detection of vitamin D3 towards point-of-care application. Appl Surf Sci. 2020;521:146427.
  44. Lee BH, Nguyen VT, Gu MB. Highly sensitive detection of 25-Hydroxyvitamin D3 by using a target-induced displacement of aptamer. Biosens Bioelectron. 2017;88:174-178.
  45. Liu B, Liu J. Accelerating peroxidase mimicking nanozymes using DNA. Nanoscale. 2015;7:13831-13835.
  46. Hizir MS, Top M, Balcioglu M, Rana M, Robertson NM, Shen F, Sheng J, Yigit MV. Multiplexed activity of perAuxidase: DNA-capped AuNPs act as adjustable peroxidase. Anal Chem. 2016;88:600-605.

 

Nanomedicine Journal

Articles in Press, Accepted Manuscript
Available Online from 08 July 2025

Files

Share

How to cite

Statistics