ENZYME-FREE COLOURIMETRIC SENSOR BASED ON ZIRCONIUM-BASED MOF FOR THE DETECTION OF MALATHION
Keywords:
Zirconium-based MOF, Nanoenzyme, Enzyme-free colourimetric sensor, MalathionAbstract
To address the issues of enzyme inactivation, complex procedures, and dependence on large-scale equipment in traditional organophosphorus pesticide detection, we developed an enzyme-free colourimetric sensor employing Hemin@UiO-66 composite nanomaterials for the rapid and accurate detection of malathion. The UiO-66 metal-organic framework was synthesised using the solvothermal method, while the Hemin@UiO-66nanoenzyme was produced through post-modification with hemin chloride (Hemin). The structural characteristics of the material were analysed using Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS).Thisnanoenzyme demonstrated remarkable peroxidase-like activity, effectively catalysing the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to blue-coloured oxidised TMB (oxTMB) in the presence of hydrogen peroxide (H₂O₂). Malathion interacts with the active sites of the material via Zr-O-P coordination, which significantly inhibits the catalytic activity of thenanoenzyme. As a result, the colour of the system gradually diminished with increasing malathion concentration. Under optimised conditions, a strong linear correlation was established between the change in absorbance of the sensor and malathion concentration, spanning from 5 to 270 ng/mL, with a detection limit as low as 4.23 ng/mL (3σ) and a detection time of 20 minutes. This method exhibited excellent selectivity, stability, repeatability, and anti-interference capabilities. The recovery rates of malathion in spiked real samples ranged from 95.3% to 104.7%, accompanied by a relative standard deviation (RSD) of less than 4.2%. By eliminating the need for biological enzymes, this sensor facilitates straightforward operation and rapid response, thereby providing an innovative approach for the on-site rapid detection of malathion residues in food.References
[1] Mali H, Shah C, Raghunandan BH. Organophosphate pesticides an emerging environmental contaminant: pollution, toxicity, bioremediation progress, and remaining challenges. J. Environ. Sci., 2023, 127: 234-250.
[2] Serafim LF, Wang L, Rathee P, et al. Remediation of environmentally hazardous organophosphates by artificial metalloenzymes. Curr. Opin. Green Sustain. Chem., 2021, 32: 100529.
[3] Durgadevi P, Girigoswami K, Harini K, et al. Silent threats of organophosphates: surging pollutants, harmful impacts, remediation strategies, and viable eco-friendly alternatives. Toxicol. Environ. Health Sci., 2024, 17(1): 23-49.
[4] Lamb RW, McAlexander H, Woodley CM, et al. Towards a comprehensive understanding of malathion degradation: theoretical investigation of degradation pathways and related kinetics under alkaline conditions. Environ. Sci. Process. Impacts, 2021, 23(8): 1231-1241.
[5] Kaushal J, Khatri M, Arya SK. A treatise on organophosphate pesticide pollution: current strategies and advancements in their environmental degradation and elimination. Ecotoxicol. Environ. Saf., 2021, 207: 111483.
[6] Lamb RW, McAlexander H, Woodley CM, et al. Towards a comprehensive understanding of malathion degradation: comparison of degradation reactions under alkaline and radical conditions. Environ. Sci. Process. Impacts, 2022, 24(7): 1026-1036.
[7] Liu Y, Xu Y, Yuan B, et al. Bioaccumulation mediated by water solubility leads to differences in the acute toxicity of organophosphorus insecticides to zebrafish (Danio rerio). Ecotoxicology, 2024, 33(7): 750-761.
[8] Ramadori GP. Organophosphorus poisoning: acute respiratory distress syndrome (ARDS) and cardiac failure as cause of death in hospitalized patients. Int. J. Mol. Sci., 2023, 24(8): 7168.
[9] Rafacho ML. Acute and chronic effects of the organophosphate malathion on the pancreatic α and β cell viability, cell structure, and voltage-gated K+ currents. Environ. Toxicol. Pharmacol., 2022, 94: 103929.
[10] Kaur R, Choudhary D, Bali S, et al. Pesticides: an alarming detrimental to health and environment. Sci. Total Environ., 2024, 915: 113-170.
[11] Wang C. Selective and sensitive detection of malathion pesticide with CdSe quantum dots as ligand-exchange probes. Environ. Eng. Sci., 2021, 38(10): 920-928.
[12] Khatoon R, Uddin R, Khurshid S, et al. Determination of lambda-cyhalothrin and malathion residues in locust by gas chromatography with electron capture detection. J. Anal. Chem., 2022, 77(5): 611-617.
[13] Michlig N, Amirav A, Neumark B, et al. Comparison of different fast gas chromatography-mass spectrometry techniques (Cold EI, MS/MS, and HRMS) for the analysis of pyrethroid insecticide residues in food. Anal. Methods, 2024, 16(32): 5599-5618.
[14] Garvey J, Margalit A, Kelly M, et al. A method for the quantitative analysis of polar anionic pesticides in milk/infant formula, cereals and fruit and vegetables using ion chromatography coupled to tandem mass spectrometry. Anal. Methods, 2024, 16(23): 3692-3700.
[15] Ziani I, Bouakline H, Guerraf AE, et al. Integrating AI and advanced spectroscopic techniques for precision food safety and quality control. Trends Food Sci. Technol., 2025, 156: 104850.
[16] Sestak J, Guttman A, Lavicka J. Fluorescence detection setups in capillary electrophoresis and microscale liquid chromatography: developments over the past decade. TrAC, Trends Anal. Chem., 2024, 181: 118001.
[17] Patel M, Agrawal M, Srivastava A. Signal amplification strategies in electrochemical biosensors via antibody immobilization and nanomaterial-based transducers. Mater. Adv., 2022, 3(24): 8864-8885.
[18] Marrazza G, Ramalingam M, Jaisankar A, et al. Advancements and emerging technologies in biosensors for rapid and accurate virus detection. TrAC, Trends Anal. Chem., 2024, 172: 117609.
[19] Altan EA. Nanomaterial interfaces designed with different biorecognition elements for biosensing of key foodborne pathogens. Compr. Rev. Food Sci. Food Saf., 2023, 22(4): 3151-3184.
[20] Zlobin A, Smirnov I, Golovin A. Dynamic interchange between two protonation states is characteristic of active sites of cholinesterases. Protein Sci., 2024, 33(2): e4894.
[21] Alves AA, Furtado DA, de Oliveira RR, et al. Mechanisms of organophosphate toxicity and the role of acetylcholinesterase inhibition. Toxics, 2023,11(6): 512.
[22] Holyavka MG, Artyukhov VG. Prospects and problems in enzyme immobilization methodology: comprehensive review. Biophys. Rev., 2025.
[23] Shahbaz A, Hussain N, Intisar A, et al. Immobilized enzymes-based biosensing cues for strengthening biocatalysis and biorecognition. Catal. Lett., 2021, 152(9): 2637-2649.
[24] Bolivar JM, Woodley JM, Fernandez-Lafuente R. Is enzyme immobilization a mature discipline? Some critical considerations to capitalize on the benefits of immobilization. Chem. Soc. Rev., 2022, 51(15): 6251-6290.
[25] Jiang Y, Zheng J, Wang M, et al. Pros and cons in various immobilization techniques and carriers for enzymes. Appl. Biochem. Biotechnol., 2024, 196(9): 5633-5655.
[26] Bie J, Sepodes B, Ribeiro MHL. Enzyme immobilization and co-immobilization: main framework, advances and some applications. Processes, 2022, 10(1): 1-20.
[27] Karadurmus L, Kaya IS, Ozkan SA. Recent advances of enzyme biosensors for pesticide detection in foods. J. Food Meas. Charact., 2021, 15(5): 4582-4595.
[28] Abedeen MZ, Sharma M, Kushwaha HS, et al. Sensitive enzyme-free electrochemical sensors for the detection of pesticide residues in food and water. TrAC, Trends Anal. Chem., 2024, 176: 117729.
[29] Majdinasab M, Daneshi M, Marty JL. Recent developments in non-enzymatic (bio)sensors for detection of pesticide residues: focusing on antibody, aptamer and molecularly imprinted polymer. Talanta, 2021, 232: 122397.
[30] Praharaj C, Nara S. Nanotechnology driven biorecognition element and label free sensing of pesticides. J. Environ. Chem. Eng., 2024, 12(2): 112168.
[31] Jeyachandran S, Srinivasan R, Ramesh T, et al. Recent development and application of nanozyme artificial enzymes-a review. Biomimetics, 2023, 8(5): 1-20.
[32] Bilal M, Khaliq N, Ashraf M, et al. Enzyme mimic nanomaterials as nanozymes with catalytic attributes. Colloids Surf. B: Biointerfaces, 2023, 221: 112950.
[33] Abdullah KA, Tahir TF, Qader AF, et al. Nanozymes: classification and analytical applications - a review. J. Fluoresc., 2024, 34(5): 1987-2004.
[34] Cedrun-Morales M, Ceballos M, Polo E, et al. Nanosized metal-organic frameworks as unique platforms for bioapplications. Chem. Commun., 2023, 59(20): 2887-2896.
[35] Wang D, Yao H, Ye J, et al. Metal-organic frameworks (MOFs): classification, synthesis, modification, and biomedical applications. Small, 2024, 20(47): 2404350.
[36] Udourioh GA, Solomon MM, Matthews-Amune CO, et al. Current trends in the synthesis, characterization and application of metal-organic frameworks. React. Chem. Eng., 2022, 8(2): 278-310.
[37] Zhao R, Scott TR, Schmid J, et al. Cross-aldol condensation on missing linker sites of metal-organic framework UiO-66. J. Catal., 2025, 448: 116204.
[38] Guo J, Liu Y, Zha J, et al. Enhancing the peroxidase-mimicking activity of hemin by covalent immobilization in polymer nanogels. Polym. Chem., 2021, 12(6): 866-858.
[39] Sun H, Wu H, Teng Q, et al. Enzyme-mimicking materials from designed self-assembly of lysine-rich peptides and G-quadruplex DNA/hemin DNAzyme: charge effect of the key residues on the catalytic functions. Biomacromolecules, 2022, 23(8): 3476-3489.
[40] Lai S, Yang D, Wang Y, et al. Artificial peroxidase of short peptide and hemin co-assemblies with selective chiral catalytic activity in DOPA oxidation. Colloids Surf. A: Physicochem. Eng. Asp., 2023, 665: 131257.
[41] Cunha-Silva AV, de Sousa IR, de Oliveira LG, et al. Influence of UiO-66(Zr) preparation strategies in its catalytic efficiency for desulfurization process. Materials, 2019, 12(10): 1610.
[42] Richezzi M, Donnarumma RP, Howarth AJ. Developments in the discovery, synthesis, and characterization of RE(III)-UiO-66 and its structural analogues. CrystEngComm, 2025, 27(8): 1062-1075.
[43] Li Z, Deng X, Hong X, et al. Nanozyme based on dispersion of hemin by graphene quantum dots for colourimetric detection of glutathione. Molecules, 2022, 27(15): 1-12.
[44] Chen S, Guo J, Sun Q, et al. Enhanced electrochemical sensor based on Uio-66-NH2 carbon nanotubes hybrid for selective detection of ofloxacin. Mater. Today Chem., 2024, 42: 102441.
[45] Li X, Liu P, Niu X, et al. Tri-functional Fe-Zr bi-metal-organic frameworks enable high-performance phosphate ion ratiometric fluorescent detection. Nanoscale, 2020, 12(37): 19383-19389.
[46] Aledo JC. Enzyme kinetic parameters estimation: a tricky task? Biochem. Mol. Biol. Educ., 2021, 49(4): 514-515.
[47] Dong J. On catalytic kinetics of enzymes. Processes, 2021, 9(5): 1-8.