Copyright (c) 2026 Chamaraja N A, Praveen Kumar C B, Mahesh B, Kumara Swamy N, Kavya H V
This work is licensed under a Creative Commons Attribution 4.0 International License.
Sensitive Colorimetric Determination of Hydrogen Peroxide and Glucose using Ammonium Metavanadate as Chromogen: Applications in Human Urine Samples
Corresponding Author(s) : N.A. Chamaraja
Asian Journal of Chemistry,
Vol. 38 No. 10 (2026): Vol 38, Issue 10, 2026
Abstract
A rapid and precise enzymatic method was developed for the determination of hydrogen peroxide (H2O2) and glucose using acidified ammonium metavanadate (NH4VO3) as a chromogenic reagent. In this method, H2O2 is generated in situ through the glucose oxidase (GOD)-catalyzed oxidation of glucose and subsequently reacts with vanadate to produce a red-orange coloured complex exhibiting maximum absorbance at 450 nm. The method showed a linear response for H2O2 over the concentration range of 0.67-86 µmol L–1. For glucose determination, linear ranges of 20-1040 µmol L–1 and 5-925 µmol L–1 were obtained using rate-based and single-point measurement methods, respectively. The Michaelis-Menten kinetic parameters (Km and Vmax) were determined to be 3453 µmol L–1 and 0.9582 µmol min–1, respectively. The within-day precision ranged from 0.25-0.76% (n = 10), whereas the between-day precision was 0.47-1.98% (n = 20). Glucose recoveries of 98.9-100.4% were obtained in the analyzed samples, with no appreciable influence from commonly encountered urinary constituents. The measured accuracy fell within the range of 90-102%. The method provided limits of detection and quantification of 0.268 and 0.894 µmol L–1, respectively, for glucose. The proposed scheme is a quick and effective for quantifying glucose in human urine samples with a single enzyme, hence the method is possibly intended for use by clinical laboratories.
Keywords
Download Citation
Endnote/Zotero/Mendeley (RIS)BibTeX
- D. Sim, M.C. Brothers, J.M. Slocik, A.E. Islam, B. Maruyama, C.C. Grigsby, R.R. Naik and S.S. Kim, Adv. Sci., 9, 2104426 (2022); https://doi.org/10.1002/advs.202104426
- P. González, P. Lozano, G. Ros and F. Solano, Int. J. Mol. Sci., 24, 9352 (2023); https://doi.org/10.3390/ijms24119352
- T. Yang, F. Qi, F. Guo, M. Shao, Y. Song, G. Ren, Z. Linlin, G. Qin and Y. Zhao, Mol. Med., 30, 71 (2024); https://doi.org/10.1186/s10020-024-00824-9
- R.M. Carrillo‐Larco, W.C. Guzman‐Vilca, X. Xu and A. Bernabe‐Ortiz, Diabet. Med., 41, e15174 (2024); https://doi.org/10.1111/dme.15174
- N.A. Chamaraja, M. Basavaraju and N.K. Swamy, Anal. Biochem., 590, 113536 (2020); https://doi.org/10.1016/j.ab.2019.113536
- V. Vallon and T. Nakagawa, Compr. Physiol., 12, 2995 (2022); https://doi.org/10.1002/j.2040-4603.2022.tb00207.x
- H. Sohrabi, F. Maleki, P. Khaaki, M. Kadhom, N. Kudaibergenov and A. Khataee, Biosensors, 13, 347 (2023); https://doi.org/10.3390/bios13030347
- M. Harun-Or-Rashid, M.N. Aktar, V. Preda and N. Nasiri, Sens. Diagn., 3, 893 (2024); https://doi.org/10.1039/D4SD00086B
- Z. Li, W. Zeng and Y. Li, Molecules, 28, 4891 (2023); https://doi.org/10.3390/molecules28134891
- H. Khosravi Ardakani, M. Gerami, M. Chashmpoosh, N. Omidifar and A. Gholami, Biochem. Res. Int., 2022, 2964705 (2022); https://doi.org/10.1155/2022/2964705
- S. Fang, S. Li, P. Yin, G. Yao, H. Yu, Y. He, X. Li, M. Yang and W. Tan, Chemosphere, 364, 143171 (2024); https://doi.org/10.1016/j.chemosphere.2024.143171
- Z. Yuan, M. Fu, X. Wang, M. Wang, Y. Wei, Y. Sun, Q. Zhang, Y. Zhang and B. Zhang, Anal. Methods, 17, 320 (2025); https://doi.org/10.1039/D4AY01878H
- W. Yao, X. Zhang and Z. Lin, Spectrochim. Acta A Mol. Biomol. Spectrosc., 265, 120401 (2022); https://doi.org/10.1016/j.saa.2021.120401
- I. Rahmawati, Y. Einaga, T.A. Ivandini and A. Fiorani, ChemElectroChem, 9, e202200175 (2022); https://doi.org/10.1002/celc.202200175
- J. Xu, M. Back and S. Tanabe, in eds.: R.-S. Liu and X.-J. Wang, Near-Infrared Phosphors with Persistent Luminescence over 1000 nm for Optical Imaging, In: Phosphor Handbook: Experimental Methods for Phosphor Evaluation and Characterization, CRC Press, edn 3, pp. 363-418 (2022).
- N.R. Jalal, F. Mollarasouli, M.R. Jalali Sarvestani, S. Khalili, S. Asadi, Z. Derakhshan, T. Madrakian, A. Afkhami and M. Ahmadi, in eds.: M. Thompson and Z. Ramezani, Quantum Dots in Medical Detection/ Diagnosis, In: Quantum Dots in Bioanalytical Chemistry and Medicine, Royal Society of Chemistry: Cambridge, U.K., vol. 22, pp. 75-106 (2023).
- W. Zheng, P. Huang, D. Tu, E. Ma, H. Zhu and X. Chen, Chem. Soc. Rev., 44, 1379 (2015); https://doi.org/10.1039/C4CS00178H
- L. Tong, L. Wu, Y. Zai, Y. Zhang, E. Su and N. Gu, Biosens. Bioelectron., 219, 114787 (2023); https://doi.org/10.1016/j.bios.2022.114787
- P.C. Lee, N.S. Li, Y.P. Hsu, C. Peng and H.W. Yang, Analyst, 144, 3038 (2019); https://doi.org/10.1039/C8AN02440E
- M. Schröder and F.J. Meyer-Almes, ChemTexts., 11, 14 (2025); https://doi.org/10.1007/s40828-025-00211-9
- S.Ü. Pektaş, M. Keskin, O.C. Bodur and F. Arslan, J. Food Compos. Anal., 129, 106133 (2024); https://doi.org/10.1016/j.jfca.2024.106133
- M.H. Hadwan and S. Ali, Anal. Biochem., 542, 29 (2018); https://doi.org/10.1016/j.ab.2017.11.013
- S. Naveen Prasad, P. Weerathunge, M.N. Karim, S. Anderson, S. Hashmi, P.D. Mariathomas, V. Bansal and R. Ramanathan, Anal. Bioanal. Chem., 413, 1279 (2021); https://doi.org/10.1007/s00216-020-03090-w
- S. Ma, C. Wei, Y. Bao, Y. Liu, H. Jiang, W. Tong, D. Chen and X. Huang, Mikrochim. Acta, 191, 107 (2024); https://doi.org/10.1007/s00604-023-06150-x
- D.-M. Kim, J.-M. Moon, W.-C. Lee, J.-H. Yoon, C.S. Choi and Y.-B. Shim, Biosens. Bioelectron., 91, 276 (2017); https://doi.org/10.1016/j.bios.2016.12.046
- Y. Yang, C. Yi, J. Luo, R. Liu, J. Liu, J. Jiang and X. Liu, Biosens. Bioelectron., 26, 2607 (2011); https://doi.org/10.1016/j.bios.2010.11.015
- Riyanto and M.S. Hakim, IOP Conf. Ser.: Mater. Sci. Eng., 299, 012001 (2018); https://doi.org/10.1088/1757-899X/299/1/012001
- M. Liang, Y. Ren, H. Zhang, Y. Ma, X. Niu and X. Chen, Luminescence, 32, 1031 (2017); https://doi.org/10.1002/bio.3287
- M. Chen, X. Cao, K. Chang, H. Xiang and R. Wang, Electrochim. Acta, 368, 137603 (2021); https://doi.org/10.1016/j.electacta.2020.137603
- L. Dong, R. Li, L. Wang, X. Lan, H. Sun, Y. Zhao and L. Wang, Int. J. Biol. Macromol., 172, 289 (2021); https://doi.org/10.1016/j.ijbiomac.2021.01.049
- D. Zheng, S.K. Vashist, K. Al-Rubeaan, E. Lam, S. Hrapovic, J.H. Luong and F.S. Sheu, J. Nanopharm. Drug Deliv., 1, 64 (2013); https://doi.org/10.1166/jnd.2013.1017
- Z. Wang, F. Liu and C. Lu, Biosens. Bioelectron., 38, 284 (2012); https://doi.org/10.1016/j.bios.2012.06.003
References
D. Sim, M.C. Brothers, J.M. Slocik, A.E. Islam, B. Maruyama, C.C. Grigsby, R.R. Naik and S.S. Kim, Adv. Sci., 9, 2104426 (2022); https://doi.org/10.1002/advs.202104426
P. González, P. Lozano, G. Ros and F. Solano, Int. J. Mol. Sci., 24, 9352 (2023); https://doi.org/10.3390/ijms24119352
T. Yang, F. Qi, F. Guo, M. Shao, Y. Song, G. Ren, Z. Linlin, G. Qin and Y. Zhao, Mol. Med., 30, 71 (2024); https://doi.org/10.1186/s10020-024-00824-9
R.M. Carrillo‐Larco, W.C. Guzman‐Vilca, X. Xu and A. Bernabe‐Ortiz, Diabet. Med., 41, e15174 (2024); https://doi.org/10.1111/dme.15174
N.A. Chamaraja, M. Basavaraju and N.K. Swamy, Anal. Biochem., 590, 113536 (2020); https://doi.org/10.1016/j.ab.2019.113536
V. Vallon and T. Nakagawa, Compr. Physiol., 12, 2995 (2022); https://doi.org/10.1002/j.2040-4603.2022.tb00207.x
H. Sohrabi, F. Maleki, P. Khaaki, M. Kadhom, N. Kudaibergenov and A. Khataee, Biosensors, 13, 347 (2023); https://doi.org/10.3390/bios13030347
M. Harun-Or-Rashid, M.N. Aktar, V. Preda and N. Nasiri, Sens. Diagn., 3, 893 (2024); https://doi.org/10.1039/D4SD00086B
Z. Li, W. Zeng and Y. Li, Molecules, 28, 4891 (2023); https://doi.org/10.3390/molecules28134891
H. Khosravi Ardakani, M. Gerami, M. Chashmpoosh, N. Omidifar and A. Gholami, Biochem. Res. Int., 2022, 2964705 (2022); https://doi.org/10.1155/2022/2964705
S. Fang, S. Li, P. Yin, G. Yao, H. Yu, Y. He, X. Li, M. Yang and W. Tan, Chemosphere, 364, 143171 (2024); https://doi.org/10.1016/j.chemosphere.2024.143171
Z. Yuan, M. Fu, X. Wang, M. Wang, Y. Wei, Y. Sun, Q. Zhang, Y. Zhang and B. Zhang, Anal. Methods, 17, 320 (2025); https://doi.org/10.1039/D4AY01878H
W. Yao, X. Zhang and Z. Lin, Spectrochim. Acta A Mol. Biomol. Spectrosc., 265, 120401 (2022); https://doi.org/10.1016/j.saa.2021.120401
I. Rahmawati, Y. Einaga, T.A. Ivandini and A. Fiorani, ChemElectroChem, 9, e202200175 (2022); https://doi.org/10.1002/celc.202200175
J. Xu, M. Back and S. Tanabe, in eds.: R.-S. Liu and X.-J. Wang, Near-Infrared Phosphors with Persistent Luminescence over 1000 nm for Optical Imaging, In: Phosphor Handbook: Experimental Methods for Phosphor Evaluation and Characterization, CRC Press, edn 3, pp. 363-418 (2022).
N.R. Jalal, F. Mollarasouli, M.R. Jalali Sarvestani, S. Khalili, S. Asadi, Z. Derakhshan, T. Madrakian, A. Afkhami and M. Ahmadi, in eds.: M. Thompson and Z. Ramezani, Quantum Dots in Medical Detection/ Diagnosis, In: Quantum Dots in Bioanalytical Chemistry and Medicine, Royal Society of Chemistry: Cambridge, U.K., vol. 22, pp. 75-106 (2023).
W. Zheng, P. Huang, D. Tu, E. Ma, H. Zhu and X. Chen, Chem. Soc. Rev., 44, 1379 (2015); https://doi.org/10.1039/C4CS00178H
L. Tong, L. Wu, Y. Zai, Y. Zhang, E. Su and N. Gu, Biosens. Bioelectron., 219, 114787 (2023); https://doi.org/10.1016/j.bios.2022.114787
P.C. Lee, N.S. Li, Y.P. Hsu, C. Peng and H.W. Yang, Analyst, 144, 3038 (2019); https://doi.org/10.1039/C8AN02440E
M. Schröder and F.J. Meyer-Almes, ChemTexts., 11, 14 (2025); https://doi.org/10.1007/s40828-025-00211-9
S.Ü. Pektaş, M. Keskin, O.C. Bodur and F. Arslan, J. Food Compos. Anal., 129, 106133 (2024); https://doi.org/10.1016/j.jfca.2024.106133
M.H. Hadwan and S. Ali, Anal. Biochem., 542, 29 (2018); https://doi.org/10.1016/j.ab.2017.11.013
S. Naveen Prasad, P. Weerathunge, M.N. Karim, S. Anderson, S. Hashmi, P.D. Mariathomas, V. Bansal and R. Ramanathan, Anal. Bioanal. Chem., 413, 1279 (2021); https://doi.org/10.1007/s00216-020-03090-w
S. Ma, C. Wei, Y. Bao, Y. Liu, H. Jiang, W. Tong, D. Chen and X. Huang, Mikrochim. Acta, 191, 107 (2024); https://doi.org/10.1007/s00604-023-06150-x
D.-M. Kim, J.-M. Moon, W.-C. Lee, J.-H. Yoon, C.S. Choi and Y.-B. Shim, Biosens. Bioelectron., 91, 276 (2017); https://doi.org/10.1016/j.bios.2016.12.046
Y. Yang, C. Yi, J. Luo, R. Liu, J. Liu, J. Jiang and X. Liu, Biosens. Bioelectron., 26, 2607 (2011); https://doi.org/10.1016/j.bios.2010.11.015
Riyanto and M.S. Hakim, IOP Conf. Ser.: Mater. Sci. Eng., 299, 012001 (2018); https://doi.org/10.1088/1757-899X/299/1/012001
M. Liang, Y. Ren, H. Zhang, Y. Ma, X. Niu and X. Chen, Luminescence, 32, 1031 (2017); https://doi.org/10.1002/bio.3287
M. Chen, X. Cao, K. Chang, H. Xiang and R. Wang, Electrochim. Acta, 368, 137603 (2021); https://doi.org/10.1016/j.electacta.2020.137603
L. Dong, R. Li, L. Wang, X. Lan, H. Sun, Y. Zhao and L. Wang, Int. J. Biol. Macromol., 172, 289 (2021); https://doi.org/10.1016/j.ijbiomac.2021.01.049
D. Zheng, S.K. Vashist, K. Al-Rubeaan, E. Lam, S. Hrapovic, J.H. Luong and F.S. Sheu, J. Nanopharm. Drug Deliv., 1, 64 (2013); https://doi.org/10.1166/jnd.2013.1017
Z. Wang, F. Liu and C. Lu, Biosens. Bioelectron., 38, 284 (2012); https://doi.org/10.1016/j.bios.2012.06.003
