Abstract
Detecting pyruvic acid and alanine aminotransferase (ALT) is crucial for diagnosing and managing clinical conditions. Pyruvic acid plays a vital role in metabolic pathways and is linked to metabolic disorders, diabetes, and cancer, while ALT, primarily found in the liver, serves as a critical indicator of hepatic function and damage. Accurate monitoring of these biomarkers is essential for early diagnosis, tracking disease progression, and evaluating therapeutic interventions, with convenient detection methods also facilitating home-based monitoring. Herein, we report the novel application of a previously described BSA-Os nanozyme as the foundation for a cost-effective colorimetric sensing platform. Leveraging its intrinsic peroxidase-like activity, the BSA-Os nanozyme catalyzes hydrogen peroxide (H2O2) to generate hydroxyl radicals, which oxidize 3,3’,5,5’-tetramethylbenzidine (TMB) to its blue oxidized form (ox-TMB), detectable at 652 nm. For ALT detection, the enzyme catalyzes the reaction of α-ketoglutarate and L-alanine to produce pyruvate, which is subsequently oxidized by pyruvate oxidase to generate H2O2, initiating the same BSA-Os/TMB colorimetric cascade. The method demonstrated a detection range of 2–200 μM for pyruvic acid, with a limit of detection (LOD) of 208.55 nM, and 10–125 U/L for ALT, with a LOD of 6.4 U/L. The platform was successfully applied to human serum samples, for pyruvic acid detection, the relative standard deviation (RSD) was < 3.43%, and recoveries varied from 101.2% to 106.5%, while for ALT detection, the RSD was < 4.03% and recoveries varied from 90.8% to 96.1%, demonstrating good accuracy and precision. This study highlights the promising potential of BSA-Os in clinical diagnostics, offering a reliable and cost-effective tool for monitoring pyruvic acid and ALT, facilitating early disease detection, personalized treatment, improved patient outcomes, and supporting home-based monitoring for patient convenience.
Similar content being viewed by others
References
Banakar M, Hamidi M, Khurshid Z, Zafar MS, Sapkota J, Azizian R, Rokaya D (2022) Electrochemical biosensors for pathogen detection: an updated review. Biosensors (Basel) 12(11):927. https://doi.org/10.3390/bios12110927
Bansal V, Seal S, Wei H (2024) Introduction to nanozymes. Anal Bioanal Chem 416(27):5855–5857. https://doi.org/10.1007/s00216-024-05543-y
Chen M, Rong L, Chen X (2015) A simple and sensitive detection of glutamic-pyruvic transaminase activity based on fluorescence quenching of bovine serum albumin. RSC Adv 5(125):103557–103562. https://doi.org/10.1039/c5ra24162f
Fernandez-Caggiano M, Eaton P (2021) Heart failure-emerging roles for the mitochondrial pyruvate carrier. Cell Death Differ 28(4):1149–1158. https://doi.org/10.1038/s41418-020-00729-0
Gao L, Zhuang J, Nie L, Zhang J, Zhang Y, Gu N, Wang T, Feng J, Yang D, Perrett S, Yan X (2007) Intrinsic peroxidase-like activity of ferromagnetic nanoparticles. Nat Nanotechnol 2(9):577–583. https://doi.org/10.1038/nnano.2007.260
Gao L, Wei H, Dong S, Yan X (2024) Nanozymes. Adv Mater 36(10):e2305249. https://doi.org/10.1002/adma.202305249
Guo L, Zeng W, Xu S, Zhou J (2020) Fluorescence-activated droplet sorting for enhanced pyruvic acid accumulation by Candida glabrata. Bioresour Technol 318:124258. https://doi.org/10.1016/j.biortech.2020.124258
He SB, Lin MT, Yang L, Noreldeen HAA, Peng HP, Deng HH, Chen W (2021) Protein-assisted osmium nanoclusters with intrinsic peroxidase-like activity and extrinsic antifouling behavior. ACS Appl Mater Interfaces 13(37):44541–44548. https://doi.org/10.1021/acsami.1c11907
He SB, Yang L, Lin MT, Balasubramanian P, Peng HP, Kuang Y, Deng HH, Chen W (2024) Platinum group element-based nanozymes for biomedical applications: an overview. Biomed Mater 16(3):032001. https://doi.org/10.1088/1748-605X/abc904
He S, Guo X, Zheng Q, Shen H, Xu Y, Lin F, Chen J, Deng H, Zeng Y, Chen W (2025) Engineering nickel-supported osmium bimetallic nanozymes with specifically improved peroxidase-like activity for immunoassay. Chin Chem Lett 36(04):110096. https://doi.org/10.1016/j.cclet.2024.110096
Le XH, Lee CP, Millar AH (2021) The mitochondrial pyruvate carrier (MPC) complex mediates one of three pyruvate-supplying pathways that sustain Arabidopsis respiratory metabolism. Plant Cell 33(8):2776–2793. https://doi.org/10.1093/plcell/koab148
Li W, Pan C, Hou T, Wang X, Li F (2014) Selective and colorimetric detection of pyruvic acid using conformational switch of i-motif DNA and unmodified gold nanoparticles. Anal Methods 6(6):1645. https://doi.org/10.1039/c3ay41883a
Li YS, Li QJ, Yang W, Gao XF (2017) Research on a new micro-volume fluorescence capillary biosensor assay for sequentially quantifying pyruvate and lactate. J Fluoresc 27(3):883–894. https://doi.org/10.1007/s10895-017-2024-3
Li M, Li X, Chen L, Li X, Liu C (2025) An “off-on” fluorescent probe for imaging pyruvic acid in living systems. Talanta 284:127225. https://doi.org/10.1016/j.talanta.2024.127225
Malik M, Chaudhary R, Pundir C (2019) An improved enzyme nanoparticles based amperometric pyruvate biosensor for detection of pyruvate in serum. Enzyme Microb Technol 123:30–38. https://doi.org/10.1016/j.enzmictec.2019.01.006
Moed S, Zaman MH (2019) Towards better diagnostic tools for liver injury in low-income and middle-income countries. BMJ Glob Health 4(4):e001704. https://doi.org/10.1136/bmjgh-2019-001704
Mora-Sanz V, Saa L, Briz N, Möller M, Pavlov V (2020) Antibody-directed synthesis of catalytic nanoclusters for bioanalytical assays. ACS Appl Mater Interfaces 12(26):28993–28999. https://doi.org/10.1021/acsami.0c05229
Noreldeen HAA, Yang L, Guo XY, He SB, Peng HP, Deng HH, Chen W (2021) A peroxidase-like activity-based colorimetric sensor array of noble metal nanozymes to discriminate heavy metal ions. Analyst 147(1):101–108. https://doi.org/10.1039/d1an01895g
Pan J, He Q, Lao Z, Zou Y, Su J, Li Q, Chen Z, Cui X, Cai Y, Zhao S (2022) A bifunctional immunosensor based on osmium nano-hydrangeas as a catalytic chromogenic and tinctorial signal output for folic acid detection. Analyst 147(1):55–65. https://doi.org/10.1039/d1an01432c
Peng P, Liu C, Li Z, Xue Z, Mao P, Hu J, Xu F, Yao C, You M (2022) Emerging ELISA derived technologies for in vitro diagnostics. TrAC, Trends Anal Chem 152:116605. https://doi.org/10.1016/j.trac.2022.116605
Quan C, Quan L, Wen Q, Yang M, Li T (2024) Alanine aminotransferase electrochemical sensor based on graphene@ MXene composite nanomaterials. Mikrochim Acta 191(1):45. https://doi.org/10.1007/s00604-023-06131-0
Samy MA, Abdel-Tawab AH, Abdel-Ghani NT, El Nashar R (2023) Application of molecularly imprinted microelectrode as a promising point-of-care biosensor for alanine aminotransferase enzyme. Chemosensors 11(5):262. https://doi.org/10.3390/chemosensors11050262
Sellami K, Couvert A, Nasrallah N, Maachi R, Abouseoud M, Amrane A (2022) Peroxidase enzymes as green catalysts for bioremediation and biotechnological applications: a review. Sci Total Environ 806(Pt 2):150500. https://doi.org/10.1016/j.scitotenv.2021.150500
Tang G, He J, Liu J, Yan X, Fan K (2021) Nanozyme for tumor therapy: Surface modification matters. Exploration (Beijing) 1(1):75–89. https://doi.org/10.1002/EXP.20210005
Tang C, Fang T, Chen S, Zhang D, Yin J, Wang H (2023) Citrate-functionalized osmium nanoparticles with peroxidase-like specific activity for highly efficient degradation of phenolic pollutants. Chem Eng J. https://doi.org/10.1016/j.cej.2023.142726
Valenti L, Pelusi S, Bianco C, Ceriotti F, Berzuini A, Iogna Prat L, Trotti R, Malvestiti F, D’Ambrosio R, Lampertico P, Colli A, Colombo M, Tsochatzis EA, Fraquelli M, Prati D (2021) Definition of healthy ranges for alanine aminotransferase levels: A 2021 update. Hepatol Commun 5(11):1824–1832. https://doi.org/10.1002/hep4.1794
Verma SK, Huang J, Hutchinson HG, Estevez I, Kuang K, Reynolds SL, Schneeweiss S (2023) Statin use and severe acute liver injury among patients with elevated alanine aminotransferase. Clin Epidemiol 14:1535–1545. https://doi.org/10.2147/CLEP.S385712
Wang J, Xu Z, Zou H, Hu S, Wang D, Liu J, Wang L (2017) Electrochemical determination of glutamic pyruvic transaminase using a microfluidic chip. Microfluidics Nanofluidics 21(2):27.1-27.7. https://doi.org/10.1007/s10404-017-1869-8
Wei P, Bott AJ, Cluntun AA, Morgan JT, Cunningham CN, Schell JC, Ouyang Y, Ficarro SB, Marto JA, Danial NN, DeBerardinis RJ, Rutter J (2022) Mitochondrial pyruvate supports lymphoma proliferation by fueling a glutamate pyruvate transaminase 2-dependent glutaminolysis pathway. Sci Adv 8(39):eabq0117. https://doi.org/10.1126/sciadv.abq0117
Yan F, Nie G, Zhou N, Zhang M, Peng W (2023) Combining fat-to-muscle ratio and alanine aminotransferase/aspartate aminotransferase ratio in the prediction of cardiometabolic risk: a cross-sectional study. Diabetes Metab Syndr Obes 16:795–806. https://doi.org/10.2147/DMSO.S401024
Yip TC, Wong VW, Wong GL (2022) Alanine aminotransferase level: the road to normal in 2021. Hepatol Commun 5(11):1807–1809. https://doi.org/10.1002/hep4.1788
Yu L, Teoh ST, Ensink E, Ogrodzinski MP, Yang C, Vazquez AI, Lunt SY (2019) Cysteine catabolism and the serine biosynthesis pathway support pyruvate production during pyruvate kinase knockdown in pancreatic cancer cells. Cancer Metab 7:13. https://doi.org/10.1186/s40170-019-0205-z
Zhang YY, Meng ZJ (2022) Definition and classification of acute-on-chronic liver diseases. World J Clin Cases 10(15):4717–4725. https://doi.org/10.12998/wjcc.v10.i15.4717
Zhuang QQ, Deng HH, He SB, Peng HP, Lin Z, Xia XH, Chen W (2019) Immunoglobulin G-encapsulated gold nanoclusters as fluorescent tags for dot-blot immunoassays. ACS Appl Mater Interfaces 11(35):31729–31734. https://doi.org/10.1021/acsami.9b11599
Acknowledgements
The authors gratefully acknowledge financial support from the Joint Funds for the Innovation of Science and Technology, Fujian Province (2023Y9226), the Natural Science Foundation of Fujian Province (2022J01271), the Introduced High-Level Talent Team Project of Quanzhou City (2023CT008), the Startup Fund for Scientific Research, Fujian Medical University (2023QH1117), and the Science and Technology Program of Quanzhou (2024NY069).
Author information
Authors and Affiliations
Contributions
Shao-Bin He and Yin Zhang conceiving the idea, designing the experiments, and co-supervising this work. Jin-Cheng Chen and Feng-Lin Lin carrying out the experiments, characterizations, and writing the manuscript. Yin-Feng Xiao, Jian-Qing Liu, Qiao-Ling Liu, and Qiu-Xia Xu was performed data curation, discussing the results and commenting.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Chen, JC., Lin, FL., Xiao, YF. et al. Osmium nanozyme-based colorimetric assay for pyruvic acid and alanine aminotransferase detection. Chem. Pap. 79, 6879–6885 (2025). https://doi.org/10.1007/s11696-025-04230-1
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1007/s11696-025-04230-1