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A new colorimetric assay for amylase based on starch-supported Cu/Au nanocluster peroxidase-like activity

  • Zahra Dehghani
  • Javad Mohammadnejad
  • Morteza HosseiniEmail author
Research Paper
  • 30 Downloads

Abstract

In this paper, we present a new colorimetric technique as a novel assay for the easy and direct detection of α-amylase activity. This detection system utilizes the interaction of α-amylase with starch that is supporting copper/gold (Cu/Au) nanoclusters. The Cu/Au nanoclusters are synthesized using starch as a stabilizing agent at room temperature. These nanoclusters show robust peroxidase-like activity and are able to catalyze the oxidation of TMB (3,3,5,5-tetramethylbenzidine) in the presence of hydrogen peroxide (H2O2), leading to the generation of a blue-colored solution. The α-amylase detection mechanism is based on the digestion of the starch by α-amylase, which results in nanocluster aggregation, leading to increased nanoparticle size and thus decreased peroxidase-like activity of the Cu/Au NCs. Experiments showed that the gradual addition of α-amylase causes the peroxidase activity to decrease step by step in a linear fashion. Using this method, colorimetric sensing of α-amylase was achieved with a detection limit (LOD) of 0.04 U/mL and a linear range of 0.1–10 U/mL. This method is significantly selective for α-amylase and could be affordably and conveniently applied to the detection of α-amylase in blood serum.

Graphical Abstract

Keywords

Copper/gold Nanocluster Peroxidase-like activity α-Amylase 

Notes

Acknowledgements

This research was sponsored by the research council of the University of Tehran, and the authors are grateful for their commercial support.

Funding

Financial support was provided by the Faculty of New Sciences and Technologies, Tehran University, Tehran (Iran).

Compliance with ethical standards

Conflict of interest

Zahra Dehghani, Morteza Hosseini, and Javad Mohammadnejad declare no conflict of interest.

Human plasma samples were obtained from the Emam Khomeini Hospital (Tehran, Iran). All experiments on the human plasma samples were performed in accordance with the Declaration of Helsinki and approved by the ethics committee at the University of Tehran.

Informed consent

Not applicable.

Supplementary material

216_2019_1844_MOESM1_ESM.pdf (275 kb)
ESM 1 (PDF 237 kb)

References

  1. 1.
    Zhang J, Cui J, Liu Y, Chen Y, Li G. A novel electrochemical method to determine α-amylase activity. Analyst. 2014;139:3429–33.CrossRefGoogle Scholar
  2. 2.
    Attia MS, Zoulghena H, Abdel-Mottaleb MSA. A new nano-optical sensor thin film cadmium sulfide doped in sol–gel matrix for assessment of α-amylase activity in human saliva. Analyst. 2014;139:793–800.CrossRefGoogle Scholar
  3. 3.
    Karun T, Choodum A, Sa-E B, Hayee U. Smart phone: a popular device supports amylase activity assay in fisheries research. Food Chem. 2014;163:87–91.CrossRefGoogle Scholar
  4. 4.
    Rizliya V, Jayathilake C, Liyanage R. A simple microplate-based method for the determination of α-amylase activity using the glucose assay kit (GOD method). Food Chem. 2016;211:853–9.CrossRefGoogle Scholar
  5. 5.
    Wang Q, Wang H, Yang X, Wang K, Liu R, Li Q, et al. A sensitive one-step method for quantitative detection of α-amylase in serum and urine using a personal glucose meter. Analyst. 2015;140:1161–5.CrossRefGoogle Scholar
  6. 6.
    Holmes MJ, Southworth T, Watson NG, Povey MJW. Enzyme activity determination using ultrasound. J Phys Conf Ser. 2014;498:012003.CrossRefGoogle Scholar
  7. 7.
    Wang Z, Lu Y. Functional DNA directed assembly of nanomaterials for biosensing. J Mater Chem. 2009;19:1788–98.CrossRefGoogle Scholar
  8. 8.
    Arancon RD, Lin SHT, Chen G, Lin CSK, Lai J, Xud G, et al. Nanoparticle tracking analysis of gold nanomaterials stabilized by various capping agents. RSC Adv. 2014;4:17114–9.CrossRefGoogle Scholar
  9. 9.
    Burnside ER, Winter FD, Didangelos A, James ND, Andreica EC, Layard-Horsfall H, et al. Immune-evasive gene switch enables regulated delivery of chondroitinase after spinal cord injury. Analyst. 2016;141:2362–81.CrossRefGoogle Scholar
  10. 10.
    Borghei YS, Hosseini M, Dadmehr M, Hosseinkhani S, Ganjali MR, Sheikhnejad R. Visual detection of cancer cells by colorimetric aptasensor based on aggregation of gold nanoparticles induced by DNA hybridization. Anal Chim Acta. 2016;904:92–7.CrossRefGoogle Scholar
  11. 11.
    Dadmehr M, Hosseini M, Hosseinkhani S, Ganjali MR, Sheikhnejad R. Label free colorimetric and fluorimetric direct detection of methylated DNA based on silver nanoclusters for cancer early diagnosis. Biosens Bioelectron. 2015;73:108–13.CrossRefGoogle Scholar
  12. 12.
    Fakhri N, Hosseini M, Tavakoli O. Aptamer-based colorimetric determination of Pb2+ using a paper-based microfluidic platform. Anal Methods. 2018;10:4438–44.Google Scholar
  13. 13.
    Shokri E, Hosseini M, Davari MD, Ganjali MR, Peppelenbosch MP, Rezaee F. Disulfide-induced self-assembled targets: a novel strategy for the label free colorimetric detection of DNAs/RNAs via unmodified gold nanoparticles. Sci Rep. 2017;7:45837.CrossRefGoogle Scholar
  14. 14.
    Shahsavar K, Hosseini M, Shokri E, Ganjali MR, Jue H. A sensitive colorimetric aptasensor with a triple-helix molecular switch based on peroxidase-like activity of a DNAzyme for ATP detection. Anal Methods. 2017;9:4726–31.CrossRefGoogle Scholar
  15. 15.
    Hosseini M, Sabet FS, Khabbaz H, Aghazadeh M, Mizani F, Ganjali MR. Enhancement of the peroxidase-like activity of cerium-doped ferrite nanoparticles for colorimetric detection of H2O2 and glucose. Anal Methods. 2017;9:3519–24.Google Scholar
  16. 16.
    Farka Z, Cundelova V, Horackova V, Pastucha M, Mikusova Z, Hlavacek A, et al. Prussian blue nanoparticles as a catalytic label in a sandwich nanozyme-linked immunosorbent assay. Anal Chem. 2018;90:2348–54.CrossRefGoogle Scholar
  17. 17.
    Wu LL, Wang LY, Xie ZJ, Pan N, Peng CF. Colorimetric assay of l-cysteine based on peroxidase-mimicking DNA-Ag/Pt nanoclusters. Sensors Actuators B Chem. 2016;235:110–6.Google Scholar
  18. 18.
    Dehghani Z, Hosseini M, Mohammadnejad J, Bakhshi B, Rezayan AH. Colorimetric aptasensor for Campylobacter jejuni cells by exploiting the peroxidase like activity of Au@Pd nanoparticles. Microchim Acta. 2018;185:448.Google Scholar
  19. 19.
    Borghei YS, Hosseini M, Ganjali MR, Ju H. Colorimetric and energy transfer based fluorometric turn-on method for determination of microRNA using silver nanoclusters and gold nanoparticles. Microchim Acta. 2018;185:286.CrossRefGoogle Scholar
  20. 20.
    Sun Y, Wang J, Li W, Zhang J, Zhang Y, Fu Y. DNA-stabilized bimetallic nanozyme and its application on colorimetric assay of biothiols. Biosens Bioelectron. 2015;74:1038–46.CrossRefGoogle Scholar
  21. 21.
    Palza H. Antimicrobial polymers with metal nanoparticles. Int J Mol Sci. 2015;16:2099–116.CrossRefGoogle Scholar
  22. 22.
    Gholinejad M, Saadati F, Shaybanizadeh S, Pullithadathil B. Copper nanoparticles supported on starch micro particles as a degradable heterogeneous catalyst for three-component coupling synthesis of propargylamines. RSC Adv. 2016;6:4983.CrossRefGoogle Scholar
  23. 23.
    Suramwar NV, Thakare SR, Khaty NT. One pot synthesis of copper nanoparticles at room temperature and its catalytic activity. Arab J Chem. 2016;9:S1807–12.CrossRefGoogle Scholar
  24. 24.
    Bashir O, Hussain S, Al-Thabaiti SA, Khan Z. Synthesis, optical properties, stability, and encapsulation of Cu-nanoparticles. Spectrochim Acta Part A. 2015;140:265–73.Google Scholar
  25. 25.
    Khachatryan K, Khachatryan G, Fiedorowicz M. Silver and gold nanoparticles embedded in potato starch gel films. J Mater Sci Chem Eng. 2016;4:22.Google Scholar
  26. 26.
    Vasileva P, Donkova B, Karadjova I, Dushkin C. Synthesis of starch-stabilized silver nanoparticles and their application as a surface plasmon resonance-based sensor of hydrogen peroxide. Colloids Surf A. 2011;382:203–10.CrossRefGoogle Scholar
  27. 27.
    Vigneshwaran N, Nachane RP, Balasubramanya RH, Varadarajan PV. A novel one-pot ‘green’ synthesis of stable silver nanoparticles using soluble starch. Carbohydr Res. 2006;341:2012–8.Google Scholar
  28. 28.
    Engelbrekt C, Sorensen KH, Zhang J, Welinder AC, Jensen PS, Ulstrup J. Green synthesis of gold nanoparticles with starch–glucose and application in bioelectrochemistry. J Mater Chem. 2009;19:7839–7.CrossRefGoogle Scholar
  29. 29.
    Jiang C, Zhu J, Li Z, Luo J, Wang J, Sun Y. Chitosan–gold nanoparticles as peroxidase mimic and their application in glucose detection in serum. RSC Adv. 2017;7:44463–9.CrossRefGoogle Scholar
  30. 30.
    Fu Y, Zhao X, Zhang J, Li W. DNA-based platinum nanozymes for peroxidase mimetics. J Phys Chem C. 2014;118:18116–25.CrossRefGoogle Scholar
  31. 31.
    Valodkar M, Singh Rathore P, Jadeja RN, Thounaojam M, Devkar RV, Thakore S. Cytotoxicity evaluation and antimicrobial studies of starch capped water soluble copper nanoparticles. J Hazard Mater. 2012;201:244–9.CrossRefGoogle Scholar
  32. 32.
    Borghei YS, Hosseini M, Ganjali MR. Visual detection of miRNA using peroxidase-like catalytic activity of DNA-CuNCs and methylene blue as indicator. Clin Chim Acta. 2018;483:119–25.CrossRefGoogle Scholar
  33. 33.
    Borghei YS, Hosseini M, Ganjali MR. Oxidase-like catalytic activity of Cys-AuNCs upon visible light irradiation and its application for visual miRNA detection. Sensors Actuators B Chem. 2018;273:1618–26.Google Scholar
  34. 34.
    Ahmadzade Kermani H, Hosseini M, Miti A, Dadmehr M, Zuccheri G, Hosseinkhani S, et al. A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters. Anal Bio Anal. 2018;410:4943–52.Google Scholar
  35. 35.
    Butterworth PJ, Warren FJ, Ellis PR. Human α-amylase and starch digestion: an interesting marriage. Starch-Stärke. 2011;63:395–405.CrossRefGoogle Scholar
  36. 36.
    Zheng C, Zheng AX, Liu B, Zhang XL, He Y, Li J, et al. One-pot synthesized DNA-templated Ag/Pt bimetallic nanoclusters as peroxidase mimics for colorimetric detection of thrombin. Chem Commun. 2014;50:13103.CrossRefGoogle Scholar
  37. 37.
    Gao L, Zhuang J, Nie L, Zhang J, Zhang Y, Gu N, et al. Intrinsic peroxidase-like activity of ferromagnetic nanoparticles. Nat Nanotechnol. 2007;2:577–83.CrossRefGoogle Scholar
  38. 38.
    Garcia PT, Guimaraes LN, Dias AA, Ulhoa C, Coltro WKT. Amperometric detection of salivary α-amylase on screen-printed carbon electrodes as a simple and inexpensive alternative for point-of-care testing. Sensors Actuators B Chem. 2018;258:342–8.CrossRefGoogle Scholar
  39. 39.
    Sakac N, Regusic L, Sak-Bosnar M, Horvat M, Breslauer N. Direct potentiometric determination of ptyalin in saliva. Int J Electrochem Sci. 2014;9:7097–109.Google Scholar
  40. 40.
    Mahosenaho M, Caprio F, Micheli L, Secay AM, Palleschi G, Virtanen V. A disposable biosensor for the determination of alpha-amylase in human saliva. Microchim Acta. 2010;170:243–9.CrossRefGoogle Scholar
  41. 41.
    ThermoFisher Scientific. EnzChek™ Ultra Amylase Assay Kit. https://www.thermofisher.com/order/catalog/product/E33651.
  42. 42.
    Ahmadzade Kermani H, Hosseini M, Dadmehr M, Hosseinkhani S, Ganjali MR. DNA methyltransferase activity detection based on graphene quantum dots using fluorescence and fluorescence anisotropy. Sensors Actuators B Chem. 2017;241:217–23.Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Zahra Dehghani
    • 1
  • Javad Mohammadnejad
    • 1
  • Morteza Hosseini
    • 1
    Email author
  1. 1.Department of Life Science Engineering, Faculty of New Sciences & TechnologiesUniversity of TehranTehranIran

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