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Urease Immobilized Fluorescent Gold Nanoparticles for Urea Sensing

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Abstract

We report a surfactant-free synthesis of monodispersed gold nanoparticles (AuNPs) with average size of 15 nm. An approach for visual and fluorescent sensing of urea in aqueous solution based on shift in surface plasmon band (SPB) maxima as well as quench in fluorescence intensity. To enable the urea detection, we functionalized the thiol-capped gold nanoparticles with urease, the enzyme specific to urea using carbodiimide chemistry. The visible color changed of the gold colloidal solution from red to blue (or purple); this was evident from quenching in absorbance and fluorescence intensity, is the principle applied here for the sensing of urea. The solution turns blue when the urea concentration exceeds 8 mg/dL which reveals visual lower detection limit. The lower detection limits governed by the fluorescence quenching were found 5 mg/dL (R 2 = 0.99) which is highly sensitive and selective compared to shift in SPB maxima. The approach depicted here seems to be important in clinical diagnosis.

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Acknowledgments

Author UKP thankfully acknowledges the financial support from the University Grant Commission (UGC), Govt. of India. Author NRN acknowledges the financial support from UGC (RJNF-JRF-SC-252). Author AS acknowledges the DST and CAS of Department of Physics, B.H.U. Varanasi, India, Author PSS acknowledges CAS of Department of Zoology, B.H.U. Varanasi, India. Author UKP is specially thankful to Dr. Gauthama (IIT Kanpur) for TEM analysis and Dr. A. C. Pandey (NAC, Allahabad) for SEM facilities.

Compliance with Ethical Standards

On behalf of all the authors in the present manuscript, I wish to declare that:

  • This manuscript has not been submitted to more than one journal for simultaneous consideration.

  • No data have been fabricated or manipulated (including images) to support our conclusions.

  • No data, text, or theories by others have been presented in manuscript. The statements in the text have been given with the study of the cited literature.

  • Consent to submit the manuscript has been received explicitly from all coauthors before the work is submitted.

  • Authors whose names appear on the submission have contributed sufficiently to the scientific work and therefore share collective responsibility and accountability for the results.

  • Research grants received from various funding agencies for all the authors have been acknowledged, respectively, in the manuscript.

  • The authors declare that they have no conflict of interest.

  • The authors declare that there is no experiment that has been done on any human or animal during the study of the manuscript.

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    Authors and Affiliations

    1. Department of Physics, Indian Institute of Technology, Kanpur, 208016, India

      Upendra Kumar Parashar

    2. Department of Physics, Banaras Hindu University, Varanasi, 221005, India

      Upendra Kumar Parashar & Anchal Srivastava

    3. Department of Zoology, Banaras Hindu University, Varanasi, 221005, India

      Narsingh R. Nirala & P. S. Saxena

    4. School of Material Science, Indian Institute of Technology (B.H.U.), Varanasi, 221005, India

      Chandan Upadhyay

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    1. Upendra Kumar Parashar
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    Correspondence to Anchal Srivastava.

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    Upendra Kumar Parashar and Narsingh R. Nirala contributed equally to this work.

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    Parashar, U.K., Nirala, N.R., Upadhyay, C. et al. Urease Immobilized Fluorescent Gold Nanoparticles for Urea Sensing. Appl Biochem Biotechnol 176, 480–492 (2015). https://doi.org/10.1007/s12010-015-1589-z

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