Skip to main content
SpringerLink
Log in

Monitoring the activity and inhibition of alkaline phosphatase via quenching and restoration of the fluorescence of carbon dots

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

We report that the fluorescence of carbon dots (C-dots) in water is quenched by the addition of Cu2+ ions, and that the subsequent addition of pyrophosphate (PPi) restores fluorescence. This is likely to be due to the coordination of Cu2+ by PPi. This effect forms the basis for a method to determine the activity and inhibition of the enzyme alkaline phosphatase (ALP). If ALP is added to a system composed of C-dots, Cu2+ and PPi, fluorescence will decrease over time because ALP catalyzes the hydrolysis of PPi to form orthophosphate (Pi). This results in a release of the quencher Cu2+. The decrease in fluorescence is related to the activity of ALP. The method is simple and displays good sensitivity (with a limit of detection of 1 units per L) and selectivity. The method was successfully applied to the determination of ALP in serum samples. We also have studied the inhibitory effect of Pi on the activity of ALP. We presume that this method holds a large potential in terms of diagnosis of ALP-related diseases, to evaluate the function of ALP in biological systems and in screening for potential inhibitors of ALP.

The activity and inhibition of the enzyme alkaline phosphatase (ALP) can be assayed with a system composed of C-dots, Cu(II) and pyrophosphate (PPi). The fluorescence of C-dots is quenched by Cu(II) ions but restored by PPi. If, however, ALP catalyzes the hydrolysis of PPi, Cu(II) is released. The resulting decrease in fluorescence is directly related to the activity of ALP

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Siddique A, Kowdley KV (2012) Approach to a patient with elevated serum alkaline phosphatase. Clin Liver Dis 16(2):199–229

    Article  Google Scholar 

  2. Liu S, Wang X, Pang S, Na W, Yan X, Su X (2014) Fluorescence detection of adenosine-5′-triphosphate and alkaline phosphatase based on the generation of CdS quantum dots. Anal Chim Acta 827:103–110

    Article  CAS  Google Scholar 

  3. Ooi K, Shiraki K, Morishita Y, Nobori T (2007) High-molecular intestinal alkaline phosphatase in chronic liver diseases. J Clin Lab Anal 21(3):133–139

    Article  CAS  Google Scholar 

  4. Kong R-M, Fu T, Sun N-N, Qu F-L, Zhang S-F, Zhang X-B (2013) Pyrophosphate-regulated Zn2 + -dependent DNAzyme activity: an amplified fluorescence sensing strategy for alkaline phosphatase. Biosens Bioelectron 50:351–355

    Article  CAS  Google Scholar 

  5. Wu N, Lan J, Yan L, You J (2014) A sensitive colorimetric and fluorescent sensor based on imidazolium-functionalized squaraines for the detection of GTP and alkaline phosphatase in aqueous solution. Chem Commun 50(34):4438–4441

    Article  CAS  Google Scholar 

  6. Liu S, Pang S, Na W, Su X (2014) Near-infrared fluorescence probe for the determination of alkaline phosphatase. Biosens Bioelectron 55:249–254

    Article  CAS  Google Scholar 

  7. Wolfbeis OS, Koller E (1985) Photometric and fluorimetric assay of alkaline phosphatase with new coumarin-derived substrates. Microchim Acta 85:389–395

  8. Schrenkhammer P, Rosnizeck I, Duerkop A, Wolfbeis OS, Schaeferling M (2008) Time-resolved fluorescence-based assay for the determination of alkaline phosphatase activity, and application to the screening of its inhibitors. J Biomol Screen 13:9–16

  9. Sasamoto K, Deng G, Ushijima T, Ohkura Y, Ueno K (1995) Benzothiazole derivatives as substrates for alkaline phosphatase assay with fluorescence and chemiluminescence detection. Analyst 120:1709–1714

    Article  CAS  Google Scholar 

  10. Murata T, Yasukawa T, Shiku H, Matsue T (2009) Electrochemical single-cell gene-expression assay combining dielectrophoretic manipulation with secreted alkaline phosphatase reporter system. Biosens Bioelectron 25(4):913–919

    Article  CAS  Google Scholar 

  11. Chen J, Jiao H, Li W, Liao D, Zhou H, Yu C (2013) Real-time fluorescence turn-on detection of alkaline phosphatase activity with a novel perylene probe. Chem Asian J 8(1):276–281

    Article  CAS  Google Scholar 

  12. Ingram A, Moore BD, Graham D (2009) Simultaneous detection of alkaline phosphatase and β-galactosidase activity using SERRS. Bioorg Med Chem Lett 19:1569–1571

    Article  CAS  Google Scholar 

  13. Chien CY, Lai WT, Chang YJ, Wang CC, Kuo MH, Li PW (2014) Size tunable Ge quantum dots for near-ultraviolet to near-infrared photosensing with high figures of merit. Nanoscale 6(10):5303–5308

    Article  CAS  Google Scholar 

  14. Ma Y, Huang S, Zeng C, Zhou T, Zhong Z, Zhou T, Fan Y, Yang X, Xia J, Jiang Z (2014) Towards controllable growth of self-assembled SiGe single and double quantum dot nanostructures. Nanoscale 6(8):3941–3948

    Article  CAS  Google Scholar 

  15. Peng B, Li Z, Mutlugun E, Hernandez Martinez PL, Li D, Zhang Q, Gao Y, Demir HV, Xiong Q (2014) Quantum dots on vertically aligned gold nanorod monolayer: plasmon enhanced fluorescence. Nanoscale 6(11):5592–5598

    Article  CAS  Google Scholar 

  16. Zhai X, Zhang P, Liu C, Bai T, Li W, Dai L, Liu W (2012) Highly luminescent carbon nanodots by microwave-assisted pyrolysis. Chem Commun 48(64):7955–7957

    Article  CAS  Google Scholar 

  17. Yazid S, Chin S, Pang S, Ng S (2013) Detection of Sn(II) ions via quenching of the fluorescence of carbon nanodots. Microchim Acta 180:137–143

    Article  Google Scholar 

  18. Du F, Zeng F, Ming Y, Wu S (2013) Carbon dots-based fluorescent probes for sensitive and selective detection of iodide. Microchim Acta 180:453–460

    Article  CAS  Google Scholar 

  19. Zhu S, Meng Q, Wang L, Zhang J, Song Y, Jin H, Zhang K, Sun H, Wang H, Yang B (2013) Highly photoluminescent carbon dots for multicolor patterning, sensors, and bioimaging. Angew Chem Int Ed 52(14):3953–3957

    Article  CAS  Google Scholar 

  20. Hou J, Yan J, Zhao Q, Li Y, Ding H, Ding L (2013) A novel one-pot route for large-scale preparation of highly photoluminescent carbon quantum dots powders. Nanoscale 5(20):9558–9561

    Article  CAS  Google Scholar 

  21. Kong W, Liu J, Liu R, Li H, Liu Y, Huang H, Li K, Liu J, Lee S-T, Kang Z (2014) Quantitative and real-time effects of carbon quantum dots on single living HeLa cell membrane permeability. Nanoscale 6(10):5116–5120

    Article  CAS  Google Scholar 

  22. Peng H, Travas SJ (2009) Simple aqueous solution route to luminescent carbogenic dots from carbohydrates. Chem Mater 21:5563–5565

    Article  CAS  Google Scholar 

  23. Sun H, Gao N, Wu L, Ren J, Wei W, Qu X (2013) Highly photoluminescent amino-functionalized graphene quantum dots used for sensing copper ions. Chem Eur J 19(40):13362–13368

    Article  CAS  Google Scholar 

  24. Dong Y, Wang R, Li G, Chen C, Chi Y, Chen G (2012) Polyamine-functionalized carbon quantum dots as fluorescent probes for selective and sensitive detection of copper ions. Anal Chem 84(14):6220–6224

    Article  CAS  Google Scholar 

  25. Badarau A, Dennison C (2011) Copper trafficking mechanism of CXXC-containing domains: insight from the pH-dependence of their Cu(I) affinities. J Am Chem Soc 133(9):2983–2988

    Article  CAS  Google Scholar 

  26. Deng J, Jiang Q, Wang Y, Yang L, Yu P, Mao L (2013) Real-time colorimetric assay of inorganic pyrophosphatase activity based on reversibly competitive coordination of Cu2+ between cysteine and pyrophosphate ion. Anal Chem 85(19):9409–9415

    Article  CAS  Google Scholar 

  27. Zhang L, Zhao J, Duan M, Zhang H, Jiang J, Yu R (2013) Inhibition of dsDNA-templated copper nanoparticles by pyrophosphate as a label-free fluorescent strategy for alkaline phosphatase assay. Anal Chem 85(8):3797–3801

    Article  CAS  Google Scholar 

  28. Wang H, Mu L, She G, Xu H, Shi W (2014) Fluorescent biosensor for alkaline phosphatase based on fluorescein derivatives modified silicon nanowires. Sensors Actuators B Chem 203:774–781

    Article  CAS  Google Scholar 

  29. Lin JH, Tseng WL (2013) A method for fluorescence sensing of adenosine and alkaline phosphatase based on the inhibition of S-adenosylhomocysteine hydrolase activity. Biosens Bioelectron 41:379–385

    Article  CAS  Google Scholar 

  30. Jia L, Xu JP, Li D, Pang SP, Fang Y, Song ZG, Ji J (2010) Fluorescence detection of alkaline phosphatase activity with beta-cyclodextrin-modified quantum dots. Chem Commun 46(38):7166–7168

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank the support of work by National Key Basic Research Program of China (2014CB744502) and the National Natural Science Foundation of China (NO. 21105128).

Author information

Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China

    Wenjing Kang, Yingying Ding, Hui Zhou, Qiuyue Liao, Xiao Yang, Yugui Yang, Jingshu Jiang & Minghui Yang

Authors
  1. Wenjing Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yingying Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hui Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qiuyue Liao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiao Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yugui Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jingshu Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Minghui Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Minghui Yang.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kang, W., Ding, Y., Zhou, H. et al. Monitoring the activity and inhibition of alkaline phosphatase via quenching and restoration of the fluorescence of carbon dots. Microchim Acta 182, 1161–1167 (2015). https://doi.org/10.1007/s00604-014-1439-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00604-014-1439-7

Keywords

Search

Navigation