Skip to main content
Log in

Studies towards an amperometric phosphate ion biosensor for urine and water analysis

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

Abstract

An amperometric biosensor for phosphate ion is described that is based on a cobalt phthalocyanine modified screen-printed carbon electrode (CoPC-SPCE). The biosensor operation is based on the enzyme pyruvate oxidase (PyOd) which catalyses the oxidative decarboxylation of pyruvate, in the presence of inorganic phosphate and O2, to acetyl phosphate, hydrogen peroxide (H2O2) and CO2. The transducer allows the electrocatalytic oxidation of H2O2 in order to generate the analytical signal. The enzyme was immobilised onto the CoPC-SPCE using a sandwich format. The inner membrane was formed in situ by depositing an acetone solution containing cellulose acetate first onto the transducer surface. The enzyme and cofactors were then deposited onto this layer and allowed to dry; finally a second aliquot of the cellulose acetate solution was deposited onto the enzyme layer and allowed to dry. The biosensor was characterised by amperometry in stirred solution to produce current-voltage curves and for calibration studies. From these it was deduced that a reliable electrocatalytic response was obtained for phosphate ion; an operating potential of +0.4 V was selected for the analysis of urine samples. The precision of the response for urine analysis and recovery data for potable water suggests that the biosensor could have applications in clinical and environmental monitoring.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Engblom SO (1998) The phosphate sensor. Biosens Bioelectron 13:981–994

    Article  CAS  Google Scholar 

  2. Palleschi G, Amine A (2004) Phosphate, nitrate and sulphate biosensors. Anal Lett 37:1–19

    Article  Google Scholar 

  3. Villalba MM, McKeegan KJ, Vaughan DH, Cardosi MF, Davis J (2009) Bioelectroanalytical determination of phosphate: a review. J Mol Catal B: Enzym 59:1–8

    Article  CAS  Google Scholar 

  4. Harper D (1992) Eutrophication of freshwaters principles, problems and restoration. Chapman & Hall, London

    Google Scholar 

  5. Kawasaki H, Sato K, Ogawa J, Hasegawa Y, Yuki H (1989) Determination of inorganic phosphate by flow injection method with immobilized enzymes and chemiluminescence detection. Anal Biochem 182:366–370

    Article  CAS  Google Scholar 

  6. Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorus. J Biol Chem 66:375–400

    CAS  Google Scholar 

  7. Murphy J, Riley JP (1962) A modified single-solution method for the determination of phosphate in natural water. Anal Chim Acta 27:31–36

    Article  CAS  Google Scholar 

  8. Hart JP, Serban S, Jones LJ, Biddle N, Pittson R, Drago GA (2006) Selective and rapid biosensor integrated into a commercial hand-held instrument for the measurement of ammonium ion in sewage effluent. Anal Lett 39:1657–1667

    Article  CAS  Google Scholar 

  9. Guilbault GG, Nanjo M (1975) A phosphate selective electrode based on immobilised alkaline phosphatase and glucose oxidase. Anal Chim Acta 78:69–80

    Article  CAS  Google Scholar 

  10. Rahman Md A, Park DS, Chang SC, McNeil CJ, Shim YB (2006) The biosensor based on the pyruvate oxidase modified conducting polymer for phosphate ions determinations. Biosens Bioelectron 21:1116–1124

    Article  Google Scholar 

  11. Kwan RCH, Leung HF, Hon PYT, Barford JP, Renneberg R (2005) A screen-printed biosensor using pyruvate oxidase for rapid determination of phosphate in synthetic wastewater. Appl Microbiol Biotechnol 66:377–383

    Article  CAS  Google Scholar 

  12. Gajovic N, Habermuller K, Warsinke A, Schuhmann W, Scheller FW (1999) A pyruvate oxidase electrode based on an electrochemically deposited redox polymer. Electroanalysis 11:1377–1383

    Article  CAS  Google Scholar 

  13. Nakamura H, Ikebukuro K, McNiven S, Karube I, Yamamoto H, Hayashi K, Suzuki M, Kubo I (1997) A chemiluminescent FIA biosensor for phosphate ion monitoring using pyruvate oxidase. Biosens Bioelectron 12:959–966

    Article  CAS  Google Scholar 

  14. Schubert F, Renneberg R, Scheller FW, Kirstein L (1984) Plant tissue hybrid electrode for determination of phosphate and fluoride. Anal Chem 56:1677–1682

    Article  CAS  Google Scholar 

  15. d’Urso EM, Coulet PR (1990) Phosphate sensitive enzyme electrode: a potential sensor for environmental control. Anal Chim Acta 239:1–5

    Article  Google Scholar 

  16. Su Y, Mascini M (1995) AP-GOD biosensor based on a modified poly(phenol) film electrode and its determination of low levels of phosphate. Anal Lett 28:1359–1378

    CAS  Google Scholar 

  17. Mousty C, Cosnier S, Shan D, Mu S (2001) Trienzymatic biosensor for the determination of inorganic phosphate. Anal Chim Acta 443:1–8

    Article  CAS  Google Scholar 

  18. Fernandez JJ, Lopez JR, Correig X, Katakis I (1998) Reagentless carbon paste phosphate biosensors: preliminary studies. Sens Actuators, A 47:13–20

    Google Scholar 

  19. Conrath N, Grundig B, St. Huwel K, Cammann K (1995) A novel enzyme sensor for the determination of inorganic phosphate. Anal Chim Acta 309:47–52

    Article  CAS  Google Scholar 

  20. Hart JP, Crew A, Crouch E, Honeychurch KC, Pemberton RM (2005) Some recent designs and developments of screen-printed carbon electrochemical biosensors for biomedical, environmental, and industrial analyses. Anal Lett 37:789–830

    Article  Google Scholar 

  21. Galan-Vidal CA, Munoz J, Dominguez C, Alegret S (1998) Glucose biosensor strip in a three electrode configuration based on composite and biocomposite materials applied by planar thick film technology. Sens Actuators B 52:257–263

    Article  Google Scholar 

  22. Young SJ, Hart JP, Dowman AA, Cowell DC (2001) The non-specific inhibition of enzymes by environmental pollutants: a study of a model system towards the development of electrochemical biosensor arrays. Biosens Bioelectron 16:887–894

    Article  CAS  Google Scholar 

  23. Crew A, Hart JP, Wedge R, Marty JL, Fournier D (2004) A screen-printed, amperometric biosensor array for the detection of organophosphate pesticides based on inhibition of wild type, and mutant acetylcholinesterases from Drosophilia melanogaster. Anal Lett 37:1601–1610

    Article  CAS  Google Scholar 

  24. Gilmartin MAT, Hart JP (1994) Novel, reagentless, amperometric biosensor for uric acid based on a chemically modified screen-printed carbon electrode coated with cellulose acetate and uricase. Analyst 119:833–840

    Article  CAS  Google Scholar 

  25. Gilbert L, Browning S, Jenkins ATA, Hart JP (2009) Development of an amperometric assay for phosphate ions in urine based on a chemically modified screen-printed carbon electrode. Anal Biochem 393:242–247

    Article  CAS  Google Scholar 

  26. Wang J (2006) Analytical electrochemistry. Wiley-VCH, USA

    Book  Google Scholar 

  27. Santos LM, Baldwin RP (1987) Liquid chromatography/electrochemical detection of carbohydrates at a cobalt phthalocyanine containing chemically modified electrode. Anal Chem 59:1766–1770

    Article  CAS  Google Scholar 

  28. Wallach JB (2006) Interpretation of diagnostic tests, 8th edn. Lippincott, Williams and Wilkins, Philidelphia

    Google Scholar 

  29. Penny C (1988) Sensory perception. Medical Laboratory World June: 41–45

Download references

Acknowledgements

The authors wish to thank Great Western Research (GWR), The Environment Agency and UWE, Bristol for funding. Gwent Electronic Materials (GEM) is thanked for the provision of the CoPC-SPCEs.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to John P. Hart.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gilbert, L., Browning, S., Jenkins, A.T.A. et al. Studies towards an amperometric phosphate ion biosensor for urine and water analysis. Microchim Acta 170, 331–336 (2010). https://doi.org/10.1007/s00604-010-0316-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00604-010-0316-2

Keywords

Navigation