AMPEROMETRIC BIOSENSOR FOR HISTAMINE DETERMINATION

• US 20100096277A1
• Filed: 03/19/2008
• Published: 04/22/2010
• Est. Priority Date: 03/21/2007
• Status: Active Grant

• Current Assignee: Universiti Putra Malaysia (Government of Malaysia)
• Original Assignee: Universiti Putra Malaysia (Government of Malaysia)
• Inventors: Ching, Mai Keow, Salleh, Abu Backar, Wagiran, Rahman, Lee, Yook Heng, Ahmad, Anuar, Abu Bakar, Fatimah, Mustaffa, Rosnin
• Granted Patent: US 8,551,309 B2
• Time in Patent Office: Days
• US Class Current: 205/777.5
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: G01N 2333/90633 with oxygen as acceptor (1.4.3) in general
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48
• CPC Class Codes: G01N 33/12 Meat; Fish

Abstract

There is provided by this invention a simple and rapid amperometric biosensor for determining the level of histamine in seafood or fish. The biosensor combines the technology of screen-printing with immobilized diamine oxidase as the bioreceptor. IQ one embodiment of the present invention, the biosensor incorporates potassium hexacyanoferrate (III) as a mediator.

Claims

1. An amperometric biosensor for histamine determination comprising a working electrode, a counter electrode and a reference electrode, wherein the working electrode is a screen-printed electrode, characterized in that diamine oxidase is immobilized via drop-coating with poly (2-hydroxyethyl methacrylate) (photoHEMA) on the surface of the screen-printed working electrode.

2. The biosensor of claim 1, wherein the screen-printed working electrode is carbon paste based.

3. The biosensor of claim 1, wherein the counter electrode is a platinum rod.

4. The biosensor of claim 1, wherein the reference electrode is a silver/silver chloride (Ag/AgCl) electrode.

9. A method for histamine determination, comprising the steps of:

—

a) applying a sample suspected to contain histamine to the biosensor of claim 1; and

b) measuring the current to provide an output signal indicative of the presence of histamine.

10. The method of claim 9, wherein the voltage range for histamine determination is from 0.30 volt to 0.50 volt.

11. The method of claim 10, wherein the voltage for histamine determination is preferably 0.35 volt.

12. The method according to claim 9, wherein the pH range for histamine determination is from 6.4 to 8.4.

13. The method of claim 12, wherein the pH for histamine determination is preferably 7.4.

14. The method according to claim 9, wherein the reaction time for histamine determination is from 20 seconds.

15. The method of claim 14, wherein the reaction time for histamine determination is preferably 50 seconds.

5. A method of immobilizing diamine oxidase via drop-coating with poly (2-hydroxyethyl methacrylate) (photoHEMA) on the surface of a screen-printed working electrode, characterized in that the method comprises the following steps:

—

a) mixing diamine oxidase solution with poly (2-hydroxyethyl methacrylate) (photoHEMA);

b) drop-coating the mixture onto the surface of the screen-printed working electrode; and

c) photo-curing the electrode.

6. The method of claim 5, wherein diamine oxidase is dissolved in phosphate buffer before mixing with poly (2-hydroxyethyl methacrylate) (photoHEMA).

7. The method of claim 5, wherein diamine oxidase solution is mixed with poly (2-hydroxyethyl methacrylate) (photoHEMA) according to the ratio of 1:

4.

8. The method according to claim 5, wherein the electrode is photo-cured in a UV-exposure unit under nitrogen gas flow for 300 seconds.

16. An amperometric biosensor for histamine determination comprising a working electrode, a counter electrode and a reference electrode, wherein all of the electrodes are screen-printed onto a substrate, characterized in that diamine oxidase is immobilized via drop-coating with poly (2-hydroxyethyl methacrylate) (photoHEMA) and potassium hexacyanoferrate (III) is electrodeposited on the surface of the screen-printed working electrode.

17. The biosensor of claim 16, wherein the screen-printed working and counter electrodes are carbon paste based.

18. The biosensor of claim 16, wherein the screen-printed reference electrode is a silver chloride (AgCl) paste based screen-printed electrode.

19. The biosensor of claim 16, wherein the electrodes are screen-printed onto a polyester substrate.

20. The biosensor of claim 16, wherein the biosensor further comprises a silver layer printed as a basal track layer.

21. The biosensor of claim 16, wherein diamine oxidase is immobilized via drop-coating with poly (2-hydroxyethyl methacrylate) (photoHEMA) according to the following steps:

—

a) mixing diamine oxidase solution with poly (2-hydroxyethyl methacrylate) (photoHEMA);

b) drop-coating the mixture onto the surface of the screen-printed working electrode; and

c) photo-curing the electrode.

22. The biosensor of claim 21, wherein diamine oxidase is dissolved in phosphate buffer before mixing with poly (2-hydroxyethyl methacrylate) (photoHEMA).

23. The biosensor of claim 21, wherein diamine oxidase solution is mixed with poly (2-hydroxyethyl methacrylate) (photoHEMA) according to the ratio of 1:

4.

24. The biosensor according to claim 21, wherein the electrode is photo-cured in a UV-exposure unit under nitrogen gas flow for 300 seconds.

25. The biosensor of claim 16, wherein potassium hexacyanoferrate (III) is electrodeposited by cyclic voltammetry method, comprising the step of cycling the screen-printed working electrode in a solution of potassium hexacyanoferrate (III) with stirring.

26. The biosensor of claim 25, wherein the electrode is cycled at least fifteen times in a solution of 0.1 M potassium hexacyanoferrate (III) dissolved in deionized water at 0.2 vs^″

1 with stirring.

27. A method for histamine determination, comprising the steps of:

—

a) applying a sample suspected to contain histamine to the biosensor of claim 16; and

b) measuring the current to provide an output signal indicative of the presence of histamine.

28. The method of claim 27, wherein the voltage range for histamine determination is from 0.30 volt to 0.50 volt.

29. The method of claim 28, wherein the voltage for histamine determination is preferably 0.35 volt.

30. The method according to claim 27, wherein the pH range for histamine determination is from 6.4 to 8.4.

31. The method of claim 30, wherein the pH for histamine determination is preferably 7.4.

32. The method according to claim 27, wherein the reaction time for histamine determination is from 20 seconds.

33. The method of claim 32, wherein the reaction time for histamine determination is preferably 50 seconds.