Absorption-emission optrode and methods of use thereof

• US 4,929,561 A
• Filed: 08/08/1985
• Issued: 05/29/1990
• Est. Priority Date: 08/08/1985
• Status: Expired due to Fees

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Hirschfeld, Tomas B.
• Primary Examiner(s): Hill, Jr., Robert J.
• Application Number:
  US06/763,557
• Time in Patent Office: 1,755 Days
• Field of Search: 250/373, 250/458.1, 250/459.1, 250/461.1, 436/74, 436/116-118, 436/121, 436/172, 436/537, 436/800, 436/79, 435/808, 422/55-58, 422/68
• US Class Current: 436/116
• CPC Class Codes: G01N 2021/6434 Optrodes
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/643 non-biological material
• CPC Class Codes: G01N 21/783 for analysing gases
• CPC Class Codes: Y10T 436/177692 Oxides of nitrogen
• CPC Class Codes: Y10T 436/184 Only hydrogen sulfide

Abstract

A method and apparatus for monitoring the physical and chemical properties of a sample fluid by measuring an optical signal generated by a fluorescent substance and modulated by an absorber substance. The emission band of the fluorescent substance overlaps the absorption band of the absorber substance, and the degree of overlap is dependent on the physical and chemical properties of the sample fluid. The fluorescent substance and absorber substance are immobilized on a substrate so that an effective number of molecules thereof are sufficiently close for resonant energy transfer to occur, thereby providing highly efficient modulation of the fluorescent emissions of the fluorescent substance by the absorber substance.

Claims

1. A method for monitoring physical and chemical parameters of a sample substance and for generating a signal related to said physical and chemical parameters, the method comprising the steps of:

immobilizing on a substrate, a fluorescent substance having an emission band and an absorption band;

immobilizing on the same substrate, an absorber substance having an absorption band overlapping the emission band of the fluorescent substance, the degree of overlap being responsive to the physical and chemical parameters of the sample substance, and the absorber substance being immobilized such that an effective number of molecules of the absorber substance are closely adjacent to an effective number of molecules of the fluorescent substance to produce detectable resonant energy transfer therebetween;

contacting the substrate with the sample substance;

illuminating the fluorescent substance with an illumination beam having a frequency within the absorption band of the fluorescent substance, so that excited fluorescent states are generated in the fluorescent substance;

detecting a signal resulting from the decay of the excited fluorescent states;

providing a fiber optic through which said illumination beam is transmitted from a first end of the fiber optic to a second end of the fiber optic;

attaching said substrate to the second end of the fiber optic such that light from said illumination beam emanating from the second end of the fiber optic illuminates said fluorescent substance and such that said resulting signal is collected by the second end of the fiber optic and transmitted to the first end of the fiber optic;

separating at the first end of the fiber optic said illumination beam from said resulting signal;

measuring the intensity of backscattered Raman emissions from the fiber;

forming a ratio of the resulting signal to the backscattered Raman intensity;

extracting information concerning the physical and chemical parameters of the sample substance from the ratio.

2. The method of claim 1 wherein said effective number of molecules of said absorber substance are within about 10-100 Angstroms of said effective number of molecules of said fluorescent substance.

3. The method of claim 2 wherein said effective number of molecules of said absorber substance are within about 10-70 Angstroms of said effective number of molecules of said fluorescent substance.

4. The method of claim 1 wherein said substrate is porous glass.

5. The method of claim 4 wherein said effective number of molecules of said absorber substance are within about 10-70 Angstroms of said effective number of molecules of said fluorescent substance.

6. The method of claim 1 wherein said substrate is sintered glass.

7. The method of claim 6 wherein said effective number of molecules of said absorber substance are within about 10-70 Angstroms of said effective number of molecules of said fluorescent substance.

8. An apparatus for monitoring physical and chemical parameters of a sample substance and for generating an optical signal related to said physical and chemical parameters, the apparatus comprising:

a fluorescent substance having an absorption band and an emission band;

an absorber substance having an absorption band overlapping the emission band of the fluorescent substance, the degree of overlap being responsive to the physical and chemical parameters of the sample substance;

a substrate on which the fluorescent substance and the absorber substance are immobilized such that an effective number of molecules of the absorber substance are closely adjacent to an effective number of molecules of the fluorescent substance to produce detectable resonant energy transfer therebetween;

means for illuminating the fluorescent substance with light having a wavelength within the absorption band of the fluorescent substance so that excited fluorescent states are generated wherein said means for illuminating includes a fiber optic through which an illumination beam from an associated light source is transmitted from a first end of the fiber optic to a second end of the fiber optic, the fiber optic having a core, and wherein said substrate is attached to the second end of the fiber optic such that light from the illumination beam emanating from the second end of the fiber optic illuminates said fluorescent substance, and such that said resulting optical signal is collected by the second end of the fiber optic and transmitted to the first end of the fiber optic, and wherein said means for illuminating further includes separation means adjacent to the first end of the fiber optic for separating the illuminating beam from said resulting optical signal;

means for detecting an optical signal resulting from the decay of the excited fluorescent states;

means for measuring the intensity of backscattered Raman emissions from the fiber; and

means for forming a ratio of the optical signal to the backscattered Raman intensity.

9. The apparatus of claim 8 wherein said effective number of molecules of said absorber substance are within about 10-100 Angstroms of said effective number of molecules of said fluorescent substance.

10. The apparatus of claim 8 wherein said means for detecting an optical signal comprises means for detecting fluorescent emissions of said fluorescent substance modulated by said absorber substance.

11. The apparatus of claim 10 wherein said effective number of molecules of said absorber substance are within about 10-70 Angstroms of said effective number of molecules of said fluorescent substance.

12. The apparatus of claim 11 wherein said substrate is in the form of a carrier particle.

13. The apparatus of claim 12 wherein said fiber optic core has a diameter and said carrier particle has an average diameter between about 1-3 times the diameter of said core of said fiber optic.

14. The apparatus of claim 13 wherein said carrier particle is made of porous glass.

15. The apparatus of claim 14 wherein said porous glass is doped with said fluorescent substance which is selected from the group consisting of ions of fluorescent lanthanides or fluoresscent actinides.

16. The apparatus of claim 13 wherein said carrier particle is made of sintered glass.

17. The apparatus of claim 16 wherein said sintered glass is doped with said fluorescent substance which is selected from the group consisting of ions of fluorescent lanthanides or fluorescent actinides.

18. The apparatus of claim 17 wherein said sintered glass is doped with said ions of fluorescent lanthanides or fluorescent actinides by ion implantation so that the implanted ions of fluorescent lanthanides or fluorescent actinides are within about 10-100 Angstroms from the surface of the glass.

19. The apparatus of claim 17 wherein said doped sintered glass is uranium glass.

20. A method for detecting nitrogen oxides in a sample fluid, the method comprising the steps of:

providing a fiber optic through which an illumination beam from an associated light source is transmitted from a first end of the fiber optic to a second end of the fiber optic, and a carrier particle attached to the second end of the fiber optic such that light from the illumination beam emanating from the second end of the fiber optic illuminates the carrier particle;

immobilizing on the carrier particle Saltzman'"'"'s reagent, the Saltzman'"'"'s reagent reacting with any nitrogen oxides in the sample fluid when in contact therewith to produce an absorber substance on the particle, the absorber substance having an absorption band;

immobilizing on the carrier particle a fluorescent substance having an emission band overlapping the absorption band of any absorber substance formed, such that an effective number of molecules of the fluorescent substance are closely adjacent to an effective number of molecules of the absorber to produce detectable resonant energy transfer therebetween whenever the Saltzman'"'"'s reagent reacts with nitrogen oxides in the sample fluid;

contacting the carrier particle with the sample fluid;

illuminating the fluorescent substance with the illumination beam so that excited fluorescent states are generated in the fluorescent substance;

collecting at the second end of the fiber optic fluorescent emissions of the fluorescent substance;

separating at the first end of the fiber optic the collected fluorescent emissions from the illumination beam;

measuring the intensity of backscattered Raman emissions from the fiber;

forming a ratio of the fluorescent emissions to the backscattered Raman intensity; and

relating the ratio to the concentration of nitrogen oxides in the sample fluid.

21. A method for detecting hydrogen sulfide in a sample fluid, the method comprising the steps of:

providing a fiber optic through which an illumination beam from an associated light source is transmitted from a first end of the fiber optic to a second end of the fiber optic, and a carrier particle attached to the second end of the fiber optic such that light from the illumination beam emanating from the second end of the fiber optic illuminates the carrier particle;

immobilizing on the carrier particle lead acetate, the lead acetate reacting with any hydrogen sulfide in the sample fluid when in contact therewith to produce lead sulfide having an absorption band;

immobilizing on the carrier particle a fluorescent substance having an emission band overlapping the absorption band of any lead sulfide formed, such that an effective number of molecules are the fluorescent substance are closely adjacent to an effective number of lead sulfide molecules to produce detectable resonant energy transfer therebetween whenever the lead acetate reacts with hydrogen sulfide in the sample fluid;

contacting the carrier particle with the sample fluid;

illuminating the fluorescent substance with the illumination beam so that excited fluorescent states are generated in the fluorescent substance;

collecting at the second end of the fiber optic fluorescent emissions of the fluorescent substance;

separating at the first end of the fiber optic the collected fluorescent emissions from the illumination beam;

measuring the intensity of backscattered Raman emissions from the fiber;

forming a ratio of the fluorescent emissions to the backscattered Raman intensity; and

relating the ratio to the concentration of hydrogen sulfide in the sample fluid.

22. The method of claim 21 wherein said fluorescent substance is fluorescein.

23. The method of claim 21 wherein said fluorescent substance is acridine orange.

24. A method for detecting alkali metal ions in a sample fluid, the method comprising the steps of:

providing a fiber optic through which an illumination beam from an associated light source is transmitted from a first end of the fiber optic to a second end of the fiber optic;

providing a carrier particle attached to the second end of the fiber optic such that light from the illumination beam emanating from the second end of the fiber optic illuminates the carrier particle;

immobilizing on the carrier particle a chromogenic crown ether, the chromogenic crown ether interacting with any alkali metal ions in the sample fluid when in contact therewith to produce a crown ether-alkali metal ion complex having an absorption band;

immobilizing on the carrier particle a fluorescent substance having an emission band overlapping the absorption band of any crown ether-alkali metal ion complex formed, such that an effective number of molecules of the fluorescent substance are closely adjacent to an effective number of crown ether-alkali metal ion complex molecules to produce detectable resonant energy transfer therebetween whenever such complex molecules are formed;

contacting the carrier particle with the sample fluid wherein the contacting step is carried out at a pH sufficient for the crown ether to display chromogenic characteristics in the presence of alkali metal ions;

illuminating the fluorescent substance with the illumination beam so that excited fluorescent states are generated in the fluorescent substance;

collecting at the second end of the fiber optic fluorescent emissions of the fluorescent substance;

separating at the first end of the fiber optic the collected fluorescent emissions from the illumination beam; and

relating the intensity of the collected and separated fluorescent emissions to the concentration of alkali metal ions in the sample fluid.

25. The method of claim 24 wherein said chromogenic crown ether is selected from the group consisting of crown-7, crown-6, crown-5, or crown-4 ethers.

26. The method of claim 25 wherein said chromogenic crown ether is selected from the group consisting of crown-6 or crown-5 ethers.

27. The method of claim 26 wherein said crown-6 and crown-5 ethers are 18-crown-6 and 15-crown-5 ethers, respectively.

28. The method of claim 27 wherein said alkali metal ions are potassium and/or sodium.

29. The method of claim 28 wherein said fluorescent substance is fluorescein.

30. The method of claim 27 wherein said 18-crown-6 and 15-crown-5 ethers are 4'"'"'-(substituted-phenyl)-amino-5'"'"'-substituted-benzo-18-crown-6 and 4'"'"'-(substituted-phenyl)-amino-5'"'"'-substituted-benzo-15-crown-5 ethers, respectively.

31. The method of claim 30 wherein said 18-crown-6 and 15-crown-5 ethers are 4'"'"'-(substituted-phenyl)-aminobenzo-18-crown-6 and 4'"'"'-(substituted-phenyl)-aminobenzo-15-crown-5 ethers, respectively.

32. The method of claim 31 wherein said 18-crown-6 and 15-crown-5 ethers are selected from the group consisting of

4'"'"'-picrylaminobenzo-18-crown-6,

4'"'"'-picrylaminobenzo-15-crown-5,

4'"'"'-picrylamino-5'"'"'-nitrobenzo-18-crown-6,

4'"'"'-picrylamino-5'"'"'-nitrobenzo-15-crown-5,

4'"'"'-picrylamino-5'"'"'-bromo-18-crown-6,

4'"'"'-picrylamino-5'"'"'-bromo-15-crown 5,

4'"'"'-(2",6"-dinitro-4"-trifluoromethylphenyl)aminobenzo-18-crown-6

4'"'"'-(2",6"-dinitro-4"-trifluoromethylphenyl)aminobenzo-15crown-5,

4'"'"'-(2",4"-dinitro-6"-trifluoromethylphenyl)aminobenzo-18-crown-6,

'"'"' -(2",4"-dinitro-6"-trifluoromethylphenyl)aminobenzo-15-crown-5,

4'"'"'-(4"-cyano-2",6"-dinitrophenyl)-aminobenzo-18-crown-6, and

4'"'"'-(4"-cyano-2",6"-dinitrophenyl)-aminobenzo-15-crown-5.

33. The method of claim 32 wherein said 18-crown-6 and 15-crown-5 ethers are selected from the group consisting of:

4'"'"'-picrylaminobenzo-18-crown-6,

4'"'"'-picrylaminobenzo-15-crown-5,

4'"'"'-(2",6"-dinitro-4"-trifluoromethylphenyl)aminobenzo-15-crown-5,

4'"'"'-(2"4"-dinitro-6"-trifluoremethylphenyl)aminobenzo-15-crown-5, and

4'"'"'-(4"-cyano-2",6"-dinitrophenyl)-aminobenzo-15-crown-5.

34. The method of claim 33 wherein said 18-crown-6 and 15-crown-5 ethers are respectively 4'"'"'-picrylaminobenzo-18-crown-6 and 4'"'"'-picrylaminobenzo-15-crown-5.

35. An apparatus for monitoring alkali metal ions in a sample substance, the apparatus comprising:

a fiber optic through which an illumination beam from an associated light source is transmitted from a first end of the fiber optic to a second end of the fiber optic;

a carrier particle attached to the second end of the fiber optic such that light from the illumination beam emanating from the second end of the fiber optic illuminates the carrier particle;

a fluorescent substance immobilized on the carrier particle, the fluorescent substance having an emission band; and

a chromogenic crown ether immobilized on the carrier particle, the chromogenic crown ether having a first absorption band, and the chromogenic crown ether being capable of forming a crown ether-alkali metal ion complex having a second absorption band, and the first absorption band and the second absorption band overlapping the emission band of the fluorescent substance to different degrees;

means for detecting a signal from the fluorescent substance modulated by the ether-alkali metal complex.

36. The apparatus of claim 35 wherein said chromogenic crown ether is selected from the group consisting of 18-crown-6 and 15-crown-5 ethers.

37. The apparatus of claim 36 wherein said carrier particle is porous glass.

38. The apparatus of claim 37 wherein said fluorescent substance is fluorescein.

39. The apparatus of claim 36 wherein said 18-crown-6 and 15-crown-5 ethers are respectively 4'"'"'-picrylaminobenzo-18-crown-6 and 4'"'"'-picrylaminobenzo-15-crown-5.