Simultaneous dual excitation/single emission fluorescent sensing method for PH and pCO.sub.2

• US 5,672,515 A
• Filed: 09/12/1995
• Issued: 09/30/1997
• Est. Priority Date: 09/12/1995
• Status: Expired due to Fees

• Current Assignee: Optical Sensors Incorporated
• Original Assignee: Optical Sensors Incorporated
• Inventors: Furlong, Steven C.
• Primary Examiner(s): SNAY, JEFFREY R
• Application Number:
  US08/526,950
• Time in Patent Office: 749 Days
• Field of Search: 422/82.06, 422/82.07, 422/82.08, 436/172, 436/133, 436/163, 250/227.21, 250/458.1, 250/459.1
• US Class Current: 436/133
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 2021/772 Tip coated light guide
• CPC Class Codes: G01N 2021/7759 Dipstick; Test strip
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/643 non-biological material
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 2201/062 LED's
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH
• CPC Class Codes: Y10T 436/204998 Inorganic carbon compounds

Abstract

A simultaneous dual excitation/single emission ratiometric method using fiber optic sensors is provided for detecting or measuring a parameter of interest in a sample, including pH, concentration of gases such as CO₂, and the like. The method involves simultaneously providing modulated optical light corresponding to first and second regions of an indicator species'"'"' absorption or excitation spectrum, detecting modulated emission signals from the indicator species, demodulating the emission signals, calculating the apparent quantity of analyte present in the sample from the indicator emission signals and correcting the apparent quantity of analyte present for variations resulting from external factors, by determining the ratio of the demodulated indicator emission signals. An apparatus is provided for performing the method as well.

Claims

1. A method for quantitating an analyte in a sample, comprising:

(a) providing an optical sensor comprising an optical fiber means having a distal end portion for contacting the fluid sample, and a proximal end portion for communication with a means for receiving a signal from the distal end portion, and wherein the distal end portion has an analyte sensing means comprising an indicator species having an absorption or excitation spectrum that includes a first region and a second region and wherein the first and second regions do not overlap substantially, and an emission spectrum that is distinct from the absorption or excitation spectrum;

(b) contacting the sample with the distal end portion of the optical sensor;

(c) simultaneously exciting the indicator species using modulated radiation of a first optical wavelength corresponding to the first region, thereby producing a first modulated indicator emission signal, and modulated radiation of a second optical wavelength corresponding to the second region, thereby producing a second modulated indicator emission signal, wherein the radiation of first and second optical wavelengths are respectively transmitted at first and second electrical frequencies;

(d) demodulating the first and second modulated indicator emission signals;

(e) calculating the apparent quantity of analyte present in the sample from the first and second indicator emission signals; and

(f) correcting the apparent quantity of analyte present for variations resulting from external factors, by determining the ratio of the first and second indicator emission signals.

2. The method of claim 1, wherein the sample is a fluid sample.

3. The method of claim 1, wherein the analyte sensing means comprises an indicator species having an excitation spectrum that includes a first region and a second region.

4. The method of claim 1, wherein the analyte sensing means comprises an indicator species having an absorption spectrum that includes a first region and a second region.

5. The method of claim 1, wherein the first and second regions respectively correspond to the acid and base forms of the indicator species.

6. The method of claim 3, wherein the first and second regions respectively correspond to the acid and base forms of the indicator species.

7. The method of claim 3, wherein the first region of the excitation spectrum is sensitive to the analyte and the second region of the excitation spectrum is insensitive to the analyte.

8. The method of claim 7, wherein the second region is the isosbestic point.

9. The method of claim 1, wherein the analyte is selected from the group consisting of pH and pCO₂.

10. The method of claim 9, wherein the analyte is pH.

11. The method of claim 9, wherein the analyte is pCO₂.

12. The method of claim 1, wherein the indicator species is selected from the group consisting of fluorescein, carboxyfluorescein, seminaphthorhodafluor, seminaphthofluorescein, naphthofluorescein, hydroxypyrene trisulfonic acid and dichlorofluorescein.

13. The method of claim 12, wherein the indicator species is hydroxypyrene trisulfonic acid.

14. The method of claim 12, wherein the indicator species is fluorescein.

15. An apparatus for quantitating an analyte in a sample, comprising:

(a) an optical sensor having an indicator species having an absorption or excitation spectrum that includes a first region and a second region such that the first and second regions do not overlap substantially, and an emission spectrum that is distinct from the absorption or excitation spectrum;

(b) a means for simultaneously generating radiation of first and second modulated optical wavelengths by which the indicator species can be excited;

(c) a means for modulating the first and second optical wavelengths;

(d) a means to detect the emission signal from the excited indicator; and

(e) a means to demodulate the emission signal.

16. The apparatus of claim 15, wherein the analyte sensing means comprises an indicator species having an excitation spectrum that includes a first region and a second region.

17. The apparatus of claim 15, wherein the analyte sensing means comprises an indicator species having an absorption spectrum that includes a first region and a second region.

18. The apparatus of claim 15, wherein the first and second regions respectively correspond to the acid and base forms of the indicator species.

19. The apparatus of claim 16, wherein the first and second regions respectively correspond to the acid and base forms of the indicator species.

20. The apparatus of claim 16, wherein the first region of the excitation spectrum is sensitive to the analyte and the second region of the excitation spectrum is insensitive to the analyte.

21. The apparatus of claim 20, wherein the second region is the isosbestic point.

22. The apparatus of claim 15, wherein the analyte is selected from the group consisting of pH and pCO₂.

23. The apparatus of claim 22, wherein the analyte is pH.

24. The apparatus of claim 22, wherein the analyte is pCO₂.

25. The apparatus of claim 15, wherein the indicator species is selected from the group consisting of fluorescein, carboxyfluorescein, seminaphthorhodafluor, seminaphthofluorescein, naphthofluorescein, hydroxypyrene trisulfonic acid and dichlorofluorescein.

26. The apparatus of claim 25, wherein the indicator species is hydroxypyrene trisulfonic acid.

27. The apparatus of claim 25, wherein the indicator species is fluorescein.

28. The apparatus of claim 15, wherein the means for simultaneously generating radiation is selected from the group consisting of an incandescent lamp, an arc lamp, a flash lamp, a light emitting diode and a laser.

29. The apparatus of claim 28, wherein the means for simultaneously generating radiation is a light emitting diode.