Method for quantitative determination of at least one chemical parameter of a sample medium

• US 5,232,858 A
• Filed: 03/04/1992
• Issued: 08/03/1993
• Est. Priority Date: 04/25/1989
• Status: Expired due to Term

• Current Assignee: Avl Medical Instruments AG (Roche Holding AG)
• Original Assignee: Avl Medical Instruments AG (Roche Holding AG)
• Inventors: Wolfbeis, Otto S., Leiner, Marco J.
• Primary Examiner(s): Johnston, Jill A.
• Application Number:
  US07/844,938
• Time in Patent Office: 517 Days
• Field of Search: 436/77, 436/79, 436/80, 436/81, 436/144, 436/172
• US Class Current: 436/77
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/80 Indicating pH value
• CPC Class Codes: Y10T 436/22 Hydrogen, per se

Abstract

A method for the quantitative determination of one or more chemical parameters of a sample medium employs a fluorescent agent which is not responsive to the parameter to be determined and which has an excitation spectrum and an emission spectrum, and a closely adjacent chromogenic agent which responds to the parameter of the sample medium to be determined by a change in its absorption spectrum the emission spectrum of the fluorescent agent at least partically overlapping the absorption spectrum of the chromogenic agent. The absorption maximum of chromogenic agent is shifted in accordance with the parameter to be determined, causing a reduction of fluorescence decay time t in the fluorescent agent, thereby enabling quantitative determination of the chemical parameter with improved long-term stability and infrequent calibrations.

Claims

1. A method for the quantitative determination of at least one chemical parameter of a sample medium, comprising the steps of:

(a) providing a fluorescent substance with an excitation spectrum and an emission spectrum and which displays a fluorescent decay time (t), said fluorescent substance being non-responsive to said chemical parameter to be determined;

(b) providing a chromogenic substance having an absorption maximum which can be shifted in wavelength in accordance with said chemical parameter to be determined;

(c) immobilizing said fluorescent and chromogenic substances in close vicinity to each other on a substrate so that the emission spectrum of said fluorescent substance at least partially overlaps the absorption spectrum of said chromogenic substance, influencing the fluorescence decay time of said fluorescent substance;

(d) contacting said fluorescent and chromogenic substances with a sample medium defining at least one chemical parameter,

(e) exposing said fluorescent substance to excitation radiation, and

(f) measuring a reduction of said fluorescence decay time (t) caused by energy transfer between said chromogenic and said fluorescent substances for the quantitative determination of said chemical parameter.

2. A method according to claim 1, wherein said fluorescent substance is 7-diethylaminocoumarin-3-carboxylic acid, said chromogenic substance is methyl orange and said chemical parameter is hydrogen ion concentration.

3. A method according to claim 1, wherein said fluorescent substance is 8-aminopyrene-1,3,6-trisulphonate, said chromogenic substance is phenol red, and said chemical parameter is hydrogen ion concentration.

4. A method according to claim 1, wherein said fluorescent substance is sulphorhodamine 101, said chromogenic substance is alizarin-complexon, and said chemical parameter is barium ion concentration.

5. A method according to claim 1, wherein said fluorescent substance is rhodamine 6G, said chromogenic substance is beryllone, and said chemical parameter is magnesium ion concentration.

6. A method according to claim 1, wherein said fluorescent substance is 2,7-dichloro-fluorescein, said chromogenic substance is catechol violet, and said chemical parameter is cadmium ion concentration.

7. A method according to claim 1, wherein said fluorescent substance is fluorescein, said chromogenic substance is chrome azurol S, and said chemical parameter is aluminium ion concentration.

8. A method according to claim 1, wherein said fluorescent substance is coumarin 7, said chromogenic substance is eriochrome black T, and said chemical parameter is calcium or lead ion concentration.

9. A method according to claim 1, wherein said fluorescent substance is coumarin 343, said chromogenic substance is murexide, and said chemical parameter is copper ion concentration.

10. A method according to claim 1, wherein said fluorescent substance is coumarin 334, said chromogenic substance is zincone, and said chemical parameter is zinc ion concentration.

11. A method according to claim 1, wherein said fluorescent substance (donor) and said chromogenic substance (acceptor) are covalently attached to each other.