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US4565790A
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Method for fluorescence spectroscopic determination of a biologically active substance
Assignee: Wallac Oy
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Filing: 1983-07-14
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Granted: 1986-01-21
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Expires: 2003-07-14
Abstract
Abstract Method for fluorescence spectroscopic determination of a biological substance provided with a marker consisting of a lanthanide chelate complex formed by a lanthanide coupled to the substance via a chelate forming compound, such as an EDTA-analogue, the lanthanide ion before the detection is dissociated from the active substance for instance by adding a buffer with a pH value below 3.5 and a splitting detergent, whereafter the excitation in the determination takes place in the solution in the presence of a β-diketone by using a short radiation pulse and the fluorescence from the marker is detected when the fluorescence from the noise sources has substantially ceased.
Inventors
- 👤 Ilkka Hemmila
- 👤 Salifu Dakubu
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 07:07
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/539; G01N33/58; G01N33/582; G01N2458/00; G01N2458/40; Y; Y10; Y10S; Y10S436/00; Y10S436/80; Y10S436/805; Y10S436/825
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
111 words
In a method for fluorescence spectroscopic determination of a biological substance provided with a marker consisting of a lanthanide chelate complex formed of a lanthanide coupled to the substance via a chelate forming compound, by excitation by a short radiation pulse, and detection of the fluorescence of the marker when the fluorescence from any noise source has substantially ceased, wherein the improvement comprises, adding, before the determination, a solution that brings the pH to 3.5 or below and contains a detergent and a β-diketone to dissociate the lanthanide ion from the chelate complex and to transfer the dissociated lanthanide ion into a fluorescent form to enable the determination in the solution.
Claim 2
Dependent Claim
111 words
In a method for fluorescence spectroscopic determination of a biological substance provided with a marker consisting of a lanthanide chelate complex formed of a lanthanide coupled to the substance via a chelate forming compound, by excitation by a short radiation pulse, and detection of the fluorescence of the marker when the fluorescence from any noise source has substantially ceased, wherein the improvement comprises, adding, before the determination, a solution that brings the pH to 3.5 or below and contains a detergent and a β-diketone to dissociate the lanthanide ion from the chelate complex and to transfer the dissociated lanthanide ion into a fluorescent form to enable the determination in the solution.
Claim 3
Dependent Claim
12 words
Method according to claim 1 wherein the solution is an aqueous solution.
Claim 4
Dependent Claim
10 words
Method according to claim 1 wherein the lanthanide is europium.
Claim 5
Dependent Claim
14 words
Method according to claim 1 wherein the detergent is an alkyl aryl polyether alcohol.
Claim 6
Dependent Claim
12 words
Method according to claim 1 wherein the β-diketone is 2-naphthoyltrifluoroacetone or 1-naphthoyltrifluoroacetone.
Claim 7
Dependent Claim
19 words
Method according to claim 1 wherein the β-diketone is 2-naphthoyltrifluoroacetone, 1-naphthoyltrifluoroacetone, p-methoxybenzoyltrifluoroacetone, p-fluoribenzoyltrifluoroacetone, benzoyltrifluoroacetone, furoyltrifluoroacetone, naphthoylfuroylmethane, dithenoylmethane and dibenzoylmethane.
Claim 8
Dependent Claim
12 words
Method according to claim 1 wherein the solution contains a Lewis base.
Claim 9
Dependent Claim
11 words
Method according to claim 3 wherein the Lewis base is trioctylphosphineoxide.
Claim 10
Dependent Claim
13 words
Method according to claim 1 wherein the chelate forming compound is an EDTA-analogue.
Claim 11
Dependent Claim
43 words
Method according to claim 1 wherein the solution is a buffer, having a pH of 2.8 to 35, which includes 10 to 100 μM of 2-naphthoyltrifluroacetone, 10 to 100 μm of trioctylphosphineoxide and 0.1 to 0.5 percent of an alkyl aryl polyether alcohol.
References Cited (Backward Citations)
US4341957A
Cited
US4283382A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-01-20 | 1989-07-22 | 1990-01-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-01-20 | 1993-07-22 | 1994-01-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-01-20 | 1997-07-22 | 1998-01-21 | $7700.00 | Window Closed / Lapsed |