EP0371253A2
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EP
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Method and reagents for detecting amphetamine and/or d-methamphetamine in biological samples
Assignee: Abbott Laboratories
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Filing: 1989-10-24
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Granted: —
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Expires: 2009-10-24
Abstract
Abstract This disclosure relates to a method and reagents for determining amphetamine and d-methamphetamine in a biological fluid, such as urine. In particular, this disclosure relates to improvements in a fluorescence polarization immunoassay procedure for determining the presence of amphetamine and d-methamphetamine in a single assay and to a novel class of tracer compounds employed as reagents in such procedures. The procedure described includes pretreatment of the biological sample to eliminate cross-reactants such as beta -hydroxyphenethylamine by preincubating the sample solely with an aqueous periodate solution having a pH from about 4.0 to about 7.5 without adjustment to an alkaline pH, and contacting the sample with riboflavin binding protein to reduce interference from fluorescent components in the sample. The procedure also maintains the cross reactivity of the immunoassay for tyramine at about 0.4% and for l-methamphetamine below about 5.1% and eliminates the necessity of using controlled substances as starting materials.
Inventors
- 👤 Daniel Feulner Heiman
- 👤 Yang Hu Hsiang-Yun
- 👤 Sharon Ann Johnson
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-03 09:54
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/542; G01N33/94; G01N33/946
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
119 words
A method for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid by fluorescence polarization assay comprising the steps of: (a) intermixing a sample with: (1) a salt of a first tracer of Formula 1; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (b) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample.
Claim 2
Independent Claim
119 words
A method for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid by fluorescence polarization assay comprising the steps of: (a) intermixing a sample with: (1) a salt of a first tracer of Formula 1; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (b) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample.
Claim 3
Independent Claim
119 words
A method for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid by fluorescence polarization assay comprising the steps of: (a) intermixing a sample with: (1) a salt of a first tracer of Formula 2; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (b) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample.
Claim 4
Independent Claim
119 words
A method for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid by fluorescence polarization assay comprising the steps of: (a) intermixing a sample with: (1) a salt of a first tracer of Formula 2; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (b) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample.
Claim 5
Dependent Claim
44 words
The method according to Claim 1 wherein said sample is pretreated with an effective amount of an aqueous periodate solution having a pH from about 4.0 to about 7.5 for a period of time sufficient to eliminate β-hydroxyphenethylamine cross reactants to amphetamine and d-methamphetamine.
Claim 6
Dependent Claim
44 words
The method according to Claim 2 wherein said sample is pretreated with an effective amount of an aqueous periodate solution having a pH from about 4.0 to about 7.5 for a period of time sufficient to eliminate β-hydroxyphenethylamine cross reactants to amphetamine and d-methamphetamine.
Claim 7
Dependent Claim
27 words
The method according to Claim 1 wherein an amount of riboflavin binding protein which is effective to reduce fluorescence interference by riboflavin is added to said sample.
Claim 8
Dependent Claim
27 words
The method according to Claim 2 wherein an amount of riboflavin binding protein which is effective to reduce fluorescence interference by riboflavin is added to said sample.
Claim 9
Dependent Claim
27 words
The method according to Claim 3 wherein an amount of riboflavin binding protein which is effective to reduce fluorescence interference by riboflavin is added to said sample.
Claim 10
Dependent Claim
27 words
The method according to Claim 4 wherein an amount of riboflavin binding protein which is effective to reduce fluorescence interference by riboflavin is added to said sample.
Claim 11
Independent Claim
180 words
A method for determining amphetamine and d-methamphetamine in test samples of biological fluid by fluorescence polarization assay utilizing an automatic assay apparatus having a sample and reagent dispensing means comprising the steps of: (a) pretreating a sample to eliminate β-hydroxyphenethylamine cross-reactivity using only an effective amount of an aqueous periodate solution having a pH from about 4.0 to 7.5 for a period of time sufficient to eliminate said β-hydroxyphenethylamine cross-reactivity; (b) intermixing a sample with: (1) a salt of a first tracer of Formula 1; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (c) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample; and (d) washing said dispensing means in a solution of 5 solution to minimize sample adhesion to said dispensing means.
Claim 12
Independent Claim
180 words
A method for determining amphetamine and d-methamphetamine in test samples of biological fluid by fluorescence polarization assay utilizing an automatic assay apparatus having a sample and reagent dispensing means comprising the steps of: (a) pretreating a sample to eliminate β-hydroxyphenethylamine cross-reactivity using only an effective amount of an aqueous periodate solution having a pH from about 4.0 to 7.5 for a period of time sufficient to eliminate said β-hydroxyphenethylamine cross-reactivity; (b) intermixing a sample with: (1) a salt of a first tracer of Formula 1; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (c) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample; and (d) washing said dispensing means in a solution of 5 solution to minimize sample adhesion to said dispensing means.
Claim 13
Independent Claim
180 words
A method for determining amphetamine and d-methamphetamine in test samples of biological fluid by fluorescence polarization assay utilizing an automatic assay apparatus having a sample and reagent dispensing means comprising the steps of: (a) pretreating a sample to eliminate β-hydroxyphenethylamine cross-reactivity using only an effective amount of an aqueous periodate solution having a pH from about 4.0 to 7.5 for a period of time sufficient to eliminate said β-hydroxyphenethylamine cross-reactivity; (b) intermixing a sample with: (1) a salt of a first tracer of Formula 2; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (c) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample; and (d) washing said dispensing means in a solution of 5 solution to minimize sample adhesion to said dispensing means.
Claim 14
Independent Claim
180 words
A method for determining amphetamine and d-methamphetamine in test samples of biological fluid by fluorescence polarization assay utilizing an automatic assay apparatus having a sample and reagent dispensing means comprising the steps of: (a) pretreating a sample to eliminate β-hydroxyphenethylamine cross-reactivity using only an effective amount of an aqueous periodate solution having a pH from about 4.0 to 7.5 for a period of time sufficient to eliminate said β-hydroxyphenethylamine cross-reactivity; (b) intermixing a sample with: (1) a salt of a first tracer of Formula 2; (2) a salt of a second tracer of Formula 3; (3) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (4) a second antibody capable of specifically recognizing and binding d-methamphetamine and said second tracer thereof; and (c) determining the amount of each tracer bound to said first and second antibodies by fluorescence polarization techniques as a measure of the amount of amphetamine and d-methamphetamine in the sample; and (d) washing said dispensing means in a solution of 5 solution to minimize sample adhesion to said dispensing means.
Claim 15
Dependent Claim
68 words
A reagent kit useful in the determination of amphetamine and d-methamphetamine in biological samples comprising: (a) a salt of a first tracer of Formula 1; (b) a salt of a second tracer of Formula 3; (c) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (d) a second antibody capable of specifically recognizing and binding methamphetamine and said second tracer thereof.
Claim 16
Dependent Claim
68 words
A reagent kit useful in the determination of amphetamine and d-methamphetamine in biological samples comprising: (a) a salt of a first tracer of Formula 1; (b) a salt of a second tracer of Formula 3; (c) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (d) a second antibody capable of specifically recognizing and binding methamphetamine and said second tracer thereof.
Claim 17
Dependent Claim
68 words
A reagent kit useful in the determination of amphetamine and d-methamphetamine in biological samples comprising: (a) a salt of a first tracer of Formula 2; (b) a salt of a second tracer of Formula 3; (c) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (d) a second antibody capable of specifically recognizing and binding methamphetamine and said second tracer thereof.
Claim 18
Dependent Claim
68 words
A reagent kit useful in the determination of amphetamine and d-methamphetamine in biological samples comprising: (a) a salt of a first tracer of Formula 2; (b) a salt of a second tracer of Formula 3; (c) a first antibody capable of specifically recognizing and binding amphetamine and said first tracer thereof; and (d) a second antibody capable of specifically recognizing and binding methamphetamine and said second tracer thereof.
Claim 19
Dependent Claim
40 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Formula 1.
Claim 20
Dependent Claim
40 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Formula 1.
Claim 21
Dependent Claim
40 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Formula 2.
Claim 22
Dependent Claim
40 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Formula 2.
Claim 23
Dependent Claim
40 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Formula 3.
Claim 24
Dependent Claim
40 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Formula 3.
Claim 25
Dependent Claim
59 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Figure 1, Figure 2, Figure 3, Figure 4, Figure 5, Figure 6, Figure 7, Figure 8, Figure 9 or Figure 10.
Claim 26
Dependent Claim
59 words
A tracer reagent useful in a fluorescence polarization immunoassay for for detecting the presence, or determining the amount, of amphetamine and/or d-methamphetamine in a test sample of biological fluid comprising a tracer which has the structure shown in Figure 1, Figure 2, Figure 3, Figure 4, Figure 5, Figure 6, Figure 7, Figure 8, Figure 9 or Figure 10.
Claim 27
Dependent Claim
20 words
A tracer precursor compound comprising a compound which has the structure shown in Formula 4, Formula 5 or Formula 6.
Claim 28
Dependent Claim
20 words
A tracer precursor compound comprising a compound which has the structure shown in Formula 4, Formula 5 or Formula 6.
Claim 29
Dependent Claim
19 words
An immunogen compound comprising an immunogen which has the structure shown in Formula 7, Formula 8 or Formula 9.
Claim 30
Dependent Claim
19 words
An immunogen compound comprising an immunogen which has the structure shown in Formula 7, Formula 8 or Formula 9.
Claim 31
Dependent Claim
21 words
An immunogen compound according to Claim 17 wherein said immunogen compound has the structure shown in Figure 20 or Figure 21.
Claim 32
Dependent Claim
17 words
An immunogen precursor compound comprising a compound which has the structure shown in Figure 22 or 23.
Claim 33
Dependent Claim
17 words
An immunogen precursor compound comprising a compound which has the structure shown in Figure 22 or 23.
References Cited (Backward Citations)
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