US3871825A
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Fluorometric reagents and methods
Assignee: F Hoffmann La Roche AG
•
Filing: 1974-03-04
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Granted: 1975-03-18
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Expires: 1994-03-04
Abstract
Abstract A novel class of reagents react with primary amine-containing compounds, preferably polypeptides and amino acids to form highly fluorescent products. This reaction serves as a basis for a rapid and highly sensitive assay method for primary amine-containing compounds.
Inventors
- 👤 Willy Leimgburger
- 👤 Manfred Weigele
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 09:15
CPC Classifications
C; C07; C07D; C07D493/00; C07D493/02; C07D493/10; C07C; C07C229/00; C07C229/38; C07C45/00; C07C45/61; C07C45/67; C07C45/68; C07C45/72; C07C45/74; C07C49/00; C07C49/587; C07C49/657; C07C49/683; C07C49/687; C07C49/697; C07C49/753; C07C49/755; C07C65/00; C07C65/32; C07C65/40; C07D303/00; C07D303/02; C07D303/12; C07D303/32; C07D311/00; C07D311/02; C07D311/76; C07D317/00; C07D317/08; C07D317/44; C07D317/46; C07D317/48; C07D317/50; C07D317/60; G; G01; G01N; G01N31/00; G01N31/22; G01N33/00; G01N33/48; G01N33/50; G01N33/52; G01N33/68; G01N33/6803; G01N33/6806; G01N33/6809; Y; Y10; Y10T; Y10T436/00; Y10T436/17; Y10T436/173845
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
25 words
A METHOD FOR DETERMINING A PRIMARY AMINE-CONTAINING COMPOUND IN A MIXTURE CONTAINING SAME WHICH METHOD COMPRISES: TREATING SAID MIXTURE WITH A COMPOUND OF FORMULA I
Claim 2
Independent Claim
25 words
A METHOD FOR DETERMINING A PRIMARY AMINE-CONTAINING COMPOUND IN A MIXTURE CONTAINING SAME WHICH METHOD COMPRISES: TREATING SAID MIXTURE WITH A COMPOUND OF FORMULA I
Claim 3
Dependent Claim
42 words
The method of claim 1 wherein the primary amine-containing compound is present on at least a portion of a surface of a paper or thin-layer chromatographic medium and the compound of formula I is applied to said surface of said chromatographic medium.
Claim 4
Dependent Claim
13 words
The method of claim 2 wherein R is hydrogen and R'' is phenyl.
Claim 5
Dependent Claim
18 words
The method of claim 3 wherein the pH of the chromatographic medium is kept between 8 and 9.
Claim 6
Dependent Claim
31 words
The method of claim 3 wherein the compound of formula I is present in a molar excess of between 10 and 5000 as compared with the amount of primary amine-containing compound.
Claim 7
Dependent Claim
15 words
The method of claim 3 wherein the primary amine-containing compound is an Alpha -amino acid.
Claim 8
Dependent Claim
13 words
The method of claim 3 wherein the primary amine-containing compound is a peptide.
Claim 9
Dependent Claim
70 words
The method of claim 1 wherein the primary amine containing compound is determined by an automated procedure which comprises: a. continuously treating a sample stream containing said primary amine-containing compound in solution with an excess of a compound of formula I, and b. passing the mixture obtained in step (a) at a constant rate through a flow cell of a fluorophotometer whereby any fluorescent material present is fluoresced and detected.
Claim 10
Dependent Claim
21 words
The method of claim 8 wherein the sample stream is at least a portion of the effluent of a chromatographic column.
Claim 11
Dependent Claim
13 words
The method of claim 8 wherein R is hydrogen and R'' is phenyl.
Claim 12
Dependent Claim
31 words
The method of claim 8 wherein the compound of formula I is present in a molar excess of between 10 and 5000 as compared with the amount of primary amine-containing compound.
Claim 13
Dependent Claim
19 words
The method of claim 8 wherein the pH of the solution being fluoresced is maintained between 8 and 9.
Claim 14
Dependent Claim
15 words
The method of claim 8 wherein the primary amine-containing compound is an Alpha -amino acid.
Claim 15
Dependent Claim
13 words
The method of claim 8 wherein the primary amine-containing compound is a peptide.
Claim 16
Dependent Claim
84 words
The method of claim 8 wherein the total amount of a specific known primary amine-containing compound present in the sample stream is quantitatively determined by a. determining the total fluorescence produced by a known quantity of said primary amine-containing compound, b. determining the total fluorescence produced by the unknown quantity of said primary amine-containing compound under the same conditions as step (a), and c. obtaining the unknown amount by multiplying the known quantity by the ratio of the unknown to the known total fluorescence.
Claim 17
Dependent Claim
12 words
The method of claim 1 wherein said treatment is done in solution.
Claim 18
Dependent Claim
13 words
The method of claim 16 wherein R is hydrogen and R'' is phenyl.
Claim 19
Dependent Claim
16 words
The method of claim 16 wherein the pH of the solution is between 8 and 9.
Claim 20
Dependent Claim
31 words
The method of claim 16 wherein the compound of formula I is present in a molar excess of between 10 and 5000 as compared with the amount of primary amine-containing compound.
Claim 21
Dependent Claim
15 words
The method of claim 16 wherein the primary amine-contAining compound is an Alpha -amino acid.
Claim 22
Dependent Claim
13 words
The method of claim 16 wherein the primary amine-containing compound is a peptide.
Claim 23
Dependent Claim
14 words
The method of claim 16 wherein a fluorophotometer is utilized for fluorescing said substance.
Claim 24
Dependent Claim
66 words
The method of claim 16 wherein the concentration in solution of a known primary amine-containing compound is quantitatively determined by a. determining the fluorescence produced by a measured concentration of said amine, b. determining the fluorescence produced by the unknown concentration of said amine, and c. obtaining the unknown concentration by multiplying the known concentration by the ratio of the unknown to the known fluorescence values.
References Cited (Backward Citations)
US3689221A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-03-17 | 1978-09-16 | 1979-03-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-03-17 | 1982-09-16 | 1983-03-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-03-17 | 1986-09-16 | 1987-03-18 | $7700.00 | Window Closed / Lapsed |