Rapid fluorescence tagging of glycans and other biomolecules with enhanced MS signals

Patent metadata

Field	Value
Publication number	US-9772333-B2
Application number	US-201414193418-A
Country	US
Kind code	B2
Filing date	Feb 28, 2014
Priority date	Sep 28, 2011
Publication date	Sep 26, 2017
Grant date	Sep 26, 2017

Abstract

Mass spectrometry (MS) active, fluorescent rapid tagging reagent is provided having three substituent groups: (a) a tertiary amino group or other MS active atom; (b) a highly fluorescent moiety; and (c) a reactive group that can react with an amine. The reactive group provides rapid tagging of desired bio-molecules. The fluorescent moiety provides the fluorescent signal. The tertiary amino group provides the MS signal.

First claim

We claim:

1. A compound of the structural formula:

2. A compound of the structural formula:

3. A compound of Formula IV:

wherein
X=C or N
R 1 is O═C═N— or

R 2 is independently selected from —H, —C 1 -C 8 alkyl, —C 1 -C 8 cycloalkyl, halo, dialkylamino, CH 2 -dialkylamino, aminocarbonyl, alkoxycarbonyl, or alkoxy, but not Cl or O═C═N—; and
R 3 and R 4 are independently selected from —H, alkyl, alkyl amino, alkylsulfonic acid, alkyl phosphonic acid, wherein either R 3 or R 4 is alkylamino, alkyl phosphonic acid, or alkylsulfonic acid.

4. A compound of Formula V:

wherein
m=0-9;
n=0-9;
X=C or N
R 1 is O═C═N—, S═C═N—, or

R 2 is independently selected from methylene, substituted nitrogen, oxygen, carbonyl, amide, ester, sulfur, sulfoxide, or sulfone;
R 3 and R 4 are independently selected from —H, alkyl, alkyl amino, alkylsulfonic acid, alkyl phosphonic acid, wherein either R 3 or R 4 is alkylamino, alkyl phosphonic acid, or alkylsulfonic acid; and
R 5 is independently selected from —H, —C 1 -C 8 alkyl, —C 1 -C 8 cycloalkyl, halo, dialkylamino, CH 2 -dialkylamino, aminocarbonyl, alkoxycarbonyl, or alkoxy, but not Cl when R 1 is O═C═N—, and when R 1 is S═C═N, R 5 is H.

5. A method for analyzing a glycan, an amino acid or an amine containing compound in a sample by means of liquid chromatography and mass spectrometry comprising labeling the glycan, the amino acid or the amine containing compound in the sample by reacting with a compound of the structural formula:

for a time and under conditions suitable to facilitate the labeling; ; and subjecting the labeled compound to liquid chromatograph and mass spectrometry.

6. A method for analyzing a glycan, an amino acid or an amine containing compound in a sample by means of liquid chromatography and mass spectrometry comprising labeling the glycan, the amino acid or the amine containing compound in the sample by reacting with a compound of the structural formula

for a time and under conditions suitable to facilitate the labeling; ; and subjecting the labeled compound to liquid chromatograph and mass spectrometry.

7. A method for analyzing a glycan, an amino acid or an amine containing compound in a sample by means of liquid chromatography and mass spectrometry comprising labeling the glycan, the amino acid or the amine containing compound in the sample by reacting with a compound according to claim 3 for a time and under conditions suitable to facilitate the labeling and subjecting the labeled compound to liquid chromatograph and mass spectrometry.

8. A method for analyzing a glycan, an amino acid or an amine containing compound in a sample by means of liquid chromatography and mass spectrometry comprising labeling the glycan, the amino acid or the amine containing compound in the sample by reacting with a compound according to claim 4 for a time and under conditions suitable to facilitate the labeling and subjecting the labeled compound to liquid chromatograph and mass spectrometry.

9. A method for rapid tagging of a glycan, an amino acid or an amine containing compound comprising reacting the glycan, the amino acid or the amine containing compound with the compound of the structural formula:

10. A method for rapid tagging of a glycan, an amino acid or an amine containing compound comprising reacting the glycan, the amino acid or the amine containing compound with the compound of the structural formula:

11. A method for rapid tagging of a glycan, an amino acid or an amine containing compound comprising reacting the glycan, the amino acid or the amine containing compound with the compound of claim 3 .

12. A method for rapid tagging of a glycan, an amino acid or an amine containing compound comprising reacting the glycan, the amino acid or the amine containing compound with the compound of claim 4 .

Assignees

• Waters Technologies Corp
• Water Tech Corp

Inventors

• Brousmiche Darryl W
• Yu Ying-Qing

Classifications

• C07D405/14

  containing three or more hetero rings · CPC title

• C07D413/12

  linked by a chain containing hetero atoms as chain links · CPC title

• G01N2400/12

  Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar · CPC title

• G01N2458/20

  Labels for detection by gas chromatography, e.g. haloaryl systems · CPC title

• C07D207/46 Primary

  with hetero atoms directly attached to the ring nitrogen atom · CPC title