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