Method for analysis of compounds with amino group and analytical reagent therefor

Patent metadata

Field	Value
Publication number	US-9274123-B2
Application number	US-51413006-A
Country	US
Kind code	B2
Filing date	Sep 1, 2006
Priority date	Feb 14, 2002
Publication date	Mar 1, 2016
Grant date	Mar 1, 2016

Abstract

The present invention provides a method for the analysis of a compound with amino group (e.g., an amino acid or peptide) contained in a sample and convenient manner with a high sensitivity. The compound with amino group in a sample containing the compound with amino group is labeled with a specific carbamate compound such as p-trimethylammonium anilyl-N-hydroxysuccinimidyl carbamate iodide to enhance the selectivity and sensitivity. The present invention is preferably used in conjunction with mass spectrometry such as MS/MS method to facilitate quantitative analysis. The present invention further provides labeling reagents for mass spectrometry.

First claim

The invention claimed is:

1. A method for analyzing a compound having an amino group, an imino group, or mixture thereof, in a sample by tandem mass spectrometry, comprising:
(a) labeling the compound having an amino group, an imino group, or mixture thereof with a carbamate compound represented by formula (Ia) or a salt thereof to form a labelled compound having a bond between a nitrogen of the amino group, imino group, or mixture thereof and a labeling a portion of the carbamate compound represented by formula (Ia) or a salt thereof:

wherein Ar is an aromatic carbocyclic group or an aromatic heterocyclic group, wherein the aromatic carbocyclic group or the aromatic heterocyclic group may have one or more substituents,
(b) selecting or scanning, in a first mass analyzer in positive ion mode, m/z values derived from pre-selected ions of the labeled compound or ranges of m/z values which include ions of the labeled compounds;
(c) collisionally cleaving the ions of the labeled compound at the bond between the nitrogen of the amino group or the imino group of the labelled compound and the labeling portion of the carbamate compound represented by formula (Ia) or a salt thereof to produce a fragment ion and a neutral fragment; and
(d) selecting or scanning, in a second mass analyzer:
(1) pre-selected m/z values of a fragment ion derived from the labeling portion of the carbamate compound represented by formula (Ia) or a salt thereof,
(2) pre-selected m/z values of a fragment ion derived from the compound having an amino group, an imino group, or a mixture thereof, or
(3) ranges of m/z values at a pre-selected offset value from the first mass analyzer, wherein the offset is derived from loss of the labeling portion of the carbamate compound represented by formula (Ia) or a salt thereof.

2. The method of claim 1 , wherein Ar is selected from the group consisting of a phenyl group, naphthyl group, anthryl group, pyridyl group, pyrazyl group, quinolyl group, acridyl group and coumaryl group.

3. The method of claim 2 , wherein Ar is substituted and the substituent is selected from the group consisting of an alkyl group, aromatic group, halogen atom, carboxyl group, hydroxyl group, nitro group, diazo group, cyano group, alkoxy group, acyl group, sulfonic acid group, phosphoric acid group, guanidyl group, dialkylamino group and trialkylammonium group.

4. The method of claim 1 , wherein Ar is substituted and the substituent is selected from the group consisting of an alkyl group, aromatic group, halogen atom, carboxyl group, hydroxyl group, nitro group, diazo group, cyano group, alkoxy group, acyl group, sulfonic acid group, phosphoric acid group, guanidyl group, dialkylamino group and trialkylammonium group.

5. The method of claim 1 , wherein the compound having an amino group, an imino group, or mixture thereof is a compound having an amino group.

6. The method of claim 1 , wherein the compound having an amino group, an imino group, or mixture thereof is an amino acid.

7. The method of claim 1 , wherein the compound having an amino group, an imino group, or mixture thereof is a peptide.

8. The method of claim 1 , wherein the compound having an amino group, an imino group, or mixture thereof is a protein.

9. The method of claim 1 , wherein the carbamate compound is 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) or p-trimethylammoniumanilyl-N-hydroxysuccinimidyl carbamate iodide (TAHS).

10. The method of claim 1 , wherein the carbamate compound is selected from the group consisting of p-dimethylaminoanilyl-N-hydroxysuccinimidyl carbamate, 3-aminopyridyl-N-hydroxysuccinimidyl carbamate, p-trimethylammonium anilyl-N-hydroxysuccinimidyl carbamate iodide, and aminopyrazyl-N-hyroxysuccinimidyl carbamate.

11. The method of claim 1 , wherein the carbamate compound is 1-naphthylamino-N-hydroxysuccinimide carbamate (NAHS).

12. The method of claim 1 , comprising selecting or scanning (1) in (d).

13. The method of claim 1 , comprising selecting or scanning (2) in (d).

14. The method of claim 1 , comprising selecting or scanning (3) in (d).

15. The method of claim 1 , further comprising determining an isotope ratio of the compound having an amino group, an imino group, or mixture thereof.

16. The method of claim 1 , further comprising, after (a) and prior to (b),
(a′) mixing the labeled compound with at least one additional sample containing the same compound having an amino group, an imino group, or mixture thereof labeled with an isotope labeled carbamate compound represented by formula (Ib) or a salt thereof:

wherein
Ar is an aromatic carbocyclic group or an aromatic heterocyclic group, wherein the aromatic carbocyclic group or an aromatic heterocyclic group may have one or more substituents,
the compound represented by formula (Ib) has been enriched in one or more isotopes selected from the group consisting of 13 C, 2 H, 15 N and 18 O, and
wherein the carbamate compound represented by formula (Ia) or a salt thereof has the same general formula as the carbamate compound represented by formula (Ib) or a salt thereof.

17. The method of claim 16 , wherein the isotope-labeled carbamate compound represented by formula (Ib) or a salt thereof is enriched in 13 C.

18. The method of claim 16 , wherein the isotope-labeled carbamate compound represented by formula (Ib) or a salt thereof is enriched in 2 H.

19. The method of claim 16 , wherein the isotope-labeled carbamate compound represented by formula (Ib) or a salt thereof is enriched in 15 N.

20. The method of claim 16 , wherein the isotope-labeled carbamate compound represented by formula (Ib) or a salt thereof is enriched in 18 O.

Assignees

• Miyano Hiroshi
• Yahashi Akihisa
• Shimbo Kazutaka
• Nakazawa Masakazu
• Hirayama Kazuo
• Ajinomoto Kk

Inventors

• Miyano Hiroshi
• Yahashi Akihisa
• Shimbo Kazutaka
• Nakazawa Masakazu
• Hirayama Kazuo

Classifications

• G01N2458/15

  Non-radioactive isotope labels, e.g. for detection by mass spectrometry · CPC title

• Y10T436/13

  Tracers or tags · CPC title

• Y10T436/14

  Heterocyclic carbon compound [i.e. , O, S, N, Se, Te, as only ring hetero atom] · CPC title

• Y10T436/145555

  Hetero-N · CPC title

• C07D207/46

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