Mass spectrometric identification and/or quantitation of catecholamines using aminopyrazoles

Patent metadata

Field	Value
Publication number	US-9964552-B2
Application number	US-201415032121-A
Country	US
Kind code	B2
Filing date	Oct 22, 2014
Priority date	Oct 30, 2013
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

A method is described for mass spectrometric analysis, detection and quantification of catecholamines. The methods can comprise reacting the catecholamines with a 4-aminoantipyrine reagent and detecting and/or quantifying the adduct produced by the reaction. The methods can also allow for multiplexing. Compounds formed by the reactions are also provided.

First claim

The invention claimed is:

1. A method of detecting the presence of one or more catecholamines in a sample, the method comprising:
a) reacting the sample with a 4-aminoantipyrine reagent, or a heavy atom derivative thereof, under conditions sufficient to react with a catecholamine, if any, present in the sample to form a 4-aminoantipyrine-reagent-catecholamine adduct, or a heavy atom derivative thereof;
b) obtaining a mass spectrum of said reacted sample; and
c) analyzing the mass spectrum to determine whether the sample contains a 4-aminoantipyrine-reagent-catecholamine adduct or an ionic fragment thereof, wherein the 4-aminoantipyrine reagent is a compound of Formula I or a heavy atom derivative thereof:

wherein
R 1

y is 1-6; and
X is an anion.

2. The method of claim 1 , wherein the one or more catecholamines is epinephrine, isoprenaline, norepinephrine, nordefrin, adrenalone hydrochloride, epinine hydrochloride, 2-(dimethylamino)-3′,4′-dihydroxy-(8Cl), 2-(Dimethylamino)-3′,4′-dihydroxyacetophenone, dioxethedrin, etyprenaline, N-methylepinephrine, dihydroxyephedrine hydrochloride, butylnoradrenaline, ethylnoradrenaline, 3,4-dihydroxy-phenylalanine, dopamine, or a salt or hydrates thereof, or any combination thereof.

3. The method of claim 1 , wherein the analyzing step comprises detecting one or more mass signals associated with any of said adduct and one or more fragments thereof and quantitating the amount of the catecholamine in the sample.

4. The method of claim 1 , wherein the heavy atom derivative of Formula I is a compound of Formula Ia, Ib, or Ic.

5. A method of detecting the presence of one or more catecholamines in a sample, the method comprising:
a) reacting the sample with a 4-aminoantipyrine reagent, or a heavy atom derivative thereof, under conditions sufficient to react with a catecholamine, if any, present in the sample to form a 4-aminoantipyrine-reagent-catecholamine adduct, or a heavy atom derivative thereof;
b) obtaining a mass spectrum of said reacted sample; and
c) analyzing the mass spectrum to determine whether the sample contains a 4-aminoantipyrine-reagent-catecholamine adduct or an ionic fragment thereof, wherein the aminoantipyrine-catecholamine adduct is a compound of Formula II, or a heavy atom derivative thereof:

wherein
R 4 is H or

R 5 is

z is 1-6; and
Q is a halogen.

6. The method of claim 5 , wherein the heavy atom derivative of Formula II is a compound of Formula IIa or IIb

7. The method of claim 5 , wherein the ionic fragment of the aminoantipyrine-catecholamine adduct is a compound of Formula III, or a heavy atom derivative thereof:

wherein
R 6 is

8. The method of claim 1 , wherein reacting the sample with 4-aminoantipyrine under conditions to form a 4-aminoantipyrine-catecholamine adduct comprises contacting the sample with 4-aminoantipyrine in the presence of ammonium bicarbonate and K 2 [Fe(CN) 6 ] to produce the 4-aminoantipyrine-catecholamine adduct.

Assignees

• Dh Technologies Dev Pte Ltd
• Dh Technologies Dev Pte Ltd

Inventors

• Dey Subhakar
• Purkayastha Subhasish

Classifications

• C07B59/001

  Acyclic or carbocyclic compounds · CPC title

• C07C251/22

  Quinone imines · CPC title

• C07D231/46

  Oxygen atom in position 3 or 5 and nitrogen atom in position 4 · CPC title

• C07B59/002

  Heterocyclic compounds · CPC title

• C07B2200/05

  Isotopically modified compounds, e.g. labelled · CPC title