Materials and methods for detection and quantification of β-N-methylamino-L-alanine

Patent metadata

Field	Value
Publication number	US-11370812-B2
Application number	US-201716321091-A
Country	US
Kind code	B2
Filing date	Jul 31, 2017
Priority date	Jul 29, 2016
Publication date	Jun 28, 2022
Grant date	Jun 28, 2022

Abstract

Provided herein are isotopically labeled reagents, including isotopically labeled small molecules and peptides, that can be used to detect and/or quantify β-N-methylamino-L-alanine (BMAA) in a sample. The reagents can be used as stable isotope labeled standards in analytical methods, including in conjunction with mass spectrometry, to detect and/or quantify BMAA in a sample, such as a protein sample from a subject.

First claim

What is claimed is:

1. A composition comprising an isotopically labeled compound defined by the formula below

wherein R represents hydrogen or an amine protecting group; and
wherein at least four of a C, b C, c C, d C, e N, and f N, are isotopically labeled with a stable isotope.

2. The composition of claim 1 , wherein a C, b C, c C, d C, e N, and f N are isotopically labeled with a stable isotope.

3. The composition of claim 1 , wherein the isotopically labeled compound is defined by the formula below

wherein R represents hydrogen or an amine protecting group;
wherein the 13 C isotopic enrichment factor for a C, b C, c C, and d C is at least 25; and
wherein the 15 N isotopic enrichment factor for e N and f N is at least 100.

4. The composition of claim 3 , wherein the 13 C isotopic enrichment factor for a C, b C, c C, and d C is at least 80 and the 15 N isotopic enrichment factor for e N and f N is at least 200.

5. The composition of claim 1 , wherein all eight of a C, b C, c C, d C, e N, f N, g O, and h O are isotopically labeled with a stable isotope.

6. The composition of claim 1 , wherein R is hydrogen.

7. The composition of claim 1 , wherein R represents a 9-fluorenylmethyloxycarbonyl group.

8. The composition of claim 1 , wherein the isotopically labeled compound is at least 0.5% by weight of the composition.

9. The composition of claim 1 , wherein the isotopically labeled compound is 13 C 3 15 N 2 β-N-methylamino-L-alanine.

10. The composition of claim 1 , wherein the isotopically labeled compound is defined by the formula below

11. A method for quantifying the amount of β-N-methylamino-L-alanine (BMAA) in a sample comprising:
(i) analyzing a test sample by mass spectrometry;
(ii) spiking the test sample with a defined amount of the composition of claim 1 ;
(iii) analyzing the spiked sample by mass spectrometry; and
(iv) determining the amount of BMAA in the test sample by isotope dilution analysis.

Assignees

• Univ North Carolina State

Inventors

• Muddiman David C
• Loziuk Philip L
• Pierce Joshua G
• Williams Gavin
• Moazami Yasamin

Classifications

• C12N9/0089

  acting on superoxide as acceptor (1.15) · CPC title

• G01N2800/2835

  Movement disorders, e.g. Parkinson, Huntington, Tourette · CPC title

• C07K1/13 Primary

  Labelling of peptides · CPC title

• G01N33/52

  Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)} · CPC title

• C07B59/008

  Peptides; Proteins · CPC title