# US 10,768,251 B2 · Method for analyzing an NMR spectrum of a lipoprotein-containing sample

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- Number: US 10,768,251 B2
- Document ID: US10768251B2
- Status: Granted
- Kind: B2
- Application number: 16/341,994
- Filed: 2017-12-12
- Published: 2019-08-15
- Granted: 2020-09-08
- Earliest claimed: 2016-12-12
- Assignees: numares AG
- Applicants: numares AG
- Inventors: Daniela Baumstark
- Web page: https://patentgrep.com/patent/US10768251B2

## CPC

- G01R 33/465: applied to biological material, e.g. in vitro testing
- G01N 24/08: by using nuclear magnetic resonance (G01N24/12 takes precedence)
- G01R 33/4625: {Processing of acquired signals, e.g. elimination of phase errors, baseline fitting, chemometric analysis}
- G01R: MEASURING ELECTRIC VARIABLES; MEASURING MAGNETIC VARIABLES (indicating correct tuning of resonant circuits H03J3/12)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Disclosed is a method for analyzing an NMR spectrum of a lipoprotein-containing sample. The method includes the following steps: a) defining a spectral range to be analyzed of at least 0.5 ppm of an NMR spectrum of a lipoprotein-containing sample; b) fitting a first spectral function of a first individual lipid component and at least one further spectral function of a further individual lipid component into the whole spectral range to be analyzed; and c) determining a concentration or an amount of the first individual lipid component and the further individual lipid component and therefrom a concentration or an amount of the first defined lipoprotein class or lipoprotein subclass in the lipoprotein-containing sample and optionally of each other defined lipoprotein class or lipoprotein subclass based on the fitting operations of step b).

## Claims

### Claim 1

A method for analyzing an NMR spectrum of a lipoprotein-containing sample, comprising the following steps:
a) defining a spectral range to be analyzed of at least 0.5 ppm of an NMR spectrum of a lipoprotein-containing sample,
b) fitting a first spectral function of a first individual lipid component and at least one further spectral function of a further individual lipid component into the whole spectral range to be analyzed,
wherein the first spectral function represents at least a part of an NMR spectrum of the first individual lipid component taking into account the behavior of the first lipid component in the surroundings of a lipoprotein particle of a first defined lipoprotein class or lipoprotein subclass, and
wherein the further spectral function represents at least a part of an NMR spectrum of the further individual lipid component taking into account the behavior of the further lipid component in the surroundings of a lipoprotein particle of the same defined lipoprotein class or lipoprotein subclass, and
c) determining a concentration or an amount of the first individual lipid component and the further individual lipid component and therefrom a concentration or an amount of the first defined lipoprotein class or lipoprotein subclass in the lipoprotein-containing sample and optionally of each other defined lipoprotein class or lipoprotein subclass based on the fitting operations of step b).

### Claim 2 (depends on 1)

The method according to claim 1, wherein step b) is additionally performed with further spectral functions of individual lipid components, wherein each spectral function represents at least a part of an NMR spectrum of an individual lipid component taking into account the behavior of this lipid component in the surroundings of a lipoprotein particle of another defined lipoprotein class or lipoprotein subclass.

### Claim 3 (depends on 2)

The method according to claim 2, wherein in step c) a concentration or an amount of the individual lipid components is determined and is used to determine a concentration or an amount of each other defined lipoprotein class or lipoprotein subclass.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the spectral range to be analyzed encompasses the whole spectral range measured in which spectral lines are observed.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the lipoprotein-containing sample is a blood serum or a blood plasma sample.

### Claim 6 (depends on 1)

The method according to claim 1, wherein an NMR spectrum of compounds other than lipoproteins or lipids, or a function of such an NMR spectrum, is subtracted from the NMR spectrum of the lipoprotein-containing sample prior to the fitting operation.

### Claim 7 (depends on 5)

The method according to claim 5, wherein an NMR spectrum of compounds other than lipoproteins or lipids, or a function of such an NMR spectrum, is subtracted from the NMR spectrum of the lipoprotein-containing sample prior to the fitting operation, and wherein the NMR spectrum to be subtracted is an NMR spectrum of lipoprotein deficient serum or another protein-depending spectrum or mixtures thereof.

### Claim 8 (depends on 1)

The method according to claim 1, wherein at least one of the first lipid component and the further lipid component is chosen from the group consisting of cholesterol, cholesterol esters, triglycerides, phospholipids, sphingomyelins and fatty acids.

### Claim 9 (depends on 1)

The method according to claim 1, wherein at least one of the first spectral function of the first individual lipid component, the further spectral function of the further individual lipid component, and the spectral functions of individual lipid components is obtained on the basis of an NMR spectrum of the respective individual lipid component, and wherein the lines of the NMR spectrum or a derivative of it are optionally shifted, broadened or adjusted in height to represent at least a part of an NMR spectrum of the respective lipid component in the surroundings of a lipoprotein particle.

### Claim 10 (depends on 1)

The method according to claim 1, wherein at least one of the first spectral function of the first individual lipid component, the further spectral function of the further individual lipid component, and the spectral functions of individual lipid components comprises at least two lines.

### Claim 11 (depends on 1)

The method according to claim 1, wherein at least one of the first spectral function of the first individual lipid component, the further spectral function of the further individual lipid component, and the spectral functions of individual lipid components serves for fitting more than one signal of the NMR spectrum of the lipoprotein-containing sample.

### Claim 12 (depends on 1)

The method according to claim 1, wherein at least one of the first spectral function of the first individual lipid component, the further spectral function of the further individual lipid component, and the spectral functions of individual lipid components are aligned on the basis of a defined diffusion behavior of a lipoprotein class or lipoprotein subclass.