Mass spectrometric determination of testosterone in multiplexed patient samples

Patent metadata

Field	Value
Publication number	US-2025035651-A1
Application number	US-202418914420-A
Country	US
Kind code	A1
Filing date	Oct 14, 2024
Priority date	Mar 16, 2018
Publication date	Jan 30, 2025
Grant date	—

Abstract

A method for determining the amount of testosterone in each of at least two human samples by a single tandem mass spectrometry assay includes subjecting each of a plurality of human samples to a different derivatizing agent to generate a differently derivatized testosterone in each of the plurality of samples, wherein the derivatization agents comprise hydroxylamine chloride and methoxylamine chloride; combining the plurality of samples to form a multiplex sample; and quantifying the amount of testosterone in each sample by mass spectrometry by measuring the amount of a precursor ion having a mass-to-charge ratio of 304.18±0.5 and/or 318.21±0.5 and measuring the amount of a fragment ion having a mass-to-charge ratio of 112.05±0.5, 124.05±0.5, 126.07±0.5, 138.07±0.5, and/or 152.08±0.5.

First claim

What is claimed is:

1 . A method for determining the amount of testosterone in each of at least two human samples by a single tandem mass spectrometry assay, the method comprising:
subjecting each of a plurality of human samples to a different derivatizing agent to generate a differently derivatized testosterone in each of the plurality of samples, wherein the derivatization agents comprise hydroxylamine chloride and methoxylamine chloride; combining the plurality of samples to form a multiplex sample; and quantifying the amount of testosterone in each sample by mass spectrometry by measuring the amount of a precursor ion(s) having a mass-to-charge ratio selected from the group consisting of 304.18±0.5 and 318.21±0.5 and measuring the amount of a fragment ion(s) having a mass-to-charge ratio selected from the group consisting of 112.05±0.5, 124.05±0.5, 126.07±0.5, 138.07±0.5, and 152.08±0.5.

2 . The method of claim 1 further comprising purifying the multiplex sample with liquid chromatography.

3 . The method of claim 2 , wherein the liquid chromatography comprises high performance liquid chromatography.

4 . The method of claim 3 , wherein the liquid chromatography comprises high turbulence liquid chromatography.

5 . The method of claim 1 further comprising adding an internal standard.

6 . The method of claim 5 , wherein the internal standard is isotopically labeled.

7 . The method of claim 5 further comprising measuring the amount of an internal stand precursor ion having a mass-to-charge ratio selected from the group consisting of 307.19±0.5 and 321.21±0.5, or a product ion having a mass-to-charge ratio selected from the group consisting of 127.06±0.5 and 129.07±0.5.

8 . The method of claim 1 , wherein a limit of quantitation of the methods is less than or equal to 1 ng/dL.

9 . The method of claim 1 , wherein the method has a linearity of quantitation across a range between 1 ng/dL to 2,000 ng/dL.

10 . The method of claim 1 , wherein the method is fully automated.

11 . The method of claim 1 , wherein the method is antibody-free.

12 . The method of claim 1 , wherein the sample is a serum sample.

Assignees

• Quest Diagnostics Invest Llc

Inventors

• Goldman Mildred M
• Goldman Scott
• Colletti Julia
• Clarke Nigel

Classifications

• G01N30/50

  Conditioning of the sorbent material or stationary liquid · CPC title

• G01N33/6848

  Methods of protein analysis involving mass spectrometry · CPC title

• G01N33/743 Primary

  Steroid hormones · CPC title