Methods and systems for the detection of 11-oxo androgens by LC-MS/MS
AssigneeLaboratory Corporation of America Holdings
InventorsBrett Holmquist, Donald Walt Chandler, Russell Philip Grant, William Curtin
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Abstract
Disclosed are methods, systems, and computer program products for using liquid chromatography/tandem mass spectrometry (LC-MS/MS) for the analysis of endogenous biomarkers, such as 11-oxo androgens, in a sample. The 11-oxo androgens may comprise at least one of 11-hydroxyandrostendione (11OHA), 11-hydroxytestosterone (11OHT) or 11-ketotestosterone (11KT). More specifically, the methods, systems, and computer program products are described for detecting and quantifying the amount of an 11-oxo-androgen in a sample.
Claims18
A method for determining the presence or amount of at least one 11-oxo androgen in a sample by tandem mass spectrometry, comprising:
- (a) obtaining a sample from a subject;
- (b) optionally adding a stable isotope labeled 11-oxo androgen to the sample as an internal standard;
- (c) performing liquid chromatography to purify the sample;
- and
- (d) measuring the 11-oxo androgen by tandem mass spectrometry, wherein the tandem mass spectrometry comprises the steps of:
- (i) generating a precursor ion of the 11-oxo androgen;
- (ii) generating one or more fragment ions of the precursor ion;
- and (ii) detecting the presence or amount of the precursor ion generated in step (i) and/or the at least one or more fragment ions generated in step (ii), or both, and relating the detected ions to the presence or amount of the 11-oxo androgen in the sample, and wherein the tandem mass spectrometry further comprises selecting one or more fragment ions for quantitation of the at least one 11-oxo androgen and selecting one or more additional qualifier fragment ions as a qualitative standard.
A system for determining the presence or amount of at least one biomarker of interest in a test sample, the system comprising:
- a station for providing a test sample suspected of containing one or more 11-oxo androgens;
- a station for partially purifying the one or more 11-oxo androgens from other components in the sample;
- a station for chromatographically separating one or more 11-oxo androgen from other components in the sample;
- and
- a station for analyzing the chromatographically separated one or more 11-oxo androgens by mass spectrometry to determine the presence or amount of the one or more 11-oxo androgens in the test sample, wherein the tandem mass spectrometry comprises the steps of:
- (i) generating a precursor ion of the 11-oxo androgen;
- (ii) generating one or more fragment ions of the precursor ion;
- and (ii) detecting the presence or amount of the precursor ion generated in step (i) and/or the at least one or more fragment ions generated in step (ii), or both, and relating the detected ions to the presence or amount of the 11-oxo androgen in the sample, and wherein the tandem mass spectrometry comprises selecting one or more fragment ions for quantitation of the at least one 11-oxo androgen and selecting one or more additional qualifier fragment ions as a qualitative standard.
A computer-program product tangibly embodied in a non-transitory machine-readable storage medium, including instructions configured to cause one or more computers to perform actions to measure the presence or amount of at least one 11-oxo androgen in a sample comprising at least one of the following steps:
- (a) obtaining a sample from a subject;
- (b) optionally adding a stable isotope-labeled 11-oxo androgen to the sample as an internal standard;
- (c) performing liquid chromatography;
- and
- (d) measuring the 11-oxo androgen by tandem mass spectrometry, wherein the tandem mass spectrometry comprises the steps of:
- (i) generating a precursor ion of the 11-oxo androgen;
- (ii) generating one or more fragment ions of the precursor ion;
- and (ii) detecting the presence or amount of the precursor ion generated in step (i) and/or the at least one or more fragment ions generated in step (ii), or both, and relating the detected ions to the presence or amount of the 11-oxo androgen in the sample, and wherein the tandem mass spectrometry comprises selecting one or more fragment ions for quantitation of the at least one 11-oxo androgen and selecting one or more additional qualifier fragment ions as a qualitative standard.
Dependencies are inferred from the claim text.