Multiplexed Testosterone Quantitation via Derivatization
Solution Overview
Problem
Current methods for measuring testosterone levels are inefficient and lack the accuracy needed for high-throughput quantitation, particularly in clinical settings where rapid and precise analysis of multiple samples is required.
Innovation Solution
The use of at least two different derivatizing agents of varying masses to differentiate testosterone in multiple patient samples within a single mass spectrometric assay, allowing for the simultaneous quantitation of testosterone in multiple samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional single-sample mass spectrometric methods are used, then measurement accuracy is maintained, but throughput is low and analysis time is long
Solution Approach 1:
The patent divides the analysis by processing each patient sample with a different derivatizing agent (e.g., hydroxylamine for sample 1, methoxyamine for sample 2), which segments the testosterone signals into distinct mass spectrometric profiles. This allows multiple samples to be analyzed simultaneously in a single run while maintaining individual sample identification and quantification accuracy.
Solution Approach 2:
The patent combines multiple processed samples into a single multiplex sample for simultaneous mass spectrometric analysis. By merging samples that have been differentially derivatized, the method achieves high throughput without sacrificing measurement precision, as each sample's testosterone can be distinguished by its unique derivatization mass signature.
2Productivity
If multiple samples are analyzed simultaneously, then throughput increases, but sample differentiation and measurement precision may be compromised
Solution Approach 1:
The patent uses derivatizing agents as intermediary substances that chemically modify testosterone in each sample to produce unique mass spectrometric signatures. These intermediaries (derivatized testosterone molecules) serve as mediators that allow simultaneous analysis of multiple samples while maintaining the ability to precisely distinguish and quantify testosterone from each individual sample based on their distinct mass profiles.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enables rapid and accurate quantitation of testosterone in multiple samples, significantly increasing throughput while maintaining high precision and sensitivity, with a limit of quantitation as low as 1 ng/dL and linearity across a range of 1 ng/dL to 2,000 ng/dL.
Implementation Method 1
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
Implementation Method 2
subjecting the multiplex sample to an ionization source under conditions suitable to generate one or more ions detectable by mass spectrometry
Implementation Method 3
detecting the amount of one or more ions from testosterone from each processed sample by mass spectrometry
Data Source
AI summary
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.