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UHPLC-HRMS Analysis for Steroid Profiling in Serum (Steroidomics)
Federico Ponzetto1, Julien Boccard2,3, Raul Nicoli1
1Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland.
Methods in Molecular Biology (Clifton, N.J.)
|April 15, 2018
Summary
This protocol details a method for extracting and analyzing endogenous steroids in serum using UHPLC-HRMS. The technique effectively identifies key steroids like mineralocorticoids, progestogens, and androgens in blood samples.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Endocrinology
Background:
- Endogenous steroids play crucial roles in various physiological processes.
- Accurate quantification and identification of these steroids in biological matrices are essential for clinical diagnostics and research.
- Existing methods may face challenges in sensitivity or scope for comprehensive steroid profiling.
Purpose of the Study:
- To describe a protocol for the extraction and untargeted ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) analysis of endogenous steroids in serum.
- To establish a sensitive method for identifying major endogenous steroids in human blood.
- To provide guidance on technical aspects and data analysis for reliable steroid profiling.
Main Methods:
- Serum samples undergo extraction to isolate endogenous steroids.
- Untargeted analysis is performed using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS).
- Full-scan acquisition mode is employed for sensitive detection of a wide range of steroids.
Main Results:
- The protocol enables sensitive detection of major endogenous steroids in serum, including mineralocorticoids, progestogens, and androgens.
- Detailed technical parameters for chromatography and mass spectrometry are provided.
- A framework for sample sequencing and data analysis is proposed, aiding in data quality assessment.
Conclusions:
- The described UHPLC-HRMS protocol offers a sensitive and comprehensive approach for endogenous steroid profiling in serum.
- The method facilitates the identification of key steroid classes relevant to human health.
- The provided guidelines support robust data acquisition and analysis for steroid metabolomics.
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