Comprehensive Steroid Assay with Non-Targeted Analysis Using Liquid Chromatography Ion Mobility Mass Spectrometry

Area of Science:

• Biochemistry
• Analytical Chemistry
• Endocrinology

Background:

• Aldosterone-producing adenomas (APAs) exhibit diverse steroid profiles correlated with specific genetic mutations.
• Accurate characterization of these profiles is crucial for understanding disease pathogenesis and identifying biomarkers.

Purpose of the Study:

• To develop and validate a sensitive liquid chromatography-ion mobility-mass spectrometry (LC/IM/MS) assay for analyzing serum steroid profiles in patients with APAs.
• To investigate the potential of LC/IM/MS in classifying patients based on their steroid profiles and identifying novel biomarkers.

Main Methods:

• Serum samples were purified and analyzed using liquid-liquid extraction followed by LC/IM/MS.
• A method was established to separate and quantify 20 targeted steroids based on retention time, m/z, and drift time.
• Principal component analysis was employed to classify serum samples from different patient groups.

Main Results:

• The LC/IM/MS assay demonstrated high sensitivity and selectivity, enabling clear classification of patients with primary aldosteronism, essential hypertension, and healthy controls.
• The method successfully identified targeted steroids and also detected non-targeted compounds like phosphatidylcholine and phosphatidylethanolamine.
• Distinct steroid profiles were observed, correlating with different genetic mutations underlying APAs.

Conclusions:

• LC/IM/MS is a powerful tool for analyzing complex serum steroid profiles and classifying patients with APAs.
• This analytical approach has the potential to elucidate APA pathogenesis and discover novel steroid biomarkers for genetically distinct APAs.