Magnetic Micro-Solid-Phase Extraction Using a Novel Carbon-Based Composite Coupled with HPLC-MS/MS for Steroid...

Area of Science:

• Analytical Chemistry
• Materials Science
• Biochemistry

Background:

• Steroid hormone analysis in human plasma is crucial for diagnosing various endocrine disorders.
• Existing methods for steroid extraction often involve complex procedures like protein precipitation, increasing analysis time and cost.
• Development of efficient, sensitive, and selective extraction techniques is essential for routine clinical diagnostics.

Purpose of the Study:

• To develop and characterize a novel magnetic sorbent, Magn-Humic, for simultaneous solid-phase extraction of steroid hormones.
• To optimize and validate the magnetic solid-phase extraction (MSPE) method for quantifying glucocorticoids, estrogens, progestogens, and androgens in human plasma.
• To evaluate the performance of the developed MSPE-HPLC-MS/MS method in terms of recovery, sensitivity, selectivity, and precision.

Main Methods:

• Preparation of Magn-Humic sorbent via humic acid pyrolysis onto silica-coated magnetite.
• Characterization of the sorbent using SEM, TEM, EDS, TGA, and BET surface area analysis.
• Simultaneous magnetic solid-phase extraction (MSPE) of target steroids from diluted human plasma, followed by HPLC-MS/MS analysis.

Main Results:

• Magn-Humic sorbent demonstrated effective simultaneous extraction of multiple steroid classes from human plasma with quantitative recovery (65-122%) and good clean-up.
• The MSPE method achieved low detection limits (0.02-0.3 ng mL⁻¹), suitable for analyzing steroids at ng mL⁻¹ levels.
• The method exhibited satisfactory inter-day precision (RSD <14%) and was successfully applied to real human plasma samples, showing minimal matrix effects and no significant carry-over.

Conclusions:

• The developed Magn-Humic based MSPE method offers a simplified, efficient, and sensitive approach for simultaneous steroid hormone analysis in human plasma.
• The sorbent's low affinity for proteins eliminates the need for protein precipitation, streamlining the analytical workflow.
• This technique holds promise for routine clinical monitoring and research applications requiring accurate steroid hormone quantification.