A Targeted Mass Spectrometric Assay for Reliable Sensitive Hepcidin Quantification.

Area of Science:

• Biochemistry
• Clinical Chemistry
• Oncology

Background:

• Hepcidin is a liver-secreted peptide hormone crucial for iron metabolism regulation.
• Disordered iron metabolism is linked to increased cancer risk.
• Current mass spectrometry (MS)-based hepcidin assays face challenges in precise quantification due to low fragmentation efficiency and standard synthesis difficulties.

Purpose of the Study:

• To develop a reliable and sensitive targeted MS assay for hepcidin quantification in clinical samples.
• To address the limitations of existing hepcidin assays by using a fully alkylated hepcidin internal standard.

Main Methods:

• Developed a targeted MS assay utilizing fully alkylated hepcidin as the internal standard for quantification.
• Determined the limits of detection (<0.5 ng/mL) and quantification (1 ng/mL).
• Applied the assay to analyze hepcidin concentrations in 70 clinical plasma samples (42 non-cancerous, 28 ovarian cancer).

Main Results:

• The assay reliably detected endogenous hepcidin in plasma and culture media.
• Hepcidin concentrations ranged from 0.0 to 95.6 ng/mL.
• Ovarian cancer patients exhibited significantly higher hepcidin concentrations (mean 20.6 ng/mL) compared to non-cancerous individuals (mean 5.94 ng/mL) (p < 0.001).

Conclusions:

• This study presents the first clinical application of the alkylated hepcidin assay.
• The developed assay offers improved sensitivity and quantification accuracy over existing MS-based methods.
• The findings suggest a potential link between elevated hepcidin levels and ovarian cancer.