Provisional standardization of hepcidin assays: creating a traceability chain with a primary reference material,...

Area of Science:

• Clinical Chemistry
• Biomarker Standardization
• Laboratory Medicine

Background:

• Hepcidin concentration measurements vary significantly across different methods, hindering accurate interpretation.
• A novel serum-based secondary reference material (sRM) was developed from a plasma-based candidate.
• The sRM aims to improve the equivalence and standardization of hepcidin assays internationally.

Purpose of the Study:

• To validate the functionality of a serum-based secondary reference material (sRM) for hepcidin standardization.
• To assess the commutability and harmonization potential of the sRM across various measurement procedures.
• To establish traceability for the sRM using a primary reference material (pRM) and isotope dilution mass spectrometry.

Main Methods:

• Development and validation of a two-leveled serum-based reference material (sRM).
• Round robin studies (RR1 and RR2) involving multiple measurement procedures and native human samples.
• Value assignment of the sRM using a candidate reference method calibrated by a certified primary reference material (pRM).

Main Results:

• Without harmonization, inter-assay coefficients of variation (CV) were high (42.1% in RR1, 52.8% in RR2).
• Simulated harmonization with the sRM reduced inter-assay CV to 11.0% (RR1), and calibration achieved 19.1% (RR2).
• The sRM and pRM demonstrated international homogeneity and long-term stability, with assigned values for low and middle calibrators.

Conclusions:

• Standardization of hepcidin measurements is achievable using the developed serum-based reference material (sRM).
• The sRM, value-assigned by a primary reference material (pRM), shows significant potential for harmonizing hepcidin assays.
• Implementation of this sRM as an international calibrator is proposed to enhance hepcidin measurement equivalence.