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Automated assay for non-transferrin-bound iron in serum samples
1Institute of Clinical Chemistry and Biochemistry, University Medical Centre Ljubljana, Ljubljana, Slovenia. marija.prezelj@kclj.si
Clinical Chemistry and Laboratory Medicine
|August 14, 2010
Summary
Non-transferrin-bound iron (NTBI), a toxic agent causing oxidative damage, can now be measured at very low concentrations. Exercise significantly increases NTBI levels in rowers, demonstrating the assay's practical application.
Area of Science:
- Biochemistry
- Clinical Chemistry
- Toxicology
Background:
- Non-transferrin-bound iron (NTBI) is highly toxic, promoting free radical damage to cellular components.
- NTBI contributes to oxidative stress, damaging proteins, lipids, and DNA.
Purpose of the Study:
- To develop and validate a sensitive assay for measuring serum NTBI.
- To assess changes in NTBI levels following physical exertion.
Main Methods:
- Serum NTBI was measured using a modified biochemical assay involving nitrilotriacetic acid (NTA) chelation and ferrozine reagent.
- Assay parameters were optimized for detecting extremely low NTBI concentrations.
Main Results:
- The assay demonstrated linearity up to 15 μmol/L with high analytical recovery (93%-103%).
- The limit of detection was 0.076 μmol/L, with good precision (CVs ranging from 0.812% to 2.37%).
- NTBI concentrations significantly increased after exercise in both older and younger rowers.
Conclusions:
- The modified assay provides a sensitive and specific method for quantifying serum NTBI.
- The assay is applicable for studying NTBI dynamics, such as post-exercise changes.
