ArticleMethods and protocols2026
An Improved Ferrozine-Based Protocol for Safe, Reproducible, and Accurate Quantification of Iron in Biological and Chemical Samples.
Article in Methods and protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Accurate quantification of iron is essential in biological, chemical, and nanomaterial research, yet commonly used ferrozine-based assays suffer from safety hazards, inconsistent reduction efficiency, and unstable absorbance readings. To address these issues, we systematically optimized the classical protocol and validated improvements that enhance both operational safety and analytical reproducibility. In this work, samples were digested using perchloric acid and hydrogen peroxide, reduced with hydroxylamine, and complexed with ferrozine, with all steps quantitatively evaluated to identify conditions that minimize variability. The optimized assay introduces three key refinements: combining the two traditional hydroxylamine additions into a single reduction step, extending the post-complexation incubation to 2 h to ensure complete formation of the Fe
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