Bathophenanthroline Sulfonate-Based Colorimetric Assay: A Simple and Rapid Method for Quantitation of Non-Heme Iron in Mouse Liver Tissue

Published: April 30, 2023

Abstract

Source: Duarte, T. L., Neves, J. V. Measurement of Tissue Non-Heme Iron Content using a Bathophenanthroline-Based Colorimetric Assay. J. Vis. Exp. (2022)

In this video, we describe a colorimetric-based method to determine non-heme iron content in mouse liver tissue. Bathophenanthroline sulfonate is used as the chromogenic substrate in the presence of a reducing agent, which converts ferric iron to chelatable ferrous iron.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board 1. Solution preparation NOTE: Handle and prepare all reagents and solutions with iron-free glassware or disposable plasticware. Do not allow metallic laboratory materials (e.g., stainless steel spatulas) to come in contact with any reagent or solution, due to the risk of iron contamination. Ma…

開示

The authors have nothing to disclose.

Materials

96 well UV transparent plate Sarstedt 82.1581.001
Analytical balance Kern ABJ 220-4M
Anhydrous sodium acetate Merck 106268
Bathophenanthroline sulfonate (4,7-Diphenyl-1,10-phenantroline dissulfonic acid) Sigma-Aldrich B1375
C57BL/6 mice (Mus musculus) Charles River Laboratories
Carbonyl iron powder, ≥99.5% Sigma-Aldrich 44890
Disposable cuvettes in polymethyl methacrylate (PMMA) VWR 634-0678P
Double distilled, sterile water B. Braun 0082479E
Fluorescence microplate reader BioTek Instruments FLx800
Hydrochloric acid, 37% Sigma-Aldrich 258148
Microwave digestion oven and white teflon cups CEM MDS-2000
Nitric acid Fisher Scientific 15687290
Oven Binder ED115
Rodent chow Harlan Laboratories 2014S Teklad Global 14% Protein Rodent Maintenance Diet containing 175 mg/kg iron
Sea bass (Dicentrarchus labrax) Sonrionansa
Sea bass feed Skretting L-2 Alterna 1P
Single beam UV-Vis spectrophotometer Shimadzu UV mini 1240
Thioglycolic acid Merck 100700
Trichloroacetic acid Merck 100807

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記事を引用
Bathophenanthroline Sulfonate-Based Colorimetric Assay: A Simple and Rapid Method for Quantitation of Non-Heme Iron in Mouse Liver Tissue. J. Vis. Exp. (Pending Publication), e21159, doi: (2023).

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