• Home
  • Search Results
  • High-performance liquid chromatography method for ferric iron chelators using a post-column reaction with Calcein Blue.

High-performance liquid chromatography method for ferric iron chelators using a post-column reaction with Calcein Blue.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2015-02-07)
Tomoko Ariga, Kyoko Ito, Yuki Imura, Etsuro Yoshimura
ABSTRACT

Iron (Fe) is an essential element for higher plants, which take it up from the soil at the root surface and transport it to shoots through the xylem. Fe(III) chelators, such as organic acids and phytosiderophores, play important roles in the acquisition and transportation of Fe(III). Therefore, a selective and sensitive method for analyzing Fe(III) chelators is required to study the many Fe-related physiological mechanisms in plants. A novel analytical approach employing a high-performance liquid chromatography post-column method with fluorescence detection was developed to separate and detect Fe(III) chelators. This method takes advantage of the quenching of the fluorescence of Calcein Blue (CB) that occurs with the formation of an Fe(III)-CB complex and the dequenching that occurs with the release of CB as a result of competition for Fe(III) between CB and an Fe(III) chelator. This simple experimental method does not require complicated pretreatments and can selectively detect Fe(III) chelators according to their Fe(III)-chelating ability. The detection limit for citric acid using this method was 72pmol. Furthermore, this method can also detect unknown Fe(III) chelators that exhibit a high affinity for Fe(III). The method was evaluated with xylem sap of barley, which was shown to contain several Fe(III) chelators.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Sigma-Aldrich
Acetic acid, glacial, ReagentPlus®, ≥99%
Sigma-Aldrich
Acetonitrile, HPLC Plus, ≥99.9%
Sigma-Aldrich
Acetic acid, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
Potassium chloride, ACS reagent, 99.0-100.5%
Sigma-Aldrich
Citric acid monohydrate, ACS reagent, ≥99.0%
Sigma-Aldrich
Boric acid, ACS reagent, ≥99.5%
Sigma-Aldrich
Potassium chloride, for molecular biology, ≥99.0%
Sigma-Aldrich
L-Histidine, ReagentPlus®, ≥99% (TLC)
Sigma-Aldrich
Acetonitrile, ACS reagent, ≥99.5%
Sigma-Aldrich
Boric acid, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
(+)-Sodium L-ascorbate, crystalline, ≥98%
Sigma-Aldrich
Acetic acid, glacial, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, 99.8-100.5%
Sigma-Aldrich
Propionic acid, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Potassium chloride, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
Sigma-Aldrich
Oxalic acid, 98%
Supelco
Potassium chloride solution, for Ag/AgCl electrodes, ~3 M KCl, saturated with silver chloride
Sigma-Aldrich
Malonic acid, ReagentPlus®, 99%
Sigma-Aldrich
Ammonium molybdate tetrahydrate, ACS reagent, 81.0-83.0% MoO3 basis
Sigma-Aldrich
Oxalic acid, puriss. p.a., anhydrous, ≥99.0% (RT)
Sigma-Aldrich
Ammonium molybdate, 99.98% trace metals basis
Sigma-Aldrich
Potassium chloride, BioXtra, ≥99.0%
Sigma-Aldrich
L-Histidine, suitable for cell culture, meets EP, USP testing specifications, from non-animal source
Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC
Sigma-Aldrich
Acetic acid, glacial, ≥99.99% trace metals basis
Sigma-Aldrich
Potassium chloride solution, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
(+)-Sodium L-ascorbate, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Acetic acid, natural, ≥99.5%, FG