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EDTA Standards Solution A:
For the preparation of the EDTA standard solution, 5 mg of EDTA was accurately weighed and transferred into a 5 mL volumetric flask and dissolved in 0.001N NaOH water solution with sonication or magnetic stirrer mixing. Filtered The EDTA stock solution (1.0 mg/mL) should be stored in a cold dark place and can be used for a week to prepare standards of required concentration.

Iron(III) chloride Solution B:
The standard stock solution of Iron(III) chloride (10 mg/ml) was prepared in water. 50 mg of FeCl3 was accurately weighed and transferred into a 5 mL volumetric flask and dissolved in water, with sonication if needed.

General procedure for Ferric EDTA complex analysis:
To make a sample for analysis mix 100 µL Solution A (or unknown sample) with 100 µL Solution B and 800 µL of water.  Place this mixture in a plastic HPLC vial for analysis. Setup instrument and column according to the method provided.

Ethylenediaminetetraacetic acid (EDTA) is a very common chelating agent, used particularly for collecting Iron and Calcium ions. It has a wide range of applications, including in the textile industry, paper industry, pharmaceutical industry, and in cosmetics, where it is used to capture unwanted metal ions in solution. An Iron (III)-EDTA complex can be retained and analyzed on a mixed-mode Newcrom B column with a mobile phase consisting of (mostly) water, Acetonitrile (MeCN), and Sulfuric acid (H2SO4). This analytical method can be UV detected at 260 nm with high resolution and peak symmetry.

New more reliable method for EDTA analysis is available