NTA Standards Solution A: For the preparation of the NTA standard solution, 5 mg of NTA was accurately weighed and transferred into a 5 mL volumetric flask and dissolved in water with sonication. The NTA stock solution (1 mg/mL) should be stored in a cold, dark place and can be used for upto a week to prepare standards of required concentration.
Nitrilotriacetic acid (NTA) and Ethylenediaminetetraacetic acid (EDTA) are two common chelating agents used often in laboratory and medical testing. They are often used to bind to, identify, and analyze metal ions. Below is a method we developed to retain and measure Fe (III) using NTA and EDTA complexes on a Newcrom BH mixed-mode column, with a mobile phase of acetonitrile (MeCN), water, and a buffer of either sulfuric acid (H2SO4) or formic acid:
EDTA Standards Solution B: 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.
NTA Standard – Solution A: For the preparation of the NTA standard solution, 5 mg of NTA was accurately weighed and transferred into a 5 mL volumetric flask and dissolved in water with sonication. The NTA stock solution (1 mg/mL) should be stored in a cold, dark place and can be used for upto a week to prepare standards of required concentration.
EDTA Standard – Solution B: 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. Filter 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 C: 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 NTA and EDTA complex analysis: To make a sample for analysis, mix 100 µL each Solution A and Solution B (or unknown sample) with 100 µL Solution C and 700 µL of water. Place this mixture in a plastic HPLC vial for analysis. Setup instrument and column according to the method provided.
Iron(III) chloride Solution C: 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 NTA and EDTA complexes analysis: To make a sample for analysis mix 100 µL each Solution A and Solution B (or unknown sample) with 100 µL Solution C and 700 µL of water. Place this mixture in a plastic HPLC vial for analysis. Setup instrument and column according to the method provided.
| Column | Newcrom BH, 4.6×150 mm, 100A |
| Mobile Phase | MeCN/H2O – 2/98% |
| Buffer | H2SO4 or Formic Acid |
| Flow Rate | 1.0 ml/min |
| Detection | UV 275nm |
| Class of Compounds | Acid, Hydrophilic |
| Analyzing Compounds | EDTA, NTA |
Application Column
Newcrom BH
The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.
Select optionsNitrilotriacetic acid
