Setup of Capillary Electrophoresis-Inductively Coupled Plasma Mass Spectrometry (CE-ICP-MS) for Quantification of Iron Redox Species (Fe(II), Fe(III))
| Autor: | Desiree Willkommen, Vivek Venkataramani, Bernhard Michalke |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Analyte Iron General Chemical Engineering Analytical chemistry 01 natural sciences Redox Mass Spectrometry General Biochemistry Genetics and Molecular Biology Ferrous 03 medical and health sciences Electrolytes Neuroblastoma Capillary electrophoresis medicine Tumor Cells Cultured Humans Inductively coupled plasma mass spectrometry Detection limit General Immunology and Microbiology Chemistry General Neuroscience 010401 analytical chemistry Electrophoresis Capillary 0104 chemical sciences 030104 developmental biology Standard addition Ferric Oxidation-Reduction medicine.drug |
| Zdroj: | J. Vis. Exp.:e61055 (2020) |
| ISSN: | 1940-087X |
| Popis: | Dyshomeostasis of iron metabolism is accounted in the pathophysiological framework of numerous diseases, including cancer and several neurodegenerative conditions. Excessive iron results in free redox-active Fe(II) and can cause devastating effects within the cell like oxidative stress (OS) and death by lipid peroxidation known as ferroptosis (FPT). Therefore, quantitative measurements of ferrous (Fe(II)) and ferric (Fe(III)) iron rather than total Fe-determination is the key for closer insight into these detrimental processes. Since Fe(II)/(III) determinations can be hampered by fast redox-state shifts and low concentrations in relevant samples, like cerebrospinal fluid (CSF), methods should be available that analyze quickly and provide low limits of quantification (LOQ). Capillary electrophoresis (CE) offers the advantage of fast Fe(II)/ Fe(III) separation and works without a stationary phase, which could interfere with the redox balance or cause analyte sticking. CE combined with inductively coupled plasma mass spectrometry (ICP-MS) as a detector offers further improvement of detection sensitivity and selectivity. The presented method uses 20 mM HCl as a background electrolyte and a voltage of +25 kV. Peak shapes and concentration detection limits are improved by conductivity-pH-stacking. For reduction of (56)[ArO](+), ICP-MS was operated in the dynamic reaction cell (DRC) mode with NH3 as a reaction gas. The method achieves a limit of detection (LOD) of 3 mu g/L. Due to stacking, higher injection volumes were possible without hampering separation but improving LOD. Calibrations related to peak area were linear up to 150 mu g/L. Measurement precision was 2.2% (Fe(III)) to 3.5% (Fe(II)). Migration time precision was |
| Databáze: | OpenAIRE |
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