An optimised spectrophotometric assay for convenient and accurate quantitation of intracellular iron from iron oxide nanoparticles
| Autor: | Michele Wabler, Haoming Zhou, Arsalan Nejad, Mohammad Hedayati, Robert Ivkov, Cordula Gruettner, Allen Razavi, Jana N. Mihalic, Isa Mohammed, Theodore L. DeWeese, Mohamed H. Khattab, Bedri Abubaker-Sharif |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cancer Research Physiology Iron Nanoparticle Metal Nanoparticles 02 engineering and technology Mass spectrometry Ferric Compounds Nitric Acid Colorimetry (chemical method) Mass Spectrometry Article Absorbance 03 medical and health sciences chemistry.chemical_compound Iron oxide nanoparticles Nitric acid Physiology (medical) Spectrophotometry Cell Line Tumor medicine Humans Chromatography UV/Vis spectrophotometry medicine.diagnostic_test Chromogenic Triazines 021001 nanoscience & nanotechnology 030104 developmental biology chemistry iron quantitation assay intracellular iron Biological Assay Colorimetry 0210 nano-technology |
| Zdroj: | International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group |
| ISSN: | 1464-5157 |
| Popis: | We report the development and optimisation of an assay for quantitating iron from iron oxide nanoparticles in biological matrices by using ferene-s, a chromogenic compound. The method is accurate, reliable and can be performed with basic equipment common to many laboratories making it convenient and inexpensive. The assay we have developed is suited for quantitation of iron in cell culture studies with iron oxide nanoparticles, which tend to manifest low levels of iron. The assay was validated with standard reference materials and with inductively coupled plasma-mass spectrometry (ICP-MS) to accurately measure iron concentrations ∼1 × 10−6 g in about 1 × 106 cells (∼1 × 10−12 g Fe per cell). The assay requires preparation and use of a working solution to which samples can be directly added without further processing. After overnight incubation, the absorbance can be measured with a standard UV/Vis spectrophotometer to provide iron concentration. Alternatively, for expedited processing, samples can be digested with concentrated nitric acid before addition to the working solution. Optimization studies demonstrated significant deviations accompany variable digestion times, highlighting the importance to ensure complete iron ion liberation from the nanoparticle or sample matrix to avoid underestimating iron concentration. When performed correctly, this method yields reliable iron ion concentration measurements to ∼2 × 10−6 M (1 × 10−7 g/ml sample). |
| Databáze: | OpenAIRE |
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