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Determination of the Cu/Fe ratio in various cytochrome c oxidases by total reflection X-ray fluorescence (TXRF) spectrometry (CAT#: STEM-ST-0195-WXH)

Introduction

The enzyme cytochrome c oxidase or Complex IV, is a large transmembrane protein complex found in bacteria, archaea, and mitochondria of eukaryotes.<br />The oxidized form of cytochrome c oxidase contains two CuII and two FeIII heme centers. It can be fully reduced to give a form of the enzyme containing two CuI and two FeII heme centers. The heme found in cytochrome c oxidase is different from that found in other heme proteins.




Principle

XRF describes the process where some high-energy radiation excites atoms by shooting out electrons from the innermost orbitals. When the atom relaxes, that is, when outer electrons fill inner shells, X-Ray fluorescence radiation is emitted.
Total-reflection X-ray fluorescence (TXRF) is a multi-element simultaneous analysis technology developed based on X-ray fluorescence (XRF). An aircooled X-ray tube generates an X-ray beam, which is reduced to a narrow energy range by a multi-layer monochromator. The fine beam impinges on a polished sample carrier at a very small angle and is totally reflected.

Applications

XRF is widely used as a fast characterization tool in many analytical labs across the world, for applications as diverse as metallurgy, forensics, polymers, electronics, archaeology, environmental analysis, geology and mining.

Procedure

1. Primary X-rays knock out an electron from one of the orbitals surrounding the nucleus within an atom of the material.
2. A hole is produced in the orbital, resulting in a high energy, unstable configuration for the atom.
3. To restore equilibrium, an electron from a higher energy, outer orbital falls into the hole. Since this is a lower energy position, the excess energy is emitted in the form of fluorescent X-rays.
The energy difference between the expelled and replacement electrons is characteristic of the element atom in which the fluorescence process is occurring – thus, the energy of the emitted fluorescent X-ray is directly linked to a specific element being analyzed.

Materials

XRF spectrometer (including X-ray source, sample chamber, analysing crystal, detector and signal processing computer)