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Mass Spectrometry explained – how it works
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Mass spectrometry AKA Mass Spec is an analytical technique that measures mass to charge ratio in this video you will learn how this method works and why it is such a powerful tool in research before we jump right into it I would kindly ask you to subscribe to the channel to stay updated it is free of charge but supports me a lot scientists apply Mass spectrometry if they opt to measure the m mass of different molecules within a sample Mass back helps to quantify molecules of interest and identify them based on their Mass quite often the technique is used for protein analysis which makes Mass back an important tool in the field of proteomics but how does it work well as for many laboratory techniques there are different devices to accomplish Mass back with which work all a bit differently so be aware that some steps shown in this video might deviate a bit depending on the mass spectrometer and analyzer used let's have a look at a simplified Mass spectrometer consisting of an iron Source an analyzer and a detector before inserted into the mass analyzer complex samples are often separated for example by liquid chromatography the first two steps in Mass spectrometry are called desorption and ionization and they usually occur simultaneously this refers to the process of transferring molecules into the gas phase during which singly or multiply charged ions are generated a common method is called electrospray ionization short ESI the sample is passed through a capillary with an applied electric field electrostatic repulsion of ions and other physical effects causes a fine aerosol that eventually evaporates resulting in charged gas phase ions after ionization the ions pass through tiny slits in the mass spectrometer and are accelerated in a focused iron beam using an electric field there are various analyzers in Mass spectrometry all of which aim to determine the mass to charge ratio of ions one commonly used analyzer type is the time of flight analyzer the principle is straightforward the ions are accelerated in an electric field with a known voltage and ions with the same charge gain the same kinetic energy consquently lighter ions travel faster through the fly tube than heavier ions with the same charge and more highly charged ions travel faster compared to singly charged ions with the same mass the time of flight through the tube to the detector is then measured since the the distance is known the mass to charge ratio can be determined the ions are also Quantified and the mass spe result will be featured as a chart looking like this which is referred to as a Mass Spectrum the mass to charge ratio is displayed on the xaxis and the relative intensity is shown on the Y AIS the most abundant ion for example will be the one with the highest peak the relative intensity is set to 100% for that ion in this example the iron Peaks at 24 MZ and some lower Peaks are found with an isotopic mass of 25 and 26 MZ playing detective while taking a look at the periodic table one would discover that this looks pretty much like magnesium with an atomic mass of 24.3 U and indeed it is the smaller Peaks are some heavier isotopes of the element m spectrometry is usually used for more complex molecules such as protein samples databases help to identify the molecules of interest during mass spectrometry a sample of interest is first transferred to the gas phase and ionized which means charge is added to the sample these charged ions are then accelerated which helps to separate them inside the analyzer according to their Mass to charge ratio a common way to accomplish that is by measuring time it takes for the irons to hit the detector the result will be shown as a Mass Spectrum which in combination with databases is used to identify the sample of interest you want to know how to interprete or analyze your mass back results take a look at this clip here if you need a break from studying check out this video where I explain whether and maybe even how we could bring back dinosaurs or other extinct species if this video could introduce you a bit to mass spectrometry please leave a like thanks for watching
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