Please Sceintists
am new Master Students and am finding alot of diffeculties in understanding the Mass Spectrophotometry
Can anyone help me in explaining these Peaks?
why it reads number 80 or 85 , as far as I learned from theory these represents the Mass of Protiens and protiens are theoretically having way higher Molecular weight
Hello Nabeel Ahmed
Mahsa Lotfi Marchoubeh is correct, the figures show chromatograms. You can see this in the labeling on the right. It says TIC, which means Total Ion Chromatogram.
The mass spectra are usually perpendicular to the time axis (x-axis) and can be accessed by left or right mouse click (your chromatogram window looks like a Thermo Fisher Xcalibur version, you maybe need to activate the spectra window).
You will then get the spectrum with the x-axis of the mass/charge ratio, versus the intensity.
Many greetings
Joachim Horst
yes true its thermofischer
but why we have to kind of spectrum i mean once with AUC and one as peaks
can you (sceintists) tell me where can I find trypsin spikes ?
is the lower one is the HPLC ? or they are the same yield of the same machine ?
L.G
You have in your case a three-dimensional information.
Time versus intensity and versus mass/charge ratio.
First of all, the X-axis is the time it takes for a substance to travel from the beginning of the separation system to the detector. This diagram is then called a chromatogram and can be seen in both of your figures.
Your detector is the mass spectrometer. There this substance is ionized separated by mass/charge ratio to and guided to the particle detector. As a result you get a second diagram, the mass spectrum, with the X-axis mass/charge ratio versus intensity.
You have two pieces of information:
1. the chomatogram with the separation of the substances contained in your sample.
2. the mass spectrum of these substances.
The mass spectra can be called up by clicking on the peaks in the chromatogram and appear in a separate window.
To find e.g. trypsins in your results you should look at the molecular mass and sum formula of trypsins first. I recommend to do this via the databases Pubchem or Chemspider. There you will get the molecular formula, the exact mass and maybe also an image of the mass spectrum. Since trypsins are enzymes, i.e. proteins, they will probably carry multiple charges. This is shown in the mass spectrum in the distance of the 13C isotope peak in the mass spectrum, 0.5 u distance = "two times charge, 0.3 U = three times, etc.
Otherwise you can deconvolve mass spectra with the Excalibur software (from version 2.1 and higher). You will then get the exact mass with single charge.
I hope I could help you
Many greetings
Joachim Horst
one more question Sceintists:
how can I find the effect of INF on my cells ?
ist possible from these figures?
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