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  1. Determine which two neighboring peaks will be analyzed from the MS ( [link] ) as the m / z = 5 and m / z = 10 peaks.
    Hypothetical mass spectrometry data; not based off of any particular compound. The example steps are based off of this spectrum.
  2. Establish the first peak (the one farthest left in [link] ) as the z + 1 peak (i.e., z + 1 = 5).
  3. Establish the adjacent peak to the right of the first peak as the z peak (i.e., z = 10).
  4. Establish the peak ratios, [link] and [link] .
  5. Solve the ratios for m : [link] and [link] .
  6. Substitute one equation for m : [link] .
  7. Solve for z : [link] .
  8. Find z + 1: [link] .
  9. Find average molecular mass by subtracting the mass by 1 and multiplying by the charge: [link] and [link] . Hence, the average mass = 7.2.
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Sample preparation

Samples for ESI-MS must be in a liquid state. This requirement provides the necessary medium to easily charge the macromolecules or proteins into a fine aerosol state that can be easily fragmented to provide the desired outcomes. The benefit to this technique is that solid proteins that were once difficult to analyze, like metallothionein, can dissolved in an appropriate solvent that will allow analysis through ESI-MS. Because the sample is being delivered into the system as a liquid, the capillary can easily charge the solution to begin fragmentation of the protein into smaller fractions Maximum charge of the capillary is approximately 4 kV. However, this amount of charge is not necessary for every macromolecule. The appropriate charge is dependent on the size and characteristic of the solvent and each individual macromolecule. This has allowed for the removal of the molecular weight limit that was once held true for simple mass spectrometry analysis of proteins. Large proteins and macromolecules can now easily be detected and analyzed through ESI-MS due to the facility with which the molecules can fragment.

A related technique that was developed at approximately the same time as ESI-MS is matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). This technique that was developed in the late 1980’s as wells, serves the same fundamental purpose; allowing analysis of large macromolecules via mass spectrometry through an alternative route of generating the necessary gas phase for analysis. In MALDI-MS, a matrix, usually comprised of crystallized 3,5-dimethoxy-4-hydroxycinnamic acid ( [link] ), water, and an organic solvent, is used to mix the analyte, and a laser is used to charge the matrix. The matrix then co-crystallizes the analyte and pulses of the laser are then used to cause desorption of the matrix and some of the analyte crystals with it, leading to ionization of the crystals and the phase change into the gaseous state. The analytes are then read by the tandem mass spectrometer. [link] directly compares some attributes between ESI-MS and MALDI-MS. It should be noted that there are several variations of both ESI-MS and MALDI-MS, with the methods of data collection varying and the piggy-backing of several other methods (liquid chromatography, capillary electrophoresis, inductively coupled plasma mass spectrometry, etc.), yet all of them have the same fundamental principles as these basic two methods.

Structure of 3,5-dimethoxy-4-hydroxycinnamic acid.
Comparison of the general experimental details of ESI-MS and MALDI-MS.
Experimental details ESI-MS MALDI-MS
Starting analyte state Liquid Liquid/solid
Method of ionization Charged capillary needle Matrix laser desorption
Final analyte state Gas Gas
Quantity of protein needed 1 μL 1 μL
Spectrum method Mass spectrometry Mass spectrometry

Problems with esi-ms

ESI-MS has proven to be useful in determination of tertiary structure and molecular weight calculations of large macromolecules. However, there are still several problems incorporated with the technique and macromolecule analysis. One problem is the isolation of the desired protein for analysis. If the protein is unable to be extracted from the cell, this is usually done through gel electrophoresis, there is a limiting factor in what proteins can be analyzed. Cytochrome c ( [link] ) is an example of a protein that can be isolated and analyzed, but provides an interesting limitation on how the analytical technique does not function for a completely effective protein analysis. The problem with cytochrome c is that even if the protein is in its native confirmation, it can still show different charge distribution. This occurs due to the availability of basic sites for protonation that are consistently exposed to the solvent. Any slight change to the native conformation may cause basic sites, such as in cytochrome c to be blocked causing different m / z ratios to be seen. Another interesting limitation is seen when inorganic elements, such as in metallothioneins proteins that contain zinc, are analyzed using ESI-MS. Metallothioneins have several isoforms that show no consistent trend in ESI-MS data between the varied isoforms. The marked differences occur due to the metallation of each isoform being different, which causes the electrospraying and as a result protonation of the protein to be different. Thus, incorporation of metal atoms in proteins can have various effects on ESI-MS data due to the unexpected interactions between the metal center and the protein itself.

The 3-D structure of human cytochrome P450 2A13, a sub class of human cytochrome c .


  • L. Konermann and D. J. Douglas, Biochemistry , 1997, 36 , 12296.
  • S. Pérez-Rafael, S. Atrian, M. Capdevila, and Ò. Palacios, Talanta , 2011, 83 , 1057.
  • D. L. Nelson and M. M. Cox, Lehninger Principles of Biochemistry , 5 th Ed, W. H. Freeman and Company, New York (2008).
  • A. Prange and D. Profrock, Anal. Bioanal. Chem. , 2005, 383 , 372.
  • (External Link)

Questions & Answers

Is there any normative that regulates the use of silver nanoparticles?
Damian Reply
what king of growth are you checking .?
What fields keep nano created devices from performing or assimulating ? Magnetic fields ? Are do they assimilate ?
Stoney Reply
why we need to study biomolecules, molecular biology in nanotechnology?
Adin Reply
yes I'm doing my masters in nanotechnology, we are being studying all these domains as well..
what school?
biomolecules are e building blocks of every organics and inorganic materials.
anyone know any internet site where one can find nanotechnology papers?
Damian Reply
sciencedirect big data base
Introduction about quantum dots in nanotechnology
Praveena Reply
what does nano mean?
Anassong Reply
nano basically means 10^(-9). nanometer is a unit to measure length.
do you think it's worthwhile in the long term to study the effects and possibilities of nanotechnology on viral treatment?
Damian Reply
absolutely yes
how to know photocatalytic properties of tio2 nanoparticles...what to do now
Akash Reply
it is a goid question and i want to know the answer as well
characteristics of micro business
for teaching engĺish at school how nano technology help us
Do somebody tell me a best nano engineering book for beginners?
s. Reply
there is no specific books for beginners but there is book called principle of nanotechnology
what is fullerene does it is used to make bukky balls
Devang Reply
are you nano engineer ?
fullerene is a bucky ball aka Carbon 60 molecule. It was name by the architect Fuller. He design the geodesic dome. it resembles a soccer ball.
what is the actual application of fullerenes nowadays?
That is a great question Damian. best way to answer that question is to Google it. there are hundreds of applications for buck minister fullerenes, from medical to aerospace. you can also find plenty of research papers that will give you great detail on the potential applications of fullerenes.
what is the Synthesis, properties,and applications of carbon nano chemistry
Abhijith Reply
Mostly, they use nano carbon for electronics and for materials to be strengthened.
is Bucky paper clear?
carbon nanotubes has various application in fuel cells membrane, current research on cancer drug,and in electronics MEMS and NEMS etc
so some one know about replacing silicon atom with phosphorous in semiconductors device?
s. Reply
Yeah, it is a pain to say the least. You basically have to heat the substarte up to around 1000 degrees celcius then pass phosphene gas over top of it, which is explosive and toxic by the way, under very low pressure.
Do you know which machine is used to that process?
how to fabricate graphene ink ?
for screen printed electrodes ?
What is lattice structure?
s. Reply
of graphene you mean?
or in general
in general
Graphene has a hexagonal structure
On having this app for quite a bit time, Haven't realised there's a chat room in it.
what is biological synthesis of nanoparticles
Sanket Reply
advantages of NAA
Sai Reply
how I can reaction of mercury?
Sham Reply

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