Jump to content
Main menu
Main menu
move to sidebar
hide
Navigation
Main page
Recent changes
Random page
Help about MediaWiki
Special pages
Niidae Wiki
Search
Search
Appearance
Create account
Log in
Personal tools
Create account
Log in
Pages for logged out editors
learn more
Contributions
Talk
Editing
Size-exclusion chromatography
(section)
Page
Discussion
English
Read
Edit
View history
Tools
Tools
move to sidebar
hide
Actions
Read
Edit
View history
General
What links here
Related changes
Page information
Appearance
move to sidebar
hide
Warning:
You are not logged in. Your IP address will be publicly visible if you make any edits. If you
log in
or
create an account
, your edits will be attributed to your username, along with other benefits.
Anti-spam check. Do
not
fill this in!
==Advantages== The advantages of this method include good separation of large molecules from the small molecules with a minimal volume of eluate,<ref name=":0">{{Cite book|chapter-url=http://zeus.qui.ufmg.br/~valmir/livros/analitica/Principles%20of%20Instrumental%20Analysis%20(6ed)%20-%20Skoog,%20Holler%20&%20Crouch.pdf|title=Principles of instrumental analysis|last1=Skoog|first1=Douglas A.|last2=Holler|first2=F. James|last3=Crouch|first3=Stanley R.|publisher=Thomson Brooks/Cole|year=2006|isbn=9780495012016|edition=6th|location=Belmont, CA|pages=816|chapter=Ch. 28. Liquid Chromatography|lccn=2006926952|oclc=77224390|name-list-style=vanc}}</ref> and that various solutions can be applied without interfering with the filtration process, all while preserving the biological activity of the particles to separate. The technique is generally combined with others that further separate molecules by other characteristics, such as acidity, basicity, charge, and affinity for certain compounds. With size exclusion chromatography, there are short and well-defined separation times and narrow bands, which lead to good sensitivity. There is also no sample loss because solutes do not interact with the stationary phase. The other advantage to this experimental method is that in certain cases, it is feasible to determine the approximate molecular weight of a compound. The shape and size of the compound (eluent) determine how the compound interacts with the gel (stationary phase). To determine approximate molecular weight, the elution volumes of compounds with their corresponding molecular weights are obtained and then a plot of βK<sub>av</sub>β vs βlog(Mw)β is made, where <math>K_{av} = (V_e-V_o)/(V_t-V_o)</math> and Mw is the molecular mass. This plot acts as a calibration curve, which is used to approximate the desired compound's molecular weight. The V<sub>e</sub> component represents the volume at which the intermediate molecules elute such as molecules that have partial access to the beads of the column. In addition, V<sub>t</sub> is the sum of the total volume between the beads and the volume within the beads. The V<sub>o</sub> component represents the volume at which the larger molecules elute, which elute in the beginning.<ref>{{cite book|title=Chemical analysis: modern instrumental methods and techniques|url=https://archive.org/details/chemicalanalysis00roue_541|url-access=limited|last1=Rouessac|first1=Annick|last2=Rouessac|first2=Francis|date=2000|publisher=Wiley|isbn=978-0471972617|edition=Engl.|location=Chichester|pages=[https://archive.org/details/chemicalanalysis00roue_541/page/n122 101]β103|oclc=635171657|name-list-style=vanc}}</ref><ref name=":1">{{cite book|title=Fundamental laboratory approaches for biochemistry and biotechnology|last1=Ballou|first1=David P.|last2=Benore|first2=Marilee|last3=Ninfa|first3=Alexander J.|date=2008|publisher=Wiley|isbn=9780470087664|edition=2nd|location=Hoboken, N.J.|pages=127β129|name-list-style=vanc}}</ref> Disadvantages are, for example, that only a limited number of bands can be accommodated because the time scale of the chromatogram is short, and, in general, there must be a 10% difference in molecular mass to have a good resolution.<ref name=":0" />
Summary:
Please note that all contributions to Niidae Wiki may be edited, altered, or removed by other contributors. If you do not want your writing to be edited mercilessly, then do not submit it here.
You are also promising us that you wrote this yourself, or copied it from a public domain or similar free resource (see
Encyclopedia:Copyrights
for details).
Do not submit copyrighted work without permission!
Cancel
Editing help
(opens in new window)
Search
Search
Editing
Size-exclusion chromatography
(section)
Add topic