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HILIC v/s RP HPLC
Posted: Sat May 15, 2010 8:46 pm
by doc21
Hi all,
I am a very basic user of LC. My experiment aim is nucleotide separation from serum. What basic difference is required for me to decide if I should use HILC or RP HPLC? I guess they are almost opposites of each other, but what kind of properties should my analyte have?
Cheers!
Posted: Sun May 16, 2010 3:16 pm
by Uwe Neue
Are you sure you are looking for nucleotides? Or are you looking for the bases, or nucleosides (bases with sugar)?
Posted: Mon May 17, 2010 8:14 am
by doc21
Are you sure you are looking for nucleotides? Or are you looking for the bases, or nucleosides (bases with sugar)?
Yes, I am targeting nucleotides and looking for adenine, guanine etc. However targeting and getting bases + sugar, won't harm either.
Posted: Mon May 17, 2010 2:23 pm
by Uwe Neue
10 mM ammonium acetate, pH 5.0 on Atlantis T3 from Waters (a water wettable C18 with maximum retention), gradient to the same with 6% acetonitrile.
http://www.waters.com/webassets/cms/lib ... a60208.pdf
You can manipulate the pH for getting a suitable separation for the specific analytes that you are targeting.
For the bases with sugar, you will be better off using a HILIC technique. I will see what I find...
Posted: Mon May 17, 2010 2:38 pm
by Uwe Neue
Here is the applications booklet for the Atlantis columns, including Atlantis HILIC. You will find a starting point for your application.
http://www.waters.com/webassets/cms/lib ... 0472en.pdf
This is the applications book for the BEH Amide column, again with some applications that can bet you started. This column is the best choice for HILIC applications, since it is most retentive.
http://www.waters.com/webassets/cms/lib ... A64081.pdf
http://www.waters.com/webassets/cms/lib ... a64069.pdf
Posted: Mon May 24, 2010 7:02 pm
by unmgvar
doc 21
here is a link for an article on your subject:
http://www.dionex.com/en-us/webdocs/864 ... s-hplc.pdf
Posted: Tue May 25, 2010 3:04 pm
by doc21
This is a very useful article!
Thank you so much.
Posted: Thu Jun 10, 2010 1:35 pm
by smithdwsn
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It describe very deeply so there is no problem about any query. It is really useful to me.
Great!
Posted: Mon Jul 26, 2010 2:45 pm
by alexdwsn12
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Posted: Wed Jul 28, 2010 4:29 pm
by PhotonicGuy
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Posted: Wed Jul 28, 2010 5:09 pm
by bisnettrj2