HILIC v/s RP HPLC

[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!

[Uwe Neue]

Are you sure you are looking for nucleotides? Or are you looking for the bases, or nucleosides (bases with sugar)?

[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.

[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...

[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

[unmgvar]

doc 21
here is a link for an article on your subject:
http://www.dionex.com/en-us/webdocs/864 ... s-hplc.pdf

[doc21]

doc 21
here is a link for an article on your subject:
http://www.dionex.com/en-us/webdocs/864 ... s-hplc.pdf

This is a very useful article!
Thank you so much.

[smithdwsn]

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[alexdwsn12]

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[PhotonicGuy]

I was trying to access the link and read the article, because I am also interested in the topic, but it says “Page not foundâ€

[bisnettrj2]

I think this is the same article...

http://www.dionex.com/en-us/webdocs/864 ... mology.pdf