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HPLC or HILIC for Blood metabolite analysis - Pretreatment
Posted: Tue May 25, 2010 3:07 pm
by doc21
Hi all!
I am wondering if anyone in the forum is using Blood samples for chromatography. If so, do you know the easiest method that will remove all the huge chunky proteins and leave only small metabolites - so that they don't block my column! I will be diluting this blood later on after treatment but what should I use to treat them?
Cheers!
Posted: Tue May 25, 2010 4:36 pm
by tom jupille
dump in a 3x excess of acetonitrile.
Do a Google search on "protein precipitation" HPLC and you'll get lots of information.
Posted: Wed May 26, 2010 1:38 am
by Uwe Neue
Tom's suggestion is a quick and dirty treatment that leaves lots of junk behind. For better results, SPE techniques are used. Go to the Waters website and look for the Oasis Sample Preparation Handbook, which gives you plenty of ideas and examples.
Posted: Thu May 27, 2010 6:16 pm
by Bryan Evans
I agree with Tom
Protein crash is low cost and used frequently for HT LC-MS assays. Google will bring up a lot of info - here's an article:
Journal of Chromatography B
Volume 785, Issue 2, 5 March 2003, Pages 263-275
If you still have interferences, suppression, ect. - than you can look at the more expensive alternatives (such as SPE, ect.)
Re:
Posted: Wed Feb 23, 2011 8:18 pm
by doc21
I agree with Tom
Protein crash is low cost and used frequently for HT LC-MS assays. Google will bring up a lot of info - here's an article:
Journal of Chromatography B
Volume 785, Issue 2, 5 March 2003, Pages 263-275
If you still have interferences, suppression, ect. - than you can look at the more expensive alternatives (such as SPE, ect.)
Hi all!
Thanks a lot for suggestions!
I have just finished a project with blood samples and dumping in access of ACN - did work!!
I was able to remove most of the unwanted chunks - thanks once again