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- Posts: 15
- Joined: Sun Oct 04, 2009 10:54 pm
Hi,
I am trying to separate some intracellular metabolites like fumarate, malte , succinate , isoleucine and lecuine by luna NH2 column in HILIC mode. Initial phase is 85 % ACN and 15% water/ACN(95:5, 20mM ammonium acetate buffer with pH 8.5). Whatever I changed gradient or adjusted buffer pH, malate and fumarate , leucine and isoleucine couldn't get a baseline separation. It is so hard for me to make an accurate quantification of them. I am wondering if HILIC is able to resolve them since they have so similar polar characteristic. Thanks !
Jack
I am trying to separate some intracellular metabolites like fumarate, malte , succinate , isoleucine and lecuine by luna NH2 column in HILIC mode. Initial phase is 85 % ACN and 15% water/ACN(95:5, 20mM ammonium acetate buffer with pH 8.5). Whatever I changed gradient or adjusted buffer pH, malate and fumarate , leucine and isoleucine couldn't get a baseline separation. It is so hard for me to make an accurate quantification of them. I am wondering if HILIC is able to resolve them since they have so similar polar characteristic. Thanks !
Jack
