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GRADIENT HPLC METHOD DEVELOPEMENT

http://www.chromforum.org/viewtopic.php?t=5651

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GRADIENT HPLC METHOD DEVELOPEMENT

Posted: Thu Mar 15, 2007 6:12 am
by RAMESH JANGA
HI ANY ONE ANSWER THIS QUESTION

The question is

When we developing a gradient method by HPLC , we observed baseline comes to negative mode . Not only for single product and we observed for some other products also. This problem is observed when the time program during reaching high % of solvent

Posted: Fri Mar 16, 2007 5:41 pm
by Mark Tracy
Your aqueous phase absorbs more light than the organic phase. This could be a contamination problem with your water supply or buffer ingredients, or a poorly designed buffer system.
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