GRADIENT HPLC METHOD DEVELOPEMENT

[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

[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.