I used a new column, C18, but there are peaks in the blank. I use ammonium acetate (o.1M pH 5) and acetonitrile buffers A and B respectively for dye separation. and did several washings but the column have distinguish peaks? what should I do?
Hi Rehana,
There are a lot reasons for "Ghost-Peaks". Could be a lot of reasons. Contamination of your solvents, your LC-System or a dead-volume in the system.
There is a really nice troubleshooting guide for HPLC available. For example:
http://ccc.chem.pitt.edu/wipf/Web/LCMS%20trouble%20shooting.pdf
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