Question: I am analyzing phosphorylated sugars and obtain poor HPLC peak shapes with very broad tailing. What are some possible solutions?
Answer: We have a number of strategies for fixing problems with peak shape for phosphorylated compounds:
• Ensure the column is not overloaded. We recommend an injection volume of 1 µL or less with ANP methods.
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• Use 5–10 microM EDTA in the mobile phase and sample diluent. This will help chelate metals that cause problems for phosphates.
• Try adding 0.01% ammonia to the sample diluent.