How to fix poor peak shape for phosphorylated compounds in HPLC analysis.

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AppNotes Cogent RP C8
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April 14, 2020
jimmy

ARTICLES, APPNOTES, FAQ / HPLC Tips and Suggestions

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.
• Use Teflon bottles instead of borosilicate glass for your mobile phase reservoirs. Sodium from the glass can cause peak shape problems for anionic compounds.
• 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.

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Related Articles

1) Injection volume suggestions for Cogent TYPE-C HPLC columns

2) Add 10 micro molar EDTA to Mobile Phase for Better Peak Shapes.

3) When to use Teflon Bottles for Your Mobile Phase Reservoirs.

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