How can I fix poor peak shape for phosphorylated compounds in aqueous normal phase ANP methods – FAQ

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ARTICLES, APPNOTES, FAQ
INTERNAL ONLY – CONFIDENTIAL
ONLINE TOOLS
SALES POLICIES

April 14, 2020
Bill Ciccone

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ANP Aqueous Normal Phase Articles / ARTICLES, APPNOTES, FAQ / HPLC Tips and Suggestions / Metal Free HPLC Columns Information / TYPE-C in General FAQ

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 aqueous normal pase 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 or medronic acid 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.
Try using a Cogent, metal free, coated stainless steel column.

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