HPLC Method Development For Acidic Molecules: A Case Study - PharmaGuru | Global Pharma Training, Certifications & Expert Blogs

Question 1

How to select the HPLC column and Mobile Phase for the separation of Acidic Compounds?

Accepted Answer

Keep the mobile phase pH acidic and use columns like C8 and C18

Question 2

What is the principle of separation of Acidic Compounds by HPLC?

Accepted Answer

To separate acidic molecules. Try to make these molecules less polar/non-polar by suppressing their proton donating ability. Keep the mobile phase pH acidic. These compounds can then be separated in RPC mode using non-polar or moderately polar columns such as C18 and C8.

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By Dr Pramod Kumar Pandey - June 15, 2025

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