What criteria do you use for selecting a column during HPLC method development?

Question

Accepted Answer

Selecting an HPLC column involves considering factors like analyte properties, separation goals, and method conditions.

1. Analyte Properties: Consider polarity, size, and charge of the analyte. For example, use C18 columns for non-polar compounds.
2. Separation Goals: Determine if you need resolution, speed, or sensitivity. For instance, use a shorter column for faster analysis.
3. Mobile Phase Compatibility: Ensure the column material is compatible with the chosen mobile phase. For example, silica-based columns for aqueous solvents.
4. Column Dimensions: Select appropriate length and diameter based on sample volume and desired resolution. A longer column can improve resolution.
5. Temperature Stability: Consider the thermal stability of the column material, especially for high-temperature applications.

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