In this video I have explained
Methanol can significantly affect HPLC peaks depending on how it's used. Here are the main ways it influences the chromatogram:
1. As a Mobile Phase Component:
Polarity: Methanol is less polar than water but more polar than acetonitrile. Increasing methanol concentration generally reduces retention times for non-polar compounds in reverse-phase HPLC.
Viscosity: Methanol is more viscous than acetonitrile, which can increase backpressure and possibly broaden peaks.
Peak Shape: Some compounds may exhibit better or worse peak symmetry depending on methanol's interaction with both analyte and column.
2. As a Sample Solvent:
If methanol is used to dissolve the sample but is stronger than the mobile phase, it can cause peak fronting or distortion.
Incompatibility between the sample solvent and mobile phase may result in splitting, tailing, or poor resolution.
3. UV Absorbance:
Methanol has UV absorbance at low wavelengths, so it can cause baseline noise or drift if detection is set at low UV ranges.
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