Peak Splitting in Chromatography

This Article contains the information about Peak Splitting which is commonly observed in the Chromatographic analysis..

Peak Splitting in Chromatography :

Peak splitting in chromatography refers to the appearance of shoulders or twin peaks within a single Gaussian peak. It can occur due to various factors, impacting the quality of separation. Let’s explore some common reasons for the same…..

1) Improper Connections:

Improper connection in the flow path between the injector and the detector,

Below Are the some examples :

• Tubing slippage when using PEEK finger tight fittings
• Improper ferrule depth on stainless steel fittings or other fittings
• Small void inside the connection
• If all peaks are impacted, the dead volume is often after the column

2) Particulates lodged in the inlet of the column :

• Particulates in the samples
• Particulates in the mobile phase
• Microbial contamination in Samples, Mobile Phase or in column.

3) Sample Solvent Effect:

The sample solvent composition can impact peak shapes.

Examples Includes ,

• If the sample diluent contains a higher percentage of nonpolar solvent (reverse-phase) compared with the starting mobile phase
• If the sample diluent contains a higher percentage of polar solvent (normal phase; HILIC) compared with the starting mobile phase

4) Dirty Emitter (for Mass Spectrometry):

In mass spectrometry, a dirty emitter can cause peak shape issues. Additionally, dead volumes in the probe may contribute to splitting.

5) Sugars in HILIC Chromatography :

When analyzing sugars, they can form anomers (open chain vs. closed chain), leading to peak splitting.

6) Sample matrix buildup in column :

Accumulation of sample matrix in the column or Dirty column  can result in peak splitting.

Remember that peak splitting indicates deficiencies in Analytical Techniques  and addressing these factors is important for achieving accurate and reliable chromatographic results.

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