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The Wrench
Vol. 6 Issue 1
Winter 2009
Problems with Peak Shape
Poor peak shape can cause a loss of resolution between adjoining peaks, it can lead to poor injection to injection reproducibility, or it may cause an injection to fail method acceptance criteria. Poor peak shape includes issues such as tailing, fronting, split peaks, and shouldered peaks.
The causes of poor peak shape are varied. They include everything from poorly developed methods, to contaminated columns or in other parts of the system, to extra system volume or poor system connections.
The following chart covers some of these common problems and possible remedies.
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Problem |
Indication |
Cause |
Solution |
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Peak Tailing
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One or a few peaks tailing |
Unexpected analyte interaction with solid support or stationary phase |
Use base deactivated, or hybrid silica materials. Old style silicas have active sites on the surface that can have unintended interactions with sample analyte |
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Add a mobile phase modifier to the system such as TEA |
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All peaks tailing |
Poor Connections in system |
Ensure that all tubing ends are cleanly and squarely cut.
Make sure that all nuts and ferrules are of the correct shape and tubing depth is correct for all fittings. Different manufacturers have different arrangements, and they are not all interchangeable.
Use PEEK or universal spring loaded connecting tubing. |
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Void in Column |
Replace Column |
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Dirty guard column |
Replace Guard |
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Dirty column frit |
Replace Frit |
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| Peak Fronting
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Initial Peaks fronting |
Too strong a sample solvent |
Samples should be dissolved in mobile phase or solvent weaker than mobile phase |
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All peaks fronting |
Column overload |
Reduce injection volume |
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Reduce sample concentrations |
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Use a column with a larger i.d. |
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Split Peaks
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Void or channel in column |
Replace column |
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Dirty column frit |
Replace frit |
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Dirty Column |
Back flush or replace |
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Sample solvent not suitable for use with mobile phase |
Ensure that sample and mobile phase solvents are compatible – make any necessary adjustments |
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Peak Shoulders
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Shoulder on 1 or 2 peaks |
Mobile phase is at a pH near the pKa of the analyte |
Adjust pH of mobile phase to be at least 2 pH units away from the pKa of the sample analytes
Adjust mobile phase to separate the 2 peaks |
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Shoulder on all peaks |
Dirty Inlet frit |
Replace |
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Dirty Guard Column |
Replace |
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Dirty Column |
Back flush or replace |
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Historical Wrench Articles
| Title | Issue | |
| Plunger Seals | Vol. 2 Issue 1 | Plunger Seal .pdf |
| HPLC Preventative Maintenance |
Vol. 2 Issue 2 |
Prevt. Maint .pdf |
| System Backpressure |
Vol 2 Issue 3 |
System Backpressure .pdf |
| Choosing a Lamp Vendor |
Vol 3 Issue 1 |
Lamp choices .pdf |
| Instrument Qualification and Aftermarket Parts |
Vol.3 Issue 2 |
IQ and aftermatket .pdf |
| Material Solvent Compatibility |
Vol.4 Issue 1 |
Solvent Compatibility .pdf |
| Mobile Phase Buffers |
Vol.4 Issue 2 |
Mobile Phase Buffers .pdf |
| Sample and Mobile Phase Filtering |
Vol.5 Issue 1 |
Filtering.pdf |
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