GPC/SEC Troubleshooting Guide
Your guide to solving common problems and staying productive
Places to Start Peak Tailing Possible Cause
Excessive dead volumes
Solution
Minimize tubing length
Partial Peak Possible Cause
Dead space of separation
Solution
Add columns with higher molecular
Exclusion
around half of total weight-resolving range
Solvents Day to Day Tighten injection seal elution volume
– Always choose HPLC-grade solvents and filter through – Don’t shut the system down with the columns connected;
Check connector fittings Peaks eluting close to this volume Possibly increase to 5–10 μm
0.02–0.45 μm filters reduce the flow rate to 0.1 mL/min (recycle if required)
may be partially excluded particle size
– Use stabilized solvent when appropriate (e.g., THF stabilized with – If removing the columns for any length of time, make sure they Column degradation Replace or repair column
butyl hydroxylated toluene (BHT) to extend column lifetimes) are stored in the recommended solvent and that the end caps Look for sharp peaks at the front Only reduce number of columns
are correctly fitted Column interaction Use mobile phase additives of your chromatograms if faster run times are important
– Use solvent additives as recommended to suppress any and lower resolution is acceptable
interaction between the sample and the columns – If using buffers, flush the column into pure solvent when analysis Shear degradation Replace standard solutions
is complete every two weeks
Q: Possible Cause Solution
– Make sure that any solvent change is to (or via) a miscible solvent; Increased Retention
)\GPYWMSR Modify dissolution process Flow rate reduction Check for bubbles in pump head
flush with at least three column volumes or preferably overnight (no excessive shaking) Time
8SXEP
4IVQIEXMSR
Sample Preparation Degas solvent
– Dissolution time and temperature will depend on molecular Possible Cause Solution
Peak Broadening Interaction with packing Use modifiers/additives
2S4IEOW weight and crystallinity of polymer; stirring may aid dissolution but Large dead volume Minimize tubing/check fittings
avoid sample degradation Sample adsorption Change eluent polarity
Eluent too viscous Use column oven
7ITEVEXMSR – Always use an aliquot of the mobile phase to prepare the solution
– Filter samples to remove any insoluble material (0.5–1.0 μm filters) Incorrect Molecular Possible Cause Solution
7]WXIQ 7SPZIRX Detector cell too large If possible, use smaller cell volume
4IEOW
– Remember: sample concentration is critical if using light Weights Improper selection of baseline Analyze whole peak for true
and integration limits reflection of sample
scattering or viscometry detection; ensure there is no solvent
QMR evaporation from the vial prior to analysis Select integration limits down to
Possible Cause Solution baseline on either side of peak
Typical regions of a GPC chromatogram Coeluting Peaks
Select set of columns with wider (solid black line is correct)
resolving range
Excluding components of peak
Sample is copolymer or blend If peaks cannot be separated, always will give incomplete MWD and
analyse sample as though single peak incorrect molecular weight values
Possible Cause Solution Peak Splitting Possible Cause Solution
Baseline Drift/Noise
Column/detector contamination Flush column/detector to waste Sample loading too large Reduce loading/loop size
Clean eluent Blocked/partially blocked frit Replace frit; use 2 μm inline filter
to stop clogging
Use better quality solvents
Void in column Replace column
Bubbles in detector Degas solvent
Partially blocked injection valve Replace rotor seal
Temperature variations Use column heater/insulate tubing
Molecular Weight Possible Cause Solution
Resolution Possible Cause Solution Pump drift; column aging; Mark end of run with small molecule
Variations
Analyzing polymers Choose correct column new capillaries; reswaged fittings; such as toluene, BHT, acetone,
or monomers? pore size new column connections or ethylene glycol
Narrow peak marker (internal
If main peak is above the If main polymer peak is main focus,
standard) to measure column
resolving range of the then increase column pore size
efficiency and identify degradation,
columns, sample is to cover molecular weight range
tailing, and retention shifts not
“excluded” from columns of sample
obvious with broad sample peaks
Recalibrate columns regularly
Ghost Peaks Possible Cause Solution
Peaks carried over from Let previous sample fully elute Check pump flow rate is consistent
previous injection before next injection
Check any fittings for leaks
If absorption occurs, some Either add buffer to mobile phase
material may elute after total or switch to a different solvent If symptoms continue,
permeation limit replace columns
DE3200231481
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