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SPE Troubleshooting
Need to improve solid phase extraction (SPE)?
This page might help to solve your solid phase extraction (SPE) challenges.
On this page
| Cause | Solution |
|---|---|
| Improper conditioning. | Condition column according to phase: Reversed phase: use methanol, acetonitrile or isopropanol followed by solvent used for sample solution. Ion exchange: use methanol or isopropanol followed by a buffer at a pH which charges both sorbent and analyte. |
| Analytes have a greater affinity for the sample solution than for the column. | Choose a column having a greater selectivity for analytes. Change pH of sample to increase affinity of analytes to sorbent. Change polarity of sample to produce lower affinity for the analytes. |
| Improper washing. | Wash column using appropriate wash solution. |
| Poor elution. | Increase eluent volume, increase eluent strength. Decrease elution strength of washing solvent. |
| Cause | Solution |
|---|---|
| Analytes have stronger interaction with column sorbent than with eluting solvent. | Choose a column that is less retentive for the analytes. |
| Elution volume is too low. | Increase the elution volume. |
| Elution solvent is too weak to disrupt analyte interaction with the column. | Change pH of eluting solvent to ensure greater affinity for the analytes. Change polarity of eluting solvent to ensure greater affinity for the analytes. |
| Cause | Solution |
|---|---|
| Column dries before sample added. | Re-condition column. |
| Capacity of column is exceeded. | Decrease sample volume. |
| Sample loading flow rate is too high. | Use a column with a larger amount of sorbent. Decrease flow rate. |
| Elution flow rate is too fast. | Allow elution solvent to seep into column before aspirating or forcing through column. Apply in two separate aliquots versus a single aliquot. |
| Using too strong a wash solution to remove interferences. | Reduce strength of wash solvent. |
| Elution volume is too small. | Increase volume of elution solvent. |
| Cause | Solution |
|---|---|
| Interferences are extracted at the same time as analytes. | Selectively wash interferences from column prior to elution. Use a column which retains the analytes more than the interferences. |
| Interference is due to leachables from column. | Wash column with eluting solvent prior to conditioning. |
| Cause | Solution |
|---|---|
| Excessive particulate matter in sample. | Filter or centrifuge sample. |
| Sample solution is too viscous. | Dilute sample with a weak solvent. |
| Inadequate vacuum. | Increase vacuum. |
| Cause | Solution |
|---|---|
| Sample volumes larger than extraction column capacity. | Use larger extraction column volumes. |
| Cause | Solution |
|---|---|
| Improper methodology. | Consider a different extraction mechanism, sorbent, or format. |
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