When there is a deterioration of chromatography, due to some form of contamination you will need to regenerate the column.
1. You can either send your column back to Biochrom and have the column regenerated by us. Please contact your local distributors for more information regarding this service. If you have any problems contacting your local distributor please contact us direct by email at support@biochrom.co.uk. If you do send back your column please ensure that you enclose any spare resin.2. Alternatively you may wish to regenerate the column yourself using the following procedure which has been found to be effective in recovering chromatographic performance.
The following materials and equipment are required for resin cleaning:
100ml conc. HCl AR grade
To clean the resin proceed as follows:
Pour 100ml of Distilled water and 100ml of Methanol into a flask with a
PTFE circular magnetic stirrer. Add the contaminated resin. Using the hotplate
in a fume hood start stirring VERY slowly and heat the liquid until it is just boiling.
Continue to boil for 30 mins. A funnel can be placed over the flask to act as
a condenser.
CAUTION: Do not leave the mixture unattended. Do not use anti-bumping granules.
Remove the conical flask from the hotplate and add 100ml of distilled water to the flask to dilute and cool down the mixture. Filter the mixture over Whatman No. 50 paper in a Buchner funnel with vacuum, then wash the resin with 100ml of distilled water 3 times. The filter paper must be seated correctly.
Pour 100ml of distilled water into a flask and add 2g of EDTA heat, stir
until the EDTA has dissolved, then add 100ml of Conc HCl. Transfer the resin into
the flask. Boil and stir as before for at least 1 hour.
CAUTION: Do not leave the mixture unattended. Do not use anti-bumping
granules.
Remove the conical flask from the hotplate and add 100ml of distilled water to the flask to dilute and cool down the mixture. Filter the mixture over Whatman No. 50 paper in a Buchner funnel with vacuum, then wash the resin with 100ml of distilled water 3 times. The filter paper must be seated correctly.
If the resin is required in sodium form then suspend the resin in the 200ml aliquot of 4M sodium hydroxide in the conical flask. If the resin is required in lithium form, suspend it in the 200ml aliquot of 3M lithium hydroxide. Use a small quantity of distilled water to rinse the remaining resin particles from the filter paper into the hydroxide solution.
Using the hotplate in the fume hood, boil the mixture for 30 minutes, stirring occasionally to prevent spattering (especially with the lithium form).
Remove the conical flask from the hotplate and add 100ml of distilled water to the flask to dilute and cool down the mixture. Filter the hot mixture over Whatman No. 50 paper in a Buchner funnel with aspirator, ensuring that the filter paper is seated correctly.
Wash the resin with 3 x 100ml aliquots of distilled water and leave the suction on until the resin cake develops cracks on the surface. Then transfer it to the resin bottle using a plastic spatula. The resin adhered to the filter paper and funnel should be washed into the remaining resin bottle after packing the column.
Suspend the resin in the minimal volume (3ml) of either 0.4M sodium hydroxide (sodium system) or 0.3M lithium hydroxide (lithium system).
Pack the column as described in AAA FAQ25 (How do I pack a column?).
If this does not answer your question or solve your problem, then please contact us at support@biochrom.co.uk detailing your question or the problem you are experiencing. Please list if relevant the instrument, software and version numbers you are running, and enclose any relevant data files, etc.