What You Need to Do Before You Make a Decision to Replace Your Column
You all know it can be frustrating when you run a precious sample on a column that you have used before for the same applications, only to get terrible performance leading to compromising your experiment. There are mixtures that need to be optimized to get the best results in a months’ time, it would not be right when you get results that cannot be interpreted it can be discouraging. There are high chances that you have a contaminated HPLC column. We are going to learn some of the basic ideas that can help you in troubleshooting your HPLC column before you make a decision to replace.
Check out if the cause may be due to contamination, you need to clean with the right reagents so that you do not result to contamination. In many cases the reversed-phase columns are typically stored in an organic solvent that is miscible like methanol-water. Although it is normally good practice to ensure that you wash it with running solvent like methanol or acetonitrile. Equilibrating the columns are significant in getting the best results as it has been seen to offer the best results this time around for the best results.
Check out if the column has been clogged with insoluble materials. You may either choose to use a dilution of the sample or just use an external solubility to see if it dissolves. You will find that despite the washing and even equilibration, the column will still contain the contaminants that are insoluble, they can negatively affect the performance of your experiments.
Before you make a conclusion to replace the HPLC column, you need to first clean with a strong solvent that is compatible with your mobile phase. Take a moment and get to feature ways that you can be able to flush the insoluble materials using the backward form procedure, it can be delicate as it may result to destruction of the integrity of the bed and it matters much in the results that you get after separation.
If you do not take the issues in time, you may result to ruining all the results that you may be posting, you need to ensure that monitor your HPLC column levels of pressures from time to time so that you can know if there are issues. You need to ensure that you boost performance all the time and if the troubleshooting does not bear any fruits, you need to ensure that you buy another HPLC column.
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