I am doing hplc analysis and recently purchased a new column because the old one has degraded. Can one simply install the new column and start straight away with running standards or is there a procedure to do first before I start using the new column.
I am using a C18 150mm x 4.6mm, 5um Fortis hplc column.
You can start straight away, but you might expect shifts in RT and passivation issues with the first injections.
Dear Damola,
Any column manufacturer provides column certificate with column efficiency report. check storage solvent given on the certificate and first give the flushing with the same solvent along with water in the ration 90:10. Second step is to check column performance. Refer CoA and perform the test. Compare the result of performance test with certificate results for Theoretical Plate, Asymmetry and resolution. If all parameters are matching closely with column manufacturer certificate results then you can take column for use.
before initializing analysis give the flushing with mobile phase for good baseline then you can initiate analysis. Read Column care and usage provided by manufacturer.
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If need to change solvent, please make sure that it is a compatible one. Also slowly increase the ratio by 10:90 and increase by 10 each time. Each flushing should be at least 30 minutes. Also depends on your flow rate… how fast does it take for your solvent to pass through the whole column?
Review the column information supplied by the manufacturer before use. Insure the column is safe to use with the proposed method you have (chemical compatibility, pH, pressure, flow rate etc). If all is OK, follow good chromatography practices and wash the column (using a solvent that is stronger than your mobile phase to remove any retained material or fouling) flush for about 20x column volumes with mobile phase, equilibrate the column and use.
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