Which is the best solvent used for washing of HPLC columns and in what concentration?
Hello Abhishek,
A number of solvents can be used in terms of cleaning and regeneration of HPLC columns. Heptane, isopropanol, acetonitrile, water etc.
A good detailed description can be found in the following links. Hope it helps you.
http://www.chromacademy.com/chromatography-HPLC-Column-Cleaning-Regeneration.html
http://www.hichrom.com/assets/HichromCat9-pdfs/APPENDICES-HPLC_Column_Cleaning_p372-373.pdf
Each HPLC column is different and if you use the wrong solvent you can damage or destroy the column so there is NO universal wash solution for chromatography columns. Column manufacturers can advise you better in this matter.
Start by checking with the column manufacturer for your specific column. They should be able to provide you with guidelines as to which solvents can and can not not be used. Next, choose a wash solution that is stronger than your mobile phase. You want a solution which will dissolve all of the possible sample materials that the column has been exposed to. Flush the column with this solution at a low to moderate flow rate for one or two hours. Longer times should not be needed, but an over-night flush is fine. For some columns, back flushing them is another way to remove built-up material which has accumulated at the head of the column on the inlet frit (check with the manufacturer first). Once the column has been flushed, slowly re-equilibrate the column and get it ready for testing. Use a QC test standard appropriate for the column AND your application. You should have done this before you started using the column to establish a baseline result to compare it to. Run the test method and compare the results. If the results obtained meet the specification established, you may resume work. If not, you can try a stronger wash solution (sometimes running at 40C, if safe, can help) and re-test.
Usually we have two kinds of columns. Polar columns and non polar columns.
Like dissolves like. wash polar columns with non polar solvents and vice versa.
you may use hot water also(50°C)
Regards,
Bujji Reddy.
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