I prepared a column (reverse phase C18) for Medium Pressure Liquid Chromatography (MPLC).
I injected my crude extract 5 times & now the column has become pale yellowish.
My crude extract contains flavonoids and terpenes.
I used Water and Methanol gradient as the Mobile phase.
How can I clean the column?
Thanks in advance !
Wash with methanol first, followed by dichloromethane to remove less-polar compounds. Wash again with methanol. This wash is essentially doing "non-aqueous reverse phase". Wash with a water/methanol (or acetonitrile) gradient with 0.5% TFA, then wash again with plain methanol.
Columns can look rather ugly, but still work well in my experience.
HI.... for cleaning purpose wash the column with 70% water; 15% methanol; 15% acetonitrile. Divert the column eluent to waste not to contaminate your detector(s)[if it is a RP HPLC column) . Wash the column slowly over to 100% methanol and wash for at least 15 minutes. Wash the column over to 100% acetonitrile and wash for at least 15 minutes.
for more u can check these answers Abdullah Abdullah
https://www.researchgate.net/post/What_protocol_do_you_use_to_clean_the_RP_C18_HPLC_column_daily#:~:text=In%20your%20case%20wash%20the,for%20at%20least%2015%20minutes.
Please look at the following below links which may help you in your analysis:
https://www.chemass.si/ca_content/Cleaning_and_maintaining_C-18_columns.pdf
https://www.hichrom.com/assets/HichromCat9-pdfs/APPENDICES-HPLC_Column_Cleaning_p372-373.pdf
https://www.hawachhplccolumn.com/how-to-cleaning-of-protein-residues-in-hplc-column/
Thanks
https://www.chemass.si/ca_content/Cleaning_and_maintaining_C-18_columns.pdf
https://phenomenex.blog/2017/10/05/stationary-phase/
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