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proper washing of column
Posted: Tue Feb 09, 2010 1:13 pm
by shantaram
I am working with Phenyl Hypersil Column dimension 250*4.6mm (5µ). sample i am using ids Paclitaxel formulation (containing polyoxyethylated castor oil)..
I hv given washing with methanol as it solubilises castor oil, but after 2 runs only peak is splitting and pressure is building.....
please tell me hw i can restore column performance....
Can i use the following sequence
ACN
isopropyl alcohol
hexane
isopropyl alcohol
ACN
water required????
Posted: Tue Feb 09, 2010 3:52 pm
by tom jupille
That should work OK. Things to be careful about:
- isopropanol has a high viscosity, so keep an eye on the pressure (you may have to slow the flow rate). Methylene chloride might be a good alternative
- if possible, run the wash in the reverse-flow direction.
- it takes about 10 column volumes to get complete washout.
Going forward, you might want to look at doing a more thorough sample cleanup!
Posted: Wed Feb 10, 2010 2:45 pm
by bisnettrj2
Posted: Mon Feb 15, 2010 1:26 pm
by shantaram
That should work OK. Things to be careful about:
- isopropanol has a high viscosity, so keep an eye on the pressure (you may have to slow the flow rate). Methylene chloride might be a good alternative
- if possible, run the wash in the reverse-flow direction.
- it takes about 10 column volumes to get complete washout.
Going forward, you might want to look at doing a more thorough sample cleanup!
Thanks
Posted: Mon Feb 15, 2010 1:27 pm
by shantaram
http://chromatographyonline.findanalytichem.com/lcgc/Washing-Reversed-Phase-Silica-Based-Columns/ArticleStandard/Article/detail/656286?ref=25
Thanks..