proper washing of column

[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????

[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!

[bisnettrj2]

http://chromatographyonline.findanalyti ... 286?ref=25

[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

[shantaram]

http://chromatographyonline.findanalytichem.com/lcgc/Washing-Reversed-Phase-Silica-Based-Columns/ArticleStandard/Article/detail/656286?ref=25

Thanks..