In this video, we are going to teach you how to clean your column. 🧼
This is especially important to do if you add buffers or additives to your mobile phase. For instance, most people use ammonium acetate, formic acid, etc. Over time, if the column isn’t flushed out regularly, precipitates can form and can plug your column up.
We are going to show you 2 techniques to extend your column life as much as possible.
The first method is your daily maintenance method. If you have a solenoid valve in your pump, you can select up to 4 solvents. A and B can be used as your mobile phase for regular operations. C and D can then be selected to be used as your solvents for cleaning. C and D would normally consist of water and some kind of organic solvent like methanol. You can simply do a gradient after your last run of 5% to 95% methanol for over an hour.
If you feel like your column needs a deeper cleaning, it would be best to do a reverse flush. The method for reverse flush would be to
95% water and 5% methanol for 10 column columns
Do a gradient of 5 to 95% methanol for 5 column volumes
Hold 5% water and 95% for 10-column volumes
You want to run this at half your normal flow rates for your method.
The way that you know when to do deeper cleaning is to monitor the pressure build-up on your column. Always note the back pressure on your method when you first start to use your column and if pressure kicks up around 15-20% it’s probably a good idea to use the reverse flush.
When you have questions or have a topic you want to be covered let me know in the comments! 😎
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#biotech #biology #science #hplc #clanning
16 дек 2021
