How to Clean an HPLC Column - Reverse Flush Method
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- Опубликовано: 25 июн 2019
- Caring for your HPLC column might be simple, but it is exceptionally important to the health, lifetime, and productivity of your column. Your columns health can greatly affect your analysis as well.
The following video focusing solely on how to clean your HPLC column through reverse flushing. It might be time to flush your HPLC column when you're seeing high back pressure, low efficiency, or split peaks, along with the most common issue, which is particulate build-up at the head of the column. Reverse flushing your column can be done by following the steps below:
1. Take out the column and seeing how the flow direction is currently moving.
2. Go in the opposite direction at half the traditional flow rate that is currently being run on the column.
3. Once the column is flipped and reinstalled on one end, you can begin running your system with cleaning solution, which can be found in our guide in the link below.
4. Any particulates that are in your column, will be flushed out and caught by a beaker and not into the detector. Do this for about 15-20 minutes.
5. Once it is done, go in the correct flow path, install the column, and run the prodigy test mix (also known as PTM) to confirm the efficiency and pressure is back to normal conditions.
To learn more about mobile phase limitations, column storage tips, and column protection devices check out our HPLC Column Care Guide found at www.phenomenex.com/columncare 
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For those who don't know, do this:
*Do the reverse flush.
*Set the flow rate to lets say, 2 or 3 ml per minute (check your pressure limits beforehand, because this will increase it and may cause the pump to give you an error message).
*Change the concentration if you want (I use acetonitrile and change the pump concentration to 100%)
This usually helps you with broad peaks, small peaks, etc.
Thanks a Lot! I was unaware that you guys have also a troubleshooting guide like Waters
Very good and funny videos bring a great sense of entertainment!
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Wouldn't it be better to use a higher flow rate because it may require more force to wash out the particulate matter trapped in frit?
Good
Does Phenomenex sell the test mix? Are we limited to a half ml/min on backflush? Should we run a methanol mix first then IPA if the pressure is still high? Should we adjust the pH?
Hi Michael, yes we do sell test mix for C18 and Reverse Phase columns the part number is AL0-3045. And also yes, the backflush needs to be done at a lower flow rate or else it can cause damage to the media bed. And it is better to go with the lower viscosity solvent (MeOH/ACN) to the higher (IPA) - very important to use lower flow rate. And no, it is best to use only pure water instead of buffer to wash the column. You are trying to wash the buffer salts off the column when you are cleaning. We hope this helps! If you have any more questions and need an answer right away, please feel free to reach out to our technical experts via Live Chat on our website! It can be found at www.phenomenex.com/chat. Thanks!
What flow rate do you recomend for this procedure?
Hi SCanchoGonz, this depends on the ID of the column that you are washing but if it is a 4.6-3.0mm ID then 0.1-0.2 mL/min would be a good starting place.
If you have any more questions that need an answer right away, please feel free to reach out to our technical experts via Live Chat on our website. You can find it at www.phenomenex.com/chat.
How to know.the port for the column compartment.1-6 port or not?
hi sr ... sr i want to repair the dead column by changing packing material ...can you guide me...
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Can I use Reverse Flush Method with maintenance schedule (once a year)? Or only with issues/problems?
Can i use IPA for reverseflush
Yes! You can use it to flush with the column in reverse. You will just need to lower the flow rate by at least 1/2 because IPA is really viscous. If you have any more questions, reach out to our technical specialists at www.phenomenex.com/chat. They are super helpful and it is a free service we offer!
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