How to Clean an HPLC Column

How to Clean an HPLC Column

Dec 15th 2021

In this video, we are going to teach you how to clean your HPLC column.

This is especially important to do if you want to add buffers or additives to your mobile phase. For instance, most people use ammonium acetate, formic acid, etc. And over time, if the column isn’t flushed out regularly, precipitates can form and plug your column up.

That's why 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 at 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

  1. 95% water and 5% methanol for 10 column volumes
  2. Do a gradient of 5 to 95% methanol for 5 column volumes
  3. 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 your pressure build up on your column. Always note the backpressure on your method when you first start to use your column and if pressure kick up around 15-20% it’s probably a good idea to use the reverse flush.  

Band Broadening in HPLC: A Simple Overview

Band Broadening in HPLC: A Simple Overview

May 6th 2022

I’m sure in the lab when you are analyzing your result you probably have gotten something like this before… Ahh yep, the good ‘ol HPLC band broadening problem – when your results aren’ …
How to Prep a qPCR Plate

How to Prep a qPCR Plate

Dec 23rd 2021

Today we are going to talk about setting up a plate for qPCR.For a 96 well plate the first thing you want to do is to make some master mix.Each vendor has a master mix formula you want to follow.This …
How to Clean an HPLC Column

How to Clean an HPLC Column

Dec 15th 2021

In this video, we are going to teach you how to clean your HPLC column.This is especially important to do if you want to add buffers or additives to your mobile phase. For instance, most people use a …
How to Set up and Use a Balance

How to Set up and Use a Balance

Dec 13th 2021

In this video we are going to learn how to properly set up and use a balance when you’re in the lab.Before you start using the balance, you want to make sure that it’s properly calibrated to ensure th …