Chromatography Forum

hi all:
recently I am packing reverse phase HPLC column 50*250mm with C4 media,what I want to know is when I am preparing the slurry ,which solvent I should choose!or what do you choose when you are making the slurry?do you have some packing tips to get a high efficiency column?like packing pressure and so on?

what's the particle size?

3ooA,does that matter?

Packing is a complicated exercise. May I suggest that you buy a packed column from a reputable manufacturer?

3ooA,does that matter?

Somehow, I really doubt that you have 300 Ã… particles. 300 Ã… would be much more typical of the pore diameter within the particles. The particle diameter would be measured in μm.

Uwe is correct, but he was being polite. I'll be more blunt: packing columns is a specialized business (think of it as being analogous to flying an airplane). If you are new enough that you don't realize that particle size matters, then you have several years of learning and experience to obtain before you will be able to do it well. You would be far better off obtaining prepacked columns.

thanks for your kind reply!it is my fault,the particle size is 10~15um,the pore size is 300A! I really want to know is there some packing tips to get
high column efficiency when the packing media is already choosen!

Hi

If you have a packing stand with a hydraulic piston or better a DAC-column, it´s maybe not that difficult. I would try IPA for the slurry and about 80-100 bars of compression pressure.

the trick: work quick

Best regards
Chris

I just took a new HPLC column through manufacturing validation. You know what the biggest variable in yield turned out to be? Technician experience.

Perhaps you are in a situation where buying is not a reasonable option. If so, budget lots of time to gain experience. Keep your eyes open and take good notes. You might also talk to the supplier of the column hardware. Unfortunately, the really good tips on column packing are mostly trade secrets.

With that small of a particle size I'd let the professionals handle it.

I packed my own semi-prep column, 22mmx250mm, 25-40um particle size. The housing was from Alltech (PN 96521), and it came with a piston to compress the silica after running solvent through it.

I carefully filled the housing with the silica gel. Made a dome at the top and attached the piston and end fitting. I then pumped methanol through it at 75ml/min for two days. After the three days I removed the piston and end fitting and made another dome of silica gel and repeated the procedure. I did this once more and there was no depression. I then made another dome and tightened down the piston and end fitting. It works great.

Many vendors that sell bulk stationary phase will have conditions for packing their phases into dynamic axial compression (DAC) columns. These conditions should include slurry preparation as well as packing pressure. I'd suggest contacting the vendor for recommendations. I've packed a number of different phases into DAC columns and the best conditions for each phase can vary quite a bit.

-----With that small of a particle size I'd let the professionals handle it.--- --

with DAC columns, we can pack down to 5µm, but we recommend our customers 8µm .

Only with the really big columns (>10cm) it becomes a litte difficult (and expensive)

Best regards
Chris




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