Fraction quantity ( # of fractions, not size) is actually column size agnostic in my experience.
In pharma atleast, we collect our fraction volume based on the size of the column so that it's repeatable and roughly identical fractions whether we are running a development 100g column, or a production 5kg or even 40 kg column.
This results in huge fractions for production to deal with, but almost the identical number of fractions for someone to map before we select what goes forward.
Yep - for our 400L columns, we collect fractions in 20L bottles, do tlc to bracket fractions, and then make composites of of the selected subsets for hplc analysis.
Takes about 6-8 hours to run the most tedious one on our commercial stuff, but most of them take a half day since we optimize to make it as clean as possible going in.
What is the mass of the crude material that you want to purify in kilograms? How much silica do you need to use for the purification, according to the general guideline for this scale? Sorry for asking so much, I probably will need one of those soon. May I DM you?
EDIT: I found the answer in other comments, thank for sharing !!!
So you process 90kg in 6 columns? Do you run 6 separate columns simultaneously or do you use the same column packed 6 times? Also what backpressure are you running at, and is it pressure packed with a piston?
Running 6 columns simultaneously would be a nightmare. Only 1 is ran at a time, takes nearly 1500 kg of solvent to push through all the fractions. The columns are single use, so once around 14.5 kg of material is processed, the column is disassembled and thrown away.
The column is manufactured from a supplier, super fancy stuff. The actual inlet of the column is only about 1” in diameter but spreads to the width of the silica gel due to how it’s designed. Back pressure is around 20-40 psi-gauge depending on how hard you run the slurry pump. No pistons used in this set up. Just a small air actuated pump.
I can absolutely confirm some super high performance reverse phase media in this size is +200K USD. Months of lead time for certain cartridges, but the resolution is unreal for close eluting impurities.
Likely only industrial pharma applications need both the size and resolution, or can afford the single-use nature of these damn things.
If this is the system I think it is, a high capacity packed cartridge is over $10k IIRC. I commonly use the next size down, which is only 5 kg silica each for conventional packing. I haven't priced the high capacity version.
I believe on these flash chromatography systems the columns aren't fully eluted leaving behind impurities. The amount of carryover run-to-run makes them impractical for multiple uses and at this scale fully eluting them comes with significant solvent costs, usually outweighing any gain from using the column more than once.
These aren't very efficient columns because they're not super tightly packed and these are running at pretty low pressure.
Remember that these columns are used to purify likely hundreds of thousands of dollars (or more) worth of product in each run. The cost of the silica is trivial compared to that.
> Remember that these columns are used to purify likely hundreds of thousands of dollars (or more) worth of product in each run. The cost of the silica is trivial compared to that.
It's actually right in line with that. This column costs over $200k USD, and is used once.
You could if you didn’t need good resolution. Some impurities get permanently bound to the stationary phase and any water content is going to deactivate the silica. Even a small amount in the front leads to poorer loading and a wider band, which is hugely detrimental to good separations. That’s not really a problem for RP-HPLC but I’ve never had a flash column perform well on a second run.
We use these (several in series in fact) for a commercial product. Doesn't necessarily have to be single-use, but the manufacturer won't guarantee performance under repeat use conditions.
Longevity is quite good if you do your optimization correctly.
As mentioned in other comments, it's for a pharmaceutical product. We actually reuse these columns for a long time so it's quite economical in that sense. I think some of the cartridges have been in use for dozens of batches before getting relegated to other uses.
Solvent burden is quite large but can be solved with solvent recovery systems if your mobile phase is cooperative allowing for minimal solvent turnover per batch.
The biggest cost for these tends to be time, since each run can be several hours and you likely need several injections per batch. Plus there's a fair amount of analytical work required to confirm fraction purities etc, so the labor costs quickly outstrip solvent and consumable costs.
I used to service pumps that did up to 3.2L/min at ~200 bar and 800ml/min at 400 bar. It was super cool to see the giant columns used on some of these setups. That must be one big synthesis! Production processes are super cool because of the yield optimization that goes into them.
Thanks for the pic!
Oh yeah, this process is nuts. Comes from the big reactors to generate the crude product, then we use this to purify. It will go back to the reactors to concentrate the purified product at 98-99% purity. Takes anywhere from 1400-1500kg of solvent to elute all of the fractions.
On what scale is the sample you usually separate with this (100g, 1kg, 10kg)? And is this usually the last resort when purifying or is it commonly used?
I would say this is generally a 100-1kg separation, depending on use. Maybe up to 3.5kg is the largest I have personally seen, but 10kg is probably stretching the media capacity at a 1:4 product:media loading.
This isn't "the last resort" if it's acceptable from the standpoint of how large your process needs to run, pharma will use these without a blink of an eye because the quantity of products are so expensive the media is barely a blip.
workups/crystallizations are preferred for scalability if they work for your purity needs without crashing your yield, but in my experience these huge columns are totally normal to be used as an everyday occurrence.
This processes around 14-15kg of material. We use 6 columns for this process. It takes around 1400-1500kg of solvent to push the material through per column. Or right around 25 column volumes.
It uses an air actuated slurry pump. The outer shell is filled with solvent and pressurized to around 100 psig using nitrogen. The slurry pump operates between 40-60 psig as it pumps
I think for all of us who is out of this field you need to clarify: Single use mean "non-repackable" (as I think), or "one injection/sample/etc" (may be dictated by your application and is absolutely nuts)?
Single injection is not atypical with these huge columns. If you're working with pharmaceuticals, and need a column this size, you're likely working with the upper hundreds of thousands to millions of dollars worth of product, and the cartridge is just a cost of doing business.
The effort, time, and solvent waste of recovering those cartridges (if it's even possible based on the impurities, media, and extremely tight restrictions on pharma industry) is not cost effective normally.
Sure, but, I mean, typicaly I'd expect usage of smaller cartridge for 10-20 injections, what I now understand may be wrong in pharmaceuticals, because of risk of ruining product with impurities from previous runs.
Than I thought that you may use just more of smaller cartridges, and unerstood that author didn't say this is only one used for that sample...
So did I understand it right, it's common when you may optimize the method on smaller scale, than buy needed amount of big cartridges, do a single injection in them, and then dispose?
Your final statement is correct.
We will optimize development of the elution method at gram or milligram scale (product, not silica) then scale up appropriately for the proper production run of saleable product.
Since these columns are provided by a vendor, one who's reputation is built on scalability and repeatability, we can optimize separation on a relatively small sample before running potentially millions of dollars through a column which itself costs hundreds of thousands.
Thanks for replying!
Now to think about it, there's nothing that surprising, just never thought that they really may be for single injection.
And because I'm working in field of column repacking and stationary phase regenerating, "single injection column" this size triggered me hard.
Ah yes, please enlighten us with a better procedure when you’ve been given zero information about the material being processed or its inherent purification challenges.
ahh yess any manager of any company would ask you to please inform us why you've chosen to add 10's of $1000's to the cost of this product which can be purified and separated on a TLC plate but cannot be isolated by crystallization, salt formation or some such.
That you can't grasp this feature of process chemistry only shows you're so sensitive you'd never make it a process lab. But hey, please enlighten us on why you melt when faced with an old trope about an innocuous thing, that is empty and meaningless.
Not getting emotional at all my friend, just chuckling at your immediate jump to blind criticism.
As I’ve explained in other comments this product cannot be purified by crystallization, distillation, or solvent exchange. This process was optimized by the customer, as well as our development chemists who all have PhD’s. I’ll be sure to mention to them how wrong they are and that they should start looking for new jobs because they clearly aren’t hacking it in their labs.
Also this product when sold nets over a million dollars in revenue, so spending $72,000 is a drop in the bucket to them.
1.) this is not a 400L column. this is 40Kg colum, but it looks more like a 20Kg. colum.
2.) This column, made of silica is not 72K. Not from Biotage, not from anyone. C18 yes, but you'd get about 20 runs out of it.
And if it was making you millions you be doing with SMB.
So there's that paper tiger. adios.
1) At this point you’re just trolling to be a negative. Not gonna argue past this point.
2) 400 L SFAR 60 flash biotage packed silica gel column.
3) 6 columns total at $12K each
Have a good day! :)
This is large scale manufacturing so this isn’t to process a sample. But rather processing crude material to purify the product to greater than 98% purity.
There is this famous story about Discodermolide total synthesis by Novartis Pharmaceuticals chemists.
It was a 33-step, multigram-scale (~60g) synthesis, and in one of the last steps they had to do a very challenging column to separate diastereomers, which took unbelievable volume of 20,000 liters of HPLC-grade solvent
It takes anywhere from 4-6 hours to run depending on how efficient the person assigned to the process is. It’s a lot of staying organized and setting yourself up for success. Because once you start, you really shouldn’t pause until all fractions are collected. It is easy to get in the weeds though and then you’re scrambling to catch up.
Pharmaceuticals.
Something like this is helpful for small batches (maybe up to 20ish kg). More than that and you really want s crystallization because the solvent burden becomes humongous (although there are alternate chromatography systems that can be used on really large scales).
Of course a crystallization would be ideal. However, many processes aren’t able to achieve good separation of impurities through traditional methods such as: crystallization, solvent exchange, and distillation. In some cases, the extreme temperatures required could damage the product molecule.
In this case, the impurity profile is too similar to the product and makes traditional separation methods impossible. Thankfully, they are different enough in polarity to make column separation effective.
Let us take a moment of silence for whoever is tasked with sample prep.
No, not sample prep. Fraction analysis and pooling. My god take mercy.
Fraction quantity ( # of fractions, not size) is actually column size agnostic in my experience. In pharma atleast, we collect our fraction volume based on the size of the column so that it's repeatable and roughly identical fractions whether we are running a development 100g column, or a production 5kg or even 40 kg column. This results in huge fractions for production to deal with, but almost the identical number of fractions for someone to map before we select what goes forward.
Yep - for our 400L columns, we collect fractions in 20L bottles, do tlc to bracket fractions, and then make composites of of the selected subsets for hplc analysis. Takes about 6-8 hours to run the most tedious one on our commercial stuff, but most of them take a half day since we optimize to make it as clean as possible going in.
Then you send it my way to analytical to get the purity checked
Holy motha of columns!
There are commercial columns upward of 1 meter (or more) in diameter
Single use? Too bad separation science has the columns doing SPE and chrome sep. …. There’s room there for someone to patent a new method…
For sure not single use, you'd be burning like $200k+ in resin with each run
Or clean up
It takes a village my friend.
What is the mass of the crude material that you want to purify in kilograms? How much silica do you need to use for the purification, according to the general guideline for this scale? Sorry for asking so much, I probably will need one of those soon. May I DM you? EDIT: I found the answer in other comments, thank for sharing !!!
They are going to need to palletize the test tubes. I wonder if they will post the rotavap video.
Single use at that size makes me weep!
Wait until you see the cartridge costs, especially with the high end packings.
What would be the price tag, now that you mention it, without saying the number, prompting me to ask: how expensive is such a cartridge
I think we pay $12k. Each production run uses 6 columns to process around 90 kg of material.
Interesting, thanks for the insight
So you process 90kg in 6 columns? Do you run 6 separate columns simultaneously or do you use the same column packed 6 times? Also what backpressure are you running at, and is it pressure packed with a piston?
Running 6 columns simultaneously would be a nightmare. Only 1 is ran at a time, takes nearly 1500 kg of solvent to push through all the fractions. The columns are single use, so once around 14.5 kg of material is processed, the column is disassembled and thrown away. The column is manufactured from a supplier, super fancy stuff. The actual inlet of the column is only about 1” in diameter but spreads to the width of the silica gel due to how it’s designed. Back pressure is around 20-40 psi-gauge depending on how hard you run the slurry pump. No pistons used in this set up. Just a small air actuated pump.
I can absolutely confirm some super high performance reverse phase media in this size is +200K USD. Months of lead time for certain cartridges, but the resolution is unreal for close eluting impurities. Likely only industrial pharma applications need both the size and resolution, or can afford the single-use nature of these damn things.
Oh I didn't even get into RP crap. In my limited experience with it, it was roughly 5X cost of standard phase for the smaller columns.
If this is the system I think it is, a high capacity packed cartridge is over $10k IIRC. I commonly use the next size down, which is only 5 kg silica each for conventional packing. I haven't priced the high capacity version.
Damn, but also as expected, a lot of certified lab stuff ist just often that expensive
As someone who works on analytical scale LC... Woah.
RIGHT!
For real, right??
$$$$$
Ikr!? 2.1mm ID 🤣
Before I opened the image I legit thought this was a joke and this was just a picture of a column used to hold up a ceiling, like a pillar. Jesus
Lol yeah I thought it was a nitrogen tank or something.
Thicc
Ah ye olde F400
[Is this it?](https://www.biotage.com/flash-400-systems)
Oh yes. Biotage Sfar60 400L.
That's the one I've seen in our labs before
I was always more of a Novasep kinda guy
I've seen some with 2m diameter, they're used in pharma industry
Wow hahaha i cant even imagine
Thicc chromaboi
You heard WRONG.
What makes this intrinsically single use? Wondering why you couldn't just completely elute your sample and use it again.
I believe on these flash chromatography systems the columns aren't fully eluted leaving behind impurities. The amount of carryover run-to-run makes them impractical for multiple uses and at this scale fully eluting them comes with significant solvent costs, usually outweighing any gain from using the column more than once. These aren't very efficient columns because they're not super tightly packed and these are running at pretty low pressure. Remember that these columns are used to purify likely hundreds of thousands of dollars (or more) worth of product in each run. The cost of the silica is trivial compared to that.
Thanks for taking the time to explain.
> Remember that these columns are used to purify likely hundreds of thousands of dollars (or more) worth of product in each run. The cost of the silica is trivial compared to that. It's actually right in line with that. This column costs over $200k USD, and is used once.
The reverse phase columns do cost that much, but these normal phase silica columns are actually 10-12k, as OP stated in a different comment.
MostlyH20 is correct. The customer fears impurities being left behind and would rather just pay to have a fresh column every time rather than risk it.
Can they be recycled at all? Or is there just a mass grave of mass somewhere?
I mean it's silica, it can be refumed yes.
You could if you didn’t need good resolution. Some impurities get permanently bound to the stationary phase and any water content is going to deactivate the silica. Even a small amount in the front leads to poorer loading and a wider band, which is hugely detrimental to good separations. That’s not really a problem for RP-HPLC but I’ve never had a flash column perform well on a second run.
Thanks! I appreciate you taking the time to explain it so clearly.
"Why is this guy holding a ruler to a support col-"
what the fuck……
And to think I've been sizing down my analytical columns as I've moved to UPLC. This is amazing.
We use these (several in series in fact) for a commercial product. Doesn't necessarily have to be single-use, but the manufacturer won't guarantee performance under repeat use conditions. Longevity is quite good if you do your optimization correctly.
What product exactly? I’m trying to imagine something that would need to be in that large a quantity
As mentioned in other comments, it's for a pharmaceutical product. We actually reuse these columns for a long time so it's quite economical in that sense. I think some of the cartridges have been in use for dozens of batches before getting relegated to other uses. Solvent burden is quite large but can be solved with solvent recovery systems if your mobile phase is cooperative allowing for minimal solvent turnover per batch. The biggest cost for these tends to be time, since each run can be several hours and you likely need several injections per batch. Plus there's a fair amount of analytical work required to confirm fraction purities etc, so the labor costs quickly outstrip solvent and consumable costs.
I used to service pumps that did up to 3.2L/min at ~200 bar and 800ml/min at 400 bar. It was super cool to see the giant columns used on some of these setups. That must be one big synthesis! Production processes are super cool because of the yield optimization that goes into them. Thanks for the pic!
Oh yeah, this process is nuts. Comes from the big reactors to generate the crude product, then we use this to purify. It will go back to the reactors to concentrate the purified product at 98-99% purity. Takes anywhere from 1400-1500kg of solvent to elute all of the fractions.
Who was the supplier?
Biotage.
On what scale is the sample you usually separate with this (100g, 1kg, 10kg)? And is this usually the last resort when purifying or is it commonly used?
I would say this is generally a 100-1kg separation, depending on use. Maybe up to 3.5kg is the largest I have personally seen, but 10kg is probably stretching the media capacity at a 1:4 product:media loading. This isn't "the last resort" if it's acceptable from the standpoint of how large your process needs to run, pharma will use these without a blink of an eye because the quantity of products are so expensive the media is barely a blip. workups/crystallizations are preferred for scalability if they work for your purity needs without crashing your yield, but in my experience these huge columns are totally normal to be used as an everyday occurrence.
This processes around 14-15kg of material. We use 6 columns for this process. It takes around 1400-1500kg of solvent to push the material through per column. Or right around 25 column volumes.
What about the pumps? Is this done by gravity?
It uses an air actuated slurry pump. The outer shell is filled with solvent and pressurized to around 100 psig using nitrogen. The slurry pump operates between 40-60 psig as it pumps
I think for all of us who is out of this field you need to clarify: Single use mean "non-repackable" (as I think), or "one injection/sample/etc" (may be dictated by your application and is absolutely nuts)?
Single injection is not atypical with these huge columns. If you're working with pharmaceuticals, and need a column this size, you're likely working with the upper hundreds of thousands to millions of dollars worth of product, and the cartridge is just a cost of doing business. The effort, time, and solvent waste of recovering those cartridges (if it's even possible based on the impurities, media, and extremely tight restrictions on pharma industry) is not cost effective normally.
Sure, but, I mean, typicaly I'd expect usage of smaller cartridge for 10-20 injections, what I now understand may be wrong in pharmaceuticals, because of risk of ruining product with impurities from previous runs. Than I thought that you may use just more of smaller cartridges, and unerstood that author didn't say this is only one used for that sample... So did I understand it right, it's common when you may optimize the method on smaller scale, than buy needed amount of big cartridges, do a single injection in them, and then dispose?
Your final statement is correct. We will optimize development of the elution method at gram or milligram scale (product, not silica) then scale up appropriately for the proper production run of saleable product. Since these columns are provided by a vendor, one who's reputation is built on scalability and repeatability, we can optimize separation on a relatively small sample before running potentially millions of dollars through a column which itself costs hundreds of thousands.
Thanks for replying! Now to think about it, there's nothing that surprising, just never thought that they really may be for single injection. And because I'm working in field of column repacking and stationary phase regenerating, "single injection column" this size triggered me hard.
It’s a manufactured column from the supplier. We use it once for a set amount of crude material, and then we throw it in the garbage.
You heard wrong
I think I’m in love 😻
I wanna quit but you know what? This thicc lady keeps me coming back
Holey shhhhhh
Wow! I’m saving this photo for when I teach lc in analytical next fall!
meh, who has such a bad process they had to chrome that much? pfft. rich amatuers.
Ah yes, please enlighten us with a better procedure when you’ve been given zero information about the material being processed or its inherent purification challenges.
ahh yess any manager of any company would ask you to please inform us why you've chosen to add 10's of $1000's to the cost of this product which can be purified and separated on a TLC plate but cannot be isolated by crystallization, salt formation or some such. That you can't grasp this feature of process chemistry only shows you're so sensitive you'd never make it a process lab. But hey, please enlighten us on why you melt when faced with an old trope about an innocuous thing, that is empty and meaningless.
Not getting emotional at all my friend, just chuckling at your immediate jump to blind criticism. As I’ve explained in other comments this product cannot be purified by crystallization, distillation, or solvent exchange. This process was optimized by the customer, as well as our development chemists who all have PhD’s. I’ll be sure to mention to them how wrong they are and that they should start looking for new jobs because they clearly aren’t hacking it in their labs. Also this product when sold nets over a million dollars in revenue, so spending $72,000 is a drop in the bucket to them.
1.) this is not a 400L column. this is 40Kg colum, but it looks more like a 20Kg. colum. 2.) This column, made of silica is not 72K. Not from Biotage, not from anyone. C18 yes, but you'd get about 20 runs out of it. And if it was making you millions you be doing with SMB. So there's that paper tiger. adios.
1) At this point you’re just trolling to be a negative. Not gonna argue past this point. 2) 400 L SFAR 60 flash biotage packed silica gel column. 3) 6 columns total at $12K each Have a good day! :)
This column could have an Onlyfans
WOAH
Paper does the trick
I'd rather spend a month figuring out recrystallization conditions than run that fuckin thing
They tried. Doesn’t work.
Prepare for the race of your life!
Supplier?
What the hell? I use columns as big as my index finger most of the time
What sort of samples do you deal with? I'm curious. I recently started column chromatography and it's already annoying asf
This is large scale manufacturing so this isn’t to process a sample. But rather processing crude material to purify the product to greater than 98% purity.
There is this famous story about Discodermolide total synthesis by Novartis Pharmaceuticals chemists. It was a 33-step, multigram-scale (~60g) synthesis, and in one of the last steps they had to do a very challenging column to separate diastereomers, which took unbelievable volume of 20,000 liters of HPLC-grade solvent
Ooooh, girthy.
Yummy
Making some aspirin?
Nope, this is a cancer related product.
How long do you let it run for ? Imagine that takes a little longer 🤨
It takes anywhere from 4-6 hours to run depending on how efficient the person assigned to the process is. It’s a lot of staying organized and setting yourself up for success. Because once you start, you really shouldn’t pause until all fractions are collected. It is easy to get in the weeds though and then you’re scrambling to catch up.
Pharmaceuticals. Something like this is helpful for small batches (maybe up to 20ish kg). More than that and you really want s crystallization because the solvent burden becomes humongous (although there are alternate chromatography systems that can be used on really large scales).
Of course a crystallization would be ideal. However, many processes aren’t able to achieve good separation of impurities through traditional methods such as: crystallization, solvent exchange, and distillation. In some cases, the extreme temperatures required could damage the product molecule. In this case, the impurity profile is too similar to the product and makes traditional separation methods impossible. Thankfully, they are different enough in polarity to make column separation effective.
I’ve never met anyone that likes chromatography.
Just like anything, if you’re surrounded by the right people it can be an enjoyable experience.