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column maximum flow
Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.
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- tom jupille
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There is no simple answer. That said, usually you work it the other way around: you know what flow you want to use; the question is how much back pressure will that flow generate.
The pressure for a given volumetric flow rate will depend on:
- the column length (longer column = higher pressure)
- the square of the column diameter (wider column = lower pressure)
- the particle size (smaller particles = higher pressure)
- the viscosity of the solvent (higher viscosity = higher pressure)
- the temperature (higher temperature = lower viscosity = lower pressure)
http://en.wikipedia.org/wiki/Hagen%E2%8 ... e_equation
http://fr.wikipedia.org/wiki/%C3%89coul ... Poiseuille
The pressure for a given volumetric flow rate will depend on:
- the column length (longer column = higher pressure)
- the square of the column diameter (wider column = lower pressure)
- the particle size (smaller particles = higher pressure)
- the viscosity of the solvent (higher viscosity = higher pressure)
- the temperature (higher temperature = lower viscosity = lower pressure)
http://en.wikipedia.org/wiki/Hagen%E2%8 ... e_equation
http://fr.wikipedia.org/wiki/%C3%89coul ... Poiseuille
-- Tom Jupille
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
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- Posts: 16
- Joined: Mon Nov 28, 2011 8:53 pm
thanks tom!
yes! generally i use the optimum flow or i look for this optimum but this information is easy to found because company (agilent, Waters,...) give sometimes a flow optimum range in relationship with the solvent. i don't Know how they have this information but i suppose that they study the fluid mechanic (Hagen-Poiseuille equation). but i think that if you work everytime with a high flow, even if this flow allowed to a presssure wich is lower than your hplc system , this phenomen can resulted in a bad resolution of your elution.
(sorry for my english!) and sometime when i make a column maintenance, i need to know what is the maximum flow of my column.
yes! generally i use the optimum flow or i look for this optimum but this information is easy to found because company (agilent, Waters,...) give sometimes a flow optimum range in relationship with the solvent. i don't Know how they have this information but i suppose that they study the fluid mechanic (Hagen-Poiseuille equation). but i think that if you work everytime with a high flow, even if this flow allowed to a presssure wich is lower than your hplc system , this phenomen can resulted in a bad resolution of your elution.
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- tom jupille
- Site Admin
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- Posts: 4978
- Joined: Wed Aug 11, 2004 4:55 pm
Don't worry about your English. Votre anglais est meilleur que mon français!
With small-particle packings (less than about 5 micrometers), the loss of efficiency at high flow is relatively minor (at least for small-molecule samples; proteins are a different story) so that using the "optimum" flow rate is usually not cost-effective; with a 3-micron packing you might gain 10-15% in resolution at a cost of 2x or 4x the run time. In our Advanced Method Development course, I usually suggest that the flow be set to whatever generates a back pressure of about 40% of the system maximum (so, about 150 bar on a system that can operate at 350 bar). That allows some margin for column aging (back pressure usually increases over the lifetime of a column). What that flow rate actually *is* depends on all of the parameters mentioned. Most people don't calculate this, but simply note the pressure at a given flow rate and adjust accordingly. Pressure is a linear function of flow, so it's an easy adjustment.
With small-particle packings (less than about 5 micrometers), the loss of efficiency at high flow is relatively minor (at least for small-molecule samples; proteins are a different story) so that using the "optimum" flow rate is usually not cost-effective; with a 3-micron packing you might gain 10-15% in resolution at a cost of 2x or 4x the run time. In our Advanced Method Development course, I usually suggest that the flow be set to whatever generates a back pressure of about 40% of the system maximum (so, about 150 bar on a system that can operate at 350 bar). That allows some margin for column aging (back pressure usually increases over the lifetime of a column). What that flow rate actually *is* depends on all of the parameters mentioned. Most people don't calculate this, but simply note the pressure at a given flow rate and adjust accordingly. Pressure is a linear function of flow, so it's an easy adjustment.
-- Tom Jupille
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
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- Posts: 16
- Joined: Mon Nov 28, 2011 8:53 pm
Thanks a lot for your help!!!
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- Posts: 3594
- Joined: Mon Aug 30, 2004 11:43 pm
Yep, that's me.Most people don't calculate this, but simply note the pressure at a given flow rate and adjust accordingly. Pressure is a linear function of flow, so it's an easy adjustment.
I leave the equations to Tom J.
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Dear User ,
The Pressure generated by column is toterated upto 600 Bar , that is only the packing of column which will not be disturbed during the usage in mentioned pressure.
As far HPLC or the machine is concerned , normal LC system that is Waters / Agilent they can tolerate the pressure upto 285bar ( approx. 4000 psi ) , or upto max 357 bar ( approx. 5000 psi ).
Further this much back pressure may damage to the seals in machine , which may lead to leakage and malfunctioning of machine .
My suggestion is to use the column or develop the method such that it should be having the column back pressure of NMT 3000 psi (214 bar ) in HPLC
If not possible for that method then choose UPLC.
Thanks....Manoj
The Pressure generated by column is toterated upto 600 Bar , that is only the packing of column which will not be disturbed during the usage in mentioned pressure.
As far HPLC or the machine is concerned , normal LC system that is Waters / Agilent they can tolerate the pressure upto 285bar ( approx. 4000 psi ) , or upto max 357 bar ( approx. 5000 psi ).
Further this much back pressure may damage to the seals in machine , which may lead to leakage and malfunctioning of machine .
My suggestion is to use the column or develop the method such that it should be having the column back pressure of NMT 3000 psi (214 bar ) in HPLC
If not possible for that method then choose UPLC.
Thanks....Manoj
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