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- Posts: 2916
- Joined: Mon Aug 30, 2004 10:19 pm
Adam,
Everything except for the column plate count scales with column volume, if you want to get the same retention pattern. Column plate count and analysis time vary with column length. If you need to care about this (e.g. tight resolution) then scaling may have some limitations. For example, on a shorter column, the plate count will be lower, but the analysis time will be faster. Since the plate count will be lower, you can break the rule above and inject a bit more to get a higher signal. Or you can slow down the run time (which includes flow rate and gradient time) and gain some plates again. Or you can reduce the particle size to gain back the plates, but it will cost you in pressure.
However, to keep things simple, stick with the number one rule above and scale everything with the column volume.
I can send you an article on this, if you contact me. Or you can buy the new book by Ahuda and Dong called the "Handbook of HPLC in Pharmaceutical Analysis".
Everything except for the column plate count scales with column volume, if you want to get the same retention pattern. Column plate count and analysis time vary with column length. If you need to care about this (e.g. tight resolution) then scaling may have some limitations. For example, on a shorter column, the plate count will be lower, but the analysis time will be faster. Since the plate count will be lower, you can break the rule above and inject a bit more to get a higher signal. Or you can slow down the run time (which includes flow rate and gradient time) and gain some plates again. Or you can reduce the particle size to gain back the plates, but it will cost you in pressure.
However, to keep things simple, stick with the number one rule above and scale everything with the column volume.
I can send you an article on this, if you contact me. Or you can buy the new book by Ahuda and Dong called the "Handbook of HPLC in Pharmaceutical Analysis".
